Journal
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
Volume 135, Issue 3, Pages 217-223Publisher
SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1016/j.jbiosc.2022.12.010
Keywords
L-Methionine; Multibranched pathway; Crucial gene; Metabolic engineering; Escherichia coli
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This study investigates the metabolic response and crucial genes in the biosynthetic pathway of iota-methionine. The relationship between different pathways and their controlling forces on metabolic flux are also examined. The findings provide insights into the biosynthesis of iota-methionine and offer a new research method for synthesizing other metabolic products in multilevel regulated and multibranched biosynthetic pathways.
iota-Methionine biosynthesis is through multilevel regulated and multibranched biosynthetic pathway (MRMBP). Because of the complex regulatory mechanism and the imbalanced metabolic flux between branched pathways, mi-crobial production of iota-methionine has not been commercialized. In this study, local metabolic response in MRMBP of iota- methionine was investigated and various crucial genes in branched pathways were determined. In iota-serine pathway, the crucial gene was serABC. In O-succinyl homoserine (OSH) pathway, which was the C4 backbone of iota-methionine, metB and metL controlled the metabolic flux jointly. In iota-cysteine pathway, the crucial gene cysEfbr could disturb the flux distribution of local network in iota-methionine biosynthesis. However, no crucial gene for iota-methionine production in 5 -methyl tetrahydrofolate (CH3-THF) pathway was found. The relation between these pathways was also researched. iota- Serine pathway, as the upstream pathway of iota-cysteine and CH3-THF, played a crucial role in iota-methionine biosynthesis. iota- Cysteine pathway showed the strongest controlling force of the metabolic flux, and OSH pathway was second to iota- cysteine pathway. In contrast, CH3-THF pathway was the weakest, which was probably the mainly limited steps at present and had great potential in further research. In addition, constructed W3110 IJAHFEBC/pA*HAmL was able to produce 2.62 g/iota iota-methionine in flask. This study is instructive for iota-methionine biosynthesis and provides a new research method of biosynthesizing other metabolic products in MRMBPs. (c) 2023, The Society for Biotechnology, Japan. All rights reserved.
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