Molecular Docking and Site-Directed Mutagenesis of Dichloromethane Dehalogenase to Improve Enzyme Activity for Dichloromethane Degradation

Applied Biochemistry and Biotechnology
Jianming YuZhiliang Yu

Abstract

Dichloromethane (DCM) dehalogenase in bacterial cells can catalyze the degradation of deleterious DCM in environments. However, the utility of naturally occurring DCM dehalogenase is often limited due to low enzyme activity and content in living cells. In this study, the gene encoding DCM dehalogenase was cloned from Methylobacterium rhodesianum and overexpressed in Escherichia coli. Based on molecular docking analysis of DCM dehalogenase using DCM as the ligand, all of the target amino acid residues within substrate binding pocket and 10 conservative amino acid residues were individually mutated to Ala. After determination of activity, R120, L121, W128, and T146 were chosen for further saturation mutation. Results showed that dcmT146A, dcmT146R, and dcmT146Q have higher activities, whereas dcmL121A, dcmT146L, dcmL121Q, and dcmL121F have retained activities. Next, these seven mutants with a single mutation on amino acid residue were chosen for double mutation. It was found that the mutant of dcmL121A/T146R exhibits the highest activity increasing by 52.8% relative to wild type. Bioinformatic and experimental analyses revealed that the mutant variant dcmL121A/T146R bears the reduced steric hindrance in the active center with a d...Continue Reading

References

Mar 1, 1991·The Biochemical Journal·D J MeyerB Ketterer
Apr 5, 1988·European Journal of Biochemistry·V V MozhaevK Martinek
May 1, 1985·Journal of Bacteriology·D Kohler-Staub, T Leisinger
Jan 6, 1981·Biochemistry·S W Provencher, J Glöckner
Dec 1, 1994·Biodegradation·T LeisingerS Vuilleumier
Apr 12, 1994·Proceedings of the National Academy of Sciences of the United States of America·L Song, C D Poulter
Nov 12, 1998·Trends in Biochemical Sciences·F JeanmouginT J Gibson
Oct 3, 2002·Biochemical and Biophysical Research Communications·Edward J MullaneyAbul H J Ullah
Jul 2, 2003·Proteins·Ora Schueler-Furman, David Baker
Jul 5, 2003·Journal of Microbiological Methods·Valentina I KrausovaJuan M González
Nov 20, 2008·The FEBS Journal·Nerino AllocatiCarmine Di Ilio
Dec 17, 2008·Current Protocols in Bioinformatics·Garrett M MorrisArthur J Olson
Jan 20, 2009·Bioinformatics·Andrew M WaterhouseGeoffrey J Barton
Dec 17, 2009·BMC Bioinformatics·Christiam CamachoThomas L Madden
Apr 20, 2010·Journal of Computer-aided Molecular Design·Daniel Seeliger, Bert L de Groot
Feb 4, 2011·Research in Microbiology·Emilie E L MullerStéphane Vuilleumier
Dec 27, 2011·Nature Methods·Michael RemmertJohannes Söding
Jul 11, 2013·PloS One·Gordon Lemmon, Jens Meiler
Jul 17, 2013·Environmental Science and Pollution Research International·Dong-Zhi ChenJian-Meng Chen
Sep 5, 2014·Environmental Technology·R RaviT Swaminathan

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Citations

Oct 24, 2020·Journal of Biomolecular Structure & Dynamics·Habeebat Adekilekun OyewusiRoswanira Abdul Wahab
Dec 3, 2021·Journal of Biomolecular Structure & Dynamics·Habeebat Adekilekun OyewusiFahrul Huyop

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