Gene Cloning and Characterization of Tc-βFar/Lin, Tc-Lin, Tc-Zin/βOci, Tc-βFar/Ger and Tc-αFar/Ger Terpene Synthases from Taiwania cryptomerioides and Establishment of Production System in E. coli
| Autor: | Tzu-Fang Hsu, 徐子凡 |
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| Rok vydání: | 2017 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 105 Taiwania cryptomerioides Hayata is an endemic species in Taiwan. It is an important coniferous species in Taiwan, because of its timber properties and its terpenoids in the essential oil. Most previous researches about the products and applications of T. cryptomerioides Hayata have been discussed, however, the biosynthesis pathway of T. cryptomerioides Hayata has rarely been discussed in the past. This study focuses on cloning the genes and their characteristic. Through using gas chromatography mass spectrometry (GC/MS) analysis, we identified the product of the five terpene synthases. According to the main product that synthase synthesize, they are called Tc-βFar/Lin, Tc-Lin, Tc-Zin/βOci, Tc-βFar/Ger, and Tc-αFar/Ger. Tc-Lin is a monoterpene synthase, the others are sesquiterpene synthases. Except Tc-Zin/βOci synthesizes the single ring zingiberene, all of Tc-βFar/Lin, Tc-βFar/Ger, and Tc-αFar/Ger synthesize the linear monoterpenes or sesquiterpenes; the results of reverse transcription polymerase chain reaction (RT-PCR) demonstrate that these five genes can express in the young leaf and female cone, and only Tc-βFar/Ger can also express in the wood. From the analysis of the phylogenetic tree, we can find that Tc-βFar/Lin and other closer genes of sesquiterpene synthases which are known in Taiwania are all lost the trainsit peptides. This result represents that these genes may have the process which has lost γ-domain at first and lost trainsit peptides finally. Using heterologous expression of diverse synthases in the E. coli, Tc-Zin/βOci can produce large amounts of zingiberene, and Tc-βFar/Ger can produce large amounts of β-farnesene. Thus, large amounts of these sesquiterpenes production system by using microorganism were established in this study. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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