Improvement of the catalytic activity and thermostability of a hyperthermostable endoglucanase by optimizing N-glycosylation sites

Biotechnology for Biofuels
Chao HanDuochuan Li

Abstract

Endoglucanase has been extensively employed in industrial processes as a key biocatalyst for lignocellulosic biomass degradation. Thermostable endoglucanases with high catalytic activity at elevated temperatures are preferred in industrial use. To improve the activity and thermostability, site-directed mutagenesis was conducted to modify the N-glycosylation sites of the thermostable β-1,4-endoglucanase CTendo45 from Chaetomium thermophilum. In this study, structure-based rational design was performed based on the modification of N-glycosylation sites in CTendo45. Eight single mutants and one double mutant were constructed and successfully expressed in Pichia pastoris. When the unique N-glycosylation site of N88 was eliminated, a T90A variant was active, and its specific activity towards CMC-Na and β-d-glucan was increased 1.85- and 1.64-fold, respectively. The mutant R67S with an additional N-glycosylation site of N65 showed a distinct enhancement in catalytic efficiency. Moreover, T90A and R67S were endowed with extraordinary heat endurance after 200 min of incubation at different temperatures ranging from 30 to 90 °C. Likewise, the half-lives (t1/2) indicated that T90A and R67S exhibited improved enzyme thermostability at 80 ...Continue Reading

References

Oct 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·B W MatthewsW J Becktel
Sep 23, 1993·Nature·G J DaviesM Schülein
Nov 12, 1996·Proceedings of the National Academy of Sciences of the United States of America·D H LiveS J Danishefsky
Jan 21, 2000·FEMS Microbiology Reviews·J L Cereghino, J M Cregg
Mar 27, 2003·Acta Crystallographica. Section D, Biological Crystallography·Jarkko Valjakka, Juha Rouvinen
Jun 11, 2004·Annual Review of Biochemistry·Ari Helenius, Markus Aebi
Jun 14, 2008·Proceedings of the National Academy of Sciences of the United States of America·Dalit Shental-Bechor, Yaakov Levy
Jul 13, 2013·The Journal of Biological Chemistry·Raquel Fonseca-MaldonadoRichard John Ward
Sep 28, 2013·Biotechnology for Biofuels·Ragothaman M YennamalliTaner Z Sen
May 14, 2014·Proceedings of the National Academy of Sciences of the United States of America·Liqun ChenZhongping Tan
Jul 10, 2014·Current Opinion in Structural Biology·Wai Shun Mak, Justin B Siegel
Jan 22, 2015·Proceedings of the National Academy of Sciences of the United States of America·Chengcheng HuangTadashi Suzuki
Jan 30, 2015·Chemical Reviews·Christina M PayneGregg T Beckham
Sep 11, 2016·Current Opinion in Chemical Biology·Christian P Kubicek, Eva M Kubicek
Mar 8, 2017·Journal of Chemical Theory and Computation·Karen SargsyanCarmay Lim
Nov 14, 2017·Nature Communications·Siddharth DeshpandeChester L Drum
Dec 13, 2017·Proceedings of the National Academy of Sciences of the United States of America·Antonella AmoreLarry E Taylor
Feb 8, 2018·International Journal of Biological Macromolecules·Bibekananda KarAshwani Kumar Sharma
Mar 24, 2018·Nature Communications·Larry E TaylorGregg T Beckham
May 23, 2018·Nucleic Acids Research·Andrew WaterhouseTorsten Schwede
May 31, 2018·International Journal of Biological Macromolecules·Chengyao HuaLusheng Zhu
Aug 4, 2018·Applied Microbiology and Biotechnology·Ibrahim TorktazJafar Hemmat
Jan 28, 2019·Bioresource Technology·Anil K PatelAshok Pandey
Mar 9, 2019·Biotechnology Advances·Chen-Guang LiuFeng-Wu Bai
Apr 3, 2019·International Journal of Molecular Sciences·Anna BashirovaMehdi D Davari
May 30, 2019·Biotechnology Advances·Qian LiuYan Feng

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Methods Mentioned

BETA
glycosylation
PCR
Protein Assay

Software Mentioned

SWISS
MODEL
Clustal Omega
NetOGlyc
PyMOL
NetNGlyc

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