Metabolic Engineering of Corynebacterium glutamicum for the Production of L-Ornithine

被引:78
作者
Kim, Seo Yun [1 ,2 ]
Lee, Joungmin [1 ,2 ]
Lee, Sang Yup [1 ,2 ,3 ]
机构
[1] Korea Adv Inst Sci & Technol, Metab & Biomol Engn Natl Res Lab, Dept Chem & Biomol Engn,BioProc Engn Res Ctr, Plus Program BK21,Ctr Syst & Synthet Biotechnol, Taejon 305701, South Korea
[2] Korea Adv Inst Sci & Technol, Inst BioCentury, Taejon 305701, South Korea
[3] Korea Adv Inst Sci & Technol, Bioinformat Res Ctr, Taejon 305701, South Korea
基金
新加坡国家研究基金会;
关键词
L-ornithine; Corynebacterium glutamicum; metabolic engineering; ESCHERICHIA-COLI; L-ARGININE; AVAILABILITY;
D O I
10.1002/bit.25440
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
L-ornithine is a non-essential amino acid for various industrial applications in food industry. In this study, Corynebacterium glutamicum ATCC 13032 was metabolically engineered for the production of L-ornithine. First, the proB and argF genes were deleted to block the competitive branch pathway and to block the conversion of L-ornithine to citrulline, respectively. In addition, the argR gene encoding the regulatory repressor of the L-arginine operon was also deleted. The resulting strain produced 230 mg/L of L-ornithine from glucose in flask culture. This base strain was further engineered by the plasmid-based overexpression of the argCJBD genes from C. glutamicum ATCC 21831, which resulted in the production of 7.19 g/L of L-ornithine. To enrich the NADPH pool, the carbon flux was redirected towards the pentose phosphate pathway by changing the start codons of the pgi and zwf genes and replacing the native promoter of the tkt operon with the strong sod promoter. Fed-batch cultivation of this final strain YW06 (pSY223) allowed production of 51.5 g/L of L-ornithine in 40 h with the overall productivity of 1.29 g/L/h. The results obtained in this study demonstrate the possibility of efficiently producing L-ornithine by metabolically engineered C. glutamicum. (C) 2014 Wiley Periodicals, Inc.
引用
收藏
页码:416 / 421
页数:6
相关论文
共 20 条
[1]   Comparative 13C Metabolic Flux Analysis of Pyruvate Dehydrogenase Complex-Deficient, L-Valine-Producing Corynebacterium glutamicum [J].
Bartek, Tobias ;
Blombach, Bastian ;
Lang, Siegmund ;
Eikmanns, Bernhard J. ;
Wiechert, Wolfgang ;
Oldiges, Marco ;
Noeh, Katharina ;
Noack, Stephan .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2011, 77 (18) :6644-6652
[2]   From zero to hero-Design-based systems metabolic engineering of Corynebacterium glutamicum for L-lysine production [J].
Becker, Judith ;
Zelder, Oskar ;
Haefner, Stefan ;
Schroeder, Hartwig ;
Wittmann, Christoph .
METABOLIC ENGINEERING, 2011, 13 (02) :159-168
[3]   Enhancement of L-ornithine production by disruption of three genes encoding putative oxidoreductases in Corynebacterium glutamicum [J].
Hwang, Gui-Hye ;
Cho, Jae-Yong .
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 2014, 41 (03) :573-578
[4]  
Hwang JH, 2008, J MICROBIOL BIOTECHN, V18, P704
[5]   Reengineering of a Corynebacterium glutamicum L-Arginine and L-Citrulline Producer [J].
Ikeda, Masato ;
Mitsuhashi, Satoshi ;
Tanaka, Kenji ;
Hayashi, Mikiro .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2009, 75 (06) :1635-1641
[6]   Metabolic engineering of Corynebacterium glutamicum for increasing the production of L-ornithine by increasing NADPH availability [J].
Jiang, Ling-Yan ;
Zhang, Yuan-Yuan ;
Li, Zhen ;
Liu, Jian-Zhong .
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 2013, 40 (10) :1143-1151
[7]   Metabolic evolution of Corynebacterium glutamicum for increased production of L-ornithine [J].
Jiang, Ling-Yan ;
Chen, Shang-Guang ;
Zhang, Yuan-Yuan ;
Liu, Jian-Zhong .
BMC BIOTECHNOLOGY, 2013, 13
[8]   Manufacturing Molecules Through Metabolic Engineering [J].
Keasling, Jay D. .
SCIENCE, 2010, 330 (6009) :1355-1358
[9]   Metabolic engineering of Escherichia coli for the production of phenol from glucose [J].
Kim, Byoungjin ;
Park, Hyegwon ;
Na, Dokyun ;
Lee, Sang Yup .
BIOTECHNOLOGY JOURNAL, 2014, 9 (05) :621-629
[10]  
Kind S, 2014, METAB ENG, V13, P159