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Rapid Determination of l-Glutamine using Engineered Escherichia coli Overexpressing Glutamine Synthetase

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Abstract

A genetically engineered Escherichia coli was developed as the source of enzyme for rapidly quantifying glutamine. E. coli BL21 (DE3) cells overexpressing a glutamine synthetase from Bacillus subtilis were prepared as tube aliquots and used in a small volume of nontoxic mixture. The current method was compared to high performance liquid chromatography analysis, Sigma kit (GLN-1) and Mecke method. The method is applicable to a wide range of glutamine concentrations (0.05–2.5 mM) and correlates well to the detection results obtained from high performance liquid chromatography (Pearson correlation is 0.978 at the 0.01 level). Moreover, the whole assay procedure takes less than 15 min and uses nontoxic reagents, so it can be applied to monitor glutamine production and utilization conveniently.

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Acknowledgments

This work was financially supported by Chinese High Technology Research and Development Program (Grant No. 2006AA02Z233). The authors would like to thank Prof. Ying Li for helpful suggestions and Bin Wang for providing wild-type BHK cells. Thanks also to Fei-Fei Guan, Yong-Xing Wang for HPLC analysis, Xiang Li and Zhong-Huai He for BHK cell culture.

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Authors and Affiliations

  1. State Key Laboratories for Agrobiotechnology, China Agricultural University, Beijing, 100094, People’s Republic of China

    Ni-Ning Hong, Guang Yang, Jian Li, Yao-Ping Zhang & Ji-Lun Li

  2. Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing, 100094, People’s Republic of China

    Ni-Ning Hong, Guang Yang, Jian Li, Yao-Ping Zhang & Ji-Lun Li

  3. Department of Bacteriology, University of Wisconsin Madison, Madison, WI, 53706, USA

    Yao-Ping Zhang

  4. Center for the Study of Nitrogen Fixation, University of Wisconsin Madison, Madison, WI, 53706, USA

    Yao-Ping Zhang

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  1. Ni-Ning Hong
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  2. Guang Yang
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  3. Jian Li
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  4. Yao-Ping Zhang
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  5. Ji-Lun Li
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Correspondence to Ji-Lun Li.

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Hong, NN., Yang, G., Li, J. et al. Rapid Determination of l-Glutamine using Engineered Escherichia coli Overexpressing Glutamine Synthetase. Appl Biochem Biotechnol 158, 398–407 (2009). https://doi.org/10.1007/s12010-008-8341-x

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  • DOI: https://doi.org/10.1007/s12010-008-8341-x

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