Analysis of low-level α-D-glucose-1-phosphate in thermophilic enzyme reaction mixuture using high pH anion-exchange chromatograph = 고성능 액체 크로마토그래프를 이용한 내열성 효소반응 산물인 α-D-glucose-1-phosphate의 저농도 분석

Abstract

We have used high pH anion-exchange chromatography to analyze liw level (below 20μM) α-ㅇ-glucose-1-phosphate (g-1-P) that can be used as a cytostatic compound, an antibiotic, and immunosuppressive drug. Our chromatographic method afforded excellemt peak resolution and seletivith fro 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 μM, giving slope of 4.8X10⁴ (peak area/μM). The detection limit was 2 μM. This method was applied to the purification of thermophilic α-glucan phosphorylase from Thermus caldophilus. The technique will be extremely useful in future studies concerning carbohydrate metabolism in living organisms.