Abstract
L-methionine is an important sulfur-containing amino acid essential for humans and animals. Its biosynthesis pathway is complex and highly regulated. This study aims to explore the bottleneck limiting the improvement of L-methionine productivity and apply efficient strategies to increase L-methionine production in engineered E. coli. The enzyme O-succinylhomoserine sulfhydrylase involved in thiolation of OSH to form homocysteine was overexpressed in the engineered strain E. coli W3110 IJAHFEBC/PAm, resulting in L-methionine production increased from 2.8 to 3.22g/L in shake flask cultivation. By exogenous addition of L-glycine as the precursor of one carbon unit, the titer of L-methionine was increased to 3.68g/L. The glycine cleavage system was further strengthened for the efficient one carbon unit supply and a L-methionine titer of 3.96g/L was obtained, which was increased by 42% compared with that of the original strain. Insufficient supply of one carbon unit was found to be the issue limiting the improvement of L-methionine productivity and its up-regulation significantly promoted the L-methionine production in the engineered E. coli.
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