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

来源：

JOURNAL OF BIOTECHNOLOGY | 2012年 / 158卷 / 04期

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

页码：211 / 214

页数：4

共 16 条

[1] Plasmid DNA Production Combining Antibiotic-Free Selection, Inducible High Yield Fermentation, and Novel Autolytic Purification [J].

Carnes, Aaron E. ;

Hodgson, Clague P. ;

Luke, Jeremy M. ;

Vincent, Justin M. ;

Williams, James A. .

BIOTECHNOLOGY AND BIOENGINEERING, 2009, 104 (03) :505-515

[2] Pyruvate Kinase-Deficient Escherichia coli Exhibits Increased Plasmid Copy Number and Cyclic AMP Levels [J].

Cunningham, Drew S. ;

Liu, Zhu ;

Domagalski, Nathan ;

Koepsel, Richard R. ;

Ataai, Mohammad M. ;

Domach, Michael M. .

JOURNAL OF BACTERIOLOGY, 2009, 191 (09) :3041-3049

[3] Replacement of the glucose phosphotransferase transport system by galactose permease reduces acetate accumulation and improves process performance of Escherichia coli for recombinant protein production without impairment of growth rate [J].

De Anda, Ramn ;

Lara, Alvaro R. ;

Hernandez, Vanessa ;

Hernandez-Montalvo, Vernica ;

Gosset, Guillermo ;

Bolívar, Francisco ;

Ramírez, Octavio T. .

METABOLIC ENGINEERING, 2006, 8 (03) :281-290

[4] Overcoming acetate in Escherichia coli recombinant protein fermentations [J].

Eiteman, Mark A. ;

Altman, Elliot .

TRENDS IN BIOTECHNOLOGY, 2006, 24 (11) :530-536

[5] Analysis of carbon metabolism in Escherichia coli strains with an inactive phosphotransferase system by 13C labeling and NMR spectroscopy [J].

Flores, S ;

Gosset, G ;

Flores, N ;

de Graaf, AA ;

Bolívar, F .

METABOLIC ENGINEERING, 2002, 4 (02) :124-137

[6] Using DNA as a drug-Bioprocessing and delivery strategies [J].

Han, Ying ;

Liu, Shan ;

Ho, Jenny ;

Danquah, Michael K. ;

Forde, Gareth M. .

CHEMICAL ENGINEERING RESEARCH & DESIGN, 2009, 87 (3A) :343-348

[7] A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein [J].

Herrera, Emma ;

Barcenas, Patricia ;

Hernandez, Rubicela ;

Mendez, Alfonso ;

Perez-Ishiwara, Guillermo ;

Barron, Blanca .

VIROLOGY JOURNAL, 2010, 7

[8] High cell-density processes in batch mode of a genetically engineered Escherichia coli strain with minimized overflow metabolism using a pressurized bioreactor [J].

Knabben, Ingo ;

Regestein, Lars ;

Marquering, Frank ;

Steinbusch, Sven ;

Lara, Alvaro R. ;

Buechs, Jochen .

JOURNAL OF BIOTECHNOLOGY, 2010, 150 (01) :73-79

[9] Utility of an Escherichia coli strain engineered in the substrate uptake system for improved culture performance at high glucose and cell concentrations:: An fed-batch cultures [J].

Lara, Alvaro R. ;

Caspeta, Luis ;

Gosset, Guillermo ;

Bolivar, Francisco ;

Ramirez, Octavio T. .

BIOTECHNOLOGY AND BIOENGINEERING, 2008, 99 (04) :893-901

[10] Transcriptional and metabolic response of recombinant Escherichia coli to spatial dissolved oxygen tension gradients simulated in a scale-down system [J].

Lara, AR ;

Leal, L ;

Flores, N ;

Gosset, G ;

Bolívar, F ;

Ramírez, OT .

BIOTECHNOLOGY AND BIOENGINEERING, 2006, 93 (02) :372-385

← 1 2 →