Comparison of different strategies to reduce acetate formation in Escherichia coli

被引:42
作者
De Mey, Marjan
Lequeux, Gaspard J.
Beauprez, Joeri J.
Maertens, Jo
Van Horen, Ellen
Soetaert, Wim K.
Vanrolleghem, Peter A.
Vandamme, Erick J.
机构
[1] Univ Ghent, Lab Ind Microbiol & Biocatalysis, Dept Biochem & Microbial Technol, B-9000 Ghent, Belgium
[2] Fac Biosci Engn, Dept Appl Math Biometr & Proc Control, BIOMATH, B-9000 Ghent, Belgium
关键词
D O I
10.1021/bp070170g
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
E. coli cells produce acetate as an extracellular coproduct of aerobic cultures. Acetate is undesirable because it retards growth and inhibits protein formation. Most process designs or genetic modifications to minimize acetate formation aim at balancing growth rate and oxygen consumption. In this research, three genetic approaches to reduce acetate formation were investigated: (1) direct reduction of the carbon flow to acetate (ackA-pta, poxB knock-out); (2) anticipation on the underlying metabolic and regulatory mechanisms that lead to acetate (constitutive ppc expression mutant); and (3) both (1) and (2). Initially, these mutants were compared to the wild-type E. coli via batch cultures under aerobic conditions. Subsequently, these mutants were further characterized using metabolic flux analysis on continuous cultures. It is concluded that a combination of directly reducing the carbon flow to acetate and anticipating on the underlying metabolic and regulatory mechanism that lead to acetate, is the most promising approach to overcome acetate formation and improve recombinant protein production. These genetic modifications have no significant influence on the metabolism when growing the microorganisms under steady state at relatively low dilution rates (less than 0.4 h(-1)).
引用
收藏
页码:1053 / 1063
页数:11
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