Overexpression and characterization of a novel cold-adapted and salt-tolerant GH1 β-glucosidase from the marine bacterium Alteromonas sp. L82

Jingjing Sun; Wei Wang; Congyu Yao; Fangqun Dai; Xiangjie Zhu; Junzhong Liu; Jianhua Hao

doi:10.1007/s12275-018-8018-2

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Overexpression and characterization of a novel cold-adapted and salt-tolerant GH1 β-glucosidase from the marine bacterium Alteromonas sp. L82: Jingjing Sun ^1,2, Wei Wang ^1,2, Congyu Yao ^1,2,3, Fangqun Dai ^1,2, Xiangjie Zhu ^1,2,3, Junzhong Liu ^1,2, Jianhua Hao ^1,2,4; Journal of Microbiology 2018;56(9):656-664.
DOI: https://doi.org/10.1007/s12275-018-8018-2
Published online: August 23, 2018

¹Key Laboratory of Sustainable Development of Polar Fishery, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences Qingdao 266071, P. R. China, ²Laboratory for Marine Drugs and Bioproducts, Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology Qingdao 266071, P. R. China, ³Shanghai Ocean University, Shanghai 201306, P. R. China, ⁴Jiangsu Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resource, Lianyungang 222005, P. R. China¹Key Laboratory of Sustainable Development of Polar Fishery, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences Qingdao 266071, P. R. China, ²Laboratory for Marine Drugs and Bioproducts, Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology Qingdao 266071, P. R. China, ³Shanghai Ocean University, Shanghai 201306, P. R. China, ⁴Jiangsu Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resource, Lianyungang 222005, P. R. China

Corresponding author: Jianhua Hao , Tel: +86-0532-85841193;,

Received: 12 January 2018 • Revised: 21 June 2018 • Accepted: 22 June 2018

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Abstract

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.

Keywords: β-glucosidase;marine;Alteromonas;cold adaptation;salt tolerance

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Overexpression and characterization of a novel cold-adapted and salt-tolerant GH1 β-glucosidase from the marine bacterium Alteromonas sp. L82

J. Microbiol. 2018;56(9):656-664. Published online August 23, 2018

DOI: https://doi.org/10.1007/s12275-018-8018-2

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