TY  - JOUR
AU  - Malešević, Milka
AU  - Gardijan, Lazar
AU  - Miljković, Marija
AU  - O'Connor, Paula M
AU  - Mirković, Nemanja
AU  - Jovčić, Branko
AU  - Cotter, Paul D
AU  - Jovanovic, Goran
AU  - Kojić, Milan
PY  - 2023
UR  - https://doi.org/10.1093/lambio/ovad004
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1828
AB  - Lactic acid bacterium Lactococcus lactis BGBU1-4 produces 43 amino acids (aa) long bacteriocin, lactolisterin BU (LBU), a 5.161 kDa peptide with potent antibacterial activity against many Gram-positive pathogens. In addition, BGBU1-4 produces an additional unknown product of 3.642 kDa with antibacterial activity. Here, we determined that the significant amount of naturally produced LBU breaks down to create a 3.642 kDa truncated form of LBU bacteriocin consisting of 31 N-terminal aa (LBU1-31) that exhibits 12.5% the antibacterial activity of the full-length LBU. We showed that chemically synthesized LBU is stable and 50% less active than native LBU, and so we used the synthetic peptides of LBU and its variants to further study their activities and antibacterial potential. Deletion analysis of LBU revealed that the 24 N-terminal aa of LBU (LBU1-24) are responsible for antibacterial activity, while downstream aa (25–43) determine the species-specific effectiveness of LBU. Although LBU1-31 contains aa 1–24, the truncation at position 31 is predicted to change the structure within aa 15–31 and might impact on antibacterial activity. Intriguingly, whole genome sequencing and genome mining established that BGBU1-4 is abundant in genes that encode potential antibacterials, but produces LBU and its breakdown product LBU1-31 exclusively.
T2  - Letters in Applied Microbiology
T2  - Letters in Applied MicrobiologyLetters in Applied Microbiology
T1  - Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants
IS  - 2
SP  - ovad004
VL  - 76
DO  - 10.1093/lambio/ovad004
ER  - 

@article{
author = "Malešević, Milka and Gardijan, Lazar and Miljković, Marija and O'Connor, Paula M and Mirković, Nemanja and Jovčić, Branko and Cotter, Paul D and Jovanovic, Goran and Kojić, Milan",
year = "2023",
abstract = "Lactic acid bacterium Lactococcus lactis BGBU1-4 produces 43 amino acids (aa) long bacteriocin, lactolisterin BU (LBU), a 5.161 kDa peptide with potent antibacterial activity against many Gram-positive pathogens. In addition, BGBU1-4 produces an additional unknown product of 3.642 kDa with antibacterial activity. Here, we determined that the significant amount of naturally produced LBU breaks down to create a 3.642 kDa truncated form of LBU bacteriocin consisting of 31 N-terminal aa (LBU1-31) that exhibits 12.5% the antibacterial activity of the full-length LBU. We showed that chemically synthesized LBU is stable and 50% less active than native LBU, and so we used the synthetic peptides of LBU and its variants to further study their activities and antibacterial potential. Deletion analysis of LBU revealed that the 24 N-terminal aa of LBU (LBU1-24) are responsible for antibacterial activity, while downstream aa (25–43) determine the species-specific effectiveness of LBU. Although LBU1-31 contains aa 1–24, the truncation at position 31 is predicted to change the structure within aa 15–31 and might impact on antibacterial activity. Intriguingly, whole genome sequencing and genome mining established that BGBU1-4 is abundant in genes that encode potential antibacterials, but produces LBU and its breakdown product LBU1-31 exclusively.",
journal = "Letters in Applied Microbiology, Letters in Applied MicrobiologyLetters in Applied Microbiology",
title = "Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants",
number = "2",
pages = "ovad004",
volume = "76",
doi = "10.1093/lambio/ovad004"
}

Malešević, M., Gardijan, L., Miljković, M., O'Connor, P. M., Mirković, N., Jovčić, B., Cotter, P. D., Jovanovic, G.,& Kojić, M.. (2023). Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants. in Letters in Applied Microbiology, 76(2), ovad004.
https://doi.org/10.1093/lambio/ovad004

Malešević M, Gardijan L, Miljković M, O'Connor PM, Mirković N, Jovčić B, Cotter PD, Jovanovic G, Kojić M. Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants. in Letters in Applied Microbiology. 2023;76(2):ovad004.
doi:10.1093/lambio/ovad004 .

Malešević, Milka, Gardijan, Lazar, Miljković, Marija, O'Connor, Paula M, Mirković, Nemanja, Jovčić, Branko, Cotter, Paul D, Jovanovic, Goran, Kojić, Milan, "Exploring the antibacterial potential of Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 by genome mining, bacteriocin gene overexpression, and chemical protein synthesis of lactolisterin BU variants" in Letters in Applied Microbiology, 76, no. 2 (2023):ovad004,
https://doi.org/10.1093/lambio/ovad004 . .

TY  - JOUR
AU  - Gardijan, Lazar
AU  - Miljković, Marija
AU  - Obradović, Mina
AU  - Borović, Branka
AU  - Vukotić, Goran
AU  - Jovanović, Goran
AU  - Kojić, Milan
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1603
AB  - Aims The aim of this study was to construct the improved pMAL expression vector to increase the efficacy of purification of small native peptides and their clear-cut separation from MBP tag. The modifications we introduced can be applied to many expression vectors. Methods and Results To improve the pMAL expression vector, we introduced the His(6) tag and the enterokinase cleavage site (Ek) downstream from the MBP tag and Xa cleavage site on the original vector. For cloning of a desired peptide DNA, the enterokinase site contains a unique BsaBI restriction site adjacent to the original multi-cloning site. This redesigned pMAL vector was optimized for the purification of cytoplasmic (pMALc5HisEk) and periplasmic (pMALp5HisEk) peptides. The purification of native and active peptide (P) was obtained following two-step affinity chromatography. In the first step, the entire MBP-His(6)-Ek-P fusion protein is purified using the Ni-NTA agarose column. This fusion protein was cleaved with active His(6) tagged enterokinase. In the second step, the further purification was performed by column containing the mixture of amylose and Ni-NTA agarose resins. This removes both the MBP-His(6) and His(6)-enterokinase leaving pure native protein in solution. These new vectors and the two-step purification protocol were successfully applied in purification of active native small antimicrobial peptides (AMPs), lactococcin A and human beta-defensin. Conclusions We constructed the improved pMAL expression vectors and established the pipeline and optimal conditions for their use in efficient purification of large amounts of active native small peptides. Significance and Impact of the Study Choice of expression vector impacts on the efficiency of expression and purification of desired proteins. The idea of redesigning pMAL vector was driven by the need for rapid purification of larger amounts of active native AMPs. This newly improved pMAL vector, the cloning strategy, expression conditions and two-step purification protocol represent a unique simple approach which can be applied in every laboratory.
PB  - Wiley, Hoboken
T2  - Journal of Applied Microbiology
T1  - Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides
EP  - 1013
IS  - 2
SP  - 1001
VL  - 133
DO  - 10.1111/jam.15623
ER  - 

@article{
author = "Gardijan, Lazar and Miljković, Marija and Obradović, Mina and Borović, Branka and Vukotić, Goran and Jovanović, Goran and Kojić, Milan",
year = "2022",
abstract = "Aims The aim of this study was to construct the improved pMAL expression vector to increase the efficacy of purification of small native peptides and their clear-cut separation from MBP tag. The modifications we introduced can be applied to many expression vectors. Methods and Results To improve the pMAL expression vector, we introduced the His(6) tag and the enterokinase cleavage site (Ek) downstream from the MBP tag and Xa cleavage site on the original vector. For cloning of a desired peptide DNA, the enterokinase site contains a unique BsaBI restriction site adjacent to the original multi-cloning site. This redesigned pMAL vector was optimized for the purification of cytoplasmic (pMALc5HisEk) and periplasmic (pMALp5HisEk) peptides. The purification of native and active peptide (P) was obtained following two-step affinity chromatography. In the first step, the entire MBP-His(6)-Ek-P fusion protein is purified using the Ni-NTA agarose column. This fusion protein was cleaved with active His(6) tagged enterokinase. In the second step, the further purification was performed by column containing the mixture of amylose and Ni-NTA agarose resins. This removes both the MBP-His(6) and His(6)-enterokinase leaving pure native protein in solution. These new vectors and the two-step purification protocol were successfully applied in purification of active native small antimicrobial peptides (AMPs), lactococcin A and human beta-defensin. Conclusions We constructed the improved pMAL expression vectors and established the pipeline and optimal conditions for their use in efficient purification of large amounts of active native small peptides. Significance and Impact of the Study Choice of expression vector impacts on the efficiency of expression and purification of desired proteins. The idea of redesigning pMAL vector was driven by the need for rapid purification of larger amounts of active native AMPs. This newly improved pMAL vector, the cloning strategy, expression conditions and two-step purification protocol represent a unique simple approach which can be applied in every laboratory.",
publisher = "Wiley, Hoboken",
journal = "Journal of Applied Microbiology",
title = "Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides",
pages = "1013-1001",
number = "2",
volume = "133",
doi = "10.1111/jam.15623"
}

Gardijan, L., Miljković, M., Obradović, M., Borović, B., Vukotić, G., Jovanović, G.,& Kojić, M.. (2022). Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides. in Journal of Applied Microbiology
Wiley, Hoboken., 133(2), 1001-1013.
https://doi.org/10.1111/jam.15623

Gardijan L, Miljković M, Obradović M, Borović B, Vukotić G, Jovanović G, Kojić M. Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides. in Journal of Applied Microbiology. 2022;133(2):1001-1013.
doi:10.1111/jam.15623 .

Gardijan, Lazar, Miljković, Marija, Obradović, Mina, Borović, Branka, Vukotić, Goran, Jovanović, Goran, Kojić, Milan, "Redesigned pMAL expression vector for easy and fast purification of active native antimicrobial peptides" in Journal of Applied Microbiology, 133, no. 2 (2022):1001-1013,
https://doi.org/10.1111/jam.15623 . .

TY  - JOUR
AU  - Malešević, Milka
AU  - Stanisavljević, Nemanja
AU  - Miljković, Marija
AU  - Jovčić, Branko
AU  - Filipić, Brankica
AU  - Studholme, David J.
AU  - Kojić, Milan
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1507
AB  - Plasmids are autonomous episomally replicating genetic elements, which carry backbone genes important for the replication and maintenance within their host, and accessory genes that might confer an advantage to their host under specific selective pressure in its ecological niche. The genome of dairy isolate L. lactis subsp. lactis by. diacetylactis S50 was sequenced using the PacBio SMRT Cell Seq-RSII platform and revealed to possess one of the largest plasmidomes among L. lactis strains studied so far, harboring six plasmids: pS6 (5553 bp), pS7a (7308 bp), pS7b (7266 bp), pS19 (19,027 bp), pS74 (74,256 bp) and pS127 (127,002 bp) in total representing 8.9% of genome size (240,412 bp). Based on predicted plasmid replication proteins and origins it appears that all six plasmids replicate via the theta-type mechanism. The two the largest plasmids (pS74 and pS127), carry a number of genes known to be important for growth and survival in the dairy environment. These genes encode technological functions such as bacteriocin production, protein degradation, magnesium and cobalt/nickel transporters, selenium binding, exopolysaccharides (EPS) production, bacteriophage and stress resistance. Beside genes for replication, the small plasmids (pS6, pS7a, pS7a, and pS19) also carry genes important for mobilization and host survival such as type I restriction-modification (R-M) system, metal transporters, enzymes and transcriptional regulators. All plasmids in S50 strain are mobilizable, containing an oriT sequences, while pS127 is self-conjugative and allows for mobilization of the other plasmids. Small plasmids are prone to structural and segregational instability, while pS127 appeared to be segregationally stable thanks to the possession of two partition systems. The main characteristic of plasmid p574 is EPS production, while plasmid p5127 is characterized by proteinase and multiple bacteriocins, tra locus, phage abortive systems and metal transporters. In addition to LcnA and LcnB, plasmid p5127 encodes several bacteriocin-pheromone molecules and a new bacteriocin named LcnS50, with narrow spectrum of action limited to lactococci, that has been successfully cloned and heterologously expressed.
PB  - Elsevier, Amsterdam
T2  - International Journal of Food Microbiology
T1  - The large plasmidome of Lactococcus lactis subsp. lactis by. diacetylactis S50 confers its biotechnological properties
VL  - 337
DO  - 10.1016/j.ijfoodmicro.2020.108935
ER  - 

@article{
author = "Malešević, Milka and Stanisavljević, Nemanja and Miljković, Marija and Jovčić, Branko and Filipić, Brankica and Studholme, David J. and Kojić, Milan",
year = "2021",
abstract = "Plasmids are autonomous episomally replicating genetic elements, which carry backbone genes important for the replication and maintenance within their host, and accessory genes that might confer an advantage to their host under specific selective pressure in its ecological niche. The genome of dairy isolate L. lactis subsp. lactis by. diacetylactis S50 was sequenced using the PacBio SMRT Cell Seq-RSII platform and revealed to possess one of the largest plasmidomes among L. lactis strains studied so far, harboring six plasmids: pS6 (5553 bp), pS7a (7308 bp), pS7b (7266 bp), pS19 (19,027 bp), pS74 (74,256 bp) and pS127 (127,002 bp) in total representing 8.9% of genome size (240,412 bp). Based on predicted plasmid replication proteins and origins it appears that all six plasmids replicate via the theta-type mechanism. The two the largest plasmids (pS74 and pS127), carry a number of genes known to be important for growth and survival in the dairy environment. These genes encode technological functions such as bacteriocin production, protein degradation, magnesium and cobalt/nickel transporters, selenium binding, exopolysaccharides (EPS) production, bacteriophage and stress resistance. Beside genes for replication, the small plasmids (pS6, pS7a, pS7a, and pS19) also carry genes important for mobilization and host survival such as type I restriction-modification (R-M) system, metal transporters, enzymes and transcriptional regulators. All plasmids in S50 strain are mobilizable, containing an oriT sequences, while pS127 is self-conjugative and allows for mobilization of the other plasmids. Small plasmids are prone to structural and segregational instability, while pS127 appeared to be segregationally stable thanks to the possession of two partition systems. The main characteristic of plasmid p574 is EPS production, while plasmid p5127 is characterized by proteinase and multiple bacteriocins, tra locus, phage abortive systems and metal transporters. In addition to LcnA and LcnB, plasmid p5127 encodes several bacteriocin-pheromone molecules and a new bacteriocin named LcnS50, with narrow spectrum of action limited to lactococci, that has been successfully cloned and heterologously expressed.",
publisher = "Elsevier, Amsterdam",
journal = "International Journal of Food Microbiology",
title = "The large plasmidome of Lactococcus lactis subsp. lactis by. diacetylactis S50 confers its biotechnological properties",
volume = "337",
doi = "10.1016/j.ijfoodmicro.2020.108935"
}

Malešević, M., Stanisavljević, N., Miljković, M., Jovčić, B., Filipić, B., Studholme, D. J.,& Kojić, M.. (2021). The large plasmidome of Lactococcus lactis subsp. lactis by. diacetylactis S50 confers its biotechnological properties. in International Journal of Food Microbiology
Elsevier, Amsterdam., 337.
https://doi.org/10.1016/j.ijfoodmicro.2020.108935

Malešević M, Stanisavljević N, Miljković M, Jovčić B, Filipić B, Studholme DJ, Kojić M. The large plasmidome of Lactococcus lactis subsp. lactis by. diacetylactis S50 confers its biotechnological properties. in International Journal of Food Microbiology. 2021;337.
doi:10.1016/j.ijfoodmicro.2020.108935 .

Malešević, Milka, Stanisavljević, Nemanja, Miljković, Marija, Jovčić, Branko, Filipić, Brankica, Studholme, David J., Kojić, Milan, "The large plasmidome of Lactococcus lactis subsp. lactis by. diacetylactis S50 confers its biotechnological properties" in International Journal of Food Microbiology, 337 (2021),
https://doi.org/10.1016/j.ijfoodmicro.2020.108935 . .

TY  - JOUR
AU  - Kojić, Milan
AU  - Jovčić, Branko
AU  - Miljković, Marija
AU  - Novović, Katarina
AU  - Begović, Jelena
AU  - Studholme, David J.
PY  - 2020
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1373
AB  - Bacteria can gain resistance to antimicrobials by acquiring and expressing genetic elements that encode resistance determinants such as efflux pumps and drug-modifying enzymes, thus hampering treatment of infection. Previously we showed that acquisition of spectinomycin resistance in a lactococcal strain was correlated with a reversible genomic inversion, but the precise location and the genes affected were unknown. Here we use long read whole-genome sequencing to precisely define the genomic inversion and we use quantitative PCR to identify associated changes in gene expression levels. The boundaries of the inversion fall within two identical copies of a prophage-like sequence, located on the left and right replichores; this suggests possible mechanisms for inversion through homologous recombination or prophage activity. The inversion is asymmetrical in respect of the axis between the origin and terminus of the replication and modulates the expression of a SAM-dependent methyltransferase, whose heterologous expression confers resistance to spectinomycin in lactococci and that is up regulated on exposure to spectinomycin. This study provides one of the first examples of phase variation via large-scale chromosomal inversions that confers a switch in antimicrobial resistance in bacteria and the first outside of Staphylococcus aureus.
PB  - Elsevier Gmbh, Munich
T2  - Microbiological Research
T1  - Large-scale chromosome flip-flop reversible inversion mediates phenotypic switching of expression of antibiotic resistance in lactococci
VL  - 241
DO  - 10.1016/j.micres.2020.126583
ER  - 

@article{
author = "Kojić, Milan and Jovčić, Branko and Miljković, Marija and Novović, Katarina and Begović, Jelena and Studholme, David J.",
year = "2020",
abstract = "Bacteria can gain resistance to antimicrobials by acquiring and expressing genetic elements that encode resistance determinants such as efflux pumps and drug-modifying enzymes, thus hampering treatment of infection. Previously we showed that acquisition of spectinomycin resistance in a lactococcal strain was correlated with a reversible genomic inversion, but the precise location and the genes affected were unknown. Here we use long read whole-genome sequencing to precisely define the genomic inversion and we use quantitative PCR to identify associated changes in gene expression levels. The boundaries of the inversion fall within two identical copies of a prophage-like sequence, located on the left and right replichores; this suggests possible mechanisms for inversion through homologous recombination or prophage activity. The inversion is asymmetrical in respect of the axis between the origin and terminus of the replication and modulates the expression of a SAM-dependent methyltransferase, whose heterologous expression confers resistance to spectinomycin in lactococci and that is up regulated on exposure to spectinomycin. This study provides one of the first examples of phase variation via large-scale chromosomal inversions that confers a switch in antimicrobial resistance in bacteria and the first outside of Staphylococcus aureus.",
publisher = "Elsevier Gmbh, Munich",
journal = "Microbiological Research",
title = "Large-scale chromosome flip-flop reversible inversion mediates phenotypic switching of expression of antibiotic resistance in lactococci",
volume = "241",
doi = "10.1016/j.micres.2020.126583"
}

Kojić, M., Jovčić, B., Miljković, M., Novović, K., Begović, J.,& Studholme, D. J.. (2020). Large-scale chromosome flip-flop reversible inversion mediates phenotypic switching of expression of antibiotic resistance in lactococci. in Microbiological Research
Elsevier Gmbh, Munich., 241.
https://doi.org/10.1016/j.micres.2020.126583

Kojić M, Jovčić B, Miljković M, Novović K, Begović J, Studholme DJ. Large-scale chromosome flip-flop reversible inversion mediates phenotypic switching of expression of antibiotic resistance in lactococci. in Microbiological Research. 2020;241.
doi:10.1016/j.micres.2020.126583 .

Kojić, Milan, Jovčić, Branko, Miljković, Marija, Novović, Katarina, Begović, Jelena, Studholme, David J., "Large-scale chromosome flip-flop reversible inversion mediates phenotypic switching of expression of antibiotic resistance in lactococci" in Microbiological Research, 241 (2020),
https://doi.org/10.1016/j.micres.2020.126583 . .

TY  - JOUR
AU  - Terzić-Vidojević, Amarela
AU  - Veljović, Katarina
AU  - Tolinački, Maja
AU  - Živković, Milica
AU  - Lukić, Jovanka
AU  - Lozo, Jelena
AU  - Fira, Đorđe
AU  - Jovčić, Branko
AU  - Strahinić, Ivana
AU  - Begović, Jelena
AU  - Popović, Nikola
AU  - Miljković, Marija
AU  - Kojić, Milan
AU  - Topisirović, Ljubiša
AU  - Golić, Nataša
PY  - 2020
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1353
AB  - The aim of this review was to summarize the data regarding diversity of non-starter lactic acid bacteria (NSLAB) isolated from various artisanal dairy products manufactured in Western Balkan Countries. The dairy products examined were manufactured from raw cow's, sheep's or goat's milk or mixed milk, in the traditional way without the addition of commercial starter cultures. Dairy products such as white brined cheese, fresh cheese, hard cheese, yogurt, sour cream and kajmak were sampled in the households of Serbia, Croatia, Slovenia, Bosnia and Herzegovina, Montenegro, and North Macedonia. It has been established that the diversity of lactic acid bacteria (LAB) from raw milk artisanal dairy products is extensive. In the reviewed literature, 28 LAB species and a large number of strains belonging to the Lactobacillus, Lactococcus, Enterococcus, Streptococcus, Pediococcus, Leuconostoc and Weissella genera were isolated from various dairy products. Over 3000 LAB strains were obtained and characterized for their technological and probiotic properties including: acidification and coagulation of milk, production of aromatic compounds, proteolytic activity, bacteriocins production and competitive exclusion of pathogens, production of exopolysaccharides, aggregation ability and immunomodulatory effect. Results show that many of the isolated NSLAB strains had one, two or more of the properties mentioned. The data presented emphasize the importance of artisanal products as a valuable source of NSLAB with unique technological and probiotic features important both as a base for scientific research as well as for designing novel starter cultures for functional dairy food.
PB  - Elsevier, Amsterdam
T2  - Food Research International
T1  - Diversity of non-starter lactic acid bacteria in autochthonous dairy products from Western Balkan Countries - Technological and probiotic properties
VL  - 136
DO  - 10.1016/j.foodres.2020.109494
ER  - 

@article{
author = "Terzić-Vidojević, Amarela and Veljović, Katarina and Tolinački, Maja and Živković, Milica and Lukić, Jovanka and Lozo, Jelena and Fira, Đorđe and Jovčić, Branko and Strahinić, Ivana and Begović, Jelena and Popović, Nikola and Miljković, Marija and Kojić, Milan and Topisirović, Ljubiša and Golić, Nataša",
year = "2020",
abstract = "The aim of this review was to summarize the data regarding diversity of non-starter lactic acid bacteria (NSLAB) isolated from various artisanal dairy products manufactured in Western Balkan Countries. The dairy products examined were manufactured from raw cow's, sheep's or goat's milk or mixed milk, in the traditional way without the addition of commercial starter cultures. Dairy products such as white brined cheese, fresh cheese, hard cheese, yogurt, sour cream and kajmak were sampled in the households of Serbia, Croatia, Slovenia, Bosnia and Herzegovina, Montenegro, and North Macedonia. It has been established that the diversity of lactic acid bacteria (LAB) from raw milk artisanal dairy products is extensive. In the reviewed literature, 28 LAB species and a large number of strains belonging to the Lactobacillus, Lactococcus, Enterococcus, Streptococcus, Pediococcus, Leuconostoc and Weissella genera were isolated from various dairy products. Over 3000 LAB strains were obtained and characterized for their technological and probiotic properties including: acidification and coagulation of milk, production of aromatic compounds, proteolytic activity, bacteriocins production and competitive exclusion of pathogens, production of exopolysaccharides, aggregation ability and immunomodulatory effect. Results show that many of the isolated NSLAB strains had one, two or more of the properties mentioned. The data presented emphasize the importance of artisanal products as a valuable source of NSLAB with unique technological and probiotic features important both as a base for scientific research as well as for designing novel starter cultures for functional dairy food.",
publisher = "Elsevier, Amsterdam",
journal = "Food Research International",
title = "Diversity of non-starter lactic acid bacteria in autochthonous dairy products from Western Balkan Countries - Technological and probiotic properties",
volume = "136",
doi = "10.1016/j.foodres.2020.109494"
}

Terzić-Vidojević, A., Veljović, K., Tolinački, M., Živković, M., Lukić, J., Lozo, J., Fira, Đ., Jovčić, B., Strahinić, I., Begović, J., Popović, N., Miljković, M., Kojić, M., Topisirović, L.,& Golić, N.. (2020). Diversity of non-starter lactic acid bacteria in autochthonous dairy products from Western Balkan Countries - Technological and probiotic properties. in Food Research International
Elsevier, Amsterdam., 136.
https://doi.org/10.1016/j.foodres.2020.109494

Terzić-Vidojević A, Veljović K, Tolinački M, Živković M, Lukić J, Lozo J, Fira Đ, Jovčić B, Strahinić I, Begović J, Popović N, Miljković M, Kojić M, Topisirović L, Golić N. Diversity of non-starter lactic acid bacteria in autochthonous dairy products from Western Balkan Countries - Technological and probiotic properties. in Food Research International. 2020;136.
doi:10.1016/j.foodres.2020.109494 .

Terzić-Vidojević, Amarela, Veljović, Katarina, Tolinački, Maja, Živković, Milica, Lukić, Jovanka, Lozo, Jelena, Fira, Đorđe, Jovčić, Branko, Strahinić, Ivana, Begović, Jelena, Popović, Nikola, Miljković, Marija, Kojić, Milan, Topisirović, Ljubiša, Golić, Nataša, "Diversity of non-starter lactic acid bacteria in autochthonous dairy products from Western Balkan Countries - Technological and probiotic properties" in Food Research International, 136 (2020),
https://doi.org/10.1016/j.foodres.2020.109494 . .

TY  - JOUR
AU  - Mirković, Nemanja
AU  - Kulas, Jelena
AU  - Miloradović, Zorana
AU  - Miljković, Marija
AU  - Tucović, Dina
AU  - Miocinović, Jelena
AU  - Jovčić, Branko
AU  - Mirkov, Ivana
AU  - Kojić, Milan
PY  - 2020
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1388
AB  - In last two decades, there has been a strong trend in the application of lactic acid bacteria as adjunctive cultures to control growth of spoilage and pathogenic bacteria in food. One of the most important properties that contribute to the application of these bacteria is the production of antimicrobial molecules. Lactococcus lactis subsp. lactis BGBU1-4, isolated from traditional brined cheese, produces thermostable bacteriocin named lactolisterin BU, with broad spectrum of activity against spoilage bacteria and foodborne pathogens. In this study, effect of strain BGBU1-4, as adjunct culture, on the numbers of Listeria monocytogenes ATCC19111 and Staphylococcus aureus LMM322 in artificially contaminated Quark-type, soft acid coagulated cheese, was examined. In addition, we analyzed influence of BGBU1-4 on chemical and sensory properties of the cheese, as well as immunological response of Albino oxford rats fed with Quark-type of cheese made using BGBU1-4 as adjunct culture. Results of this study revealed antibacterial potential of strain BGBU1-4 against L. monocytogenes ATCC19111 and S. aureus LMM322 in Quark-type cheese during 21 days of storage at 4 degrees C. Also, it was noticed the ability of BGBU1-4 to control the spontaneously grown yeasts and molds. Chemical composition and pH values of cheese containing BGBU1-4 were unchanged in comparison to control. The sensory quality scores showed that there was difference between cheese with and without adjunct culture in terms of flavor and oral texture, while for the odor and appearance no differences between two cheese variants were scored. Results of the immunological response of Albino rats fed with Quark-type cheese containing BGBU1-4 indicate absence of systematic inflammation. However, increased pro-inflammatory cytokines content (IL-1 beta, IL-6, IL-17) in intestine of rats fed with cheese containing BGBU1-4, concomitantly with unchanged anti-inflammatory cytokines suggests disruption of gut homeostasis and inflammation in this tissue. The changes caused by BGBU1-4 are reversible, system returns into homeostasis seven days after cessation of feeding with cheese containing BGBU1-4.
PB  - Elsevier Sci Ltd, Oxford
T2  - Food Control
T1  - Lactolisterin BU-producer Lactococcus lactis subsp. lactis BGBU1-4: Biocontrol of Listeria monocytogenes and Staphylocococcus aureus in fresh soft cheese and effect on immunological response of rats
VL  - 111
DO  - 10.1016/j.foodcont.2019.107076
ER  - 

@article{
author = "Mirković, Nemanja and Kulas, Jelena and Miloradović, Zorana and Miljković, Marija and Tucović, Dina and Miocinović, Jelena and Jovčić, Branko and Mirkov, Ivana and Kojić, Milan",
year = "2020",
abstract = "In last two decades, there has been a strong trend in the application of lactic acid bacteria as adjunctive cultures to control growth of spoilage and pathogenic bacteria in food. One of the most important properties that contribute to the application of these bacteria is the production of antimicrobial molecules. Lactococcus lactis subsp. lactis BGBU1-4, isolated from traditional brined cheese, produces thermostable bacteriocin named lactolisterin BU, with broad spectrum of activity against spoilage bacteria and foodborne pathogens. In this study, effect of strain BGBU1-4, as adjunct culture, on the numbers of Listeria monocytogenes ATCC19111 and Staphylococcus aureus LMM322 in artificially contaminated Quark-type, soft acid coagulated cheese, was examined. In addition, we analyzed influence of BGBU1-4 on chemical and sensory properties of the cheese, as well as immunological response of Albino oxford rats fed with Quark-type of cheese made using BGBU1-4 as adjunct culture. Results of this study revealed antibacterial potential of strain BGBU1-4 against L. monocytogenes ATCC19111 and S. aureus LMM322 in Quark-type cheese during 21 days of storage at 4 degrees C. Also, it was noticed the ability of BGBU1-4 to control the spontaneously grown yeasts and molds. Chemical composition and pH values of cheese containing BGBU1-4 were unchanged in comparison to control. The sensory quality scores showed that there was difference between cheese with and without adjunct culture in terms of flavor and oral texture, while for the odor and appearance no differences between two cheese variants were scored. Results of the immunological response of Albino rats fed with Quark-type cheese containing BGBU1-4 indicate absence of systematic inflammation. However, increased pro-inflammatory cytokines content (IL-1 beta, IL-6, IL-17) in intestine of rats fed with cheese containing BGBU1-4, concomitantly with unchanged anti-inflammatory cytokines suggests disruption of gut homeostasis and inflammation in this tissue. The changes caused by BGBU1-4 are reversible, system returns into homeostasis seven days after cessation of feeding with cheese containing BGBU1-4.",
publisher = "Elsevier Sci Ltd, Oxford",
journal = "Food Control",
title = "Lactolisterin BU-producer Lactococcus lactis subsp. lactis BGBU1-4: Biocontrol of Listeria monocytogenes and Staphylocococcus aureus in fresh soft cheese and effect on immunological response of rats",
volume = "111",
doi = "10.1016/j.foodcont.2019.107076"
}

Mirković, N., Kulas, J., Miloradović, Z., Miljković, M., Tucović, D., Miocinović, J., Jovčić, B., Mirkov, I.,& Kojić, M.. (2020). Lactolisterin BU-producer Lactococcus lactis subsp. lactis BGBU1-4: Biocontrol of Listeria monocytogenes and Staphylocococcus aureus in fresh soft cheese and effect on immunological response of rats. in Food Control
Elsevier Sci Ltd, Oxford., 111.
https://doi.org/10.1016/j.foodcont.2019.107076

Mirković N, Kulas J, Miloradović Z, Miljković M, Tucović D, Miocinović J, Jovčić B, Mirkov I, Kojić M. Lactolisterin BU-producer Lactococcus lactis subsp. lactis BGBU1-4: Biocontrol of Listeria monocytogenes and Staphylocococcus aureus in fresh soft cheese and effect on immunological response of rats. in Food Control. 2020;111.
doi:10.1016/j.foodcont.2019.107076 .

Mirković, Nemanja, Kulas, Jelena, Miloradović, Zorana, Miljković, Marija, Tucović, Dina, Miocinović, Jelena, Jovčić, Branko, Mirkov, Ivana, Kojić, Milan, "Lactolisterin BU-producer Lactococcus lactis subsp. lactis BGBU1-4: Biocontrol of Listeria monocytogenes and Staphylocococcus aureus in fresh soft cheese and effect on immunological response of rats" in Food Control, 111 (2020),
https://doi.org/10.1016/j.foodcont.2019.107076 . .

TY  - JOUR
AU  - Tomović, Katarina
AU  - Ilić, Budimir S.
AU  - Smelcerović, Zaklina
AU  - Miljković, Marija
AU  - Yancheva, Denitsa
AU  - Kojić, Milan
AU  - Mavrova, Anelia Ts
AU  - Kocić, Gordana
AU  - Smelcerović, Andrija
PY  - 2020
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1376
AB  - Multiple-targeting compounds might reduce complex polypharmacy of multifactorial diseases, such as diabetes, and contribute to the greater therapeutic success. Targeting reactive oxygen species-producing enzymes, as xanthine oxidase (XO), might suppress progression of diabetes-associated vascular complications. In this study a small series of benzimidazole derivatives (1-9) was evaluated for inhibitory activity against dipeptidyl peptidase-4 (DPP-4) and XO. One 1,3-disubstituted-benzimidazole-2-imine (5) and 1,3-thiazolo [3,2-a] benzimidazolone derivative (8) were shown as effective dual DPP-4 and XO inhibitors, with IC50 values lower than 200 mu M, and predicted binding modes with both target enzymes. Both selected dual inhibitors (compounds 5 and 8) did not show cytotoxicity to a greater extent on Caco-2 cells even at concentration of 250 mu M. These structures represent new non-purine scaffolds bearing two therapeutic functionalities, being DPP-4 and XO inhibitors, more favorable in comparison to DPP-4 inhibitors with DPP-4 as a single target due to pleiotropic effects of XO inhibition.
PB  - Elsevier Ireland Ltd, Clare
T2  - Chemico-Biological Interactions
T1  - Benzimidazole-based dual dipeptidyl peptidase-4 and xanthine oxidase inhibitors
VL  - 315
DO  - 10.1016/j.cbi.2019.108873
ER  - 

@article{
author = "Tomović, Katarina and Ilić, Budimir S. and Smelcerović, Zaklina and Miljković, Marija and Yancheva, Denitsa and Kojić, Milan and Mavrova, Anelia Ts and Kocić, Gordana and Smelcerović, Andrija",
year = "2020",
abstract = "Multiple-targeting compounds might reduce complex polypharmacy of multifactorial diseases, such as diabetes, and contribute to the greater therapeutic success. Targeting reactive oxygen species-producing enzymes, as xanthine oxidase (XO), might suppress progression of diabetes-associated vascular complications. In this study a small series of benzimidazole derivatives (1-9) was evaluated for inhibitory activity against dipeptidyl peptidase-4 (DPP-4) and XO. One 1,3-disubstituted-benzimidazole-2-imine (5) and 1,3-thiazolo [3,2-a] benzimidazolone derivative (8) were shown as effective dual DPP-4 and XO inhibitors, with IC50 values lower than 200 mu M, and predicted binding modes with both target enzymes. Both selected dual inhibitors (compounds 5 and 8) did not show cytotoxicity to a greater extent on Caco-2 cells even at concentration of 250 mu M. These structures represent new non-purine scaffolds bearing two therapeutic functionalities, being DPP-4 and XO inhibitors, more favorable in comparison to DPP-4 inhibitors with DPP-4 as a single target due to pleiotropic effects of XO inhibition.",
publisher = "Elsevier Ireland Ltd, Clare",
journal = "Chemico-Biological Interactions",
title = "Benzimidazole-based dual dipeptidyl peptidase-4 and xanthine oxidase inhibitors",
volume = "315",
doi = "10.1016/j.cbi.2019.108873"
}

Tomović, K., Ilić, B. S., Smelcerović, Z., Miljković, M., Yancheva, D., Kojić, M., Mavrova, A. T., Kocić, G.,& Smelcerović, A.. (2020). Benzimidazole-based dual dipeptidyl peptidase-4 and xanthine oxidase inhibitors. in Chemico-Biological Interactions
Elsevier Ireland Ltd, Clare., 315.
https://doi.org/10.1016/j.cbi.2019.108873

Tomović K, Ilić BS, Smelcerović Z, Miljković M, Yancheva D, Kojić M, Mavrova AT, Kocić G, Smelcerović A. Benzimidazole-based dual dipeptidyl peptidase-4 and xanthine oxidase inhibitors. in Chemico-Biological Interactions. 2020;315.
doi:10.1016/j.cbi.2019.108873 .

Tomović, Katarina, Ilić, Budimir S., Smelcerović, Zaklina, Miljković, Marija, Yancheva, Denitsa, Kojić, Milan, Mavrova, Anelia Ts, Kocić, Gordana, Smelcerović, Andrija, "Benzimidazole-based dual dipeptidyl peptidase-4 and xanthine oxidase inhibitors" in Chemico-Biological Interactions, 315 (2020),
https://doi.org/10.1016/j.cbi.2019.108873 . .

TY  - JOUR
AU  - Tomović, Katarina
AU  - Ilić, Budimir S.
AU  - Miljković, Marija
AU  - Dimov, Stefan
AU  - Yancheva, Denitsa
AU  - Kojić, Milan
AU  - Mavrova, Anelia T.
AU  - Kocić, Gordana
AU  - Smelcerović, Andrija
PY  - 2019
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1396
AB  - A small library of benzo[4,5]thieno[2,3-d]pyrimidine phthalimide and amine derivatives was evaluated for inhibitory activity against dipeptidyl peptidase-4 (DPP-4). The phthalimide derivatives exhibited better activity than the amine precursors, with 2-(2-(3-chlorobenzyl)-5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidin-4-yl)isoindoline-1,3-dione (compound 14) as the most effective inhibitor (IC50 = 34.17 +/- 5.11 mu M). The five most potent selected inhibitors did not show cytotoxicity to a greater extent on Caco-2 cells, even at a concentration of 250 mu M. Compound 14 is considered as a novel representative of the rare noncompetitive DPP-4 inhibitors. Molecular docking and dynamics simulation indicated the importance of the Tyr547, Lys554, and Trp629 residues of DPP-4 in the formation of the enzyme-inhibitor complex. These observations could be potentially utilized for the rational design and optimization of novel (structurally similar, with phthalimide moiety, or different) noncompetitive DPP-4 inhibitors, which are anyway rare, but favorable in terms of the saturation of substrate competition.
PB  - Wiley-V C H Verlag Gmbh, Weinheim
T2  - Archiv Der Pharmazie
T1  - Benzo[4,5]thieno[2,3-d]pyrimidine phthalimide derivative, one of the rare noncompetitive inhibitors of dipeptidyl peptidase-4
IS  - 1
VL  - 353
DO  - 10.1002/ardp.201900238
ER  - 

@article{
author = "Tomović, Katarina and Ilić, Budimir S. and Miljković, Marija and Dimov, Stefan and Yancheva, Denitsa and Kojić, Milan and Mavrova, Anelia T. and Kocić, Gordana and Smelcerović, Andrija",
year = "2019",
abstract = "A small library of benzo[4,5]thieno[2,3-d]pyrimidine phthalimide and amine derivatives was evaluated for inhibitory activity against dipeptidyl peptidase-4 (DPP-4). The phthalimide derivatives exhibited better activity than the amine precursors, with 2-(2-(3-chlorobenzyl)-5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidin-4-yl)isoindoline-1,3-dione (compound 14) as the most effective inhibitor (IC50 = 34.17 +/- 5.11 mu M). The five most potent selected inhibitors did not show cytotoxicity to a greater extent on Caco-2 cells, even at a concentration of 250 mu M. Compound 14 is considered as a novel representative of the rare noncompetitive DPP-4 inhibitors. Molecular docking and dynamics simulation indicated the importance of the Tyr547, Lys554, and Trp629 residues of DPP-4 in the formation of the enzyme-inhibitor complex. These observations could be potentially utilized for the rational design and optimization of novel (structurally similar, with phthalimide moiety, or different) noncompetitive DPP-4 inhibitors, which are anyway rare, but favorable in terms of the saturation of substrate competition.",
publisher = "Wiley-V C H Verlag Gmbh, Weinheim",
journal = "Archiv Der Pharmazie",
title = "Benzo[4,5]thieno[2,3-d]pyrimidine phthalimide derivative, one of the rare noncompetitive inhibitors of dipeptidyl peptidase-4",
number = "1",
volume = "353",
doi = "10.1002/ardp.201900238"
}

Tomović, K., Ilić, B. S., Miljković, M., Dimov, S., Yancheva, D., Kojić, M., Mavrova, A. T., Kocić, G.,& Smelcerović, A.. (2019). Benzo[4,5]thieno[2,3-d]pyrimidine phthalimide derivative, one of the rare noncompetitive inhibitors of dipeptidyl peptidase-4. in Archiv Der Pharmazie
Wiley-V C H Verlag Gmbh, Weinheim., 353(1).
https://doi.org/10.1002/ardp.201900238

Tomović K, Ilić BS, Miljković M, Dimov S, Yancheva D, Kojić M, Mavrova AT, Kocić G, Smelcerović A. Benzo[4,5]thieno[2,3-d]pyrimidine phthalimide derivative, one of the rare noncompetitive inhibitors of dipeptidyl peptidase-4. in Archiv Der Pharmazie. 2019;353(1).
doi:10.1002/ardp.201900238 .

Tomović, Katarina, Ilić, Budimir S., Miljković, Marija, Dimov, Stefan, Yancheva, Denitsa, Kojić, Milan, Mavrova, Anelia T., Kocić, Gordana, Smelcerović, Andrija, "Benzo[4,5]thieno[2,3-d]pyrimidine phthalimide derivative, one of the rare noncompetitive inhibitors of dipeptidyl peptidase-4" in Archiv Der Pharmazie, 353, no. 1 (2019),
https://doi.org/10.1002/ardp.201900238 . .

TY  - JOUR
AU  - Miljković, Marija
AU  - Jovanović, Sofija
AU  - O'Connor, Paula M.
AU  - Mirković, Nemanja
AU  - Jovčić, Branko
AU  - Filipić, Brankica
AU  - Dinić, Miroslav
AU  - Studholme, David John
AU  - Fira, Đorđe
AU  - Cotter, Paul D.
AU  - Kojić, Milan
PY  - 2019
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1264
AB  - Bacteria active against multi-drug resistant pathogens, isolated by direct selection of colonies from clover silage samples, produce zones of inhibition against two Gram-negative (Klebsiella pneumoniae Ni9 and Pseudomonas aeruginosa MMA83) and two Gram-positive (Staphylococcus aureus ATCC25923 and Listeria monocytogenes ATCC19111) pathogens. Isolates BGSP7, BGSP9, BGSP11 and BGSP12 produced the largest zones of inhibition against all four pathogens when grown in LB broth with aeration at 37 degrees C. Isolates BGSP7, BGSP9, BGSP11 and BGSP12 were identified as Brevibacillus laterosporus and pulsed field gel electrophoresis and extracellular protein profiles showed that three different strains (BGSP7, BGSP9 and BGSP11) were isolated. A semi-native SDS-PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) gel overlay assay showed that BGSP7 and BGSP9 produce small antimicrobial molecules of about 1.5 kDa, while BGSP11 produces antimicrobial molecules of 1.5 and 6 kDa active against S. aureus ATCC25923. Amino acid analysis of two antimicrobial molecules (1583.73 Da; from BGSP7 and 1556.31 Da; from BGSP11) revealed that they have a similar composition and differ only by virtue of the presence of a methionine which is present only in BGSP11 molecule. Genome sequencing of the three isolates revealed the presence of gene clusters associated with the production of non-ribosomally synthesized peptides (brevibacillin, bogorol, gramicidin S, plipastatin and tyrocin) and bacteriocins (laterosporulin, a lactococcin 972-like bacteriocin, as well as putative linocin M18, sactipeptide, UviB and lantipeptide-like molecules). Ultimately, the purification of a number of antimicrobial molecules from each isolate suggests that they can be considered as potent biocontrol strains that produce an arsenal of antimicrobial molecules active against Gram-positive and Gram-negative multi-resistant pathogens, fungi and insects.
PB  - Public Library Science, San Francisco
T2  - PLoS One
T1  - Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials
IS  - 5
VL  - 14
DO  - 10.1371/journal.pone.0216773
ER  - 

@article{
author = "Miljković, Marija and Jovanović, Sofija and O'Connor, Paula M. and Mirković, Nemanja and Jovčić, Branko and Filipić, Brankica and Dinić, Miroslav and Studholme, David John and Fira, Đorđe and Cotter, Paul D. and Kojić, Milan",
year = "2019",
abstract = "Bacteria active against multi-drug resistant pathogens, isolated by direct selection of colonies from clover silage samples, produce zones of inhibition against two Gram-negative (Klebsiella pneumoniae Ni9 and Pseudomonas aeruginosa MMA83) and two Gram-positive (Staphylococcus aureus ATCC25923 and Listeria monocytogenes ATCC19111) pathogens. Isolates BGSP7, BGSP9, BGSP11 and BGSP12 produced the largest zones of inhibition against all four pathogens when grown in LB broth with aeration at 37 degrees C. Isolates BGSP7, BGSP9, BGSP11 and BGSP12 were identified as Brevibacillus laterosporus and pulsed field gel electrophoresis and extracellular protein profiles showed that three different strains (BGSP7, BGSP9 and BGSP11) were isolated. A semi-native SDS-PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) gel overlay assay showed that BGSP7 and BGSP9 produce small antimicrobial molecules of about 1.5 kDa, while BGSP11 produces antimicrobial molecules of 1.5 and 6 kDa active against S. aureus ATCC25923. Amino acid analysis of two antimicrobial molecules (1583.73 Da; from BGSP7 and 1556.31 Da; from BGSP11) revealed that they have a similar composition and differ only by virtue of the presence of a methionine which is present only in BGSP11 molecule. Genome sequencing of the three isolates revealed the presence of gene clusters associated with the production of non-ribosomally synthesized peptides (brevibacillin, bogorol, gramicidin S, plipastatin and tyrocin) and bacteriocins (laterosporulin, a lactococcin 972-like bacteriocin, as well as putative linocin M18, sactipeptide, UviB and lantipeptide-like molecules). Ultimately, the purification of a number of antimicrobial molecules from each isolate suggests that they can be considered as potent biocontrol strains that produce an arsenal of antimicrobial molecules active against Gram-positive and Gram-negative multi-resistant pathogens, fungi and insects.",
publisher = "Public Library Science, San Francisco",
journal = "PLoS One",
title = "Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials",
number = "5",
volume = "14",
doi = "10.1371/journal.pone.0216773"
}

Miljković, M., Jovanović, S., O'Connor, P. M., Mirković, N., Jovčić, B., Filipić, B., Dinić, M., Studholme, D. J., Fira, Đ., Cotter, P. D.,& Kojić, M.. (2019). Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials. in PLoS One
Public Library Science, San Francisco., 14(5).
https://doi.org/10.1371/journal.pone.0216773

Miljković M, Jovanović S, O'Connor PM, Mirković N, Jovčić B, Filipić B, Dinić M, Studholme DJ, Fira Đ, Cotter PD, Kojić M. Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials. in PLoS One. 2019;14(5).
doi:10.1371/journal.pone.0216773 .

Miljković, Marija, Jovanović, Sofija, O'Connor, Paula M., Mirković, Nemanja, Jovčić, Branko, Filipić, Brankica, Dinić, Miroslav, Studholme, David John, Fira, Đorđe, Cotter, Paul D., Kojić, Milan, "Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials" in PLoS One, 14, no. 5 (2019),
https://doi.org/10.1371/journal.pone.0216773 . .

TY  - JOUR
AU  - Miljković, Marija
AU  - Thomas, Muriel
AU  - Serror, Pascale
AU  - Rigottier-Gois, Lionel
AU  - Kojić, Milan
PY  - 2019
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1259
AB  - Surface properties like hydrophobicity, aggregation ability, adhesion to mucosal surfaces and epithelial cells and transit time are key features for the characterization of probiotic strains. In this study, we used two Lactobacillus paracasei subsp. paracasei strains (BGNJ1-64 and BGSJ2-8) strains which were previously described with very strong aggregation capacity. The aggregation promoting factor (AggLb) expressed in these strains showed high level of binding to collagen and fibronectin, components of extracellular matrix. The working hypothesis was that strains able to aggregate have an advantage to resist in intestinal tract. So, we assessed whether these strains and their derivatives (without aggLb gene) are able to bind or not to intestinal components and we compared the transit time of each strains in mice. In that purpose parental strains (BGNJ1-64 and BGSJ2-8) and their aggregation negative derivatives (BGNJ1-641 and BGSJ2-83) were marked with double antibiotic resistance in order to be tracked in in vivo experiments in mice. Comparative analysis of binding ability of WT and aggregation negative strains to different human intestinal cell lines and mucin revealed no significant difference among them, excluding involvement of AggLb in interaction with surface of intestinal cells and mucin. In vivo experiments showed that surviving and transit time of marked strains in mice did not drastically depend on the presence of the AggLb aggregation factor.
PB  - Springer, Dordrecht
T2  - World Journal of Microbiology & Biotechnology
T1  - Binding activity to intestinal cells and transient colonization in mice of two Lactobacillus paracasei subsp. paracasei strains with high aggregation potential
IS  - 6
VL  - 35
DO  - 10.1007/s11274-019-2663-4
ER  - 

@article{
author = "Miljković, Marija and Thomas, Muriel and Serror, Pascale and Rigottier-Gois, Lionel and Kojić, Milan",
year = "2019",
abstract = "Surface properties like hydrophobicity, aggregation ability, adhesion to mucosal surfaces and epithelial cells and transit time are key features for the characterization of probiotic strains. In this study, we used two Lactobacillus paracasei subsp. paracasei strains (BGNJ1-64 and BGSJ2-8) strains which were previously described with very strong aggregation capacity. The aggregation promoting factor (AggLb) expressed in these strains showed high level of binding to collagen and fibronectin, components of extracellular matrix. The working hypothesis was that strains able to aggregate have an advantage to resist in intestinal tract. So, we assessed whether these strains and their derivatives (without aggLb gene) are able to bind or not to intestinal components and we compared the transit time of each strains in mice. In that purpose parental strains (BGNJ1-64 and BGSJ2-8) and their aggregation negative derivatives (BGNJ1-641 and BGSJ2-83) were marked with double antibiotic resistance in order to be tracked in in vivo experiments in mice. Comparative analysis of binding ability of WT and aggregation negative strains to different human intestinal cell lines and mucin revealed no significant difference among them, excluding involvement of AggLb in interaction with surface of intestinal cells and mucin. In vivo experiments showed that surviving and transit time of marked strains in mice did not drastically depend on the presence of the AggLb aggregation factor.",
publisher = "Springer, Dordrecht",
journal = "World Journal of Microbiology & Biotechnology",
title = "Binding activity to intestinal cells and transient colonization in mice of two Lactobacillus paracasei subsp. paracasei strains with high aggregation potential",
number = "6",
volume = "35",
doi = "10.1007/s11274-019-2663-4"
}

Miljković, M., Thomas, M., Serror, P., Rigottier-Gois, L.,& Kojić, M.. (2019). Binding activity to intestinal cells and transient colonization in mice of two Lactobacillus paracasei subsp. paracasei strains with high aggregation potential. in World Journal of Microbiology & Biotechnology
Springer, Dordrecht., 35(6).
https://doi.org/10.1007/s11274-019-2663-4

Miljković M, Thomas M, Serror P, Rigottier-Gois L, Kojić M. Binding activity to intestinal cells and transient colonization in mice of two Lactobacillus paracasei subsp. paracasei strains with high aggregation potential. in World Journal of Microbiology & Biotechnology. 2019;35(6).
doi:10.1007/s11274-019-2663-4 .

Miljković, Marija, Thomas, Muriel, Serror, Pascale, Rigottier-Gois, Lionel, Kojić, Milan, "Binding activity to intestinal cells and transient colonization in mice of two Lactobacillus paracasei subsp. paracasei strains with high aggregation potential" in World Journal of Microbiology & Biotechnology, 35, no. 6 (2019),
https://doi.org/10.1007/s11274-019-2663-4 . .

TY  - JOUR
AU  - Bjelogrlić, Snežana K.
AU  - Todorović, Tamara R.
AU  - Kojić, Milan
AU  - Sencanski, Milan
AU  - Nikolić, Milan
AU  - Visnjevac, Aleksandar
AU  - Araskov, Jovana
AU  - Miljković, Marija
AU  - Muller, Christian D.
AU  - Filipović, Nenad R.
PY  - 2019
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1197
AB  - Anticancer activity of Pd complexes 1-5 with bidentate N-heteroaromatic hydrazone ligands was investigated on human acute monocytic leukemia (THP-1; cells in a suspension) and human mammary adenocarcinoma (MCF-7; two-dimensional layer and three-dimensional spheroid tumor model) cell lines. For the Pd(II) complexes with condensation products of ethyl hydrazainoacetate and quinoline-8-carboxaldehyde (complex 1) and 2-for-mylpyridine (complex 3), for which apoptosis was determined as a mechanism of anticancer activity, further investigation revealed that they arrest the cell cycle in G0/G1 phase, induce generation of reactive oxygen species and inhibit Topoisomerase I in vitro. In silico studies corroborate experimental findings that these complexes show topoisomerase inhibition activity in the micromolar range and indicate binding to a DNA's minor groove as another potential target. Based on the results obtained by circular dichroism and fluorescence spectroscopy measurements, the most active complexes are suitable to be delivered to a blood stream via human serum albumin.
PB  - Elsevier Science Inc, New York
T2  - Journal of Inorganic Biochemistry
T1  - Pd(II) complexes with N-heteroaromatic hydrazone ligands: Anticancer activity, in silico and experimental target identification
VL  - 199
DO  - 10.1016/j.jinorgbio.2019.110758
ER  - 

@article{
author = "Bjelogrlić, Snežana K. and Todorović, Tamara R. and Kojić, Milan and Sencanski, Milan and Nikolić, Milan and Visnjevac, Aleksandar and Araskov, Jovana and Miljković, Marija and Muller, Christian D. and Filipović, Nenad R.",
year = "2019",
abstract = "Anticancer activity of Pd complexes 1-5 with bidentate N-heteroaromatic hydrazone ligands was investigated on human acute monocytic leukemia (THP-1; cells in a suspension) and human mammary adenocarcinoma (MCF-7; two-dimensional layer and three-dimensional spheroid tumor model) cell lines. For the Pd(II) complexes with condensation products of ethyl hydrazainoacetate and quinoline-8-carboxaldehyde (complex 1) and 2-for-mylpyridine (complex 3), for which apoptosis was determined as a mechanism of anticancer activity, further investigation revealed that they arrest the cell cycle in G0/G1 phase, induce generation of reactive oxygen species and inhibit Topoisomerase I in vitro. In silico studies corroborate experimental findings that these complexes show topoisomerase inhibition activity in the micromolar range and indicate binding to a DNA's minor groove as another potential target. Based on the results obtained by circular dichroism and fluorescence spectroscopy measurements, the most active complexes are suitable to be delivered to a blood stream via human serum albumin.",
publisher = "Elsevier Science Inc, New York",
journal = "Journal of Inorganic Biochemistry",
title = "Pd(II) complexes with N-heteroaromatic hydrazone ligands: Anticancer activity, in silico and experimental target identification",
volume = "199",
doi = "10.1016/j.jinorgbio.2019.110758"
}

Bjelogrlić, S. K., Todorović, T. R., Kojić, M., Sencanski, M., Nikolić, M., Visnjevac, A., Araskov, J., Miljković, M., Muller, C. D.,& Filipović, N. R.. (2019). Pd(II) complexes with N-heteroaromatic hydrazone ligands: Anticancer activity, in silico and experimental target identification. in Journal of Inorganic Biochemistry
Elsevier Science Inc, New York., 199.
https://doi.org/10.1016/j.jinorgbio.2019.110758

Bjelogrlić SK, Todorović TR, Kojić M, Sencanski M, Nikolić M, Visnjevac A, Araskov J, Miljković M, Muller CD, Filipović NR. Pd(II) complexes with N-heteroaromatic hydrazone ligands: Anticancer activity, in silico and experimental target identification. in Journal of Inorganic Biochemistry. 2019;199.
doi:10.1016/j.jinorgbio.2019.110758 .

Bjelogrlić, Snežana K., Todorović, Tamara R., Kojić, Milan, Sencanski, Milan, Nikolić, Milan, Visnjevac, Aleksandar, Araskov, Jovana, Miljković, Marija, Muller, Christian D., Filipović, Nenad R., "Pd(II) complexes with N-heteroaromatic hydrazone ligands: Anticancer activity, in silico and experimental target identification" in Journal of Inorganic Biochemistry, 199 (2019),
https://doi.org/10.1016/j.jinorgbio.2019.110758 . .

TY  - JOUR
AU  - Miljković, Marija
AU  - Marinković, Pavle
AU  - Novović, Katarina
AU  - Jovčić, Branko
AU  - Terzić-Vidojević, Amarela
AU  - Kojić, Milan
PY  - 2018
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1110
AB  - The ability of lactic acid bacteria to form multi-cellular aggregates via self-aggregation is regarded as an important mechanism for stress tolerance, adhesion, colonization and genetic material exchange. The novel aggLr gene encoding for the auto-aggregation promoting protein (AggLr) of Lactococcus raffinolactis BGTRK10-1 was cloned. Heterologous expression of AggLr enabled auto-aggregation, higher hydrophobicity and collagen and fibronectin binding of the carrier strains. Domain analysis and the type of aggregates formed by cells expressing AggLr confirmed that this aggregation factor belongs to the family of high molecular weight proteins that the authors propose to be called Snow-flake Forming Collagen Binding Aggregation Factors (SFCBAF). An additional feature of SFCBAF is that they are rich in threonine and lysine and are free of cysteine in all of the aggregation factors described so far. In contrast to previously discovered SFCBAF, the gene encoding for AggLr is located on the chromosome in the strain BGTRK10-1.
PB  - Taylor & Francis Ltd, Abingdon
T2  - Biofouling
T1  - AggLr, a novel aggregation factor in Lactococcus raffinolactis BGTRK10-1: its role in surface adhesion
EP  - 698
IS  - 6
SP  - 685
VL  - 34
DO  - 10.1080/08927014.2018.1481956
ER  - 

@article{
author = "Miljković, Marija and Marinković, Pavle and Novović, Katarina and Jovčić, Branko and Terzić-Vidojević, Amarela and Kojić, Milan",
year = "2018",
abstract = "The ability of lactic acid bacteria to form multi-cellular aggregates via self-aggregation is regarded as an important mechanism for stress tolerance, adhesion, colonization and genetic material exchange. The novel aggLr gene encoding for the auto-aggregation promoting protein (AggLr) of Lactococcus raffinolactis BGTRK10-1 was cloned. Heterologous expression of AggLr enabled auto-aggregation, higher hydrophobicity and collagen and fibronectin binding of the carrier strains. Domain analysis and the type of aggregates formed by cells expressing AggLr confirmed that this aggregation factor belongs to the family of high molecular weight proteins that the authors propose to be called Snow-flake Forming Collagen Binding Aggregation Factors (SFCBAF). An additional feature of SFCBAF is that they are rich in threonine and lysine and are free of cysteine in all of the aggregation factors described so far. In contrast to previously discovered SFCBAF, the gene encoding for AggLr is located on the chromosome in the strain BGTRK10-1.",
publisher = "Taylor & Francis Ltd, Abingdon",
journal = "Biofouling",
title = "AggLr, a novel aggregation factor in Lactococcus raffinolactis BGTRK10-1: its role in surface adhesion",
pages = "698-685",
number = "6",
volume = "34",
doi = "10.1080/08927014.2018.1481956"
}

Miljković, M., Marinković, P., Novović, K., Jovčić, B., Terzić-Vidojević, A.,& Kojić, M.. (2018). AggLr, a novel aggregation factor in Lactococcus raffinolactis BGTRK10-1: its role in surface adhesion. in Biofouling
Taylor & Francis Ltd, Abingdon., 34(6), 685-698.
https://doi.org/10.1080/08927014.2018.1481956

Miljković M, Marinković P, Novović K, Jovčić B, Terzić-Vidojević A, Kojić M. AggLr, a novel aggregation factor in Lactococcus raffinolactis BGTRK10-1: its role in surface adhesion. in Biofouling. 2018;34(6):685-698.
doi:10.1080/08927014.2018.1481956 .

Miljković, Marija, Marinković, Pavle, Novović, Katarina, Jovčić, Branko, Terzić-Vidojević, Amarela, Kojić, Milan, "AggLr, a novel aggregation factor in Lactococcus raffinolactis BGTRK10-1: its role in surface adhesion" in Biofouling, 34, no. 6 (2018):685-698,
https://doi.org/10.1080/08927014.2018.1481956 . .

TY  - JOUR
AU  - Novović, Katarina
AU  - Mihajlović, Sanja
AU  - Dinić, Miroslav
AU  - Malešević, Milka
AU  - Miljković, Marija
AU  - Kojić, Milan
AU  - Jovčić, Branko
PY  - 2018
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1119
AB  - Acinetobacter baumannii has been recognized as one of the most challeging pathogens in clinical settings worldwide. Outer membrane porins play a significant role in Acinetobacter antibiotic resistance and virulence. A. baumannii carbapenem resistance and virulence factor porin Omp33-36 was the subject of this study. We investigated the omp33-36 gene transcriptional response in the growth phase, its response to carbapenems, and the effect of contact with host cells. Additionally, the cytotoxic effect of A. baumannii towards keratinocytes was assessed, as well as correlation between omp33-36 gene transcription and cytotoxicity. Further, Acinetobacter spp. Omp33-36 was classified and its characteristics relevant for vaccine candidature were determined. The level of the omp33-36 gene transcription varied between growth phases, but a common pattern could not be established among different strains. Treatment with subinhibitory concentrations of carbapenems decreased, while contact with keratinocytes increased omp33-36 expression in the analysed A. baumannii strains. Variations in omp33-36 mRNA levels did not correlate with cytotoxicity levels. Decrease of omp33-36 mRNA during treatment with subinhibitory concentrations of carbapenems, indicated the importance of transcriptional changes in reversible resistance to carbapenems due to the absence of Omp33-36. The transcription of omp33-36 increased after contact with keratinocytes, indicating the important role of de novo transcription during the initial phase of A. baumannii infection. Primary structural analysis of Acinetobacter spp. Omp33-36 revealed three distinct groups (among four A. baumannii variants). Although we have shown that Omp33-36 was highly polymorphic, we propose a potential antigen (PLAEAAFL motif) for vaccine development. According to PROVEAN analysis, the highly polymorphic structure of Omp33-36 porin should not influence its function significantly.
PB  - Public Library Science, San Francisco
T2  - PLoS One
T1  - Acinetobacter spp. porin Omp33-36: Classification and transcriptional response to carbapenems and host cells
IS  - 8
VL  - 13
DO  - 10.1371/journal.pone.0201608
ER  - 

@article{
author = "Novović, Katarina and Mihajlović, Sanja and Dinić, Miroslav and Malešević, Milka and Miljković, Marija and Kojić, Milan and Jovčić, Branko",
year = "2018",
abstract = "Acinetobacter baumannii has been recognized as one of the most challeging pathogens in clinical settings worldwide. Outer membrane porins play a significant role in Acinetobacter antibiotic resistance and virulence. A. baumannii carbapenem resistance and virulence factor porin Omp33-36 was the subject of this study. We investigated the omp33-36 gene transcriptional response in the growth phase, its response to carbapenems, and the effect of contact with host cells. Additionally, the cytotoxic effect of A. baumannii towards keratinocytes was assessed, as well as correlation between omp33-36 gene transcription and cytotoxicity. Further, Acinetobacter spp. Omp33-36 was classified and its characteristics relevant for vaccine candidature were determined. The level of the omp33-36 gene transcription varied between growth phases, but a common pattern could not be established among different strains. Treatment with subinhibitory concentrations of carbapenems decreased, while contact with keratinocytes increased omp33-36 expression in the analysed A. baumannii strains. Variations in omp33-36 mRNA levels did not correlate with cytotoxicity levels. Decrease of omp33-36 mRNA during treatment with subinhibitory concentrations of carbapenems, indicated the importance of transcriptional changes in reversible resistance to carbapenems due to the absence of Omp33-36. The transcription of omp33-36 increased after contact with keratinocytes, indicating the important role of de novo transcription during the initial phase of A. baumannii infection. Primary structural analysis of Acinetobacter spp. Omp33-36 revealed three distinct groups (among four A. baumannii variants). Although we have shown that Omp33-36 was highly polymorphic, we propose a potential antigen (PLAEAAFL motif) for vaccine development. According to PROVEAN analysis, the highly polymorphic structure of Omp33-36 porin should not influence its function significantly.",
publisher = "Public Library Science, San Francisco",
journal = "PLoS One",
title = "Acinetobacter spp. porin Omp33-36: Classification and transcriptional response to carbapenems and host cells",
number = "8",
volume = "13",
doi = "10.1371/journal.pone.0201608"
}

Novović, K., Mihajlović, S., Dinić, M., Malešević, M., Miljković, M., Kojić, M.,& Jovčić, B.. (2018). Acinetobacter spp. porin Omp33-36: Classification and transcriptional response to carbapenems and host cells. in PLoS One
Public Library Science, San Francisco., 13(8).
https://doi.org/10.1371/journal.pone.0201608

Novović K, Mihajlović S, Dinić M, Malešević M, Miljković M, Kojić M, Jovčić B. Acinetobacter spp. porin Omp33-36: Classification and transcriptional response to carbapenems and host cells. in PLoS One. 2018;13(8).
doi:10.1371/journal.pone.0201608 .

Novović, Katarina, Mihajlović, Sanja, Dinić, Miroslav, Malešević, Milka, Miljković, Marija, Kojić, Milan, Jovčić, Branko, "Acinetobacter spp. porin Omp33-36: Classification and transcriptional response to carbapenems and host cells" in PLoS One, 13, no. 8 (2018),
https://doi.org/10.1371/journal.pone.0201608 . .

TY  - JOUR
AU  - Miljković, Marija
AU  - Malešević, Milka
AU  - Filipić, Brankica
AU  - Vukotić, Goran
AU  - Kojić, Milan
PY  - 2018
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1156
AB  - Restriction enzymes are the main defence system against foreign DNA, in charge of preserving genome integrity. Lactococcus raffinolactis BGTRK10-1 expresses LraI Type II restriction-modification enzyme, whose activity is similar to that shown for EcoRI; LraI methyltransferase protects DNA from EcoRI cleavage. The gene encoding LraI endonuclease was cloned and overexpressed in E. coli. Purified enzyme showed the highest specific activity at lower temperatures (between 13 degrees C and 37 degrees C) and was stable after storage at -20 degrees C in 50% glycerol. The concentration of monovalent ions in the reaction buffer required for optimal activity of LraI restriction enzyme was 100 mM or higher. The recognition and cleavage sequence for LraI restriction enzyme was determined as 5'-G/AATTC-3', indicating that LraI restriction enzyme is an isoschizomer of EcoRI. In the reaction buffer with a lower salt concentration, LraI exhibits star activity and specifically recognizes and cuts another alternative sequence 5'-A/AATTC-3', leaving the same sticky ends on fragments as EcoRI, which makes them clonable into a linearized vector. Phylogenetic analysis based on sequence alignment pointed out the common origin of LraI restriction-modification system with previously described EcoRI-like restriction-modification systems.
PB  - Hindawi Ltd, London
T2  - Biomed Research International
T1  - LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity
VL  - 2018
DO  - 10.1155/2018/5657085
ER  - 

@article{
author = "Miljković, Marija and Malešević, Milka and Filipić, Brankica and Vukotić, Goran and Kojić, Milan",
year = "2018",
abstract = "Restriction enzymes are the main defence system against foreign DNA, in charge of preserving genome integrity. Lactococcus raffinolactis BGTRK10-1 expresses LraI Type II restriction-modification enzyme, whose activity is similar to that shown for EcoRI; LraI methyltransferase protects DNA from EcoRI cleavage. The gene encoding LraI endonuclease was cloned and overexpressed in E. coli. Purified enzyme showed the highest specific activity at lower temperatures (between 13 degrees C and 37 degrees C) and was stable after storage at -20 degrees C in 50% glycerol. The concentration of monovalent ions in the reaction buffer required for optimal activity of LraI restriction enzyme was 100 mM or higher. The recognition and cleavage sequence for LraI restriction enzyme was determined as 5'-G/AATTC-3', indicating that LraI restriction enzyme is an isoschizomer of EcoRI. In the reaction buffer with a lower salt concentration, LraI exhibits star activity and specifically recognizes and cuts another alternative sequence 5'-A/AATTC-3', leaving the same sticky ends on fragments as EcoRI, which makes them clonable into a linearized vector. Phylogenetic analysis based on sequence alignment pointed out the common origin of LraI restriction-modification system with previously described EcoRI-like restriction-modification systems.",
publisher = "Hindawi Ltd, London",
journal = "Biomed Research International",
title = "LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity",
volume = "2018",
doi = "10.1155/2018/5657085"
}

Miljković, M., Malešević, M., Filipić, B., Vukotić, G.,& Kojić, M.. (2018). LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity. in Biomed Research International
Hindawi Ltd, London., 2018.
https://doi.org/10.1155/2018/5657085

Miljković M, Malešević M, Filipić B, Vukotić G, Kojić M. LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity. in Biomed Research International. 2018;2018.
doi:10.1155/2018/5657085 .

Miljković, Marija, Malešević, Milka, Filipić, Brankica, Vukotić, Goran, Kojić, Milan, "LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity" in Biomed Research International, 2018 (2018),
https://doi.org/10.1155/2018/5657085 . .

TY  - JOUR
AU  - Lozo, Jelena
AU  - Mirković, Nemanja
AU  - O'Connor, Paula M.
AU  - Malešević, Milka
AU  - Miljković, Marija
AU  - Polović, Natalija
AU  - Jovčić, Branko
AU  - Cotter, Paul D.
AU  - Kojić, Milan
PY  - 2017
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1001
AB  - Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 produces a novel bacteriocin, lactolisterin BU, with strong antimicrobial activity against many species of Gram-positive bacteria, including important food spoilage and foodborne pathogens, such as Listeria monocytogenes, Staphylococcus aureus, Bacillus spp., and streptococci. Lactolisterin BU was extracted from the cell surface of BGBU1-4 by 2-propanol and purified to homogeneity by C18 solid-phase extraction and reversed-phase high-performance liquid chromatography. The molecular mass of the purified lactolisterin BU was 5,160.94 Da, and an internal fragment, AVSWAWQH, as determined by N-terminal sequencing, showed low-level similarity to existing antimicrobial peptides. Curing and transformation experiments revealed the presence of a corresponding bacteriocin operon on the smallest plasmid, pBU6 (6.2 kb), of strain BGBU1-4. Analysis of the bacteriocin operon revealed a leaderless bacteriocin of 43 amino acids that exhibited similarity to bacteriocin BHT-B (63%) from Streptococcus ratti, a bacteriocin with analogy to aureocin A. IMPORTANCE Lactolisterin BU, a broad-spectrum leaderless bacteriocin produced by L. lactis subsp. lactis bv. diacetylactis BGBU1-4, expresses strong antimicrobial activity against food spoilage and foodborne pathogens, such as Listeria monocytogenes, Staphylococcus aureus, Bacillus spp., and streptococci. Lactolisterin BU showed the highest similarity to aureocin-like bacteriocins produced by different bacteria. The operon for synthesis is located on the smallest plasmid, pBU6 (6.2 kb), of strain BGBU1-4, indicating possible horizontal transfer among producers.
PB  - Amer Soc Microbiology, Washington
T2  - Applied and Environmental Microbiology
T1  - Lactolisterin BU, a Novel Class II Broad-Spectrum Bacteriocin from Lactococcus lactis subsp lactis bv. diacetylactis BGBU1-4
IS  - 21
VL  - 83
DO  - 10.1128/AEM.01519-17
ER  - 

@article{
author = "Lozo, Jelena and Mirković, Nemanja and O'Connor, Paula M. and Malešević, Milka and Miljković, Marija and Polović, Natalija and Jovčić, Branko and Cotter, Paul D. and Kojić, Milan",
year = "2017",
abstract = "Lactococcus lactis subsp. lactis bv. diacetylactis BGBU1-4 produces a novel bacteriocin, lactolisterin BU, with strong antimicrobial activity against many species of Gram-positive bacteria, including important food spoilage and foodborne pathogens, such as Listeria monocytogenes, Staphylococcus aureus, Bacillus spp., and streptococci. Lactolisterin BU was extracted from the cell surface of BGBU1-4 by 2-propanol and purified to homogeneity by C18 solid-phase extraction and reversed-phase high-performance liquid chromatography. The molecular mass of the purified lactolisterin BU was 5,160.94 Da, and an internal fragment, AVSWAWQH, as determined by N-terminal sequencing, showed low-level similarity to existing antimicrobial peptides. Curing and transformation experiments revealed the presence of a corresponding bacteriocin operon on the smallest plasmid, pBU6 (6.2 kb), of strain BGBU1-4. Analysis of the bacteriocin operon revealed a leaderless bacteriocin of 43 amino acids that exhibited similarity to bacteriocin BHT-B (63%) from Streptococcus ratti, a bacteriocin with analogy to aureocin A. IMPORTANCE Lactolisterin BU, a broad-spectrum leaderless bacteriocin produced by L. lactis subsp. lactis bv. diacetylactis BGBU1-4, expresses strong antimicrobial activity against food spoilage and foodborne pathogens, such as Listeria monocytogenes, Staphylococcus aureus, Bacillus spp., and streptococci. Lactolisterin BU showed the highest similarity to aureocin-like bacteriocins produced by different bacteria. The operon for synthesis is located on the smallest plasmid, pBU6 (6.2 kb), of strain BGBU1-4, indicating possible horizontal transfer among producers.",
publisher = "Amer Soc Microbiology, Washington",
journal = "Applied and Environmental Microbiology",
title = "Lactolisterin BU, a Novel Class II Broad-Spectrum Bacteriocin from Lactococcus lactis subsp lactis bv. diacetylactis BGBU1-4",
number = "21",
volume = "83",
doi = "10.1128/AEM.01519-17"
}

Lozo, J., Mirković, N., O'Connor, P. M., Malešević, M., Miljković, M., Polović, N., Jovčić, B., Cotter, P. D.,& Kojić, M.. (2017). Lactolisterin BU, a Novel Class II Broad-Spectrum Bacteriocin from Lactococcus lactis subsp lactis bv. diacetylactis BGBU1-4. in Applied and Environmental Microbiology
Amer Soc Microbiology, Washington., 83(21).
https://doi.org/10.1128/AEM.01519-17

Lozo J, Mirković N, O'Connor PM, Malešević M, Miljković M, Polović N, Jovčić B, Cotter PD, Kojić M. Lactolisterin BU, a Novel Class II Broad-Spectrum Bacteriocin from Lactococcus lactis subsp lactis bv. diacetylactis BGBU1-4. in Applied and Environmental Microbiology. 2017;83(21).
doi:10.1128/AEM.01519-17 .

Lozo, Jelena, Mirković, Nemanja, O'Connor, Paula M., Malešević, Milka, Miljković, Marija, Polović, Natalija, Jovčić, Branko, Cotter, Paul D., Kojić, Milan, "Lactolisterin BU, a Novel Class II Broad-Spectrum Bacteriocin from Lactococcus lactis subsp lactis bv. diacetylactis BGBU1-4" in Applied and Environmental Microbiology, 83, no. 21 (2017),
https://doi.org/10.1128/AEM.01519-17 . .

TY  - JOUR
AU  - Veljović, Katarina
AU  - Popović, Nikola
AU  - Miljković, Marija
AU  - Tolinački, Maja
AU  - Terzić-Vidojević, Amarela
AU  - Kojić, Milan
PY  - 2017
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/997
AB  - The understanding of mechanisms of interactions between various bacterial cell surface proteins and host receptors has become imperative for the study of the health promoting features of probiotic enterococci. This study, for the first time, describes a novel enterococcal aggregation protein, AggE, from Enterococcus faeciurn BGGO9-28, selected from a laboratory collection of enterococcal isolates with auto aggregation phenotypes. Among them, En. faecium BGG09-28 showed the strongest auto -aggregation, adhesion to components of ECM and biofilm formation. Novel aggregation promoting factor AggE, a protein of 178.1 kDa, belongs to the collagen -binding superfamily of proteins and shares similar architecture with previously discovered aggregation factors from lactic acid bacteria (LAB). Its expression in heterologous enterococcal and lactococcal hosts demonstrates that the aggE gene is sufficient for cell aggregation. The derivatives carrying aggE exhibited the ten times higher adhesion ability to collagen and fibronectin, possess about two times higher adhesion to mucin and contribute to the increase of biofilm formation, comparing to the control strains. Analysis for the presence of virulence factors (cytolysin and gelatinase production), antibiotic resistance (antibiotic susceptibility) and genes (cylA, egg, gelE, esp, hyiN, ace, efaks, and efagn) showed that BGG09-28 was sensitive to all tested antibiotics, without hemolytic or gelatinase activity. This strain does not carry any of the tested genes encoding for known virulence factors. Results showed that BGGO9-28 was resistant to low pH and high concentrations of bile salts. Also, it adhered strongly to the Caco-2 human epithelial cell line. In conclusion, the results of this study indicate that the presence of AggE protein on the cell surface in enterococci is a desirable probiotic feature.
PB  - Frontiers Media Sa, Lausanne
T2  - Frontiers in Microbiology
T1  - Novel Aggregation Promoting Factor AggE Contributes to the Probiotic Properties of Enterococcus faecium BGGO9-28
VL  - 8
DO  - 10.3389/fmicb.2017.01843
ER  - 

@article{
author = "Veljović, Katarina and Popović, Nikola and Miljković, Marija and Tolinački, Maja and Terzić-Vidojević, Amarela and Kojić, Milan",
year = "2017",
abstract = "The understanding of mechanisms of interactions between various bacterial cell surface proteins and host receptors has become imperative for the study of the health promoting features of probiotic enterococci. This study, for the first time, describes a novel enterococcal aggregation protein, AggE, from Enterococcus faeciurn BGGO9-28, selected from a laboratory collection of enterococcal isolates with auto aggregation phenotypes. Among them, En. faecium BGG09-28 showed the strongest auto -aggregation, adhesion to components of ECM and biofilm formation. Novel aggregation promoting factor AggE, a protein of 178.1 kDa, belongs to the collagen -binding superfamily of proteins and shares similar architecture with previously discovered aggregation factors from lactic acid bacteria (LAB). Its expression in heterologous enterococcal and lactococcal hosts demonstrates that the aggE gene is sufficient for cell aggregation. The derivatives carrying aggE exhibited the ten times higher adhesion ability to collagen and fibronectin, possess about two times higher adhesion to mucin and contribute to the increase of biofilm formation, comparing to the control strains. Analysis for the presence of virulence factors (cytolysin and gelatinase production), antibiotic resistance (antibiotic susceptibility) and genes (cylA, egg, gelE, esp, hyiN, ace, efaks, and efagn) showed that BGG09-28 was sensitive to all tested antibiotics, without hemolytic or gelatinase activity. This strain does not carry any of the tested genes encoding for known virulence factors. Results showed that BGGO9-28 was resistant to low pH and high concentrations of bile salts. Also, it adhered strongly to the Caco-2 human epithelial cell line. In conclusion, the results of this study indicate that the presence of AggE protein on the cell surface in enterococci is a desirable probiotic feature.",
publisher = "Frontiers Media Sa, Lausanne",
journal = "Frontiers in Microbiology",
title = "Novel Aggregation Promoting Factor AggE Contributes to the Probiotic Properties of Enterococcus faecium BGGO9-28",
volume = "8",
doi = "10.3389/fmicb.2017.01843"
}

Veljović, K., Popović, N., Miljković, M., Tolinački, M., Terzić-Vidojević, A.,& Kojić, M.. (2017). Novel Aggregation Promoting Factor AggE Contributes to the Probiotic Properties of Enterococcus faecium BGGO9-28. in Frontiers in Microbiology
Frontiers Media Sa, Lausanne., 8.
https://doi.org/10.3389/fmicb.2017.01843

Veljović K, Popović N, Miljković M, Tolinački M, Terzić-Vidojević A, Kojić M. Novel Aggregation Promoting Factor AggE Contributes to the Probiotic Properties of Enterococcus faecium BGGO9-28. in Frontiers in Microbiology. 2017;8.
doi:10.3389/fmicb.2017.01843 .

Veljović, Katarina, Popović, Nikola, Miljković, Marija, Tolinački, Maja, Terzić-Vidojević, Amarela, Kojić, Milan, "Novel Aggregation Promoting Factor AggE Contributes to the Probiotic Properties of Enterococcus faecium BGGO9-28" in Frontiers in Microbiology, 8 (2017),
https://doi.org/10.3389/fmicb.2017.01843 . .

TY  - JOUR
AU  - Mirković, Nemanja
AU  - Polović, Natalija
AU  - Vukotić, Goran
AU  - Jovčić, Branko
AU  - Miljković, Marija
AU  - Radulović, Zorica
AU  - Diep, Dzung B.
AU  - Kojić, Milan
PY  - 2016
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/980
AB  - Bacteriocin producers normally possess dedicated immunity systems to protect themselves from their own bacteriocins. Lactococcus lactis strains LMG2081 and BGBM50 are known as lactococcin G producers. However, BGBM50 was sensitive to LMG2081, which indicated that LMG2081 might produce additional bacteriocins that are not present in BGBM50. Therefore, whole-genome sequencing of the two strains was performed, and a lantibiotic operon (called lctLMG) was identified in LMG2081 but not in BGBM50. The lctLMG operon contains six open reading frames; the first three genes, lmgA, lmgM, and lmgT, are involved in the biosynthesis and export of bacteriocin, while the other three genes, lmgF, lmgE, and lmgG, are involved in lantibiotic immunity. Mutational analysis confirmed that the lctLMG operon is responsible for the additional antimicrobial activity. Specifically, site-directed mutation within this operon rendered LMG2081 inactive toward BGBM50. Subsequent purification and electrospray ionization-time of flight mass spectrometric analysis confirmed that the lantibiotic bacteriocin called lacticin LMG is exported as a 25-amino-acid peptide. Lacticin LMG is highly similar to the lacticin 481 group. It is interesting that a bacteriocin producer produces two different classes of bacteriocins, whose operons are located in the chromosome and a plasmid.
PB  - Amer Soc Microbiology, Washington
T2  - Applied and Environmental Microbiology
T1  - Lactococcus lactis LMG2081 Produces Two Bacteriocins, a Nonlantibiotic and a Novel Lantibiotic
EP  - 2562
IS  - 8
SP  - 2555
VL  - 82
DO  - 10.1128/AEM.03988-15
ER  - 

@article{
author = "Mirković, Nemanja and Polović, Natalija and Vukotić, Goran and Jovčić, Branko and Miljković, Marija and Radulović, Zorica and Diep, Dzung B. and Kojić, Milan",
year = "2016",
abstract = "Bacteriocin producers normally possess dedicated immunity systems to protect themselves from their own bacteriocins. Lactococcus lactis strains LMG2081 and BGBM50 are known as lactococcin G producers. However, BGBM50 was sensitive to LMG2081, which indicated that LMG2081 might produce additional bacteriocins that are not present in BGBM50. Therefore, whole-genome sequencing of the two strains was performed, and a lantibiotic operon (called lctLMG) was identified in LMG2081 but not in BGBM50. The lctLMG operon contains six open reading frames; the first three genes, lmgA, lmgM, and lmgT, are involved in the biosynthesis and export of bacteriocin, while the other three genes, lmgF, lmgE, and lmgG, are involved in lantibiotic immunity. Mutational analysis confirmed that the lctLMG operon is responsible for the additional antimicrobial activity. Specifically, site-directed mutation within this operon rendered LMG2081 inactive toward BGBM50. Subsequent purification and electrospray ionization-time of flight mass spectrometric analysis confirmed that the lantibiotic bacteriocin called lacticin LMG is exported as a 25-amino-acid peptide. Lacticin LMG is highly similar to the lacticin 481 group. It is interesting that a bacteriocin producer produces two different classes of bacteriocins, whose operons are located in the chromosome and a plasmid.",
publisher = "Amer Soc Microbiology, Washington",
journal = "Applied and Environmental Microbiology",
title = "Lactococcus lactis LMG2081 Produces Two Bacteriocins, a Nonlantibiotic and a Novel Lantibiotic",
pages = "2562-2555",
number = "8",
volume = "82",
doi = "10.1128/AEM.03988-15"
}

Mirković, N., Polović, N., Vukotić, G., Jovčić, B., Miljković, M., Radulović, Z., Diep, D. B.,& Kojić, M.. (2016). Lactococcus lactis LMG2081 Produces Two Bacteriocins, a Nonlantibiotic and a Novel Lantibiotic. in Applied and Environmental Microbiology
Amer Soc Microbiology, Washington., 82(8), 2555-2562.
https://doi.org/10.1128/AEM.03988-15

Mirković N, Polović N, Vukotić G, Jovčić B, Miljković M, Radulović Z, Diep DB, Kojić M. Lactococcus lactis LMG2081 Produces Two Bacteriocins, a Nonlantibiotic and a Novel Lantibiotic. in Applied and Environmental Microbiology. 2016;82(8):2555-2562.
doi:10.1128/AEM.03988-15 .

Mirković, Nemanja, Polović, Natalija, Vukotić, Goran, Jovčić, Branko, Miljković, Marija, Radulović, Zorica, Diep, Dzung B., Kojić, Milan, "Lactococcus lactis LMG2081 Produces Two Bacteriocins, a Nonlantibiotic and a Novel Lantibiotic" in Applied and Environmental Microbiology, 82, no. 8 (2016):2555-2562,
https://doi.org/10.1128/AEM.03988-15 . .

TY  - JOUR
AU  - Miljković, Marija
AU  - Uzelac, Gordana
AU  - Mirković, Nemanja
AU  - Devescovi, Giulia
AU  - Diep, Dzung B.
AU  - Venturi, Vittorio
AU  - Kojić, Milan
PY  - 2016
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/904
AB  - The Zn-dependent membrane-located protease YvjB has previously been shown to serve as a target receptor for LsbB, a class II leaderless lactococcal bacteriocin. Although yvjB is highly conserved in the genus Lactococcus, the bacteriocin appears to be active only against the subspecies L. lactis subsp. lactis. Comparative analysis of the YvjB proteins of a sensitive strain (YvjB(MN)) and a resistant strain (YvjB(MG)) showed that they differ from each other in 31 positions. In this study, we applied site-directed mutagenesis and performed directed binding studies to provide biochemical evidence that LsbB interacts with the third transmembrane helix of YvjB in susceptible cells. The site-directed mutagenesis of LsbB and YvjB proteins showed that certain amino acids and the length of LsbB are responsible for the bacteriocin activity, most probably through adequate interaction of these two proteins; the essential amino acids in LsbB responsible for the activity are tryptophan (Trp(25)) and terminal alanine (Ala(30)). It was also shown that the distance between Trp(25) and terminal alanine is crucial for LsbB activity. The crucial region in YvjB for the interaction with LsbB is the beginning of the third transmembrane helix, particularly amino acids tyrosine (Tyr(356)) and alanine (Ala(353)). In vitro experiments showed that LsbB could interact with both YvjB(MN) and YvjB(MG), but the strength of interaction is significantly less with YvjB(MG). In vivo experiments with immunofluorescently labeled antibody demonstrated that LsbB specifically interacts only with cells carrying YvjB(MN). IMPORTANCE The antimicrobial activity of LsbB bacteriocin depends on the correct interaction with the corresponding receptor in the bacterial membrane of sensitive cells. Membrane-located bacteriocin receptors have essential primary functions, such as cell wall synthesis or sugar transport, and it seems that interaction with bacteriocins is suicidal for cells. This study showed that the C-terminal part of LsbB is crucial for the bacteriocin activity, most probably through adequate interaction with the third transmembrane domain of the YvjB receptor. The conserved Tyr(356) and Ala(353) residues of YvjB are essential for the function of this Zn-dependent membrane-located protease as a bacteriocin receptor.
PB  - Amer Soc Microbiology, Washington
T2  - Applied and Environmental Microbiology
T1  - LsbB Bacteriocin Interacts with the Third Transmembrane Domain of the YvjB Receptor
EP  - 5374
IS  - 17
SP  - 5364
VL  - 82
DO  - 10.1128/AEM.01293-16
ER  - 

@article{
author = "Miljković, Marija and Uzelac, Gordana and Mirković, Nemanja and Devescovi, Giulia and Diep, Dzung B. and Venturi, Vittorio and Kojić, Milan",
year = "2016",
abstract = "The Zn-dependent membrane-located protease YvjB has previously been shown to serve as a target receptor for LsbB, a class II leaderless lactococcal bacteriocin. Although yvjB is highly conserved in the genus Lactococcus, the bacteriocin appears to be active only against the subspecies L. lactis subsp. lactis. Comparative analysis of the YvjB proteins of a sensitive strain (YvjB(MN)) and a resistant strain (YvjB(MG)) showed that they differ from each other in 31 positions. In this study, we applied site-directed mutagenesis and performed directed binding studies to provide biochemical evidence that LsbB interacts with the third transmembrane helix of YvjB in susceptible cells. The site-directed mutagenesis of LsbB and YvjB proteins showed that certain amino acids and the length of LsbB are responsible for the bacteriocin activity, most probably through adequate interaction of these two proteins; the essential amino acids in LsbB responsible for the activity are tryptophan (Trp(25)) and terminal alanine (Ala(30)). It was also shown that the distance between Trp(25) and terminal alanine is crucial for LsbB activity. The crucial region in YvjB for the interaction with LsbB is the beginning of the third transmembrane helix, particularly amino acids tyrosine (Tyr(356)) and alanine (Ala(353)). In vitro experiments showed that LsbB could interact with both YvjB(MN) and YvjB(MG), but the strength of interaction is significantly less with YvjB(MG). In vivo experiments with immunofluorescently labeled antibody demonstrated that LsbB specifically interacts only with cells carrying YvjB(MN). IMPORTANCE The antimicrobial activity of LsbB bacteriocin depends on the correct interaction with the corresponding receptor in the bacterial membrane of sensitive cells. Membrane-located bacteriocin receptors have essential primary functions, such as cell wall synthesis or sugar transport, and it seems that interaction with bacteriocins is suicidal for cells. This study showed that the C-terminal part of LsbB is crucial for the bacteriocin activity, most probably through adequate interaction with the third transmembrane domain of the YvjB receptor. The conserved Tyr(356) and Ala(353) residues of YvjB are essential for the function of this Zn-dependent membrane-located protease as a bacteriocin receptor.",
publisher = "Amer Soc Microbiology, Washington",
journal = "Applied and Environmental Microbiology",
title = "LsbB Bacteriocin Interacts with the Third Transmembrane Domain of the YvjB Receptor",
pages = "5374-5364",
number = "17",
volume = "82",
doi = "10.1128/AEM.01293-16"
}

Miljković, M., Uzelac, G., Mirković, N., Devescovi, G., Diep, D. B., Venturi, V.,& Kojić, M.. (2016). LsbB Bacteriocin Interacts with the Third Transmembrane Domain of the YvjB Receptor. in Applied and Environmental Microbiology
Amer Soc Microbiology, Washington., 82(17), 5364-5374.
https://doi.org/10.1128/AEM.01293-16

Miljković M, Uzelac G, Mirković N, Devescovi G, Diep DB, Venturi V, Kojić M. LsbB Bacteriocin Interacts with the Third Transmembrane Domain of the YvjB Receptor. in Applied and Environmental Microbiology. 2016;82(17):5364-5374.
doi:10.1128/AEM.01293-16 .

Miljković, Marija, Uzelac, Gordana, Mirković, Nemanja, Devescovi, Giulia, Diep, Dzung B., Venturi, Vittorio, Kojić, Milan, "LsbB Bacteriocin Interacts with the Third Transmembrane Domain of the YvjB Receptor" in Applied and Environmental Microbiology, 82, no. 17 (2016):5364-5374,
https://doi.org/10.1128/AEM.01293-16 . .

TY  - JOUR
AU  - Miljković, Marija
AU  - Bertani, Iris
AU  - Fira, Đorđe
AU  - Jovčić, Branko
AU  - Novović, Katarina
AU  - Venturi, Vittorio
AU  - Kojić, Milan
PY  - 2016
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/924
AB  - AggLb is the largest (318.6 kDa) aggregation-promoting protein of Lactobacillus paracasei subsp. paracasei BGNJ1-64 responsible for forming large cell aggregates, which causes auto-aggregation, collagen binding and pathogen exclusion in vitro. It contains an N-terminus leader peptide, followed by six successive collagen binding domains, 20 successive repeats (CnaB-like domains) and an LPXTG sorting signal at the C-terminus for cell wall anchoring. Experimental information about the roles of the domains of AggLb is currently unknown. To define the domain that confers cell aggregation and the key domains for interactions of specific affinity between AggLb and components of the extracellular matrix, we constructed a series of variants of the aggLb gene and expressed them in Lactococcus lactis subsp. lactis BGKP1-20 using a lactococcal promoter. All of the variants contained a leader peptide, an inter collagen binding-CnaB domain region (used to raise an anti-AggLb antibody), an anchor domain and a different number of collagen binding and CnaB-like domains. The role of the collagen binding repeats of the N-terminus in auto-aggregation and binding to collagen and fibronectin was confirmed. Deletion of the collagen binding repeats II, III, and IV resulted in a loss of the strong auto-aggregation, collagen and fibronectin binding abilities whereas the biofilm formation capability was increased. The strong auto-aggregation, collagen and fibronectin binding abilities of AggLb were negatively correlated to biofilm formation.
PB  - Frontiers Media Sa, Lausanne
T2  - Frontiers in Microbiology
T1  - Shortening of the Lactobacillus paracasei subsp paracasei BGNJ164 AggLb Protein Switches Its Activity from Auto-aggregation to Biofilm Formation
EP  - 14
SP  - 1
VL  - 7
DO  - 10.3389/fmicb.2016.01422
ER  - 

@article{
author = "Miljković, Marija and Bertani, Iris and Fira, Đorđe and Jovčić, Branko and Novović, Katarina and Venturi, Vittorio and Kojić, Milan",
year = "2016",
abstract = "AggLb is the largest (318.6 kDa) aggregation-promoting protein of Lactobacillus paracasei subsp. paracasei BGNJ1-64 responsible for forming large cell aggregates, which causes auto-aggregation, collagen binding and pathogen exclusion in vitro. It contains an N-terminus leader peptide, followed by six successive collagen binding domains, 20 successive repeats (CnaB-like domains) and an LPXTG sorting signal at the C-terminus for cell wall anchoring. Experimental information about the roles of the domains of AggLb is currently unknown. To define the domain that confers cell aggregation and the key domains for interactions of specific affinity between AggLb and components of the extracellular matrix, we constructed a series of variants of the aggLb gene and expressed them in Lactococcus lactis subsp. lactis BGKP1-20 using a lactococcal promoter. All of the variants contained a leader peptide, an inter collagen binding-CnaB domain region (used to raise an anti-AggLb antibody), an anchor domain and a different number of collagen binding and CnaB-like domains. The role of the collagen binding repeats of the N-terminus in auto-aggregation and binding to collagen and fibronectin was confirmed. Deletion of the collagen binding repeats II, III, and IV resulted in a loss of the strong auto-aggregation, collagen and fibronectin binding abilities whereas the biofilm formation capability was increased. The strong auto-aggregation, collagen and fibronectin binding abilities of AggLb were negatively correlated to biofilm formation.",
publisher = "Frontiers Media Sa, Lausanne",
journal = "Frontiers in Microbiology",
title = "Shortening of the Lactobacillus paracasei subsp paracasei BGNJ164 AggLb Protein Switches Its Activity from Auto-aggregation to Biofilm Formation",
pages = "14-1",
volume = "7",
doi = "10.3389/fmicb.2016.01422"
}

Miljković, M., Bertani, I., Fira, Đ., Jovčić, B., Novović, K., Venturi, V.,& Kojić, M.. (2016). Shortening of the Lactobacillus paracasei subsp paracasei BGNJ164 AggLb Protein Switches Its Activity from Auto-aggregation to Biofilm Formation. in Frontiers in Microbiology
Frontiers Media Sa, Lausanne., 7, 1-14.
https://doi.org/10.3389/fmicb.2016.01422

Miljković M, Bertani I, Fira Đ, Jovčić B, Novović K, Venturi V, Kojić M. Shortening of the Lactobacillus paracasei subsp paracasei BGNJ164 AggLb Protein Switches Its Activity from Auto-aggregation to Biofilm Formation. in Frontiers in Microbiology. 2016;7:1-14.
doi:10.3389/fmicb.2016.01422 .

Miljković, Marija, Bertani, Iris, Fira, Đorđe, Jovčić, Branko, Novović, Katarina, Venturi, Vittorio, Kojić, Milan, "Shortening of the Lactobacillus paracasei subsp paracasei BGNJ164 AggLb Protein Switches Its Activity from Auto-aggregation to Biofilm Formation" in Frontiers in Microbiology, 7 (2016):1-14,
https://doi.org/10.3389/fmicb.2016.01422 . .

TY  - JOUR
AU  - Živković, Milica
AU  - Miljković, Marija
AU  - Ruas-Madiedo, Patricia
AU  - Strahinić, Ivana
AU  - Tolinački, Maja
AU  - Golić, Nataša
AU  - Kojić, Milan
PY  - 2015
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/886
AB  - Lactobacillus paraplantarum BGCG11, a putative probiotic strain isolated from a soft, white, artisanal cheese, produces a high-molecular-weight heteropolysaccharide, exopolysaccharide (EPS)-CG11, responsible for the ropy phenotype and immunomodulatory activity of the strain. In this study, a 26.4-kb region originating from the pCG1 plasmid, previously shown to be responsible for the production of EPS-CG11 and a ropy phenotype, was cloned, sequenced, and functionally characterized. In this region 16 putative open reading frames (ORFs), encoding enzymes for the production of EPS-CG11, were organized in specific loci involved in the biosynthesis of the repeat unit, polymerization, export, regulation, and chain length determination. Interestingly, downstream of the eps gene cluster, a putative transposase gene was identified, followed by an additional rfb gene cluster containing the rfbACBD genes, the ones most probably responsible for dTDP-L-rhamnose biosynthesis. The functional analysis showed that the production of the high-molecular-weight fraction of EPS-CG11 was absent in two knockout mutants, one in the eps and the other in the rfb gene cluster, as confirmed by size exclusion chromatography analysis. Therefore, both eps and rfb genes clusters are prerequisites for the production of high-molecular-weight EPS-CG11 and for the ropy phenotype of strain L. paraplantarum BGCG11.
PB  - Amer Soc Microbiology, Washington
T2  - Applied and Environmental Microbiology
T1  - Exopolysaccharide Production and Ropy Phenotype Are Determined by Two Gene Clusters in Putative Probiotic Strain Lactobacillus paraplantarum BGCG11
EP  - 1396
IS  - 4
SP  - 1387
VL  - 81
DO  - 10.1128/AEM.03028-14
ER  - 

@article{
author = "Živković, Milica and Miljković, Marija and Ruas-Madiedo, Patricia and Strahinić, Ivana and Tolinački, Maja and Golić, Nataša and Kojić, Milan",
year = "2015",
abstract = "Lactobacillus paraplantarum BGCG11, a putative probiotic strain isolated from a soft, white, artisanal cheese, produces a high-molecular-weight heteropolysaccharide, exopolysaccharide (EPS)-CG11, responsible for the ropy phenotype and immunomodulatory activity of the strain. In this study, a 26.4-kb region originating from the pCG1 plasmid, previously shown to be responsible for the production of EPS-CG11 and a ropy phenotype, was cloned, sequenced, and functionally characterized. In this region 16 putative open reading frames (ORFs), encoding enzymes for the production of EPS-CG11, were organized in specific loci involved in the biosynthesis of the repeat unit, polymerization, export, regulation, and chain length determination. Interestingly, downstream of the eps gene cluster, a putative transposase gene was identified, followed by an additional rfb gene cluster containing the rfbACBD genes, the ones most probably responsible for dTDP-L-rhamnose biosynthesis. The functional analysis showed that the production of the high-molecular-weight fraction of EPS-CG11 was absent in two knockout mutants, one in the eps and the other in the rfb gene cluster, as confirmed by size exclusion chromatography analysis. Therefore, both eps and rfb genes clusters are prerequisites for the production of high-molecular-weight EPS-CG11 and for the ropy phenotype of strain L. paraplantarum BGCG11.",
publisher = "Amer Soc Microbiology, Washington",
journal = "Applied and Environmental Microbiology",
title = "Exopolysaccharide Production and Ropy Phenotype Are Determined by Two Gene Clusters in Putative Probiotic Strain Lactobacillus paraplantarum BGCG11",
pages = "1396-1387",
number = "4",
volume = "81",
doi = "10.1128/AEM.03028-14"
}

Živković, M., Miljković, M., Ruas-Madiedo, P., Strahinić, I., Tolinački, M., Golić, N.,& Kojić, M.. (2015). Exopolysaccharide Production and Ropy Phenotype Are Determined by Two Gene Clusters in Putative Probiotic Strain Lactobacillus paraplantarum BGCG11. in Applied and Environmental Microbiology
Amer Soc Microbiology, Washington., 81(4), 1387-1396.
https://doi.org/10.1128/AEM.03028-14

Živković M, Miljković M, Ruas-Madiedo P, Strahinić I, Tolinački M, Golić N, Kojić M. Exopolysaccharide Production and Ropy Phenotype Are Determined by Two Gene Clusters in Putative Probiotic Strain Lactobacillus paraplantarum BGCG11. in Applied and Environmental Microbiology. 2015;81(4):1387-1396.
doi:10.1128/AEM.03028-14 .

Živković, Milica, Miljković, Marija, Ruas-Madiedo, Patricia, Strahinić, Ivana, Tolinački, Maja, Golić, Nataša, Kojić, Milan, "Exopolysaccharide Production and Ropy Phenotype Are Determined by Two Gene Clusters in Putative Probiotic Strain Lactobacillus paraplantarum BGCG11" in Applied and Environmental Microbiology, 81, no. 4 (2015):1387-1396,
https://doi.org/10.1128/AEM.03028-14 . .

TY  - JOUR
AU  - Terzić-Vidojević, Amarela
AU  - Veljović, Katarina
AU  - Begović, Jelena
AU  - Filipić, Brankica
AU  - Popović, Dušanka
AU  - Tolinački, Maja
AU  - Miljković, Marija
AU  - Kojić, Milan
AU  - Golić, Nataša
PY  - 2015
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/820
AB  - Enterococci represent the most controversial group of dairy bacteria. They are found to be the main constituent of many traditional Mediterranean dairy products and contribute to their characteristic taste and flavor. On the other hand, during the last 50 years antibiotic resistant enterococci have emerged as leading causes of nosocomial infections worldwide. The aim of this study was to determine the diversity, technological properties, antibiotic susceptibility and virulence traits of 636 enterococci previously isolated from 55 artisan dairy products from 12 locations in the Western Balkan countries (WBC) of Serbia, Croatia and Bosnia and Herzegovina. All strains were identified both by microbiological and molecular methods. The predominant species was Enterococcus durans, followed by Enterococcus faecalis and Enterococcus faecium. Over 44% of the isolates were resistant to ciprofloxacin and erythromycin, while 26.2% of the isolates were multi resistant to three or more antibiotics belonging to different families. 185 isolates (29.1%) were susceptible to all 13 of the antibiotics tested. The antibiotic-susceptible isolates were further tested for possible virulence genes and the production of biogenic amines. Finally, five enterococci isolates were found to be antibiotic susceptible with good technological characteristics and without virulence traits or the ability to produce biogenic amines, making them possible candidates for biotechnological application as starter cultures in the dairy industry.
PB  - Frontiers Media Sa, Lausanne
T2  - Frontiers in Microbiology
T1  - Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?
VL  - 6
DO  - 10.3389/fmicb.2015.00954
ER  - 

@article{
author = "Terzić-Vidojević, Amarela and Veljović, Katarina and Begović, Jelena and Filipić, Brankica and Popović, Dušanka and Tolinački, Maja and Miljković, Marija and Kojić, Milan and Golić, Nataša",
year = "2015",
abstract = "Enterococci represent the most controversial group of dairy bacteria. They are found to be the main constituent of many traditional Mediterranean dairy products and contribute to their characteristic taste and flavor. On the other hand, during the last 50 years antibiotic resistant enterococci have emerged as leading causes of nosocomial infections worldwide. The aim of this study was to determine the diversity, technological properties, antibiotic susceptibility and virulence traits of 636 enterococci previously isolated from 55 artisan dairy products from 12 locations in the Western Balkan countries (WBC) of Serbia, Croatia and Bosnia and Herzegovina. All strains were identified both by microbiological and molecular methods. The predominant species was Enterococcus durans, followed by Enterococcus faecalis and Enterococcus faecium. Over 44% of the isolates were resistant to ciprofloxacin and erythromycin, while 26.2% of the isolates were multi resistant to three or more antibiotics belonging to different families. 185 isolates (29.1%) were susceptible to all 13 of the antibiotics tested. The antibiotic-susceptible isolates were further tested for possible virulence genes and the production of biogenic amines. Finally, five enterococci isolates were found to be antibiotic susceptible with good technological characteristics and without virulence traits or the ability to produce biogenic amines, making them possible candidates for biotechnological application as starter cultures in the dairy industry.",
publisher = "Frontiers Media Sa, Lausanne",
journal = "Frontiers in Microbiology",
title = "Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?",
volume = "6",
doi = "10.3389/fmicb.2015.00954"
}

Terzić-Vidojević, A., Veljović, K., Begović, J., Filipić, B., Popović, D., Tolinački, M., Miljković, M., Kojić, M.,& Golić, N.. (2015). Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?. in Frontiers in Microbiology
Frontiers Media Sa, Lausanne., 6.
https://doi.org/10.3389/fmicb.2015.00954

Terzić-Vidojević A, Veljović K, Begović J, Filipić B, Popović D, Tolinački M, Miljković M, Kojić M, Golić N. Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?. in Frontiers in Microbiology. 2015;6.
doi:10.3389/fmicb.2015.00954 .

Terzić-Vidojević, Amarela, Veljović, Katarina, Begović, Jelena, Filipić, Brankica, Popović, Dušanka, Tolinački, Maja, Miljković, Marija, Kojić, Milan, Golić, Nataša, "Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?" in Frontiers in Microbiology, 6 (2015),
https://doi.org/10.3389/fmicb.2015.00954 . .

TY  - JOUR
AU  - Vukotić, Goran
AU  - Mirković, Nemanja
AU  - Jovčić, Branko
AU  - Miljković, Marija
AU  - Strahinić, Ivana
AU  - Fira, Đorđe
AU  - Radulović, Zorica
AU  - Kojić, Milan
PY  - 2015
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/895
AB  - Proteinases and bacteriocins are of great importance to the dairy industry, but their interactions have not been studied so far. Lactococcus lactis subsp. lactis BGMN1-5 is a natural isolate from homemade semi-hard cheese which produces two bacteriocins (Lactococcin B and LsbB), as well as proteinase PrtP. A medium-dependent increase in the bacteriocin LcnB activity of L. lactis BGMN1-501, a derivate of L. lactis subsp. lactis BGMN1-5, was shown to be accompanied by a decrease in its promoter activity. A similar effect of media components on gene expression was reported for proteinase PrtP whose gene is co-localized on the same plasmid as the IcnB gene. Thus, the PrtP-LcnB interplay was investigated. Single gene knockout mutants were constructed with disrupted prtP or IcnB genes. PrtP mutants showed higher bacteriocin activity that had lost its growth medium dependence, which was in contrast to the original strain. When LcnB from this mutant was combined with proteinase from the LonB(-) mutant in vitro, its activity was rendered to the original level, suggesting that proteinase reduces bacteriocin activity. We propose a new model of medium dependent expression of these genes with regard to the effects of their interaction in vivo.
PB  - Frontiers Media Sa, Lausanne
T2  - Frontiers in Microbiology
T1  - Proteinase PrtP impairs lactococcin LcnB activity in Lactococcus lactis BGMN1-501: new insights into bacteriocin regulation
VL  - 6
DO  - 10.3389/fmicb.2015.00092
ER  - 

@article{
author = "Vukotić, Goran and Mirković, Nemanja and Jovčić, Branko and Miljković, Marija and Strahinić, Ivana and Fira, Đorđe and Radulović, Zorica and Kojić, Milan",
year = "2015",
abstract = "Proteinases and bacteriocins are of great importance to the dairy industry, but their interactions have not been studied so far. Lactococcus lactis subsp. lactis BGMN1-5 is a natural isolate from homemade semi-hard cheese which produces two bacteriocins (Lactococcin B and LsbB), as well as proteinase PrtP. A medium-dependent increase in the bacteriocin LcnB activity of L. lactis BGMN1-501, a derivate of L. lactis subsp. lactis BGMN1-5, was shown to be accompanied by a decrease in its promoter activity. A similar effect of media components on gene expression was reported for proteinase PrtP whose gene is co-localized on the same plasmid as the IcnB gene. Thus, the PrtP-LcnB interplay was investigated. Single gene knockout mutants were constructed with disrupted prtP or IcnB genes. PrtP mutants showed higher bacteriocin activity that had lost its growth medium dependence, which was in contrast to the original strain. When LcnB from this mutant was combined with proteinase from the LonB(-) mutant in vitro, its activity was rendered to the original level, suggesting that proteinase reduces bacteriocin activity. We propose a new model of medium dependent expression of these genes with regard to the effects of their interaction in vivo.",
publisher = "Frontiers Media Sa, Lausanne",
journal = "Frontiers in Microbiology",
title = "Proteinase PrtP impairs lactococcin LcnB activity in Lactococcus lactis BGMN1-501: new insights into bacteriocin regulation",
volume = "6",
doi = "10.3389/fmicb.2015.00092"
}

Vukotić, G., Mirković, N., Jovčić, B., Miljković, M., Strahinić, I., Fira, Đ., Radulović, Z.,& Kojić, M.. (2015). Proteinase PrtP impairs lactococcin LcnB activity in Lactococcus lactis BGMN1-501: new insights into bacteriocin regulation. in Frontiers in Microbiology
Frontiers Media Sa, Lausanne., 6.
https://doi.org/10.3389/fmicb.2015.00092

Vukotić G, Mirković N, Jovčić B, Miljković M, Strahinić I, Fira Đ, Radulović Z, Kojić M. Proteinase PrtP impairs lactococcin LcnB activity in Lactococcus lactis BGMN1-501: new insights into bacteriocin regulation. in Frontiers in Microbiology. 2015;6.
doi:10.3389/fmicb.2015.00092 .

Vukotić, Goran, Mirković, Nemanja, Jovčić, Branko, Miljković, Marija, Strahinić, Ivana, Fira, Đorđe, Radulović, Zorica, Kojić, Milan, "Proteinase PrtP impairs lactococcin LcnB activity in Lactococcus lactis BGMN1-501: new insights into bacteriocin regulation" in Frontiers in Microbiology, 6 (2015),
https://doi.org/10.3389/fmicb.2015.00092 . .

TY  - JOUR
AU  - Miljković, Marija
AU  - Strahinić, Ivana
AU  - Tolinački, Maja
AU  - Živković, Milica
AU  - Kojić, Snežana
AU  - Golić, Nataša
AU  - Kojić, Milan
PY  - 2015
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/805
AB  - Eleven Lactobacillus strains with strong aggregation abilities were selected from a laboratory collection. In two of the strains, genes associated with aggregation capability were plasmid located and found to strongly correlate with collagen binding. The gene encoding the auto-aggregation-promoting protein (AggLb) of Lactobacillus paracasei subsp. paracasei BGNJ1-64 was cloned using a novel, wide-range-host shuttle cloning vector, pAZILSJ. The clone pALb35, containing a 11377-bp DNA fragment, was selected from the SacI plasmid library for its ability to provide carriers with the aggregation phenotype. The complete fragment was sequenced and four potential ORFs were detected, including the aggLb gene and three surrounding transposase genes. AggLb is the largest known cell-surface protein in lactobacilli, consisting of 2998 aa (318,611 Da). AggLb belongs to the collagen-binding superfamily and its C-terminal region contains 20 successive repeats that are identical even at the nucleotide level. Deletion of aggLb causes a loss of the capacity to form cell aggregates, whereas overexpression increases cellular aggregation, hydrophobicity and collagen-binding potential. PCR screening performed with three sets of primers based on the aggLb gene of BGNJ1-64 enabled detection of the same type of aggLb gene in five of eleven selected aggregation-positive Lactobacillus strains. Heterologous expression of aggLb confirmed the crucial role of the AggLb protein in cell aggregation and specific collagen binding, indicating that AggLb has a useful probiotic function in effective colonization of host tissue and prevention of pathogen colonization.
PB  - Public Library Science, San Francisco
T2  - PLoS One
T1  - AggLb Is the Largest Cell-Aggregation Factor from Lactobacillus paracasei Subsp paracasei BGNJ1-64, Functions in Collagen Adhesion, and Pathogen Exclusion In Vitro
IS  - 5
VL  - 10
DO  - 10.1371/journal.pone.0126387
ER  - 

@article{
author = "Miljković, Marija and Strahinić, Ivana and Tolinački, Maja and Živković, Milica and Kojić, Snežana and Golić, Nataša and Kojić, Milan",
year = "2015",
abstract = "Eleven Lactobacillus strains with strong aggregation abilities were selected from a laboratory collection. In two of the strains, genes associated with aggregation capability were plasmid located and found to strongly correlate with collagen binding. The gene encoding the auto-aggregation-promoting protein (AggLb) of Lactobacillus paracasei subsp. paracasei BGNJ1-64 was cloned using a novel, wide-range-host shuttle cloning vector, pAZILSJ. The clone pALb35, containing a 11377-bp DNA fragment, was selected from the SacI plasmid library for its ability to provide carriers with the aggregation phenotype. The complete fragment was sequenced and four potential ORFs were detected, including the aggLb gene and three surrounding transposase genes. AggLb is the largest known cell-surface protein in lactobacilli, consisting of 2998 aa (318,611 Da). AggLb belongs to the collagen-binding superfamily and its C-terminal region contains 20 successive repeats that are identical even at the nucleotide level. Deletion of aggLb causes a loss of the capacity to form cell aggregates, whereas overexpression increases cellular aggregation, hydrophobicity and collagen-binding potential. PCR screening performed with three sets of primers based on the aggLb gene of BGNJ1-64 enabled detection of the same type of aggLb gene in five of eleven selected aggregation-positive Lactobacillus strains. Heterologous expression of aggLb confirmed the crucial role of the AggLb protein in cell aggregation and specific collagen binding, indicating that AggLb has a useful probiotic function in effective colonization of host tissue and prevention of pathogen colonization.",
publisher = "Public Library Science, San Francisco",
journal = "PLoS One",
title = "AggLb Is the Largest Cell-Aggregation Factor from Lactobacillus paracasei Subsp paracasei BGNJ1-64, Functions in Collagen Adhesion, and Pathogen Exclusion In Vitro",
number = "5",
volume = "10",
doi = "10.1371/journal.pone.0126387"
}

Miljković, M., Strahinić, I., Tolinački, M., Živković, M., Kojić, S., Golić, N.,& Kojić, M.. (2015). AggLb Is the Largest Cell-Aggregation Factor from Lactobacillus paracasei Subsp paracasei BGNJ1-64, Functions in Collagen Adhesion, and Pathogen Exclusion In Vitro. in PLoS One
Public Library Science, San Francisco., 10(5).
https://doi.org/10.1371/journal.pone.0126387

Miljković M, Strahinić I, Tolinački M, Živković M, Kojić S, Golić N, Kojić M. AggLb Is the Largest Cell-Aggregation Factor from Lactobacillus paracasei Subsp paracasei BGNJ1-64, Functions in Collagen Adhesion, and Pathogen Exclusion In Vitro. in PLoS One. 2015;10(5).
doi:10.1371/journal.pone.0126387 .

Miljković, Marija, Strahinić, Ivana, Tolinački, Maja, Živković, Milica, Kojić, Snežana, Golić, Nataša, Kojić, Milan, "AggLb Is the Largest Cell-Aggregation Factor from Lactobacillus paracasei Subsp paracasei BGNJ1-64, Functions in Collagen Adhesion, and Pathogen Exclusion In Vitro" in PLoS One, 10, no. 5 (2015),
https://doi.org/10.1371/journal.pone.0126387 . .

TY  - JOUR
AU  - Uzelac, Gordana
AU  - Miljković, Marija
AU  - Lozo, Jelena
AU  - Radulović, Zorica
AU  - Tošić, Nataša
AU  - Kojić, Milan
PY  - 2015
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/829
AB  - The production of LsbB, leaderless class II bacteriocin, is encoded by genes (lsbB and lmrB) located on plasmid pMN5 in Lactococcus lactis BGMN1-5. Heterologous expression of the lsbB gene using the pAZIL vector (pAZIL-lsbB) in L. lactis subsp. cremoris MG7284 resulted in a significant reduction (more than 30 times) of bacteriocin LsbB expression. Subcloning and deletion experiments with plasmid pMN5 revealed that full expression of LsbB requires the presence of a complete transcription terminator located downstream of the lsbB gene. RNA stability analysis revealed that the presence of a transcription terminator increased the RNA stability by three times and the expression of LsbB by 30 times. The study of the influence of transcription terminator on the expression of other bacteriocin genes (lcnB, for lactococcin B production) indicated that this translational terminator likely functions in a lsbB-specific manner rather than in a general manner.
PB  - Elsevier Gmbh, Urban & Fischer Verlag, Jena
T2  - Microbiological Research
T1  - Expression of bacteriocin LsbB is dependent on a transcription terminator
EP  - 53
SP  - 45
VL  - 179
DO  - 10.1016/j.micres.2015.06.011
ER  - 

@article{
author = "Uzelac, Gordana and Miljković, Marija and Lozo, Jelena and Radulović, Zorica and Tošić, Nataša and Kojić, Milan",
year = "2015",
abstract = "The production of LsbB, leaderless class II bacteriocin, is encoded by genes (lsbB and lmrB) located on plasmid pMN5 in Lactococcus lactis BGMN1-5. Heterologous expression of the lsbB gene using the pAZIL vector (pAZIL-lsbB) in L. lactis subsp. cremoris MG7284 resulted in a significant reduction (more than 30 times) of bacteriocin LsbB expression. Subcloning and deletion experiments with plasmid pMN5 revealed that full expression of LsbB requires the presence of a complete transcription terminator located downstream of the lsbB gene. RNA stability analysis revealed that the presence of a transcription terminator increased the RNA stability by three times and the expression of LsbB by 30 times. The study of the influence of transcription terminator on the expression of other bacteriocin genes (lcnB, for lactococcin B production) indicated that this translational terminator likely functions in a lsbB-specific manner rather than in a general manner.",
publisher = "Elsevier Gmbh, Urban & Fischer Verlag, Jena",
journal = "Microbiological Research",
title = "Expression of bacteriocin LsbB is dependent on a transcription terminator",
pages = "53-45",
volume = "179",
doi = "10.1016/j.micres.2015.06.011"
}

Uzelac, G., Miljković, M., Lozo, J., Radulović, Z., Tošić, N.,& Kojić, M.. (2015). Expression of bacteriocin LsbB is dependent on a transcription terminator. in Microbiological Research
Elsevier Gmbh, Urban & Fischer Verlag, Jena., 179, 45-53.
https://doi.org/10.1016/j.micres.2015.06.011

Uzelac G, Miljković M, Lozo J, Radulović Z, Tošić N, Kojić M. Expression of bacteriocin LsbB is dependent on a transcription terminator. in Microbiological Research. 2015;179:45-53.
doi:10.1016/j.micres.2015.06.011 .

Uzelac, Gordana, Miljković, Marija, Lozo, Jelena, Radulović, Zorica, Tošić, Nataša, Kojić, Milan, "Expression of bacteriocin LsbB is dependent on a transcription terminator" in Microbiological Research, 179 (2015):45-53,
https://doi.org/10.1016/j.micres.2015.06.011 . .

TY  - JOUR
AU  - Živković, Milica
AU  - Cadez, Neza
AU  - Uroić, Ksenija
AU  - Miljković, Marija
AU  - Tolinački, Maja
AU  - Dousova, Petra
AU  - Kos, Blazenka
AU  - Susković, Jagoda
AU  - Raspor, Peter
AU  - Topisirović, Ljubiša
AU  - Golić, Nataša
PY  - 2015
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/872
UR  - http://www.jocmr.com/fulltext/55-1410953223.pdf?1669198006
AB  - The aim of this study was to investigate the in vitro probiotic potential of dairy yeast isolates from artisanal cheeses manufactured in Serbia and Croatia.  
Methods. Twelve yeast strains isolated in from artisanal fresh soft and white brined cheeses manufactured in Serbia and Croatia were used in the study. Survival in chemically-simulated gastrointestinal conditions, adherence to epithelial intestinal cells and proliferation of gut-associated lymphoid tissue (GALT) cells were evaluated.
Results. The results revealed that two strains of Kluyvereomyces lactis ZIM 2408 and ZIM 2453 grew above one log unit ( and #916; log CFU/ml) in the complex colonic medium during 24 h of cultivation, while Torulaspora delbrueckii ZIM 2460 was the most resistant isolate in chemically-simulated conditions of gastric juice and upper intestinal tract. It was demonstrated that the strains Kluyvereomyces lactis ZIM 2408 and ZIM2441 and Saccharomyces cerevisiae ZIM 2415 were highly adhesive to Caco-2 cells, while strains Kluyvereomyces lactis ZIM 2408 and Debaryomyces hansenii ZIM 2415 exhibit the highest adhesion percentage to HT29-MTX cells. All strains significantly (p lt 0.0001) decreased the proliferation of gut-associated lymphoid tissue (GALT) cells suggesting the possible strain-specific immunomodulatory potential of the isolates. 
Conclusion. The dairy yeast isolates exhibit the strain-specific probiotic properties. Particularly, the strain K. lactis ZIM 2408 appears to be the best probiotic candidate in terms of all three criteria. Taking into account their immunomodulatory potential, the yeast isolates could be further tested for specific probiotic applications and eventually included in functional food formulated for patients suffering from diseases associated with an increased inflammatory status.
T2  - Journal of complementary medicine research
T1  - Evaluation of probiotic potential of yeasts isolated from traditional cheeses manufactured in Serbia and Croatia
EP  - 18
IS  - 1
SP  - 12
VL  - 4
DO  - 10.5455/jice.20141128051842
ER  - 

@article{
author = "Živković, Milica and Cadez, Neza and Uroić, Ksenija and Miljković, Marija and Tolinački, Maja and Dousova, Petra and Kos, Blazenka and Susković, Jagoda and Raspor, Peter and Topisirović, Ljubiša and Golić, Nataša",
year = "2015",
abstract = "The aim of this study was to investigate the in vitro probiotic potential of dairy yeast isolates from artisanal cheeses manufactured in Serbia and Croatia.  
Methods. Twelve yeast strains isolated in from artisanal fresh soft and white brined cheeses manufactured in Serbia and Croatia were used in the study. Survival in chemically-simulated gastrointestinal conditions, adherence to epithelial intestinal cells and proliferation of gut-associated lymphoid tissue (GALT) cells were evaluated.
Results. The results revealed that two strains of Kluyvereomyces lactis ZIM 2408 and ZIM 2453 grew above one log unit ( and #916; log CFU/ml) in the complex colonic medium during 24 h of cultivation, while Torulaspora delbrueckii ZIM 2460 was the most resistant isolate in chemically-simulated conditions of gastric juice and upper intestinal tract. It was demonstrated that the strains Kluyvereomyces lactis ZIM 2408 and ZIM2441 and Saccharomyces cerevisiae ZIM 2415 were highly adhesive to Caco-2 cells, while strains Kluyvereomyces lactis ZIM 2408 and Debaryomyces hansenii ZIM 2415 exhibit the highest adhesion percentage to HT29-MTX cells. All strains significantly (p lt 0.0001) decreased the proliferation of gut-associated lymphoid tissue (GALT) cells suggesting the possible strain-specific immunomodulatory potential of the isolates. 
Conclusion. The dairy yeast isolates exhibit the strain-specific probiotic properties. Particularly, the strain K. lactis ZIM 2408 appears to be the best probiotic candidate in terms of all three criteria. Taking into account their immunomodulatory potential, the yeast isolates could be further tested for specific probiotic applications and eventually included in functional food formulated for patients suffering from diseases associated with an increased inflammatory status.",
journal = "Journal of complementary medicine research",
title = "Evaluation of probiotic potential of yeasts isolated from traditional cheeses manufactured in Serbia and Croatia",
pages = "18-12",
number = "1",
volume = "4",
doi = "10.5455/jice.20141128051842"
}

Živković, M., Cadez, N., Uroić, K., Miljković, M., Tolinački, M., Dousova, P., Kos, B., Susković, J., Raspor, P., Topisirović, L.,& Golić, N.. (2015). Evaluation of probiotic potential of yeasts isolated from traditional cheeses manufactured in Serbia and Croatia. in Journal of complementary medicine research, 4(1), 12-18.
https://doi.org/10.5455/jice.20141128051842

Živković M, Cadez N, Uroić K, Miljković M, Tolinački M, Dousova P, Kos B, Susković J, Raspor P, Topisirović L, Golić N. Evaluation of probiotic potential of yeasts isolated from traditional cheeses manufactured in Serbia and Croatia. in Journal of complementary medicine research. 2015;4(1):12-18.
doi:10.5455/jice.20141128051842 .

Živković, Milica, Cadez, Neza, Uroić, Ksenija, Miljković, Marija, Tolinački, Maja, Dousova, Petra, Kos, Blazenka, Susković, Jagoda, Raspor, Peter, Topisirović, Ljubiša, Golić, Nataša, "Evaluation of probiotic potential of yeasts isolated from traditional cheeses manufactured in Serbia and Croatia" in Journal of complementary medicine research, 4, no. 1 (2015):12-18,
https://doi.org/10.5455/jice.20141128051842 . .