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KMID : 1094720110160010097
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Biotechnology and Bioprocess Engineering
2011 Volume.16 No. 1 p.97 ~ p.106
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Cloning, expression, and characterization of β-glucosidase from Exiguobacterium sp. DAU5 and transglycosylation activity
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Chang Jie
Park In-Hye
Lee Yong-Seok
Ahn Soon-Cheol
Zhou Yi
Choi Yong-Lark
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Abstract
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A gram-negative bacterium, designated strain DAU5, was isolated from shrimp shell samples because it demonstrated high β-glucosidase activity. Through 16S rDNA gene sequence analysis the strain was identified as belonging to the genus Exiguobacterium. The β-glucosidase gene of Exiguobacterium sp. DAU5 was successfully cloned by the shotgun method. Nucleotide sequence determination by sodium dodecyl sulfate-ployacrylamide gel electrophoresis indicated that the gene for the enzyme contained 1,350 bp, was coded by 450 amino acids, and was 52 kDa. The polypeptide exhibits significant homology with other bacterial β-glucosidases and belongs to the Glycoside Hydrolase Family 1. The β-glucosidase was purified by a His-fusion purification system. The optimal pH and temperature of the enzyme were 7.0 and 45°C, respectively. The enzyme activity was strongly inhibited by Ca2+, and Li+, K+, Zn2+, Mg2+, Na2+, Ni2+, and EDTA partially inhibited the enzyme activity. The BglA showed the highest activity with p-NPG and MUG. However, strain DAU5 β-glucosidase, which is for degradation of oligosaccharides, is expected to be useful for the fermentation of cellulose degradation and the transglycosylation of saccharides.
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KEYWORD
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bglA gene, β-glucosidase, purification, transglycosylation, Exiguobacterium sp.
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