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

The Journal of Microbiology
Jingjing SunJianhua Hao

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.

References

Jul 18, 1995·Proceedings of the National Academy of Sciences of the United States of America·B HenrissatG Davies
Jul 17, 1998·Current Opinion in Chemical Biology·D H Crout, G Vic
Oct 29, 2000·Journal of Agricultural and Food Chemistry·C H DeckerP Schreier
Dec 19, 2002·Critical Reviews in Biotechnology·Yukti BhatiaV S Bisaria
Jun 26, 2003·Nucleic Acids Research·Torsten SchwedeManuel C Peitsch
Apr 1, 1964·Biochemistry·W W CLELAND
Mar 24, 2004·Nature Reviews. Microbiology·Georges Feller, Charles Gerday
Feb 6, 2008·Journal of Basic Microbiology·Héctor Antonio CristóbalCarlos Mauricio Abate
Apr 29, 2009·Journal of Computational Chemistry·Garrett M MorrisArthur J Olson
May 22, 2010·Cellular and Molecular Life Sciences : CMLS·James R Ketudat Cairns, Asim Esen
Oct 5, 2010·Journal of Microbiology and Biotechnology·Zemin FangYazhong Xiao
Jun 14, 2013·The Journal of General and Applied Microbiology·Cristiane Akemi UchimaManabu Arioka
Jun 11, 2014·Acta Crystallographica. Section D, Biological Crystallography·Priscila Oliveira de GiuseppeMario Tyago Murakami
Jan 1, 2013·Biomolecules·Annette SørensenBirgitte K Ahring
Oct 18, 2015·International Journal of Biological Macromolecules·Elaine CrespimRoberto Ruller
Jan 17, 2016·International Journal of Biological Macromolecules·Shivangi ChamoliAshok Kumar Verma
Nov 12, 2016·FEBS Open Bio·Alicia Hernández-GuzmánJulio C Villagómez-Castro

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Citations

Oct 2, 2019·The Journal of Microbiology·Jaejin LeeKangseok Lee
Jun 20, 2020·Applied Microbiology and Biotechnology·Noora Barzkar, Muhammad Sohail
Jul 6, 2020·Applied Biochemistry and Biotechnology·Jingjing SunJianhua Hao
Apr 4, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ru ZhangZhi Yong Luo

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