Enhanced production of plasmid DNA by engineered Escherichia coli strains

被引:13
|
作者
Pablos, Tania E. [1 ]
Soto, Rene [2 ]
Meza Mora, Eugenio [3 ]
Le Borgne, Sylvie [1 ]
Ramirez, Octavio T. [2 ]
Gosset, Guillermo [3 ]
Lara, Alvaro R. [1 ]
机构
[1] Univ Autonoma Metropolitana Cuajimalpa, Dept Proc & Tecnol, Mexico City 01120, DF, Mexico
[2] Univ Nacl Autonoma Mexico, Inst Biotecnol, Dept Med Mol & Bioproc, Cuernavaca 62210, Morelos, Mexico
[3] Univ Nacl Autonoma Mexico, Inst Biotecnol, Dept Ingn Celular & Biocatalisis, Cuernavaca 62210, Morelos, Mexico
关键词
Plasmid DNA; Escherichia coli; PTS; pykA; pDNA vaccines; Acetate; SYSTEM; PERFORMANCE; METABOLISM; ACETATE; GLUCOSE; GROWTH;
D O I
10.1016/j.jbiotec.2011.04.015
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 strains VH33 (PTS- GalP(+) strain displaying a strongly reduced overflow metabolism) and VH34 (additionally lacking the pyruvate kinase A) were evaluated for the production of a plasmid DNA (pDNA) vaccine. The parent (W3110) and mutant strains were cultured using 10 g of glucose/L. While the specific growth rates of the three strains were similar, they presented differences in the accumulation of acetate. W3110 accumulated up to 4 g/L of acetate, VH33 produced 1.4 g/L, and VH34 only 0.78 g/L. VH33 and VH34 produced 76% and 300% more pDNA than W3110. Moreover, VH34 demanded 33% less oxygen than VH33 and W3110, which can be advantageous for large-scale applications. (C) 2011 Elsevier B.V. All rights reserved.
引用
收藏
页码:211 / 214
页数:4
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