Rapid determination of L-glutamine using engineered Escherichia coli overexpressing glutamine synthetase.
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.
Hong
Ni-Ning
NN
State Key Laboratories for Agrobiotechnology, China Agricultural University, Beijing 100094, People's Republic of China.
Yang
Guang
G
Li
Jian
J
Zhang
Yao-Ping
YP
Li
Ji-Lun
JL
D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D005818:Genetic Engineering; D005974:Glutamate-Ammonia Ligase / Q000235:genetics / Q000378:metabolism*; D005973:Glutamine / Q000032:analysis* / Q000378:metabolism*
D005973:Glutamine; D005974:Glutamate-Ammonia Ligase