Improving the Catalytic Activity and Thermostability of MAS1 Lipase by Alanine Substitution

Review

Abstract

MAS1 is a lipase isolated from Streptomyces sp. strain W007 with potential application in biotechnology. Structural analysis of MAS1 lipase showed that eight amino acids with bulkier side located in the substrate-binding pocket may be involved in affecting catalytic performance. Alanine substitutions of those residues were conducted to reduce steric clash of catalyzed pocket and probe their functional roles. The kcat/Km of mutants H108A, F153A, and V233A increased to 2.3-, 2.1-, and 1.4-fold, respectively. Interestingly, the half-life (60 °C) of F153A had shifted to 523 min after mutagenesis, which was fivefold enhancement toward that of MAS1 wide-type. Furthermore, higher hydrolysis ability of mutants H108A and F153A toward palm stearin of high melting temperature made them potentially applicable in oil/fat modification. Our work provided an example to obtain biocatalysts with desired catalytic behaviors by protein engineering.

Keywords

MAS1 lipase Steric clash Alanine substitution Site-directed mutagenesis 

Abbreviations

WT

Wild type

PUFA

Polyunsaturated fatty acids

TAG

Triacylglycerol

pNP

p-Nitrophenol

PCR

Polymerase chain reaction

SDS-PAGE

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

FAs

Fatty acids

Notes

Acknowledgements

This work was made possible with funding provided by the National Natural Science Foundation of China (21376098, 31471690, 31601398) and Science and Technology Planning project of Guangdong Province (2016B090920082).

Compliance with Ethical Standards

Conflict of interest

The authors have declared no conflict of interests.

Supplementary material

12033_2018_62_MOESM1_ESM.docx (74 kb)
Supplementary material 1 (DOCX 73 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.School of Food Science and EngineeringSouth China University of TechnologyGuangzhouPeople’s Republic of China
  2. 2.School of Bioscience and BioengineeringSouth China University of TechnologyGuangzhouPeople’s Republic of China

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