摘要
α-L-鼠李糖苷酶对天然产物之间的转化具有重要的作用和应用价值。该研究采用毕赤酵母表达α-L-鼠李糖苷酶AnRhaE,构建pPIC9K-AnRhaE表达载体,电转化至毕赤酵母GS115和KM71H中,经诱导表达,SDS-PAGE电泳显示GS115更有利于生产AnRhaE酶。AnRhaE酶在45~55℃时酶活力高,最适温度为55℃,对pH的耐受范围较广,pH 4.5~pH 8时酶活性稳定,最适pH为5。AnRhaE能水解α-1,2、α-1,6和2个鼠李糖苷连接的底物,包括柚苷二氢查耳酮、柚苷、芦丁、新橙皮苷、橙皮苷和朝藿定C等天然产物,但是AnRhaE对α-1,2糖苷键连接的底物具有较高的活性,而对α-1,6糖苷键连接的底物活性较差,对朝藿定C的活性较好,能将朝藿定C完全水解生成淫羊藿苷,因此AnRhaE酶在天然产物之间的转化具有较好的应用价值和前景。
The α-L-rhamnosidase plays an important role in the conversion of natural products,and Pichia pastoris was used to express α-L-rhamnosidase AnRhaE.The pPIC9K-AnRhaE expression vector was constructed and transformed into Pichia pastoris GS115 and KM71H.After induced expression,SDS-PAGE electrophoresis showed that GS115 was more conducive to the production of AnRhaE.High AnRhaE activity was obtained at 45-55℃,and the optimum temperature was 55℃.AnRhaE was stable at pH 4.5-8,and the optimum pH was 5.AnRhaE could hydrolyzeα-1,2,α-1,6 and rhamnosyl-rhamnosyl glycosidic bond,including naringin dihydrogen chalcone to trilobatin,naringin to prunin,rutin to isoquercetin,neohesperidin to hesperetin-7-O-glucoside,hesperidin to hesperetin-7-O-glucoside and epimedin C to icariin.AnRhaE had high activity to α-1,2 glycosidic bond than theα-1,6 glycosidic bond,and could complete hydrolysis epimedin C to icariin.Therefore,AnRhaE is potentially applicable in enzymatic conversion of natural products.
作者
叶德晓
黄佳俊
卢宇靖
林育成
李慧灵
谭景航
周金林
YE Dexiao;HUANG Jiajun;LU Yujing;LIN Yucheng;LI Huiling;TAN Jinghang;ZHOU Jinlin(HuiTeng(Foshan)Biotechnology Co.Ltd.,Foshan 528225,China;Golden Health(Guangdong)Biotechnology Co.Ltd.,Foshan 528225,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2021年第3期25-30,35,共7页
Food and Fermentation Industries
基金
佛山市南海区“蓝海人才计划”项目。
