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KMID : 0380019990140030384
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Korean Journal of Biotechnology and Bioengineering
1999 Volume.14 No. 3 p.384 ~ p.387
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Analysis of Low-level α-D-glucose-1-phosphate in Thermophilic Enzyme Reaction Mixuture Using High pH Anion-exchange Chromatograph
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신현재/Shin, Hyun Jae
신영숙/이대실/Shin, Young sook/Lee, Dae Sil
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Abstract
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We have used high pH anion-exchange chromatography to analyze low level (below 20PM) a-D-glucose-1-phosphate (G-1-P) that can be used as a cytostatic compound, an antibiotic, and immunosuppressive drug. Our chromatographic method afforded excellent peak resolution and seletivity for glucose-6-phosphate and various maltooligosaccharides as well as G-1-P. The pulsed amperometric detector yielded linear response on G-1-P ranging from 2 - 20 uM, giving slope of 4.8x104 (peak area/pM). The detection limit was 2 uM. This method was applied to the purification of thermophilic a-glucan phosphorylase from Thermus caldophilus. The technique will be extremely useful in future studies concerning carbohydrate metabolism in living organisms.
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