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
收藏
页码：116 / 121
页数：6
相关论文
共 21 条
  • [1] High-throughput classification of yeast mutants for functional genomics using metabolic footprinting
    Allen, J
    Davey, HM
    Broadhurst, D
    Heald, JK
    Rowland, JJ
    Oliver, SG
    Kell, DB
    [J]. NATURE BIOTECHNOLOGY, 2003, 21 (06) : 692 - 696
  • [2] Bakker BM, 2001, FEMS MICROBIOL REV, V25, P15, DOI 10.1016/S0168-6445(00)00039-5
  • [3] Glutathione dysregulation and the etiology and progression of human diseases
    Ballatori, Nazzareno
    Krance, Suzanne M.
    Notenboom, Sylvia
    Shi, Shujie
    Tieu, Kim
    Hammond, Christine L.
    [J]. BIOLOGICAL CHEMISTRY, 2009, 390 (03) : 191 - 214
  • [4] Identification of small molecules from human embryonic stem cells using metabolomics
    Cezar, Gabriela G.
    Quam, Jessica A.
    Smith, Alan M.
    Rosa, Guilherme J. M.
    Piekarczyk, Marian S.
    Brown, James F.
    Gage, Fred H.
    Muotri, Alysson R.
    [J]. STEM CELLS AND DEVELOPMENT, 2007, 16 (06) : 869 - 882
  • [5] Metabolomics profiling of extracellular metabolites in recombinant Chinese Hamster Ovary fed-batch culture
    Chong, William P. K.
    Goh, Lin Tang
    Reddy, Satty G.
    Yusufi, Faraaz N. K.
    Lee, Dong Yup
    Wong, Niki S. C.
    Heng, Chew Kiat
    Yap, Miranda G. S.
    Ho, Ying Swan
    [J]. RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 2009, 23 (23) : 3763 - 3771
  • [6] Enhanced IFNγ Production in Adenosine-Treated CHOCells: A Mechanistic Study
    Chong, William P. K.
    Sim, L. C.
    Wong, Kathy T. K.
    Yap, Miranda G. S.
    [J]. BIOTECHNOLOGY PROGRESS, 2009, 25 (03) : 866 - 873
  • [7] A Study of Monoclonal Antibody-Producing CHO Cell Lines: What Makes a Stable High Producer?
    Chusainow, Janet
    Yang, Yuan Sheng
    Yeo, Yessna H. M.
    Toh, Poh Choo
    Asvadi, Parisa
    Wong, Niki S. C.
    Yap, Miranda G. S.
    [J]. BIOTECHNOLOGY AND BIOENGINEERING, 2009, 102 (04) : 1182 - 1196
  • [8] The malate-aspartate NADH shuttle components are novel metabolic longevity regulators required for calorie restriction-mediated life span extension in yeast
    Easlon, Erin
    Tsang, Felicia
    Skinner, Craig
    Wang, Chen
    Lin, Su-Ju
    [J]. GENES & DEVELOPMENT, 2008, 22 (07) : 931 - 944
  • [9] Asymmetric dimethylarginine induces apoptosis via p38 MAPK/caspase-3-dependent signaling pathway in endothelial cells
    Jiang, DH
    Jia, SJ
    Dai, Z
    Li, YJ
    [J]. JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY, 2006, 40 (04) : 529 - 539
  • [10] Metabolomics as a complementary tool in cell culture
    Khoo, Soo Hean Gary
    Al-Rubeai, Mohamed
    [J]. BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, 2007, 47 (71-84) : 71 - 84
123