NADP+-dependent glyceraldehyde-3-phosphate dehydrogenase encoding gene was expressed in Corynebacterium glutamicum SNK118 to increase intracellular NADPH level to further increase the yields of L-arginine and L-ornithine. Clostridium saccharobutylicum DSM 13864-derived NADP+-dependent glyceraldehyde-3-phosphate dehydrogenase gene (CsgapC) was selected and expressed in C. glutamicum. The recombinant strain SNK118/pXMJ19-CsgapC produced 11.55 g/L L-arginine and the conversion rate of glucose was 0.13 g/g after 70 h flask fermentation, which were 26% and 10.2% higher than that of the control, respectively. Moreover, CsgapC was also introduced into L-ornithine-producing strain SNK118ΔargFΔargR to obtain a recombinant strain SNK118ΔargFΔargR/pXMJ19-CsgapC. During 70 h flask fermentation, 27.76 g/L L-ornithine with 0.274 g/g conversion rate were achieved, which were 20.1% and 15.6% higher than those of the control, respectively. This study therefore concluded that heterologously express CsgapC promotes the production of L-arginine and L-ornithine by C. glutamicum.
KAN Baojun
,
DONG Jinjun
,
LIU Hui
,
ZHAN Milin
,
XU Guochao
,
HAN Ruizhi
,
NI Ye
. Glyceraldehyde-3-phosphate dehydrogenase promotes the production of L-arginine and L-ornithine by Corynebacterium glutamicum[J]. Food and Fermentation Industries, 2019
, 45(15)
: 9
-16
.
DOI: 10.13995/j.cnki.11-1802/ts.020661
[1] SHIN J H,LEE S Y.Metabolic engineering of microorganisms for the production of L-arginine and its derivatives[J].Microb Cell Fact,2014,13(1):166.
[2] PALMER R M,ASHTON D S,MONCADA S.Vascular endothelial cells synthesize nitric oxide from L-arginine[J].Nature,1988,333:664-666.
[3] SALVATORE F,CIMINO F,D’AYELLO-CARACCIOLO M,et al.Mechanism of the protection by L-ornithine-L-aspartate mixture and by l-arginine in ammonia intoxication[J].Arch Biochem Biophys,1964,107(3):499-503.
[4] SHI H P,FISHEL R S,EFRON D T,et al.Effect of supplemental ornithine on wound healing[J].J Surg Res,2002,106(2):299-302.
[5] ZAJAC A,POPRZECKI S,Z·EBROWSKA A,et al.Arginine and ornithine supplementation increases growth hormone and insulin-like growth factor-1 serum levels after heavy-resistance exercise in strength-trained athletes[J].J Strength Cond Res,2010,24(4): 1 082-1 090.
[6] 孙安然,宋伟,刘佳,等. 生物酶法合成L-精氨酸衍生物的研究进展[J].生物工程学报,2018,34(2):165-176.
[7] BECKER J,WITTMANN C.Advanced biotechnology: metabolically engineered cells for the bio-based production of chemicals and fuels, materials, and health-care products[J].Angew Chem Int Ed Engl,2015,54(11):3 328-3 350.
[8] PARK S H,KIM H U,KIM T Y,et al.Metabolic engineering of Corynebacterium glutamicum for L-arginine production[J].Nat Commun,2014,5:4 618.
[9] KIM S Y,LEE J,LEE S Y.Metabolic engineering of Corynebacterium glutamicum for the production of L-ornithine[J].Biotechnol Bioeng,2015,112(2):416-421.
[10] TAKENO S,HORI K,OHTANI S,et al.L-lysine production independent of the oxidative pentose phosphate pathway by Corynebacterium glutamicum with the Streptococcus mutans gapN gene[J].Metab Eng,2016,37:1-10.
[11] 郭雯,赖联贺,张晓梅,等. 加强表达3-磷酸甘油醛脱氢酶对谷氨酸棒杆菌产L-丝氨酸的影响[J].食品与发酵工业,2016,42(9):15-20.
[12] ZHAN Milin, KAN Baojun,DONG Jinjun,et al.Metabolic engineering of Corynebacterium glutamicum for improved l-arginine synthesis by enhancing NADPH supply[J].J Ind Microbiol Biotechnol,2018,46:45-54.
[13] JIANG Yu,QIAN Fenghui,YANG Junjie,et al.CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum[J].Nat.Commun,2017,8:15 179.
[14] XU Meijuan,RAO Zhiming,DOU Wenfang,et al.The role of ARGR repressor regulation on L-arginine production in Corynebacterium crenatum[J].Appl Biochem and Biotechnol,2013,170(3):587-597.
[15] JAKOBY M,NGOUOTO-NKILI C E,BURKOVSKI A.Construction and application of new Corynebacterium glutamicum vectors[J].Biotechnol Tech,1999,13(6):437-441.
[16] GIBSON D G,CHUANG R Y,VENTER J C,et al. Enzymatic assembly of DNA molecules up to several hundred kilobases[J].Nat Methods,2009,6(5):343-345.
[17] SAMBROOK J,FRITSCH E F,MANIATIS T.Molecular cloning: a laboratory manual[M].Cold Spring Harbor Laboratory Press,1989.
[18] REST M E V D,LANGE C,MOLENAAR D.A heat shock following electroporation induces highly efficient transformation of Corynebacterium glutamicum with xenogeneic plasmid DNA[J].Appl Microbiol and Biot,1999,52(4):541-545.
[19] IDDAR A,VALVERDE F,SERRANO A,et al.Expression, purification, and characterization of recombinant nonphosphorylating NADP-dependent glyceraldehyde-3-phosphate dehydrogenase from Clostridium acetobutylicum[J].Protein Expression Purif,2002,25(3):519-526.
[20] XU Meijuan,RAO Zhiming,DOU Wenfang,et al.Site-directed mutagenesis and feedback-resistant N-acetyl-L-glutamate kinase (NAGK) increase Corynebacterium crenatum L-arginine production[J].Amino Acids,2012,43(1):255-266.
[21] JIANG Lingyan, ZHANG Yuanyuan,LI Zhen,et al.Metabolic engineering of Corynebacterium glutamicum for increasing the production of L-ornithine by increasing NADPH availability[J].J Ind Microbiol Biotechnol,2013, 40(10):1 143-1 151.
[22] KABUS A,GEORGI T,WENDISCH V F,et al.Expression of the Escherichia coli pntAB genes encoding a membrane-bound transhydrogenase in Corynebacterium glutamicum improves L-lysine formation[J]. Appl Microbiol Biot,2007,75(1):47-53.
[23] YAMAMOTO S,GUNJI W,SUZUKI H,et al.Overexpression of genes encoding glycolytic enzymes in Corynebacterium glutamicum enhances glucose metabolism and alanine production under oxygen deprivation conditions[J].Appl Environ Microb,2012,78(12):4 447-4 457.
