title: Metabolic engineering of Escherichia coli W3110 for the production of L-methionine.
abstract: In this study, we constructed an L-methionine-producing recombinant strain from wild-type Escherichia coli W3110 by metabolic engineering. To enhance the carbon flux to methionine and derepression met regulon, thrBC, lysA, and metJ were deleted in turn. Methionine biosynthesis obstacles were overcome by overexpression of metAFbr(Fbr, Feedback resistance), metB, and malY under control of promoter pN25. Recombinant strain growth and methionine production were further improved by attenuation of metK gene expression through replacing native promoter by metK84p. Blocking the threonine pathway by deletion of thrBC or thrC was compared. Deletion of thrC showed faster growth rate and higher methionine production. Finally, metE, metF, and metH were overexpressed to enhance methylation efficiency. Compared with the original strain E. coli W3110, the finally obtained Me05 (pETMAFbr-B-Y/pKKmetH) improved methionine production from 0 to 0.65 and 5.62 g/L in a flask and a 15-L fermenter, respectively.
authors.affiliation: The Key Laboratory of Industrial Biotechnology of Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Hua
authors.initials: H
authors.lastname: Li
authors.affiliation: The Key Laboratory of Industrial Biotechnology of Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Bao Shi
authors.initials: BS
authors.lastname: Wang
authors.affiliation: The Key Laboratory of Industrial Biotechnology of Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: You Ran
authors.initials: YR
authors.lastname: Li
authors.affiliation: The Key Laboratory of Industrial Biotechnology of Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.affiliation: The Key Laboratory of Industrial Biotechnology of Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Zhong Yang
authors.initials: ZY
authors.lastname: Ding
authors.affiliation: The Key Laboratory of Industrial Biotechnology of Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, People's Republic of China.
authors.forename: Zheng Hua
authors.initials: ZH
authors.lastname: Gu
authors.affiliation: The Key Laboratory of Industrial Biotechnology of Ministry of Education, National Engineering Laboratory for Cereal Fermentation Technology, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, Jiangsu, People's Republic of China. gyshi@jiangnan.edu.cn.
authors.forename: Gui Yang
authors.initials: GY
authors.lastname: Shi
mesh_terms: D002244:Carbon / Q000737:chemistry; D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000378:metabolism; D005285:Fermentation; D007218:Industrial Microbiology; D060847:Metabolic Engineering / Q000379:methods*; D008715:Methionine / Q000096:biosynthesis*; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D013912:Threonine
chemical_list: D017931:DNA Primers; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D013912:Threonine; D002244:Carbon; D008715:Methionine
keywords: Biosynthesis obstacle; Escherichia coli; L-homoserine; L-methionine; Metabolic engineering