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  - Mirković, Nemanja
AU  - Radulović, Zorica
AU  - Uzelac, Gordana
AU  - Lozo, Jelena
AU  - Obradović, Dragojlo
AU  - Topisirović, Ljubiša
AU  - Kojić, Milan
PY  - 2015
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/870
AB  - Lactococcus locus ssp. lactis BGBM50, a producer of lactococcin G and aggregation-promoting factor, was isolated from selected lactic acid bacteria taken from semi-hard cheese traditionally produced in the village Zanjic, Montenegro. Strain BGBM50 harbours a number of plasmids of different sizes. Plasmid curing experiments showed that genes for bacteriocin production are located on pBM140, a plasmid 140 kb in length. PCR analysis with primers specific for lactococcin Q and G genes gave fragment of the expected size. In addition, after plasmid curing of strain BGBM50, different derivatives with altered phenotypes were obtained, among them BGBM50-34 strain, which retained bacteriocin synthesis but had enhanced aggregation ability.
PB  - University of Zagreb
T2  - Food Technology and Biotechnology
T1  - Isolation and Characterisation of Bacteriocin and Aggregation-Promoting Factor Production in Lactococcus lactis ssp lactis BGBM50 Strain
EP  - 242
IS  - 2
SP  - 237
VL  - 53
DO  - 10.17113/ftb.53.02.15.3846
ER  - 

@article{
author = "Mirković, Nemanja and Radulović, Zorica and Uzelac, Gordana and Lozo, Jelena and Obradović, Dragojlo and Topisirović, Ljubiša and Kojić, Milan",
year = "2015",
abstract = "Lactococcus locus ssp. lactis BGBM50, a producer of lactococcin G and aggregation-promoting factor, was isolated from selected lactic acid bacteria taken from semi-hard cheese traditionally produced in the village Zanjic, Montenegro. Strain BGBM50 harbours a number of plasmids of different sizes. Plasmid curing experiments showed that genes for bacteriocin production are located on pBM140, a plasmid 140 kb in length. PCR analysis with primers specific for lactococcin Q and G genes gave fragment of the expected size. In addition, after plasmid curing of strain BGBM50, different derivatives with altered phenotypes were obtained, among them BGBM50-34 strain, which retained bacteriocin synthesis but had enhanced aggregation ability.",
publisher = "University of Zagreb",
journal = "Food Technology and Biotechnology",
title = "Isolation and Characterisation of Bacteriocin and Aggregation-Promoting Factor Production in Lactococcus lactis ssp lactis BGBM50 Strain",
pages = "242-237",
number = "2",
volume = "53",
doi = "10.17113/ftb.53.02.15.3846"
}

Mirković, N., Radulović, Z., Uzelac, G., Lozo, J., Obradović, D., Topisirović, L.,& Kojić, M.. (2015). Isolation and Characterisation of Bacteriocin and Aggregation-Promoting Factor Production in Lactococcus lactis ssp lactis BGBM50 Strain. in Food Technology and Biotechnology
University of Zagreb., 53(2), 237-242.
https://doi.org/10.17113/ftb.53.02.15.3846

Mirković N, Radulović Z, Uzelac G, Lozo J, Obradović D, Topisirović L, Kojić M. Isolation and Characterisation of Bacteriocin and Aggregation-Promoting Factor Production in Lactococcus lactis ssp lactis BGBM50 Strain. in Food Technology and Biotechnology. 2015;53(2):237-242.
doi:10.17113/ftb.53.02.15.3846 .

Mirković, Nemanja, Radulović, Zorica, Uzelac, Gordana, Lozo, Jelena, Obradović, Dragojlo, Topisirović, Ljubiša, Kojić, Milan, "Isolation and Characterisation of Bacteriocin and Aggregation-Promoting Factor Production in Lactococcus lactis ssp lactis BGBM50 Strain" in Food Technology and Biotechnology, 53, no. 2 (2015):237-242,
https://doi.org/10.17113/ftb.53.02.15.3846 . .

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  - Ovchinnikov, Kirill V.
AU  - Kristiansen, Per E.
AU  - Uzelac, Gordana
AU  - Topisirović, Ljubiša
AU  - Kojić, Milan
AU  - Nissen-Meyer, Jon
AU  - Nes, Ingolf F.
AU  - Diep, Dzung B.
PY  - 2014
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/750
AB  - Background: The bacteriocin LsbB targets a membrane-bound zinc-dependent peptidase. Results: The structure of LsbB was resolved by NMR. The C-terminal unstructured domains of LsbB and several other related bacteriocins were responsible for receptor binding. Conclusion: A subgroup of leaderless bacteriocins has been found to share a similar mechanism in receptor recognition. Significance: The study highlights the structure-function relationship of LsbB. LsbB is a class II leaderless lactococcal bacteriocin of 30 amino acids. In the present work, the structure and function relationship of LsbB was assessed. Structure determination by NMR spectroscopy showed that LsbB has an N-terminal -helix, whereas the C-terminal of the molecule remains unstructured. To define the receptor binding domain of LsbB, a competition assay was performed in which a systematic collection of truncated peptides of various lengths covering different parts of LsbB was used to inhibit the antimicrobial activity of LsbB. The results indicate that the outmost eight-amino acid sequence at the C-terminal end is likely to contain the receptor binding domain because only truncated fragments from this region could antagonize the antimicrobial activity of LsbB. Furthermore, alanine substitution revealed that the tryptophan in position 25 (Trp(25)) is crucial for the blocking activity of the truncated peptides, as well as for the antimicrobial activity of the full-length bacteriocin. LsbB shares significant sequence homology with five other leaderless bacteriocins, especially at their C-terminal halves where all contain a conserved KXXXGXXPWE motif, suggesting that they might recognize the same receptor as LsbB. This notion was supported by the fact that truncated peptides with sequences derived from the C-terminal regions of two LsbB-related bacteriocins inhibited the activity of LsbB, in the same manner as found with the truncated version of LsbB. Taken together, these structure-function studies provide strong evidence that the receptor-binding parts of LsbB and sequence-related bacteriocins are located in their C-terminal halves.
PB  - Amer Soc Biochemistry Molecular Biology Inc, Bethesda
T2  - Journal of Biological Chemistry
T1  - Defining the Structure and Receptor Binding Domain of the Leaderless Bacteriocin LsbB
EP  - 23845
IS  - 34
SP  - 23838
VL  - 289
DO  - 10.1074/jbc.M114.579698
ER  - 

@article{
author = "Ovchinnikov, Kirill V. and Kristiansen, Per E. and Uzelac, Gordana and Topisirović, Ljubiša and Kojić, Milan and Nissen-Meyer, Jon and Nes, Ingolf F. and Diep, Dzung B.",
year = "2014",
abstract = "Background: The bacteriocin LsbB targets a membrane-bound zinc-dependent peptidase. Results: The structure of LsbB was resolved by NMR. The C-terminal unstructured domains of LsbB and several other related bacteriocins were responsible for receptor binding. Conclusion: A subgroup of leaderless bacteriocins has been found to share a similar mechanism in receptor recognition. Significance: The study highlights the structure-function relationship of LsbB. LsbB is a class II leaderless lactococcal bacteriocin of 30 amino acids. In the present work, the structure and function relationship of LsbB was assessed. Structure determination by NMR spectroscopy showed that LsbB has an N-terminal -helix, whereas the C-terminal of the molecule remains unstructured. To define the receptor binding domain of LsbB, a competition assay was performed in which a systematic collection of truncated peptides of various lengths covering different parts of LsbB was used to inhibit the antimicrobial activity of LsbB. The results indicate that the outmost eight-amino acid sequence at the C-terminal end is likely to contain the receptor binding domain because only truncated fragments from this region could antagonize the antimicrobial activity of LsbB. Furthermore, alanine substitution revealed that the tryptophan in position 25 (Trp(25)) is crucial for the blocking activity of the truncated peptides, as well as for the antimicrobial activity of the full-length bacteriocin. LsbB shares significant sequence homology with five other leaderless bacteriocins, especially at their C-terminal halves where all contain a conserved KXXXGXXPWE motif, suggesting that they might recognize the same receptor as LsbB. This notion was supported by the fact that truncated peptides with sequences derived from the C-terminal regions of two LsbB-related bacteriocins inhibited the activity of LsbB, in the same manner as found with the truncated version of LsbB. Taken together, these structure-function studies provide strong evidence that the receptor-binding parts of LsbB and sequence-related bacteriocins are located in their C-terminal halves.",
publisher = "Amer Soc Biochemistry Molecular Biology Inc, Bethesda",
journal = "Journal of Biological Chemistry",
title = "Defining the Structure and Receptor Binding Domain of the Leaderless Bacteriocin LsbB",
pages = "23845-23838",
number = "34",
volume = "289",
doi = "10.1074/jbc.M114.579698"
}

Ovchinnikov, K. V., Kristiansen, P. E., Uzelac, G., Topisirović, L., Kojić, M., Nissen-Meyer, J., Nes, I. F.,& Diep, D. B.. (2014). Defining the Structure and Receptor Binding Domain of the Leaderless Bacteriocin LsbB. in Journal of Biological Chemistry
Amer Soc Biochemistry Molecular Biology Inc, Bethesda., 289(34), 23838-23845.
https://doi.org/10.1074/jbc.M114.579698

Ovchinnikov KV, Kristiansen PE, Uzelac G, Topisirović L, Kojić M, Nissen-Meyer J, Nes IF, Diep DB. Defining the Structure and Receptor Binding Domain of the Leaderless Bacteriocin LsbB. in Journal of Biological Chemistry. 2014;289(34):23838-23845.
doi:10.1074/jbc.M114.579698 .

Ovchinnikov, Kirill V., Kristiansen, Per E., Uzelac, Gordana, Topisirović, Ljubiša, Kojić, Milan, Nissen-Meyer, Jon, Nes, Ingolf F., Diep, Dzung B., "Defining the Structure and Receptor Binding Domain of the Leaderless Bacteriocin LsbB" in Journal of Biological Chemistry, 289, no. 34 (2014):23838-23845,
https://doi.org/10.1074/jbc.M114.579698 . .

TY  - JOUR
AU  - Terzić-Vidojević, Amarela
AU  - Mihajlović, Sanja
AU  - Uzelac, Gordana
AU  - Veljović, Katarina
AU  - Tolinački, Maja
AU  - Živković, Milica
AU  - Topisirović, Ljubiša
AU  - Kojić, Milan
PY  - 2014
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/791
AB  - The aim of this study was to investigate the composition of lactic acid bacteria (LAB) in autochthonous young cheeses, sweet creams and sweet kajmaks produced in the Vlasic mountain region of central Bosnia and Herzegovina near the town of Travnik over a four season period. These three products were made from cow's milk by a traditional method without the addition of a starter culture. Preliminary characterization with phenotype-based assays and identification using rep-PCR with a (GTG)(5) primer and 16S rDNA sequence analysis were undertaken for 460 LAB isolates obtained from all the examined samples. Fifteen species were identified as follows: Lactococcus lactis, Lactococcus raffinolactis, Lactococcus garviae, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Enterococcus faecium, Enterococcus durans, Enterococcus faecalis, Enterococcus italicus, Leuconostoc mesenteroides, Leuconostoc pseudomesenteroides, Leuconostoc lactis, Streptococcus thermophilus and Streptococcus mitis. A wide genotypic and phenotypic heterogeneity of the species was observed, particularly within the Lc. lactis strains. In all of the tested dairy products across four seasons, a significantly positive correlation (r = 0.690) between the presence of lactococci and enterococci and a negative correlation (r = 0.722) between the presence of lactococci and leuconostocs were recorded. Forty-five percent of the lactobacilli and 54.4% of the lactococci exhibited proteolytic activity, whereas 18.7% of the total LAB isolates exhibited antimicrobial activity.
PB  - Academic Press Ltd- Elsevier Science Ltd, London
T2  - Food Microbiology
T1  - Characterization of lactic acid bacteria isolated from artisanal Travnik young cheeses, sweet creams and sweet kajmaks over four seasons
EP  - 38
SP  - 27
VL  - 39
DO  - 10.1016/j.fm.2013.10.011
ER  - 

@article{
author = "Terzić-Vidojević, Amarela and Mihajlović, Sanja and Uzelac, Gordana and Veljović, Katarina and Tolinački, Maja and Živković, Milica and Topisirović, Ljubiša and Kojić, Milan",
year = "2014",
abstract = "The aim of this study was to investigate the composition of lactic acid bacteria (LAB) in autochthonous young cheeses, sweet creams and sweet kajmaks produced in the Vlasic mountain region of central Bosnia and Herzegovina near the town of Travnik over a four season period. These three products were made from cow's milk by a traditional method without the addition of a starter culture. Preliminary characterization with phenotype-based assays and identification using rep-PCR with a (GTG)(5) primer and 16S rDNA sequence analysis were undertaken for 460 LAB isolates obtained from all the examined samples. Fifteen species were identified as follows: Lactococcus lactis, Lactococcus raffinolactis, Lactococcus garviae, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus helveticus, Enterococcus faecium, Enterococcus durans, Enterococcus faecalis, Enterococcus italicus, Leuconostoc mesenteroides, Leuconostoc pseudomesenteroides, Leuconostoc lactis, Streptococcus thermophilus and Streptococcus mitis. A wide genotypic and phenotypic heterogeneity of the species was observed, particularly within the Lc. lactis strains. In all of the tested dairy products across four seasons, a significantly positive correlation (r = 0.690) between the presence of lactococci and enterococci and a negative correlation (r = 0.722) between the presence of lactococci and leuconostocs were recorded. Forty-five percent of the lactobacilli and 54.4% of the lactococci exhibited proteolytic activity, whereas 18.7% of the total LAB isolates exhibited antimicrobial activity.",
publisher = "Academic Press Ltd- Elsevier Science Ltd, London",
journal = "Food Microbiology",
title = "Characterization of lactic acid bacteria isolated from artisanal Travnik young cheeses, sweet creams and sweet kajmaks over four seasons",
pages = "38-27",
volume = "39",
doi = "10.1016/j.fm.2013.10.011"
}

Terzić-Vidojević, A., Mihajlović, S., Uzelac, G., Veljović, K., Tolinački, M., Živković, M., Topisirović, L.,& Kojić, M.. (2014). Characterization of lactic acid bacteria isolated from artisanal Travnik young cheeses, sweet creams and sweet kajmaks over four seasons. in Food Microbiology
Academic Press Ltd- Elsevier Science Ltd, London., 39, 27-38.
https://doi.org/10.1016/j.fm.2013.10.011

Terzić-Vidojević A, Mihajlović S, Uzelac G, Veljović K, Tolinački M, Živković M, Topisirović L, Kojić M. Characterization of lactic acid bacteria isolated from artisanal Travnik young cheeses, sweet creams and sweet kajmaks over four seasons. in Food Microbiology. 2014;39:27-38.
doi:10.1016/j.fm.2013.10.011 .

Terzić-Vidojević, Amarela, Mihajlović, Sanja, Uzelac, Gordana, Veljović, Katarina, Tolinački, Maja, Živković, Milica, Topisirović, Ljubiša, Kojić, Milan, "Characterization of lactic acid bacteria isolated from artisanal Travnik young cheeses, sweet creams and sweet kajmaks over four seasons" in Food Microbiology, 39 (2014):27-38,
https://doi.org/10.1016/j.fm.2013.10.011 . .

TY  - JOUR
AU  - Terzić-Vidojević, Amarela
AU  - Mihajlović, Sanja
AU  - Uzelac, Gordana
AU  - Golić, Nataša
AU  - Fira, Đorđe
AU  - Kojić, Milan
AU  - Topisirović, Ljubiša
PY  - 2014
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/770
AB  - The aim of this study was to identify and characterize the lactic acid bacteria (LAB) of artisanal Golija raw and cooked cows' milk cheeses traditionally manufactured without the addition of starter culture. A total of 188 Gram-positive and catalase-negative isolates of Golija cheeses were obtained from seven samples of different ripening time. Phenotype-based assays as well as rep-PCR and 16S rDNA sequence analysis were undertaken for all 188 LAB strains. The most diverse species were isolated from 20-day-old BGGO8 cheese (Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus casei/paracasei, Lactobacillus sucicola, Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. lactis by. diacetylactis, Enterococcus faecium, Enterococcus durans and Leuconostoc mesenteroides). In other Golija cheeses Lactobacillus reuteri, Lactobacillus curvatus, Lactobacillus rhamnosus, Lactococcus lactis subsp. cremoris, Lactococcus garvieae, Streptococcus thermophilus and Leuconostoc pseudomesenteroides were found. Pronounced antimicrobial properties showed enterococci (13/42) and lactococci (12/31), while the good proteolytic activity demonstrated lactococci (13/31) and lactobacilli (10/29).
PB  - Srpsko biološko društvo, Beograd, i dr.
T2  - Archives of Biological Sciences
T1  - Identification and characterization of lactic acid bacteria isolated from artisanal white brined Golija cows' milk cheeses
EP  - 192
IS  - 1
SP  - 179
VL  - 66
DO  - 10.2298/ABS1401179T
ER  - 

@article{
author = "Terzić-Vidojević, Amarela and Mihajlović, Sanja and Uzelac, Gordana and Golić, Nataša and Fira, Đorđe and Kojić, Milan and Topisirović, Ljubiša",
year = "2014",
abstract = "The aim of this study was to identify and characterize the lactic acid bacteria (LAB) of artisanal Golija raw and cooked cows' milk cheeses traditionally manufactured without the addition of starter culture. A total of 188 Gram-positive and catalase-negative isolates of Golija cheeses were obtained from seven samples of different ripening time. Phenotype-based assays as well as rep-PCR and 16S rDNA sequence analysis were undertaken for all 188 LAB strains. The most diverse species were isolated from 20-day-old BGGO8 cheese (Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus casei/paracasei, Lactobacillus sucicola, Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. lactis by. diacetylactis, Enterococcus faecium, Enterococcus durans and Leuconostoc mesenteroides). In other Golija cheeses Lactobacillus reuteri, Lactobacillus curvatus, Lactobacillus rhamnosus, Lactococcus lactis subsp. cremoris, Lactococcus garvieae, Streptococcus thermophilus and Leuconostoc pseudomesenteroides were found. Pronounced antimicrobial properties showed enterococci (13/42) and lactococci (12/31), while the good proteolytic activity demonstrated lactococci (13/31) and lactobacilli (10/29).",
publisher = "Srpsko biološko društvo, Beograd, i dr.",
journal = "Archives of Biological Sciences",
title = "Identification and characterization of lactic acid bacteria isolated from artisanal white brined Golija cows' milk cheeses",
pages = "192-179",
number = "1",
volume = "66",
doi = "10.2298/ABS1401179T"
}

Terzić-Vidojević, A., Mihajlović, S., Uzelac, G., Golić, N., Fira, Đ., Kojić, M.,& Topisirović, L.. (2014). Identification and characterization of lactic acid bacteria isolated from artisanal white brined Golija cows' milk cheeses. in Archives of Biological Sciences
Srpsko biološko društvo, Beograd, i dr.., 66(1), 179-192.
https://doi.org/10.2298/ABS1401179T

Terzić-Vidojević A, Mihajlović S, Uzelac G, Golić N, Fira Đ, Kojić M, Topisirović L. Identification and characterization of lactic acid bacteria isolated from artisanal white brined Golija cows' milk cheeses. in Archives of Biological Sciences. 2014;66(1):179-192.
doi:10.2298/ABS1401179T .

Terzić-Vidojević, Amarela, Mihajlović, Sanja, Uzelac, Gordana, Golić, Nataša, Fira, Đorđe, Kojić, Milan, Topisirović, Ljubiša, "Identification and characterization of lactic acid bacteria isolated from artisanal white brined Golija cows' milk cheeses" in Archives of Biological Sciences, 66, no. 1 (2014):179-192,
https://doi.org/10.2298/ABS1401179T . .

TY  - JOUR
AU  - Uzelac, Gordana
AU  - Kojić, Milan
AU  - Lozo, Jelena
AU  - Aleksandrzak-Piekarczyk, Tamara
AU  - Gabrielsen, Christina
AU  - Kristensen, Tom
AU  - Nes, Ingolf F.
AU  - Diep, Dzung B.
AU  - Topisirović, Ljubiša
PY  - 2013
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/703
AB  - Lactococcus lactis subsp. lactis BGMN1-5 produces a leaderless class II bacteriocin called LsbB. To identify the receptor for LsbB, a cosmid library of the LsbB-sensitive strain BGMN1-596 was constructed. About 150 cosmid clones were individually isolated and transferred to LsbB-resistant mutants of BGMN1-596. Cosmid pAZILcos/MN2, carrying a 40-kb insert, was found to restore LsbB sensitivity in LsbB-resistant mutants. Further subcloning revealed that a 1.9-kb fragment, containing only one open reading frame, was sufficient to restore sensitivity. The fragment contains the gene yvjB coding for a Zn-dependent membrane-bound metallopeptidase, suggesting that this gene may serve as the receptor for LsbB. Further support for this notion derives from several independent experiments: (i) whole-genome sequencing confirmed that all LsbB-resistant mutants contain mutations in yvjB; (ii) disruption of yvjB by direct gene knockout rendered sensitive strains BGMN1-596 and IL1403 resistant to LsbB; and (iii) most compellingly, heterologous expression of yvjB in naturally resistant strains of other species, such as Lactobacillus paracasei and Enterococcus faecalis, also rendered them sensitive to the bacteriocin. To our knowledge, this is the first time a membrane-bound peptidase gene has been shown to be involved in bacteriocin sensitivity in target cells. We also demonstrated a novel successful approach for identifying bacteriocin receptors.
PB  - Amer Soc Microbiology, Washington
T2  - Journal of Bacteriology
T1  - A Zn-Dependent Metallopeptidase Is Responsible for Sensitivity to LsbB, a Class II Leaderless Bacteriocin of Lactococcus lactis subsp lactis BGMN1-5
EP  - 5621
IS  - 24
SP  - 5614
VL  - 195
DO  - 10.1128/JB.00859-13
ER  - 

@article{
author = "Uzelac, Gordana and Kojić, Milan and Lozo, Jelena and Aleksandrzak-Piekarczyk, Tamara and Gabrielsen, Christina and Kristensen, Tom and Nes, Ingolf F. and Diep, Dzung B. and Topisirović, Ljubiša",
year = "2013",
abstract = "Lactococcus lactis subsp. lactis BGMN1-5 produces a leaderless class II bacteriocin called LsbB. To identify the receptor for LsbB, a cosmid library of the LsbB-sensitive strain BGMN1-596 was constructed. About 150 cosmid clones were individually isolated and transferred to LsbB-resistant mutants of BGMN1-596. Cosmid pAZILcos/MN2, carrying a 40-kb insert, was found to restore LsbB sensitivity in LsbB-resistant mutants. Further subcloning revealed that a 1.9-kb fragment, containing only one open reading frame, was sufficient to restore sensitivity. The fragment contains the gene yvjB coding for a Zn-dependent membrane-bound metallopeptidase, suggesting that this gene may serve as the receptor for LsbB. Further support for this notion derives from several independent experiments: (i) whole-genome sequencing confirmed that all LsbB-resistant mutants contain mutations in yvjB; (ii) disruption of yvjB by direct gene knockout rendered sensitive strains BGMN1-596 and IL1403 resistant to LsbB; and (iii) most compellingly, heterologous expression of yvjB in naturally resistant strains of other species, such as Lactobacillus paracasei and Enterococcus faecalis, also rendered them sensitive to the bacteriocin. To our knowledge, this is the first time a membrane-bound peptidase gene has been shown to be involved in bacteriocin sensitivity in target cells. We also demonstrated a novel successful approach for identifying bacteriocin receptors.",
publisher = "Amer Soc Microbiology, Washington",
journal = "Journal of Bacteriology",
title = "A Zn-Dependent Metallopeptidase Is Responsible for Sensitivity to LsbB, a Class II Leaderless Bacteriocin of Lactococcus lactis subsp lactis BGMN1-5",
pages = "5621-5614",
number = "24",
volume = "195",
doi = "10.1128/JB.00859-13"
}

Uzelac, G., Kojić, M., Lozo, J., Aleksandrzak-Piekarczyk, T., Gabrielsen, C., Kristensen, T., Nes, I. F., Diep, D. B.,& Topisirović, L.. (2013). A Zn-Dependent Metallopeptidase Is Responsible for Sensitivity to LsbB, a Class II Leaderless Bacteriocin of Lactococcus lactis subsp lactis BGMN1-5. in Journal of Bacteriology
Amer Soc Microbiology, Washington., 195(24), 5614-5621.
https://doi.org/10.1128/JB.00859-13

Uzelac G, Kojić M, Lozo J, Aleksandrzak-Piekarczyk T, Gabrielsen C, Kristensen T, Nes IF, Diep DB, Topisirović L. A Zn-Dependent Metallopeptidase Is Responsible for Sensitivity to LsbB, a Class II Leaderless Bacteriocin of Lactococcus lactis subsp lactis BGMN1-5. in Journal of Bacteriology. 2013;195(24):5614-5621.
doi:10.1128/JB.00859-13 .

Uzelac, Gordana, Kojić, Milan, Lozo, Jelena, Aleksandrzak-Piekarczyk, Tamara, Gabrielsen, Christina, Kristensen, Tom, Nes, Ingolf F., Diep, Dzung B., Topisirović, Ljubiša, "A Zn-Dependent Metallopeptidase Is Responsible for Sensitivity to LsbB, a Class II Leaderless Bacteriocin of Lactococcus lactis subsp lactis BGMN1-5" in Journal of Bacteriology, 195, no. 24 (2013):5614-5621,
https://doi.org/10.1128/JB.00859-13 . .

TY  - JOUR
AU  - Filipić, Brankica
AU  - Jovčić, Branko
AU  - Uzelac, Gordana
AU  - Miljković, Marija
AU  - Antić-Stanković, Jelena
AU  - Topisirović, Ljubiša
AU  - Golić, Nataša
PY  - 2013
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/638
AB  - U ovom radu je izučavan uticaj povećane ekspresije cmbT gena, odgovornog za sintezu CmbT MDR transportera, na rast Lactococcus lactis. L. lactis pripada grupi bakterija mlečne kiseline (BMK) i ima veliku primenu u prehrambenoj industriji kao starter kultura. CmbT transporter je nedavno okarakterisan MDR protein soja L. lactis, koji doprinosi rezistenciji na različite toksične agense kao i na neke klinički značajne antibiotike. U ovom radu je cmbT gen višestruko eksprimiran u soju L. lactis NZ9000 dodavanjem nizina kao inducera. Povećana ekspresija cmbT gena je praćena metodom reverzne transkripcije (RT-PCR). Pokazano je da se nakon dodatka subinhibitornih koncentracija nizina u medijum za rast povećava količina sintetisane informacione RNK specifične za cmbT gen. Rast soja L. lactis NZ9000/pCT50, u kome je višestruko eksprimiran cmbT gen i L. lactis NZ9000 kontrolnog soja praćen je u bogatom i hemijski definisanom medijumu u prisustvu samo metionina (0.084 mM) ili kombinacije metionina i cisteina (8.4 mM i 8.2 mM). Praćene su krive rasta oba soja, a nakon izračunavanja odgovarajućih vremena generacije, rezultati su pokazali da L. lactis NZ9000/pCT50, brže raste u odnosu na kontrolni soj. Uočena razlika je najverovatnije posledica aktivnosti CmbT transportera koji doprinosi izbacivanju toksičnih agenasa iz ćelije i na taj način poboljšava adaptivne sposobnosti bakterije koja ga eksprimira i daje joj selektivnu prednost.
AB  - The influence of the over-expression of CmbT multidrug resistance transporter on the growth rate of Lactococcus lactis NZ9000 was studied. L. lactis is a lactic acid bacteria (LAB) widely used as a starter culture in dairy industry. Recently characterized CmbT MDR transporter in L. lactis confers resistance to a wide variety of toxic compounds as well as to some clinically relevant antibiotics. In this study, the cmbT gene was over-expressed in the strain L. lactis NZ9000 in the presence of nisin inducer. Over-expression of the cmbT gene in L. lactis NZ9000 was followed by RT-PCR. The obtained results showed that the cmbT gene was successfully over-expressed by addition of sub-inhibitory amounts of nisin. Growth curves of L. lactis NZ9000/pCT50 over-expressing the cmbT gene and L. lactis NZ9000 control strain were followed in the rich medium as well as in the chemically defined medium in the presence solely of methionine (0.084 mM) or mix of methionine and cysteine (8.4 mM and 8.2 mM, respectively). Resulting doubling times revealed that L. lactis NZ9000/pCT50 had higher growth rate comparing to the control strain. This could be a consequence of the CmbT efflux activity, which improves the fitness of the host bacterium through the elimination of toxic compounds from the cell.
PB  - Društvo genetičara Srbije, Beograd
T2  - Genetika-Belgrade
T1  - Uticaj povećane ekspresije CmbT MDR transportera na rast Lactococcus lactis
T1  - Over-expressed CmbT multidrug resistance transporter improves the fitness of Lactococcus lactis
EP  - 206
IS  - 1
SP  - 197
VL  - 45
DO  - 10.2298/GENSR1301197F
ER  - 

@article{
author = "Filipić, Brankica and Jovčić, Branko and Uzelac, Gordana and Miljković, Marija and Antić-Stanković, Jelena and Topisirović, Ljubiša and Golić, Nataša",
year = "2013",
abstract = "U ovom radu je izučavan uticaj povećane ekspresije cmbT gena, odgovornog za sintezu CmbT MDR transportera, na rast Lactococcus lactis. L. lactis pripada grupi bakterija mlečne kiseline (BMK) i ima veliku primenu u prehrambenoj industriji kao starter kultura. CmbT transporter je nedavno okarakterisan MDR protein soja L. lactis, koji doprinosi rezistenciji na različite toksične agense kao i na neke klinički značajne antibiotike. U ovom radu je cmbT gen višestruko eksprimiran u soju L. lactis NZ9000 dodavanjem nizina kao inducera. Povećana ekspresija cmbT gena je praćena metodom reverzne transkripcije (RT-PCR). Pokazano je da se nakon dodatka subinhibitornih koncentracija nizina u medijum za rast povećava količina sintetisane informacione RNK specifične za cmbT gen. Rast soja L. lactis NZ9000/pCT50, u kome je višestruko eksprimiran cmbT gen i L. lactis NZ9000 kontrolnog soja praćen je u bogatom i hemijski definisanom medijumu u prisustvu samo metionina (0.084 mM) ili kombinacije metionina i cisteina (8.4 mM i 8.2 mM). Praćene su krive rasta oba soja, a nakon izračunavanja odgovarajućih vremena generacije, rezultati su pokazali da L. lactis NZ9000/pCT50, brže raste u odnosu na kontrolni soj. Uočena razlika je najverovatnije posledica aktivnosti CmbT transportera koji doprinosi izbacivanju toksičnih agenasa iz ćelije i na taj način poboljšava adaptivne sposobnosti bakterije koja ga eksprimira i daje joj selektivnu prednost., The influence of the over-expression of CmbT multidrug resistance transporter on the growth rate of Lactococcus lactis NZ9000 was studied. L. lactis is a lactic acid bacteria (LAB) widely used as a starter culture in dairy industry. Recently characterized CmbT MDR transporter in L. lactis confers resistance to a wide variety of toxic compounds as well as to some clinically relevant antibiotics. In this study, the cmbT gene was over-expressed in the strain L. lactis NZ9000 in the presence of nisin inducer. Over-expression of the cmbT gene in L. lactis NZ9000 was followed by RT-PCR. The obtained results showed that the cmbT gene was successfully over-expressed by addition of sub-inhibitory amounts of nisin. Growth curves of L. lactis NZ9000/pCT50 over-expressing the cmbT gene and L. lactis NZ9000 control strain were followed in the rich medium as well as in the chemically defined medium in the presence solely of methionine (0.084 mM) or mix of methionine and cysteine (8.4 mM and 8.2 mM, respectively). Resulting doubling times revealed that L. lactis NZ9000/pCT50 had higher growth rate comparing to the control strain. This could be a consequence of the CmbT efflux activity, which improves the fitness of the host bacterium through the elimination of toxic compounds from the cell.",
publisher = "Društvo genetičara Srbije, Beograd",
journal = "Genetika-Belgrade",
title = "Uticaj povećane ekspresije CmbT MDR transportera na rast Lactococcus lactis, Over-expressed CmbT multidrug resistance transporter improves the fitness of Lactococcus lactis",
pages = "206-197",
number = "1",
volume = "45",
doi = "10.2298/GENSR1301197F"
}

Filipić, B., Jovčić, B., Uzelac, G., Miljković, M., Antić-Stanković, J., Topisirović, L.,& Golić, N.. (2013). Uticaj povećane ekspresije CmbT MDR transportera na rast Lactococcus lactis. in Genetika-Belgrade
Društvo genetičara Srbije, Beograd., 45(1), 197-206.
https://doi.org/10.2298/GENSR1301197F

Filipić B, Jovčić B, Uzelac G, Miljković M, Antić-Stanković J, Topisirović L, Golić N. Uticaj povećane ekspresije CmbT MDR transportera na rast Lactococcus lactis. in Genetika-Belgrade. 2013;45(1):197-206.
doi:10.2298/GENSR1301197F .

Filipić, Brankica, Jovčić, Branko, Uzelac, Gordana, Miljković, Marija, Antić-Stanković, Jelena, Topisirović, Ljubiša, Golić, Nataša, "Uticaj povećane ekspresije CmbT MDR transportera na rast Lactococcus lactis" in Genetika-Belgrade, 45, no. 1 (2013):197-206,
https://doi.org/10.2298/GENSR1301197F . .