INVESTIGATION OF SUBSTRATE SPECIFICITY AND KINETIC PARAMETERS OF ESTERASE FROM THE SPOROGENIC STRAIN BACILLUS PARALICHENIFORMIS T7
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Authors
A. Mussakhmetov
«National Center for Biotechnology» LLP, 13/5 Korgalzhyn Road, Astana, 01000
«L.N. Gumilyov Eurasian National University » NJSC. 2 Satbaev st., Astana 010000
«GenLab» LLP, 19/1 M. Gabdullin Street, Astana 010000
M. Astrakhanov
«National Center for Biotechnology» LLP, 13/5 Korgalzhyn Road, Astana, 010000
Z. Akishev
«National Center for Biotechnology» LLP, 13/5 Korgalzhyn Road, Astana, 010000
«GenLab» LLP, 19/1 M. Gabdullin Street, Astana 010000
B. Khassenov
«National Center for Biotechnology» LLP, 13/5 Korgalzhyn Road, Astana, 010000
«GenLab» LLP, 19/1 M. Gabdullin Street, Astana 010000
Abstract
Esterases represent an important class of hydrolase enzymes widely utilized in biotechnology due to their high catalytic activity, substrate specificity, and stability against various physico-chemical factors. The sporogenic strain Bacillus paralicheniformis T7 produces extracellular enzymes, including esterase, which are associated with sporulation processes and adaptation to stressful conditions. In this study, the substrate specificity and catalytic properties of a recombinant esterase from the sporogenic strain B. paralicheniformis T7, expressed in Escherichia coli cells, were investigated. Structural analysis revealed that the esterase belongs to the GDSL/SGNH-hydrolase family and contains a conserved Ser–Asp–His catalytic triad. Substrate specificity analysis using p-nitrophenyl esters (C2–C18) showed that the enzyme exhibits maximum activity toward short-chain substrates, specifically p-nitrophenyl acetate (250.0 ± 0.1 U/mg), classifying it as a true esterase. The study of various factors demonstrated that divalent metal ions Ca2+, Mg2+, Mn2+, Cu2+, Zn2+, Fe2+ inhibit enzyme activity, while the reducing agents DTT and β-mercaptoethanol completely suppress its catalytic function. At the same time, the enzyme shows relative stability toward non-ionic detergents and sodium dodecyl sulfate (SDS). Kinetic analysis revealed that the Km value for p-nitrophenyl acetate is 26.18 mM, Vmax is 52.37 U/mg, and the catalytic constant kcat is 1.74 s-1. The results indicate a distinct affinity of the enzyme for the substrate and moderate catalytic efficiency compared to other bacillar esterases. Thus, the esterase from B. paralicheniformis T7 is characterized by a pronounced specificity for short-chain esters and resistance to a range of surfactants, determining its potential for application in the biocatalytic synthesis of esters used in the food and fragrance industries.
Keywords
Esterase, Endospore, Activity, Bacillus
Article Details
References
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