Protein Expression and Functional Characterization of a Novel Gene Similar to Furanose Biosynthetic Gene in Sequence

• Main Authors: Hung, Chang-Yuan, 洪章元
• Other Authors: 林苕吟
• Format: Others
• Language: zh-TW
• Published: 2012
• Online Access: http://ndltd.ncl.edu.tw/handle/11567285761647897499

碩士 === 國立交通大學 === 生物科技學系 === 101 === Many natural products with biological activities, including antitumor, antibiotic and antifungal activities, contain sugar moieties. These sugar moieties often play important roles in activity of their parent compounds. The modification or removal of the sugar moieties often changes the reported activities. In this study, we choose a novel gene similar to furanose biosynthetic gene as a research target, which has been speculated to participate in the biosynthetic pathway of furanose or related glycosides. By heterologous expression in Escherichia coli, the expression condition of the protein encoded by the gene was optimized. Then, this enzyme was purified by affinity column and analyzed with SDS-PAGE. In order to characterize the enzymatic function, a spectophotometric method was used to detect enzyme activity. In this study, this enzyme was analyzed with possible cofactors, NADPH and NADH. The results thus provide information about this novel enzyme possibly participating in biosynthetic pathways of furanose or related glycosides.