Metabolomics-driven approach for the improvement of Chinese hamster ovary cell growth: Overexpression of malate dehydrogenase II

被引：71

作者：

Chong, William P. K. ^{[1
,2
]}

Reddy, Satty G. ^{[1
]}

Yusufi, Faraaz N. K. ^{[1
]}

Lee, Dong-Yup ^{[1
,3
]}

Wong, Niki S. C. ^{[1
]}

Heng, Chew Kiat ^{[2
]}

Yap, Miranda G. S. ^{[1
,3
]}

Ho, Ying Swan ^{[1
]}

机构：

[1] Agcy Sci Technol & Res, Bioproc Technol Inst, Singapore 138668, Singapore

[2] Natl Univ Singapore, Yong Loo Lin Sch Med, Singapore 119220, Singapore

[3] Natl Univ Singapore, Dept Chem & Biomol Engn, Singapore 117576, Singapore

来源：

JOURNAL OF BIOTECHNOLOGY | 2010年 / 147卷 / 02期

关键词：

Metabolomics; Chinese hamster ovary; Liquid chromatography-mass spectrometry (LC-MS); Metabolic engineering; Aspartate; Malate dehydrogenase II; SACCHAROMYCES-CEREVISIAE; METABOLISM; NADH; BIOMARKERS; APOPTOSIS; DISEASES; CULTURE; YEAST;

D O I：

10.1016/j.jbiotec.2010.03.018

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：

We have established a liquid chromatography-mass spectrometry based metabolomics platform to identify extracellular metabolites in the medium of recombinant Chinese hamster ovary (CHO) fed-batch reactor cultures. Amongst the extracellular metabolites identified, malate accumulation was the most significant. The contributing factors to malate efflux were found to be the supply of aspartate from the medium, and an enzymatic bottleneck at malate dehydrogenase II (MDH II) in the tricarboxylic acid cycle. Subsequent metabolic engineering to overexpress MDH II in CHO resulted in increases in intracellular ATP and NADH, and up to 1.9-fold improvement in integral viable cell number. (C) 2010 Elsevier B.V. All rights reserved.

引用

页码：116 / 121

页数：6