Metabolomics-driven approach for the improvement of Chinese hamster ovary cell growth: overexpression of malate dehydrogenase II.
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.
Chong
William P K
WP
Bioprocessing Technology Institute, A*STAR (Agency for Science, Technology and Research), 20 Biopolis Way, #06-01 Centros, Singapore 138668, Singapore. william_chong@bti.a-star.edu.sg
Reddy
Satty G
SG
Yusufi
Faraaz N K
FN
Lee
Dong-Yup
DY
Wong
Niki S C
NS
Heng
Chew Kiat
CK
Yap
Miranda G S
MG
Ho
Ying Swan
YS
D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D016466:CHO Cells / Q000166:cytology* / Q000378:metabolism; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D048708:Cell Growth Processes / Q000502:physiology; D002853:Chromatography, Liquid; D006224:Cricetinae; D003412:Cricetulus; D008291:Malate Dehydrogenase / Q000096:biosynthesis* / Q000378:metabolism; D008293:Malates / Q000378:metabolism; D013058:Mass Spectrometry; D053858:Metabolic Networks and Pathways; D055432:Metabolomics / Q000379:methods*
D008293:Malates; D001224:Aspartic Acid; C030298:malic acid; D008291:Malate Dehydrogenase