Conversion of a Mono‐ and Diacylglycerol Lipase into a Triacylglycerol Lipase by Protein Engineering |
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| Authors: | Dr. Dongming Lan Dr. Grzegorz Maria Popowicz Dr. Ioannis V. Pavlidis Pengfei Zhou Prof. Dr. Uwe T. Bornscheuer Prof. Dr. Yonghua Wang |
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| Affiliation: | 1. College of Light Industry and Food Sciences, South China University of Technology, Wushan Rd, Tianhe District, Guangzhou 510641 (China);2. Institute of Structural Biology, Helmholtz Zentrum München, Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH), Ingolst?dter Landstrasse 1, 85764 Oberschlei?heim (Germany);3. Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, University of Greifswald, Felix‐Hausdorff‐Strasse 4, 17487 Greifswald (Germany);4. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou Higher Education Mega Centre, Panyu District, Guangzhou 510006 (China) |
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| 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. |
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| Keywords: | enzyme catalysis lipases protein engineering steric hindrance substrate specificity |
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