@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"
}