重组里氏木霉粗酶液提取虎杖中白藜芦醇的研究  

Extraction of Resveratrol from Polygonumcuspidatum Using Crude Cellulase from Trichoderma Reesei Recombinants

作　　者：杨祎彬 龚泽华 裘海平 朱有贵 陈钦钦 俞佳军 王冰冰[3] 金欣[1] 

机构地区：[1]绍兴文理学院医学院,浙江绍兴312000 [2]浙江工业大学生物工程学院,浙江杭州310014 [3]浙江大学化学工程与生物工程学院,浙江杭州310027

出　　处：《高校化学工程学报》2017年第5期1156-1161,共6页Journal of Chemical Engineering of Chinese Universities

基　　金：浙江大学感染病诊治国家重点实验室开放课题(2016KF01);绍兴市科技局公益性技术应用研究计划项目(2014B70051);浙江省大学生科技创新活动计划暨新苗人才计划(2016R428030)

摘　　要：为从虎杖中提取白藜芦醇并同步实现高效转化,经平板初筛和48 h的试管产酶复筛,从619株里氏木霉的基因重组菌中定向筛选出3株高产β-葡萄糖苷酶的菌株(BG-2,BG-4和BG-5)。摇瓶条件下产酶48h,三者的β-葡萄糖苷酶酶活为1.75,3.50和5.71 IU×m L^(-1),分别是出发菌株的36倍、73倍和119倍。以三者的发酵粗酶液直接处理虎杖粗提物,45℃条件下反应5 h后,与出发菌株相对比,重组菌株对白藜芦醇的提取率可达14.86~16.41 mg×g^(-1),是出发菌株的2.41~2.66倍;转化率可达120.5%~138.4%,是出发菌株的6.0~6.85倍,且反应液中基本无白藜芦醇苷残留。该研究策略从根本上提高了纤维素酶法对苷元型白藜芦醇的提取效率,可推广应用于其他易糖苷化的中草药成分的提取。In order to increase extraction efficiency of resveratrol from Polygonumcuspidatum, 619 Trichoderma reesei recombinants were screened for high β-glucosidase activity via plate screening and enzyme production in tubes for 48 h, and 3 Trichoderma reesei recombinants（BG-2, BG-4, BG-5） were selected. The β-glucosidase activity of the three recombinants are 1.75, 3.50 and 5.71 IU×m L^（-1） respectively after 48 h fermentation in shake flasks, which are 36,73 and 119 times higher than the original.Polygonumcuspidatum was treated with the crude enzymes. Compared with the original strains, the resveratrol extraction ratio increases by 2.41~2.66 times（14.86~16.41 mg×g^（-1）） after 5 hours reaction in 45°C, and the transformation ratio of resveratrol increases by 6.0~6.85 times（120.5%~138.4%） with hardly any piceid remains. This strategy significantly improves the extraction efficiency of resveratrol by cellulose, which can be applied to the extraction of other glycoside of Chinese traditional herb components.

关 键 词：白藜芦醇 纤维素酶 提取 虎杖 白藜芦醇苷 

分 类 号：Q814.9[生物学—生物工程]