Engineering O‐Succinyl‐L‐Homoserine Mercaptotransferase for Efficient L‐Methionine Biosynthesis by Fermentation‐Enzymatic Coupling Route

L‐Methionine is the unique sulfur‐containing amino acid essential for humans and animals, which is widely used in pharmaceutical, food and feed industries. Its green and efficient production has attracted a great deal of attention. Fermentation‐enzymatic coupling route is regarded to be promising fo...

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Published in:Advanced synthesis & catalysis 2023-04, Vol.365 (7), p.1048-1057
Main Authors: Tang, Xiao‐Ling, Li, Ning, Liu, Yan‐Lai, Li, Jing‐Peng, Zhao, Kai‐Xuan, Liu, Zhi‐Qiang, Zheng, Yu‐Guo
Format: Article
Language:English
Subjects:
Amino acids
Biosynthesis
catalytic activity
Coupling
Fermentation
fermentation-enzymatic coupling route
L-methionine
Methionine
O-succinyl-L-homoserine mercaptotransferase
protein engineering
Substrates
Thiols
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Summary:L‐Methionine is the unique sulfur‐containing amino acid essential for humans and animals, which is widely used in pharmaceutical, food and feed industries. Its green and efficient production has attracted a great deal of attention. Fermentation‐enzymatic coupling route is regarded to be promising for L‐methionine biosynthesis, while O‐succinyl‐L‐homoserine mercaptotransferase (MetZ) is the key biocatalyst. Exploring and engineering of MetZ with ideal catalytic properties is highly desired. Herein, the MetZ from Chromobacterium violaceum (CvMetZ) was screened and through in silico analyses, potential beneficial amino acid residues both at the access channel and around the oxygen anion holes were anchored for site‐saturation mutagenesis. The positive mutants were obtained with improved activity for L‐methionine biosynthesis using O‐succinyl‐L‐homoserine (OSH) and methyl mercaptan as substrate. The best mutant was further applied for L‐methionine production and supply of methyl mercaptan was optimized in a fed‐batch approach. The conversion of 100 g/L OSH reached 92% with L‐methionine yield of 62.6 g/L, which was well coupled with the OSH fermentation level.
ISSN:1615-4150
1615-4169
DOI:10.1002/adsc.202300030