101. Novel Method for l-Methionine Production Catalyzed by the Aminotransferase ARO8 from Saccharomyces cerevisiae.
- Author
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Wu Y, Zha M, Yin S, Yang H, Boutet J, Huet R, Wang C, and Sun B
- Subjects
- Biocatalysis, Butyric Acid chemistry, Butyric Acid metabolism, Kinetics, Methionine metabolism, Propanols chemistry, Propanols metabolism, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae Proteins genetics, Saccharomyces cerevisiae Proteins isolation & purification, Saccharomyces cerevisiae Proteins metabolism, Sulfides chemistry, Sulfides metabolism, Trans-Activators genetics, Trans-Activators isolation & purification, Trans-Activators metabolism, Industrial Microbiology methods, Methionine chemistry, Saccharomyces cerevisiae enzymology, Saccharomyces cerevisiae Proteins chemistry, Trans-Activators chemistry
- Abstract
The aminotransferase ARO8 was proved to play an efficient role in conversion of l-methionine into methionol via the Ehrlich pathway in Saccharomyces cerevisiae in our previous work. In this work, the reversible transamination activity of ARO8 for conversion of α-keto-γ-(methylthio) butyric acid (KMBA) into l-methionine was confirmed in vitro. ARO8 was cloned from S. cerevisiae S288c and overexpressed in Escherichia coli BL21. A 2-fold higher aminotransferase activity was detected in the recombinant strain ARO8-BL21, and ARO8 was detected in the supernatant of ARO8-BL21 lysate with IPTG induction by SDS-PAGE analysis. The recombinant ARO8 was then purified and used for transforming KMBA into l-methionine. An approximately 100% conversion rate of KMBA into l-methionine was achieved by optimized enzymatic reaction catalyzed by ARO8. This work fulfilled l-methionine biosynthesis catalyzed by the aminotransferase ARO8 using glutamate and KMBA, which provided a novel method for l-methionine production by enzymatic catalysis with the potential application prospect in industry.
- Published
- 2018
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