Abstract
Many serine hydrolases catalyze perhydrolysis, the reversible formation of peracids from carboxylic acids and hydrogen peroxide. Recently, we showed that a single amino acid substitution in the alcohol binding pocket, L29P, in Pseudomonas fluorescens (SIK WI) aryl esterase (PFE) increased the specificity constant of PFE for peracetic acid formation >100-fold [Bernhardt et al. (2005) Angew. Chem., Int. Ed. 44, 2742]. In this paper, we extend this work to address the three following questions. First, what is the molecular basis of the increase in perhydrolysis activity? We previously proposed that the L29P substitution creates a hydrogen bond between the enzyme and hydrogen peroxide in the transition state. Here we report two X-ray structures of L29P PFE that support this proposal. Both structures show a main chain carbonyl oxygen closer to the active site serine as expected. One structure further shows acetate in the active site in an orientation consistent with reaction by an acyl-enzyme mechanism. We also detected an acyl-enzyme intermediate in the hydrolysis of epsilon-caprolactone by mass spectrometry. Second, can we further increase perhydrolysis activity? We discovered that the reverse reaction, hydrolysis of peracetic aci...Continue Reading
References
Jan 1, 1976·Journal of the Chemical Society. Perkin Transactions 1·C R Harrison, P Hodge
Apr 1, 1992·Protein Engineering·D L OllisJ Schrag
Mar 1, 1991·Acta Crystallographica. Section A, Foundations of Crystallography·T A JonesM Kjeldgaard
Feb 23, 1988·Biochemistry·S C BlacklowJ R Knowles
Aug 1, 1994·Nature Structural Biology·H J HechtK H van Pée
Feb 1, 1995·Microbiology·I Pelletier, J Altenbuchner
Aug 1, 1996·Microbiology·S KirnerK H van Pée
Jun 1, 1997·Nature Structural Biology·R C WilmouthC J Schofield
Jun 27, 1998·Journal of Molecular Biology·B HofmannH J Hecht
Jul 10, 1998·Biochimica Et Biophysica Acta·D ChipmanJ V Schloss
May 20, 1999·Nature Structural Biology·A PerrakisV S Lamzin
Oct 26, 1999·Acta Crystallographica. Section D, Biological Crystallography·J W Pflugrath
Dec 22, 1999·European Journal of Biochemistry·M KataokaS Shimizu
Jan 25, 2003·European Journal of Biochemistry·Kohsuke HondaSakayu Shimizu
Jan 31, 2003·Proteins·Simon C LovellDavid C Richardson
Mar 5, 2004·The Journal of Biological Chemistry·Karl GruberChristoph Kratky
Mar 19, 2004·Journal of Medicinal Chemistry·Richard A FriesnerPeter S Shenkin
Apr 15, 2004·Journal of Molecular Biology·Kirsty LineJennifer A Littlechild
Jun 24, 2004·Acta Crystallographica. Section D, Biological Crystallography·Jeremy D CheesemanRomas J Kazlauskas
Jul 21, 2004·Journal of Computational Chemistry·Eric F PettersenThomas E Ferrin
May 1, 1997·Acta Crystallographica. Section D, Biological Crystallography·G N MurshudovE J Dodson
Sep 9, 2004·Chemical Reviews·G-J ten BrinkR A Sheldon
Sep 24, 2004·Bioorganic Chemistry·Timothy D H Bugg
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
Apr 2, 2005·Angewandte Chemie·Peter BernhardtRomas J Kazlauskas
Jun 28, 2005·Analytical Biochemistry·Zhili LiKaren S Anderson
Dec 7, 2007·Chembiochem : a European Journal of Chemical Biology·Chen LiTimothy D H Bugg
Apr 1, 1999·Angewandte Chemie·Ole Kirk, Lars Sparre Conrad
Citations
Aug 31, 2011·PloS One·Kin-Kwan LaiClaudio F Gonzalez
Jul 10, 2013·Journal of Biotechnology·Dragana DespotovicUlrich Schwaneberg
Feb 7, 2012·Nature Chemical Biology·Sagar D KhareDavid Baker
Sep 25, 2014·International Journal of Molecular Sciences·Hua ShaoYunjun Yan
Jan 18, 2011·Journal of Molecular Biology·Jasleen BainsMartin J Boulanger
Nov 4, 2015·Frontiers in Bioengineering and Biotechnology·Jennifer A Littlechild
Jan 18, 2013·Chemistry : a European Journal·DeLu Tyler YinRomas J Kazlauskas
Feb 26, 2011·Chembiochem : a European Journal of Chemical Biology·Yun JiangRomas J Kazlauskas
Jul 2, 2010·Chembiochem : a European Journal of Chemical Biology·Robert KouristUwe T Bornscheuer
May 24, 2012·Chemistry : a European Journal·DeLu Tyler Yin, Romas J Kazlauskas
Apr 6, 2016·Journal of Biotechnology·Zeinab AssafMélanie Hall
Jan 13, 2015·New Biotechnology·Ljubica VojcicUlrich Schwaneberg
May 27, 2015·The FEBS Journal·Christopher SayerJennifer A Littlechild
Feb 25, 2011·Bioresource Technology·DeLu Tyler YinRomas J Kazlauskas
Mar 19, 2015·Biochemical and Biophysical Research Communications·Xuping WangYonghua Wang
May 11, 2016·Scientific Reports·Christopher SayerJennifer A Littlechild
Oct 28, 2014·Chemical Science·David M NedrudRomas J Kaz-Lauskas
Mar 7, 2012·Journal of Biomolecular Screening·Dragana DespotovicUlrich Schwaneberg
Jan 27, 2017·PloS One·Chang Woo LeeJun Hyuck Lee
Mar 25, 2018·Angewandte Chemie·JiaJia DongWuyuan Zhang
Oct 30, 2016·Applied and Environmental Microbiology·Xiaodan YanSheng Wu
Aug 1, 2018·Biotechnology Letters·Paula IstvanRicardo Henrique Krüger
Apr 13, 2017·Journal of Industrial Microbiology & Biotechnology·J A Littlechild
Sep 19, 2019·Biotechnology and Bioengineering·Diana M MateAurelio Hidalgo
Feb 24, 2012·Organic & Biomolecular Chemistry·Leticia L TorresAurelio Hidalgo
Oct 26, 2018·Journal of Industrial Microbiology & Biotechnology·Shaozhou Zhu, Guojun Zheng
Jan 25, 2021·Journal of Biotechnology·Wenjing JiaZhengyu Shu
Oct 2, 2015·ACS Catalysis·Alissa Rauwerdink, Romas J Kazlauskas
Oct 28, 2017·Journal of Chemical Theory and Computation·Esra BozkurtUrsula Rothlisberger