生物合成单葡糖苷酸基甘草次酸的菌株筛选与其产酶动力学的研究.pdfVIP

生物合成单葡糖苷酸基甘草次酸的菌株筛选及其产酶动力学的研究∗ 1 冯世江，李 春 ， 王小艳，韩喜勇，文先军 石河子大学化学化工学院生物化工实验室，新疆 石河子，832003 ） （ 摘 要 本文以单葡糖苷酸基甘草次酸的生物合成为目的，筛选到一株青霉菌属真菌，经甘草酸诱导后，表达的葡糖酸苷 酶属于胞内酶，具有较高底物特异性，能定向水解甘草酸后生成单葡糖苷酸基甘草次酸，其摩尔转化率可达到88.45 ％。 产酶动力学研究表明，该菌株生长与产酶为非偶联型，表达目标酶时存在着碳代谢阻遏现象，甘草酸是最佳碳源及诱导剂， 培养基初始pH4.8 、培养温度32℃及添加0.12 ％的Tween80 能显著提高产酶能力，最高酶活可达286.13 U/ml 。 关键词 单葡糖苷酸基甘草次酸 生物合成 菌株筛选 产酶动力学 SCREENING FOR β-D-MONO-GLUCURONIDE-GLYCYRRHIZIN DIRECTED BIOSYNTHESIS STRAIN AND KINETICS STUDY OF ITS ENZYME PRODUCTION 1 FENG Shijiang, LI Chun , WANG Xiaoyan, HAN xiyong , WEN xianjun (Lab. of Biochemical Engineering Biochemical engineering department of Chemistry and Chemical Engineering School, Shihezi University, 832003,Xinjiang Shihezi, P.R.China) Abstract One fungus, tentatively named Penicillium sp. Li-3, was screened to biosynthesize β-D-Mono-Glucuronide-Glycyrrhizin (GAMG), directly. Using Glycyrrhizin as elicitor and the sole carbon source, this strain was capable of expressing β-D-glucuronidase, one intracellular enzyme with high substrate specificity. And Glycyrrhizin hydrolyzing reaction was directly catalyzed by this enzyme with high GAMG production. It was found that the mol conversion ratio of this reaction was up to 88.45%. Research about kinetics of β-D-glucuronidase production showed that the growth and enzyme production of this strain was uncoupled. During the expressing of target enzyme, carbon catabolite repression existed, so only glycyrrhizin was the best carbon source as well as the elicitor. It was found that the surfactant (Tween80 0.12%) could improve the ability of enzyme production markedly. Under the co