ENGINEERING OF ESCHERICHIA-COLI CENTRAL METABOLISM FOR AROMATIC METABOLITE PRODUCTION WITH NEAR THEORETICAL YIELD

被引:170
作者
PATNAIK, R
LIAO, JC
机构
[1] TEXAS A&M UNIV,DEPT CHEM ENGN,COLLEGE STN,TX 77843
[2] TEXAS A&M UNIV,DEPT BIOCHEM & BIOPHYS,COLLEGE STN,TX 77843
关键词
D O I
10.1128/AEM.60.11.3903-3908.1994
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
Escherichia coli and many other microorganisms synthesize aromatic amino acids through the condensation reaction between phosphoenolpyruvate (PEP) and erythrose 4-phosphate to form 3-deoxy-D-arabinoheptulosonate 7-phosphate (DAHP). It has been shown that overexpression of transketolase increases the production of DAHP in an aroB mutant strain (unable to further metabolize DAHP) with elevated DAHP synthase. However, the yield (percent conversion) of DAHP from glucose is still low. Stoichiometric analysis shows that many enzymes compete for intracellular PEP. In particular, the phosphotransferase system, responsible for glucose transport in E. coli, uses PEP as a phosphate donor and converts it to pyruvate, which is less likely to recycle back to PEP. This stoichiometric limitation greatly reduces the yield of aromatic metabolites. To relieve this limitation, we overexpressed PEP synthase in the presence of glucose and showed that it increased the final concentration and the yield of DAHP by almost twofold, to a near theoretical maximum. The PEP synthase effect is not observed without overproduced transketolase, suggesting that erythrose 4-phosphate is the first limiting metabolite. This result demonstrates the utility of pathway analysis and the limitation of central metabolites in the high-level overproduction of desired metabolites.
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页码:3903 / 3908
页数:6
相关论文
共 26 条
[1]   NEW APPROACH TO TRYPTOPHAN PRODUCTION BY ESCHERICHIA-COLI - GENETIC MANIPULATION OF COMPOSITE PLASMIDS INVITRO [J].
AIBA, S ;
TSUNEKAWA, H ;
IMANAKA, T .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1982, 43 (02) :289-297
[2]   TOWARD A SCIENCE OF METABOLIC ENGINEERING [J].
BAILEY, JE .
SCIENCE, 1991, 252 (5013) :1668-1675
[3]   CELLULAR AND METABOLIC ENGINEERING - AN OVERVIEW [J].
CAMERON, DC ;
TONG, IT .
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 1993, 38 (1-2) :105-140
[4]   PLASMID INSERTION MUTAGENESIS AND LAC GENE FUSION WITH MINI-MU BACTERIOPHAGE TRANSPOSONS [J].
CASTILHO, BA ;
OLFSON, P ;
CASADABAN, MJ .
JOURNAL OF BACTERIOLOGY, 1984, 158 (02) :488-495
[5]  
CHAO YP, 1994, J BIOL CHEM, V269, P5122
[6]   ALTERATION OF GROWTH-YIELD BY OVEREXPRESSION OF PHOSPHOENOLPYRUVATE CARBOXYLASE AND PHOSPHOENOLPYRUVATE CARBOXYKINASE IN ESCHERICHIA-COLI [J].
CHAO, YP ;
LIAO, JC .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1993, 59 (12) :4261-4265
[7]   BIOCATALYTIC SYNTHESIS OF AROMATICS FROM D-GLUCOSE - THE ROLE OF TRANSKETOLASE [J].
DRATHS, KM ;
POMPLIANO, DL ;
CONLEY, DL ;
FROST, JW ;
BERRY, A ;
DISBROW, GL ;
STAVERSKY, RJ ;
LIEVENSE, JC .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1992, 114 (10) :3956-3962
[8]   EXPRESSION OF NAPHTHALENE OXIDATION GENES IN ESCHERICHIA-COLI RESULTS IN THE BIOSYNTHESIS OF INDIGO [J].
ENSLEY, BD ;
RATZKIN, BJ ;
OSSLUND, TD ;
SIMON, MJ ;
WACKETT, LP ;
GIBSON, DT .
SCIENCE, 1983, 222 (4620) :167-169
[9]   CORRELATION OF THEORETICAL AND EXPERIMENTAL YIELDS OF PHENYLALANINE FROM NON-GROWING CELLS OF A REC ESCHERICHIA-COLI STRAIN [J].
FORBERG, C ;
ELIAESON, T ;
HAGGSTROM, L .
JOURNAL OF BIOTECHNOLOGY, 1988, 7 (04) :319-332
[10]  
GOLLUB E, 1971, METHODS ENZYMOL A, V17, P349