Engineering of Corynebacterium glutamicum to Enhance L-ornithine Production by Gene Knockout and Comparative Proteomic Analysis

Chinese Journal of Chemical Engineering ›› 2012, Vol. 20 ›› Issue (4): 731-739.

Engineering of Corynebacterium glutamicum to Enhance L-ornithine Production by Gene Knockout and Comparative Proteomic Analysis

卢冬梅, 刘建忠, 毛宗万

The Key Laboratory of Gene Engineering of Ministry of Education, State Key Laboratory of Biocontrol, Sun Yat-Sen University, Guangzhou 510275, China

收稿日期:2011-03-25 修回日期:2011-10-16 出版日期:2012-08-28 发布日期:2012-09-15
通讯作者: LIU Jianzhong, E-mail:lssljz@mail.sysu.edu.cn
基金资助:
Supported by the National Natural Science Foundation of China (30970089,20876181,20831006);the Natural Science Foundation of Guangdong Province (9351027501000003)

Engineering of Corynebacterium glutamicum to Enhance L-ornithine Production by Gene Knockout and Comparative Proteomic Analysis

LU Dongmei, LIU Jianzhong, MAO Zongwan

The Key Laboratory of Gene Engineering of Ministry of Education, State Key Laboratory of Biocontrol, Sun Yat-Sen University, Guangzhou 510275, China

Received:2011-03-25 Revised:2011-10-16 Online:2012-08-28 Published:2012-09-15
Supported by:
Supported by the National Natural Science Foundation of China (30970089,20876181,20831006);the Natural Science Foundation of Guangdong Province (9351027501000003)

摘要/Abstract

摘要： Engineered Corynebacterium glutamicum was constructed for L-ornithine production by disrupting genes of argF and proB to prevent the flux away from L-ornithine.Effect of the inactivation of 2-oxoglutarate de-hydrogenase complex(ODHC) on L-ornithine production was also investigated.It was found that the inactivation of ODHC by knockout of the kgd gene enhanced L-ornithine production.The engineered C.glutamicum ATCC13032(ΔargFΔproBΔkgd) produced L-ornithine up to 4.78 g·L^-1 from 0.24 g·L^-1 of the wild-type strain.In order to understand the mechanism of L-ornithine production in C.glutamicum ATCC13032(ΔargFΔproBΔkgd) and find out new strategies for further enhancing L-ornithine production,the comparative proteome between the wild-type and the engineered strain was analyzed.L-Ornithine overproduction in the engineered strain was related to the up-regulation of the expression levels of enzymes involved in L-ornithine biosynthesis pathway and down-regulation of the expression levels of proteins involved in pentose phosphate pathway.The overexpression of genes in the upstream pathway of glutamate to increase the availability of endogenous glutamate may further in-crease ornithine production in the engineered C.glutamicum and the ornithine synthesis enzymes(ArgCJBD) may not be the limiting enzymes in the engineered C.glutamicum.

关键词: L-ornithine production, gene knockout, 2-oxoglutarate dehydrogenase, Corynebacterium glutamicum, proteomic analysis

Abstract: Engineered Corynebacterium glutamicum was constructed for L-ornithine production by disrupting genes of argF and proB to prevent the flux away from L-ornithine.Effect of the inactivation of 2-oxoglutarate de-hydrogenase complex(ODHC) on L-ornithine production was also investigated.It was found that the inactivation of ODHC by knockout of the kgd gene enhanced L-ornithine production.The engineered C.glutamicum ATCC13032(ΔargFΔproBΔkgd) produced L-ornithine up to 4.78 g·L^-1 from 0.24 g·L^-1 of the wild-type strain.In order to understand the mechanism of L-ornithine production in C.glutamicum ATCC13032(ΔargFΔproBΔkgd) and find out new strategies for further enhancing L-ornithine production,the comparative proteome between the wild-type and the engineered strain was analyzed.L-Ornithine overproduction in the engineered strain was related to the up-regulation of the expression levels of enzymes involved in L-ornithine biosynthesis pathway and down-regulation of the expression levels of proteins involved in pentose phosphate pathway.The overexpression of genes in the upstream pathway of glutamate to increase the availability of endogenous glutamate may further in-crease ornithine production in the engineered C.glutamicum and the ornithine synthesis enzymes(ArgCJBD) may not be the limiting enzymes in the engineered C.glutamicum.

Key words: L-ornithine production, gene knockout, 2-oxoglutarate dehydrogenase, Corynebacterium glutamicum, proteomic analysis

卢冬梅, 刘建忠, 毛宗万. Engineering of Corynebacterium glutamicum to Enhance L-ornithine Production by Gene Knockout and Comparative Proteomic Analysis[J]. Chinese Journal of Chemical Engineering, 2012, 20(4): 731-739.

LU Dongmei, LIU Jianzhong, MAO Zongwan. Engineering of Corynebacterium glutamicum to Enhance L-ornithine Production by Gene Knockout and Comparative Proteomic Analysis[J]. Chin.J.Chem.Eng., 2012, 20(4): 731-739.

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Engineering of Corynebacterium glutamicum to Enhance L-ornithine Production by Gene Knockout and Comparative Proteomic Analysis

Engineering of Corynebacterium glutamicum to Enhance L-ornithine Production by Gene Knockout and Comparative Proteomic Analysis

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