一种作用于花青素和甜菊醇的甜菊糖基转移酶的基因克隆和功能分析

本文对一种新的甜菊糖基转移酶进行了基因克隆和功能分析。获得的基因cDNA全长1419bp,编码473个氨基酸,蛋白质分子量约53.2K Da。与常见的糖基转移酶基因比较,相似性达44%以上,且具有糖基转移酶的保守序列。体外异源表达获得的融合蛋白,具有在花青素类和甜菊醇等糖基受体上转糖基的酶活性。在对一系列不同底物的酶活性进行比较后,推测这种糖基转移酶在体内参与了甜菊糖苷的合成。结果表明,具有广泛的底物活性的类黄酮类糖基转移酶,在甜菊体内不仅对类黄酮转糖基,而且在生成水溶性甜菊糖苷的过程中也扮演重要的角色。

MOLECULAR CLONING AND CHARACTERIZATION OF STEVIA REBA UDIANA UDP-GLUCOSYLTRANSFERASE

We report here the cloning and characterization of a UDP-glucose flavonoid glucosyltransferase (srUFGT) in Stevia rebaudiana. The isolated cDNA was 1419bp in length encoding 473 deduced amino acids with a predicted molecular mass of 53. 2 kDa. The products of in vitro translation from an expression vector had anthocyanidins and steviol glucosyltransferase activity. Comparison of the activity of the recombinant UDP-glucosyltransferase toward a range of acceptor substrates suggests that it may participate in the synthesis of steviol glycosides. The results support the hypothesis that the flavonoid glucosyltransferases, which have a broad substrate specificity,may be not only involved in flavonoid glucosylation but also play a role in producing the water-soluble steviol-glycosides in S. rebaudiana .