TY  - CONF
AU  - Atanasković, Marija
AU  - Morić, Ivana
AU  - B. Rokić, Miloš
AU  - Đokić, Anđela
AU  - Pantović, Jelena
AU  - Despotović, Dragana
AU  - Šenerović, Lidija
PY  - 2024
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2369
AB  - The formation of biofilms by foodborne pathogens
is a constant challenge in the food industry,
leading to an increased risk of contamination and
compromising food safety. Many of the chemicals
commonly used for sanitation in the food industry
are unable to remove biofilms, are harmful
to surfaces and can be toxic. The effectiveness
of disinfectants can be improved using enzymes
that specifically target biofilm components such
as exopolysaccharides, extracellular DNA, or proteins.
In this study we investigated the potential
of glycoside hydrolases originating from the
gill microbiota of freshwater fish to control biofilm
formation in the most common foodborne
pathogens. We demonstrated that β-glucosidase
from Microbacterium sp. BG28 (BglB-BG28) effectively
inhibits cellulose-rich biofilms formed by
Salmonella enteritidis, S. typhimurium, S. infantis,
and Escherichia coli. When these bacteria were cultivated overnight with 200 μL/mL enzyme, up
to 80% less biofilm was formed. By fluorescence
microscopy, we visualised the inhibition of biofilms
on plastic, glass and aluminium, materials
commonly used in the food industry. When used
as a pre-treatment, BglB-BG28 increased the
bactericidal efficacy of Oxicid®S, a commercially
available surface disinfectant. Its effectiveness at
temperatures up to 50 °C and in a pH range from
4 to 8 together with compatibility with non-ionic
detergents and high tolerance to sodium chloride
and glucose give BglB-BG28 advantages in
harsh and diverse industrial environments. Importantly,
no toxicity to Caenorhabditis elegans
was observed at enzyme concentrations of up
to 1 mg/ml. Overall, these results demonstrate
the suitability of the β-glucosidase BglB-BG28 for
the formulation of a novel enzyme-based disinfectant
to be used in food processing facilities.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
T1  - GLYCOSIDE HYDROLASES FROM FRESHWATER FISH GILL MICROBIOTA AS BIOFILM INHIBITORS FOR ENHANCED FOOD SAFETY
EP  - 42
SP  - 42
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2369
ER  - 

@conference{
author = "Atanasković, Marija and Morić, Ivana and B. Rokić, Miloš and Đokić, Anđela and Pantović, Jelena and Despotović, Dragana and Šenerović, Lidija",
year = "2024",
abstract = "The formation of biofilms by foodborne pathogens
is a constant challenge in the food industry,
leading to an increased risk of contamination and
compromising food safety. Many of the chemicals
commonly used for sanitation in the food industry
are unable to remove biofilms, are harmful
to surfaces and can be toxic. The effectiveness
of disinfectants can be improved using enzymes
that specifically target biofilm components such
as exopolysaccharides, extracellular DNA, or proteins.
In this study we investigated the potential
of glycoside hydrolases originating from the
gill microbiota of freshwater fish to control biofilm
formation in the most common foodborne
pathogens. We demonstrated that β-glucosidase
from Microbacterium sp. BG28 (BglB-BG28) effectively
inhibits cellulose-rich biofilms formed by
Salmonella enteritidis, S. typhimurium, S. infantis,
and Escherichia coli. When these bacteria were cultivated overnight with 200 μL/mL enzyme, up
to 80% less biofilm was formed. By fluorescence
microscopy, we visualised the inhibition of biofilms
on plastic, glass and aluminium, materials
commonly used in the food industry. When used
as a pre-treatment, BglB-BG28 increased the
bactericidal efficacy of Oxicid®S, a commercially
available surface disinfectant. Its effectiveness at
temperatures up to 50 °C and in a pH range from
4 to 8 together with compatibility with non-ionic
detergents and high tolerance to sodium chloride
and glucose give BglB-BG28 advantages in
harsh and diverse industrial environments. Importantly,
no toxicity to Caenorhabditis elegans
was observed at enzyme concentrations of up
to 1 mg/ml. Overall, these results demonstrate
the suitability of the β-glucosidase BglB-BG28 for
the formulation of a novel enzyme-based disinfectant
to be used in food processing facilities.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health",
title = "GLYCOSIDE HYDROLASES FROM FRESHWATER FISH GILL MICROBIOTA AS BIOFILM INHIBITORS FOR ENHANCED FOOD SAFETY",
pages = "42-42",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2369"
}

Atanasković, M., Morić, I., B. Rokić, M., Đokić, A., Pantović, J., Despotović, D.,& Šenerović, L.. (2024). GLYCOSIDE HYDROLASES FROM FRESHWATER FISH GILL MICROBIOTA AS BIOFILM INHIBITORS FOR ENHANCED FOOD SAFETY. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
Serbian Society for Microbiology., 42-42.
https://hdl.handle.net/21.15107/rcub_imagine_2369

Atanasković M, Morić I, B. Rokić M, Đokić A, Pantović J, Despotović D, Šenerović L. GLYCOSIDE HYDROLASES FROM FRESHWATER FISH GILL MICROBIOTA AS BIOFILM INHIBITORS FOR ENHANCED FOOD SAFETY. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health. 2024;:42-42.
https://hdl.handle.net/21.15107/rcub_imagine_2369 .

Atanasković, Marija, Morić, Ivana, B. Rokić, Miloš, Đokić, Anđela, Pantović, Jelena, Despotović, Dragana, Šenerović, Lidija, "GLYCOSIDE HYDROLASES FROM FRESHWATER FISH GILL MICROBIOTA AS BIOFILM INHIBITORS FOR ENHANCED FOOD SAFETY" in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health (2024):42-42,
https://hdl.handle.net/21.15107/rcub_imagine_2369 .

TY  - JOUR
AU  - Atanasković, Marija
AU  - Morić, Ivana
AU  - Rokić, Miloš
AU  - Đokić, Anđela
AU  - Pantović, Jelena
AU  - Despotović, Dragana
AU  - Šenerović, Lidija
PY  - 2024
UR  - https://www.sciencedirect.com/science/article/pii/S221242922301194X
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2331
AB  - SalmonellaEnteritidis is the most commonly reported pathogen for foodborne illness outbreaks in both underdeveloped and developed regions. S. Enteritidis biofilms, which form on various food contact surfaces, are resistant to conventional physical and chemical cleaning and disinfection procedures routinely used in food processing. The aim of this study was to identify novel, industrially applicable enzymes that are active against S. Enteritidis biofilms. We describe the properties and anti-biofilm activity of heterologously expressed β-glucosidase B derived from the environmental strain Microbacterium sp. BG28 (BglB-BG28) collected from gills of bream fish. The enzyme inhibited adhesion and the early stages of biofilm formation in clinical isolates of S. Enteritidis. At a concentration of 200 μg/mL, BglB-BG28 effectively reduced biofilm formation, by decreasing biofilm biomass by 50% and metabolic activity within biofilms by 80%. The enzyme reduced the formation of air-liquid biofilms on various surfaces, including plastic, glass and metal, as observed by fluorescence microscopy. BglB-BG28 inhibited biofilm formation in Escherichia coli, another important food pathogen that also forms cellulose-rich biofilms. Using o-NPG as substrate, the enzyme showed activity at temperatures up to 50 °C and in a pH range between 4 and 8, high tolerance to sodium chloride and glucose, and compatibility with nonionic detergents. Importantly, no toxicity was observed in the model system Caenorhabditis elegans even at an enzyme concentration of 1 mg/mL. These results suggest that the β-glucosidase BglB-BG28 is a promising candidate for the development of a new enzyme-based disinfection protocol that can be used in food processing facilities.
PB  - Elsevier
T2  - Food Bioscience
T2  - Food BioscienceFood Bioscience
T1  - Inhibition of Salmonella Enteritidis adhesion and biofilm formation by β-glucosidase B from Microbacterium sp. BG28
SP  - 103543
VL  - 57
DO  - 10.1016/j.fbio.2023.103543
ER  - 

@article{
author = "Atanasković, Marija and Morić, Ivana and Rokić, Miloš and Đokić, Anđela and Pantović, Jelena and Despotović, Dragana and Šenerović, Lidija",
year = "2024",
abstract = "SalmonellaEnteritidis is the most commonly reported pathogen for foodborne illness outbreaks in both underdeveloped and developed regions. S. Enteritidis biofilms, which form on various food contact surfaces, are resistant to conventional physical and chemical cleaning and disinfection procedures routinely used in food processing. The aim of this study was to identify novel, industrially applicable enzymes that are active against S. Enteritidis biofilms. We describe the properties and anti-biofilm activity of heterologously expressed β-glucosidase B derived from the environmental strain Microbacterium sp. BG28 (BglB-BG28) collected from gills of bream fish. The enzyme inhibited adhesion and the early stages of biofilm formation in clinical isolates of S. Enteritidis. At a concentration of 200 μg/mL, BglB-BG28 effectively reduced biofilm formation, by decreasing biofilm biomass by 50% and metabolic activity within biofilms by 80%. The enzyme reduced the formation of air-liquid biofilms on various surfaces, including plastic, glass and metal, as observed by fluorescence microscopy. BglB-BG28 inhibited biofilm formation in Escherichia coli, another important food pathogen that also forms cellulose-rich biofilms. Using o-NPG as substrate, the enzyme showed activity at temperatures up to 50 °C and in a pH range between 4 and 8, high tolerance to sodium chloride and glucose, and compatibility with nonionic detergents. Importantly, no toxicity was observed in the model system Caenorhabditis elegans even at an enzyme concentration of 1 mg/mL. These results suggest that the β-glucosidase BglB-BG28 is a promising candidate for the development of a new enzyme-based disinfection protocol that can be used in food processing facilities.",
publisher = "Elsevier",
journal = "Food Bioscience, Food BioscienceFood Bioscience",
title = "Inhibition of Salmonella Enteritidis adhesion and biofilm formation by β-glucosidase B from Microbacterium sp. BG28",
pages = "103543",
volume = "57",
doi = "10.1016/j.fbio.2023.103543"
}

Atanasković, M., Morić, I., Rokić, M., Đokić, A., Pantović, J., Despotović, D.,& Šenerović, L.. (2024). Inhibition of Salmonella Enteritidis adhesion and biofilm formation by β-glucosidase B from Microbacterium sp. BG28. in Food Bioscience
Elsevier., 57, 103543.
https://doi.org/10.1016/j.fbio.2023.103543

Atanasković M, Morić I, Rokić M, Đokić A, Pantović J, Despotović D, Šenerović L. Inhibition of Salmonella Enteritidis adhesion and biofilm formation by β-glucosidase B from Microbacterium sp. BG28. in Food Bioscience. 2024;57:103543.
doi:10.1016/j.fbio.2023.103543 .

Atanasković, Marija, Morić, Ivana, Rokić, Miloš, Đokić, Anđela, Pantović, Jelena, Despotović, Dragana, Šenerović, Lidija, "Inhibition of Salmonella Enteritidis adhesion and biofilm formation by β-glucosidase B from Microbacterium sp. BG28" in Food Bioscience, 57 (2024):103543,
https://doi.org/10.1016/j.fbio.2023.103543 . .

TY  - CONF
AU  - Atanasković, Marija
AU  - Morić, Ivana
AU  - Rokić, Miloš
AU  - Šenerović, Lidija
PY  - 2023
UR  - https://belbi.bg.ac.rs/
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2031
AB  - Biofilms are ubiquitous in nature, and the food industry is vulnerable to the risks posed by
biofilm formation. Not only do they interfere with the food production process, but they also
pose a public health threat. However, complete elimination of biofilms on food and food
contact surfaces cannot be achieved by conventional methods (cleaning and disinfection)
alone. New biofilm control strategies must be developed to prevent its formation and/or
persistence. Novel approaches may be based on enzymes that depolymerize components
of the biofilm matrix, making bacterial cells accessible to antimicrobial agents.
Environmental microorganisms are an inexhaustible source of new enzymes. In
Salmonella Enteritidis and Escherichia coli, known foodborne pathogens, cellulose is an
important component of the biofilm matrix, so our isolates from untapped environments
were tested for cellulolytic activity. Of the more than 70 isolates examined, isolate BG28
was selected as the most promising. Its genome was sequenced, annotated, and it was
identified as Gram-positive Microbacterium sp. Genome mining revealed the presence of
four complete genes for different β-glucosidases, one of three enzyme types of cellulase
complexes. To select the best candidate for heterologous expression DeepTMHMM,
ProtParam, and SoluProt were used to predict the presence/absence of signal peptide
and transmembrane domains, instability index, aliphatic index, hydrophilicity, and soluble
expression in E. coli. Based on the prediction results, the gene annotated as β-glucosidase
B was selected for recombinant expression. In addition, I-TASSER was used to model the
tertiary structure of the selected enzyme.
The β-glucosidase B was recombinantly expressed, purified, and tested for its anti-biofilm
activity. It was active and showed a 50% inhibitory effect on S. Enteritidis and E. coli biofilm
formation at a concentration of 100 μg/ml. To further evaluate this in silico approach in
the preselection of candidate enzymes for recombinant expression and purification, we
will use it to identify other enzymes of the cellulase complex.
PB  - Belgrade : Institute of molecular genetics and genetic engineering
C3  - 4th Belgrade Bioinformatics Conference
T1  - In silico pre-selection of β-glucosidase gene for heterologous recombinant expression
EP  - 86
SP  - 86
VL  - 4
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2031
ER  - 

@conference{
author = "Atanasković, Marija and Morić, Ivana and Rokić, Miloš and Šenerović, Lidija",
year = "2023",
abstract = "Biofilms are ubiquitous in nature, and the food industry is vulnerable to the risks posed by
biofilm formation. Not only do they interfere with the food production process, but they also
pose a public health threat. However, complete elimination of biofilms on food and food
contact surfaces cannot be achieved by conventional methods (cleaning and disinfection)
alone. New biofilm control strategies must be developed to prevent its formation and/or
persistence. Novel approaches may be based on enzymes that depolymerize components
of the biofilm matrix, making bacterial cells accessible to antimicrobial agents.
Environmental microorganisms are an inexhaustible source of new enzymes. In
Salmonella Enteritidis and Escherichia coli, known foodborne pathogens, cellulose is an
important component of the biofilm matrix, so our isolates from untapped environments
were tested for cellulolytic activity. Of the more than 70 isolates examined, isolate BG28
was selected as the most promising. Its genome was sequenced, annotated, and it was
identified as Gram-positive Microbacterium sp. Genome mining revealed the presence of
four complete genes for different β-glucosidases, one of three enzyme types of cellulase
complexes. To select the best candidate for heterologous expression DeepTMHMM,
ProtParam, and SoluProt were used to predict the presence/absence of signal peptide
and transmembrane domains, instability index, aliphatic index, hydrophilicity, and soluble
expression in E. coli. Based on the prediction results, the gene annotated as β-glucosidase
B was selected for recombinant expression. In addition, I-TASSER was used to model the
tertiary structure of the selected enzyme.
The β-glucosidase B was recombinantly expressed, purified, and tested for its anti-biofilm
activity. It was active and showed a 50% inhibitory effect on S. Enteritidis and E. coli biofilm
formation at a concentration of 100 μg/ml. To further evaluate this in silico approach in
the preselection of candidate enzymes for recombinant expression and purification, we
will use it to identify other enzymes of the cellulase complex.",
publisher = "Belgrade : Institute of molecular genetics and genetic engineering",
journal = "4th Belgrade Bioinformatics Conference",
title = "In silico pre-selection of β-glucosidase gene for heterologous recombinant expression",
pages = "86-86",
volume = "4",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2031"
}

Atanasković, M., Morić, I., Rokić, M.,& Šenerović, L.. (2023). In silico pre-selection of β-glucosidase gene for heterologous recombinant expression. in 4th Belgrade Bioinformatics Conference
Belgrade : Institute of molecular genetics and genetic engineering., 4, 86-86.
https://hdl.handle.net/21.15107/rcub_imagine_2031

Atanasković M, Morić I, Rokić M, Šenerović L. In silico pre-selection of β-glucosidase gene for heterologous recombinant expression. in 4th Belgrade Bioinformatics Conference. 2023;4:86-86.
https://hdl.handle.net/21.15107/rcub_imagine_2031 .

Atanasković, Marija, Morić, Ivana, Rokić, Miloš, Šenerović, Lidija, "In silico pre-selection of β-glucosidase gene for heterologous recombinant expression" in 4th Belgrade Bioinformatics Conference, 4 (2023):86-86,
https://hdl.handle.net/21.15107/rcub_imagine_2031 .

TY  - CONF
AU  - Atanasković, Marija
AU  - Morić, Ivana
AU  - Šenerović, Lidija
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1905
AB  - About one in ten people contract a foodborne illness within a year. Children under the age of five are
the most affected, with 125,000 deaths each year. Many of the foodborne illness outbreaks can be
linked to the presence of biofilms in the food industry, and Salmonella enteritidis is an extremely
important foodborne pathogen that thrives in these conditions. It has been shown that biofilms can be
resistant to physical and chemical treatments used in cleaning and disinfection procedures in food
processing. The problem with using more aggressive disinfectants is that they often violate food safety
regulations. The use of enzymes which degrade biofilm matrix structural components should facilitate
current disinfection procedures and not compromise food safety. In this study, the anti-biofilm activity
of recombinantly expressed β-glucosidase B and its potential use as a protective agent to control
Salmonella biofilm formation is investigated. The putative target of this enzyme is cellulose, the
structural component of the Salmonella biofilm matrix. β-Glucosidase B deriving from the
environmental strain Microbacterium sp. BG28 was heterologously expressed in Escherichia coli and
successfully purified by affinity chromatography. The anti-biofilm activity of the enzyme was
evaluated in in vitro assays using various clinical isolates of S. enteritidis. The toxicity of the enzyme
was studied in Caenorhabditis elegans. β-glucosidase B effectively inhibited the formation of
Salmonella biofilms grown in a temperature range of 8°C to 37°C, achieving 50% inhibition at
concentrations of 100μg/ml. Biochemical characterization showed that the optimal pH activity of the
enzyme is between 6 and 7, with the highest activity observed at temperatures between 37°C and 47°C.
The absence of toxicity and other presented results indicate that beta-glucosidase B can be used in
biofilm control in the food industry.
PB  - Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo
C3  - Genetics & Applications
T1  - Β-glucosidase b from microbacterium sp. Bg28 as a biofilm control agent In food processing environment
IS  - 2 (Special edition)
SP  - 129
VL  - 7
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1905
ER  - 

@conference{
author = "Atanasković, Marija and Morić, Ivana and Šenerović, Lidija",
year = "2023",
abstract = "About one in ten people contract a foodborne illness within a year. Children under the age of five are
the most affected, with 125,000 deaths each year. Many of the foodborne illness outbreaks can be
linked to the presence of biofilms in the food industry, and Salmonella enteritidis is an extremely
important foodborne pathogen that thrives in these conditions. It has been shown that biofilms can be
resistant to physical and chemical treatments used in cleaning and disinfection procedures in food
processing. The problem with using more aggressive disinfectants is that they often violate food safety
regulations. The use of enzymes which degrade biofilm matrix structural components should facilitate
current disinfection procedures and not compromise food safety. In this study, the anti-biofilm activity
of recombinantly expressed β-glucosidase B and its potential use as a protective agent to control
Salmonella biofilm formation is investigated. The putative target of this enzyme is cellulose, the
structural component of the Salmonella biofilm matrix. β-Glucosidase B deriving from the
environmental strain Microbacterium sp. BG28 was heterologously expressed in Escherichia coli and
successfully purified by affinity chromatography. The anti-biofilm activity of the enzyme was
evaluated in in vitro assays using various clinical isolates of S. enteritidis. The toxicity of the enzyme
was studied in Caenorhabditis elegans. β-glucosidase B effectively inhibited the formation of
Salmonella biofilms grown in a temperature range of 8°C to 37°C, achieving 50% inhibition at
concentrations of 100μg/ml. Biochemical characterization showed that the optimal pH activity of the
enzyme is between 6 and 7, with the highest activity observed at temperatures between 37°C and 47°C.
The absence of toxicity and other presented results indicate that beta-glucosidase B can be used in
biofilm control in the food industry.",
publisher = "Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo",
journal = "Genetics & Applications",
title = "Β-glucosidase b from microbacterium sp. Bg28 as a biofilm control agent In food processing environment",
number = "2 (Special edition)",
pages = "129",
volume = "7",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1905"
}

Atanasković, M., Morić, I.,& Šenerović, L.. (2023). Β-glucosidase b from microbacterium sp. Bg28 as a biofilm control agent In food processing environment. in Genetics & Applications
Sarajevo : Institute for Genetic Engineering and Biotechnology, University of Sarajevo., 7(2 (Special edition)), 129.
https://hdl.handle.net/21.15107/rcub_imagine_1905

Atanasković M, Morić I, Šenerović L. Β-glucosidase b from microbacterium sp. Bg28 as a biofilm control agent In food processing environment. in Genetics & Applications. 2023;7(2 (Special edition)):129.
https://hdl.handle.net/21.15107/rcub_imagine_1905 .

Atanasković, Marija, Morić, Ivana, Šenerović, Lidija, "Β-glucosidase b from microbacterium sp. Bg28 as a biofilm control agent In food processing environment" in Genetics & Applications, 7, no. 2 (Special edition) (2023):129,
https://hdl.handle.net/21.15107/rcub_imagine_1905 .