Akademik Veri Yönetim Sistemi
EXTREMOPHILES 2022, Korinthos, Yunanistan, 18 - 22 Eylül 2022, ss.66
This study reports a novel BglA9 gene of 1345 bp encoding β-glucosidase from Anoxybacillus ayderensis A9, which was amplified and expressed in E. coli BL21 (DE3): pLysS cells, purified with Ni-NTA column having molecular weight of 52.6 kDa and was used in the bioconversion of polydatin to resveratrol. β-glucosidase from Anoxybacillus ayderensis A9 (BglA9) is a potent enzyme for enzymatic hydrolysis of polydatin to resveratrol. Based on structural and bioinformatics analysis an area near +2 subsite of the active site pocket of BglA9 was selected and single point mutations were introduced with the aim to enhance the substrate specificity of the enzyme towards pNPG and polydatin. The changes introduced in the active site residues were L221S, N222S and G226Q. All the mutants were expressed in E. coli BL21 (DE3) cells and purified with Ni-NTA column chromatography. All the mutants retained their thermal and pH stability and the best mutant in terms of catalytic efficiency for pNPG and polydatin was N222S. The docking analysis supported the results and by comparing binding energies; the mutant N222S showed the best docked complex. This investigation suggests that +2 subsite of BglA9 is an interesting area to be mutated for enhanced catalytic efficiency for pNPG and polydatin. The deglycosylated derivate showed enhanced antioxidant potential as compared to glycoside measured by DPPH assay.