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. 2006 Aug 5;51(1):21–28. doi: 10.1007/s10616-006-9010-y

Metabolic characteristics of recombinant Chinese hamster ovary cells expressing glutamine synthetase in presence and absence of glutamine

Fang Zhang 1, Xiangming Sun 1, Xiaoping Yi 1, Yuanxing Zhang 1,✉
PMCID: PMC3449477  PMID: 19002891

Abstract

To elucidate the metabolic characteristics of recombinant CHO cells expressing glutamine synthetase (GS) in the medium with or without glutamine, the concentrations of extra- and intracellular metabolites and the activities of key metabolic enzymes involved in glutamine metabolism pathway were determined. In the absence of glutamine, glutamate was utilized for glutamine synthesis, while the production of ammonia was greatly decreased. In addition, the expression of recombinant protein was increased by 18%. Interestingly, the intracellular glutamine maintained almost constant, independent of the presence of glutamine or not. Activities of glutamate-oxaloacetate aminotransferase (GOT), glutamate-pyruvate aminotransferase (GPT), and glutamate dehydrogenase (GDH) increased in the absence of glutamine. On the other hand, intracellular isocitrate and the activities of its downstream isocitrate dehydrogenase in the TCA cycle increased also. In combination with these two factors, a 8-fold increase in the intracellular α-ketoglutarate was observed in the culture of CHO-GS cells in the medium without glutamine.

Keywords: Animal cell culture, Cell metabolism, Glutamine, Glutamine synthetase, Recombinant CHO cell

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Acknowledgements

This work was supported by the grants from the National High-Tech Research and Development Programs of China (No. 2004AA2Z3792).

Abbreviations

CHO-GS

Recombinant Chinese hamster ovary cell bearing the exogenous glutamine synthetase gene

GS

Glutamine synthetase

GOT

Glutamate-oxaloacetate aminotransferase

GPT

Glutamate-pyruvate aminotransferase

GDH

Glutamate dehydrogenase

PAG

Phosphate-activated glutaminase

CX

Viable cell density

α-KG

α-Ketoglutarate

Fum

Fumarate

Cit

Citrate

Icit

Isocitrate

Mal

Malate

MSX

Methionine sulphoximine

QP

Specific production rate of metabolite

QS

Specific consumption rate of substrate

YP/S

Apparent yield coefficient of product to substrate

Subscripts

0

Seeding time

t

Harvest time

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