| Summary: | 碩士 === 明志科技大學 === 生化工程研究所 === 99 === In this study, Taitung temperature of hot spring water for a specific strain of the screening, and successfully combined with the protein sequencing method and DNA sequencing methods to complete Portugal Sugar Sugar dehydrogenase (Glucose dehydrogenase, GDH-NAD) to confirm the source of genetic effects Sulfolobus sp. LJ01, and in order to quickly quantify enzyme required for the GDH-NAD, this study using gene recombination technology GDH-NAD E. coli gene transfer to a large number of E. coli expression system to facilitate performance analysis, based on research results, recombinant GDH-NAD enzyme by SDS-PAGE and Western-blot analysis, the molecular weight of about 39 kDa, optimum pH and temperature stability of the reaction were pH 8.0 and 70 ℃, and with a high degree of specificity to glucose can be long-term storage at 4 ℃ and will not live beneath the phenomenon; in enzyme kinetics analysis, Km is 0.0219 M, the results of this study will be further applied to second-generation intrusion type specimen of blood glucose meters on the preparation.
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