title: Conversion of a Mono- and Diacylglycerol Lipase into a Triacylglycerol Lipase by Protein Engineering.
abstract: Despite the fact that most lipases are believed to be active against triacylglycerides, there is a small group of lipases that are active only on mono- and diacylglycerides. The reason for this difference in substrate scope is not clear. We tried to identify the reasons for this in the lipase from Malassezia globosa. By protein engineering, and with only one mutation, we managed to convert this enzyme into a typical triacylglycerol lipase (the wild-type lipase does not accept triacylglycerides). The variant Q282L accepts a broad spectrum of triacylglycerides, although the catalytic behavior is altered to some extent. From in silico analysis it seems that specific hydrophobic interactions are key to the altered substrate specificity.
authors.affiliation: College of Light Industry and Food Sciences, South China University of Technology, Wushan Rd, Tianhe District, Guangzhou 510641 (China).
authors.forename: Dongming
authors.initials: D
authors.lastname: Lan
authors.affiliation: Institute of Structural Biology, Helmholtz Zentrum München, Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH), Ingolstädter Landstrasse 1, 85764 Oberschleißheim (Germany).
authors.forename: Grzegorz Maria
authors.initials: GM
authors.lastname: Popowicz
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany).
authors.forename: Ioannis V
authors.initials: IV
authors.lastname: Pavlidis
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou Higher Education Mega Centre, Panyu District, Guangzhou 510006 (China).
authors.forename: Pengfei
authors.initials: P
authors.lastname: Zhou
authors.affiliation: Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17487 Greifswald (Germany).
authors.forename: Uwe T
authors.initials: UT
authors.lastname: Bornscheuer
authors.affiliation: College of Light Industry and Food Sciences, South China University of Technology, Wushan Rd, Tianhe District, Guangzhou 510641 (China). yonghw@scut.edu.cn.
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Wang
mesh_terms: D020134:Catalytic Domain; D008049:Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008071:Lipoprotein Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008290:Malassezia / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D008994:Monoacylglycerol Lipases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017354:Point Mutation; D015202:Protein Engineering; D013379:Substrate Specificity
chemical_list: D008994:Monoacylglycerol Lipases; D008049:Lipase; D008071:Lipoprotein Lipase
keywords: enzyme catalysis; lipases; protein engineering; steric hindrance; substrate specificity