Cloning, characterization and expression of flavonoid biosynthetic genes from Antrodia cinnamomea

• Main Authors: Sheng-Syong Shih, 施勝雄
• Other Authors: Shean-Shong Tzean
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/10573960545372079910

碩士 === 國立臺灣大學 === 植物病理與微生物學研究所 === 98 === Antrodia cinnamomea （Neu-Chang-Ts） an unique endemic resupinate basidiomycete of Taiwan, inhabited on the heartwood cavity of Cinnamomum kanehirae, reputed for alleviating, hangover, inflammation, the hepatitis syndrome and oxidative stress, etc. Attempt to clarify the antioxidation activity of A. cinnamomea, we cloned five flavonoid biosynthesis genes: flavonol synthase （FLS）, flavonoid 3’ and 5’ hydroxylase （F3’5’H）, chalcone isomerase（CHI）, flavonone 3-hydorxylase （F3H） and 4-coumaryl CoA ligase （4CL） via the constructed cDNA and fosmid libraries and PCR approaches. Furthermore, we also worked out the full length of gDNA and cDNA of these genes and their properties characterized. Currently, these genes have been constructed on pKLAC1 shuttle vectors and transformed them into Kluyveromyces lactis, host were undergoing. The expressed secondary products will be assessed and assayed. In other aspects, to clarify wether the existence of flavonoid in A. cinnamomea, the methanolic extracts from mycelium were subjected to high performance liquid chromatography（HPLC）, nuclear magnetic resonance（NMR）analysis. The outcomes indicated carbohydrates, instead of flavonoid-like substance, dominated the most partially purified fraction. Given flavonoid processes major anti-inflammation and antioxidation capacity, perspectively, overexpress the genetically engineered flavonoid biosynthetic genes in pertinent hosts to produce adequate quantify of desired compounds for clinic trals will be the resolution.