Journal Article
- Overexpression and characterization of a novel cold-adapted and salt-tolerant GH1 β-glucosidase from the marine bacterium Alteromonas sp. L82
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Jingjing Sun , Wei Wang , Congyu Yao , Fangqun Dai , Xiangjie Zhu , Junzhong Liu , Jianhua Hao
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J. Microbiol. 2018;56(9):656-664. Published online August 23, 2018
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DOI: https://doi.org/10.1007/s12275-018-8018-2
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Abstract
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A novel gene (bgl) encoding a cold-adapted β-glucosidase
was cloned from the marine bacterium Alteromonas sp.
L82. Based on sequence analysis and its putative catalytic
conserved region, Bgl belonged to the glycoside hydrolase
family 1. Bgl was overexpressed in E. coli and purified by
Ni2+ affinity chromatography. The purified recombinant β-
glucosidase showed maximum activity at temperatures between
25°C to 45°C and over the pH range 6 to 8. The enzyme
lost activity quickly after incubation at 40°C. Therefore,
recombinant β-glucosidase appears to be a cold-adapted
enzyme. The addition of reducing agent doubled its activity
and 2 M NaCl did not influence its activity. Recombinant
β-glucosidase was also tolerant of 700 mM glucose and some
organic solvents. Bgl had a Km of 0.55 mM, a Vmax of 83.6
U/mg, a kcat of 74.3 s-1 and kcat/Km of 135.1 at 40°C, pH 7 with
4-nitrophenyl-β-D-glucopyranoside as a substrate. These
properties indicate Bgl may be an interesting candidate for
biotechnological and industrial applications.
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Citations
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Research Support, Non-U.S. Gov't
- Cel8H, a Novel Endoglucanase from the Halophilic Bacterium Halomonas sp. S66-4: Molecular Cloning, Heterogonous Expression, and Biochemical Characterization
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Xiaoluo Huang , Zongze Shao , Yuzhi Hong , Ling Lin , Chanjuan Li , Fei Huang , Hui Wang , Ziduo Liu
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J. Microbiol. 2010;48(3):318-324. Published online June 23, 2010
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DOI: https://doi.org/10.1007/s12275-009-0188-5
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Abstract
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A recombinant Escherichia coli clone expressing an endoglucanase was identified from a genomic library of the halophilic bacterium Halomonas sp. S66-4, and the enzyme was designated Cel8H. The cel8H gene consisted of 1,053 bp and encoded 350 amino acids sharing the highest identity of 48% to other known endoglucanases. The protein was expressed in E. coli BL21 (DE3) and purified to homogeneity. The purified recombinant enzyme had an optimal activity of 4.9 U/mg at pH 5 and 45°C toward the substrate carboxymethylcellulose. It exhibited extraordinary properties which differed from endoglucanases reported previously at the point of high salt tolerance above 5 M, simultaneously with high pH stability at pH 4-12 and high temperature stability at 40-60°C. Various substrate tests indicated that the enzyme hydrolyzes β-1,4-glucosidic bonds specifically.