与茶叶香气形成有关的β葡萄糖苷酶活性研究及其基因表达茶学专业论文.docxVIP

AbstractVolatile Abstract Volatile aroma compounds in tea leaves exist in the form of glycosidic aroma precursors based on primeverosides and glucosides．The volatile aglycones released from glucosides hydrolysis by 13-glucosidase are not only the material base of tea aroma，but also participate in the tea plant defense system and strengthen the plant disease resistance． Therefore，13-glucosidase plays a very important role on tea aroma and stress． In this paper，the activity o f 13一glucosidase in tea leaves，gene expression level and the expression of Glu in vitro were studied．The main results are as follows． Studying on the change of 13-glucosidase activities during the manufacturing process of black tea，the activity of 13-glucosidase reached the highest level at the end of withering and then decreased gradually．The content of volatiles such as(Z)一3-hexene-1-ol，benzyl alcohol，phenylethyl alcohol，linalool，methyl salicylate and geraniol increased obviously， the results showed that 13-glucosidase can effectively hydrolyzed glucosidic aroma precursors，liberate aroma constituents． Researched on lB-glucosidase activity and gene expression level of Glu in various positions of different varieties，it indicated that the leaves more tender have the higher activity and expression level．The enzyme activity and expression level of Glu in the four tested cultivars thus Anhui Yihao，Fuding Dabaicha and Shanpolv were higher than Longjin943． Bioinformatic analysis showed that Gh was hydrophilic and had two strong transmembrane helices．There were 24 phosphorylation sites and signal peptide cleavage site between pos．26 and 27 in the polypeptide chain．The recombinant expression vectors pMAL—Glu was then constructed by inserting the Glu containing no signal peptide into pMAL-c5X vector．The recombinant was reduced by IPTG at 37。C and the fusion protein mainly existed as inclusion body．The fusion protein in soluble part was obtained at 1 6。C． The eukaryotic expression system for Glu was const