Effect of odhA overexpression and odhA antisense RNA expression on Tween-40-triggered glutamate production by Corynebacterium glutamicum

被引:48
作者
Kim, Jongpill [2 ]
Hirasawa, Takashi [1 ]
Sato, Yoshiyasu [1 ]
Nagahisa, Keisuke [1 ]
Furusawa, Chikara [1 ]
Shimizu, Hiroshi [1 ]
机构
[1] Osaka Univ, Dept Bioinformat Engn, Grad Sch Informat Sci & Technol, Suita, Osaka 5650871, Japan
[2] Osaka Univ, Dept Biotechnol, Grad Sch Engn, Suita, Osaka 5650871, Japan
关键词
Corynebacterium glutamicum; Glutamate overproduction; odhA; Antisense RNA; Tween; 40; 2-Oxoglutarate dehydrogenase complex; BREVIBACTERIUM-LACTOFERMENTUM; 2-OXOGLUTARATE DEHYDROGENASE; CLOSTRIDIUM-ACETOBUTYLICUM; CORYNEFORM BACTERIA; METABOLIC FLUX; ACID FERMENTATION; BIOTIN LIMITATION; GENOME SEQUENCE; DOWN-REGULATION; AMINO-ACIDS;
D O I
10.1007/s00253-008-1743-4
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
Recent studies have suggested that a decrease in the specific activity of the 2-oxoglutarate dehydrogenase complex (ODHC) is important for glutamate overproduction by Corynebacterium glutamicum. To further investigate the role of the odhA gene and its product in this process, we constructed the recombinant strains of C. glutamicum in which the expression of the odhA and its product could be controlled by odhA overexpression and odhA antisense RNA expression. We examined changes in glutamate production and ODHC specific activity of the constructed strains during glutamate production triggered by Tween 40 addition. The ODHC specific activity increased with odhA overexpression, resulting in dramatically reduced glutamate production despite Tween 40 addition, indicating that a decrease in the specific activity of ODHC is required for glutamate production induced by Tween 40 addition. However, odhA antisense RNA expression alone did not result in glutamate overproduction in spite of the decrease in ODHC specific activity. Rather, it enhanced glutamate production triggered by Tween 40 addition due to the additional decrease in ODHC specific activity, suggesting that odhA antisense RNA expression is effective in enhancing Tween-40-triggered glutamate overproduction. Our results suggest that a change in ODHC specific activity is critical but is not the only factor responsible for glutamate overproduction by C. glutamicum.
引用
收藏
页码:1097 / 1106
页数:10
相关论文
共 38 条
[1]   Altered metabolic flux due to deletion of odhA causes L-glutamate overproduction in Corynebacterium glutamicum [J].
Asakura, Yoko ;
Kimura, Eiichiro ;
Usuda, Yoshihiro ;
Kawahara, Yoshio ;
Matsui, Kazuhiko ;
Osumi, Tsuyoshi ;
Nakamatsu, Tsuyoshi .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2007, 73 (04) :1308-1319
[2]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[3]  
Desai RP, 1999, APPL ENVIRON MICROB, V65, P936
[4]   CARRIER-MEDIATED GLUTAMATE SECRETION BY CORYNEBACTERIUM-GLUTAMICUM UNDER BIOTIN LIMITATION [J].
GUTMANN, M ;
HOISCHEN, C ;
KRAMER, R .
BIOCHIMICA ET BIOPHYSICA ACTA, 1992, 1112 (01) :115-123
[5]   EVIDENCE FOR AN EFFLUX CARRIER SYSTEM INVOLVED IN THE SECRETION OF GLUTAMATE BY CORYNEBACTERIUM-GLUTAMICUM [J].
HOISCHEN, C ;
KRAMER, R .
ARCHIVES OF MICROBIOLOGY, 1989, 151 (04) :342-347
[6]   The Corynebacterium glutamicum genome:: features and impacts on biotechnological processes [J].
Ikeda, M ;
Nakagawa, S .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2003, 62 (2-3) :99-109
[7]   The complete Corynebacterium glutamicum ATCC 13032 genome sequence and its impact on the production of L-aspartate-derived amino acids and vitamins [J].
Kalinowski, J ;
Bathe, B ;
Bartels, D ;
Bischoff, N ;
Bott, M ;
Burkovski, A ;
Dusch, N ;
Eggeling, L ;
Eikmanns, BJ ;
Gaigalat, L ;
Goesmann, A ;
Hartmann, M ;
Huthmacher, K ;
Krämer, R ;
Linke, B ;
McHardy, AC ;
Meyer, F ;
Möckel, B ;
Pfefferle, W ;
Pühler, A ;
Rey, DA ;
Rückert, C ;
Rupp, O ;
Sahm, H ;
Wendisch, VF ;
Wiegräbe, I ;
Tauch, A .
JOURNAL OF BIOTECHNOLOGY, 2003, 104 (1-3) :5-25
[8]   Relationship between the glutamate production and the activity of 2-oxoglutarate dehydrogenase in Brevibacterium lactofermentum [J].
Kawahara, Y ;
TakahashiFuke, K ;
Shimizu, E ;
Nakamatsu, T ;
Nakamori, S .
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 1997, 61 (07) :1109-1112
[9]  
Kinoshita S., 1957, J GEN MICROBIOL, V3, P193
[10]   Antisense technology in molecular and cellular bioengineering [J].
Lee, LK ;
Roth, CM .
CURRENT OPINION IN BIOTECHNOLOGY, 2003, 14 (05) :505-511