牛樟芝聚酮合成酶基因的功能界定及其相關生合成路徑探討

余浡維

doi:10.6342/NTU201701259

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牛樟芝聚酮合成酶基因的功能界定及其相关生合成路径探讨

Functional Characterization of Polyketide Synthase-encoding Genes and the Related Biosynthetic Pathway in Antrodia cinnamomea

余浡维 , 博士　　导师：曾显雄;刘瑞芬;李宗徽　　

英文 DOI： 10.6342/NTU201701259

牛樟芝；酮合成酶；生合成；苔藓酸；安卓奎诺尔； Antrodia cinnamomea ； polyketide synthase ； biosynthesis ； orsellinic acid ； antroquinonol

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摘要 │ 参考文献 (212) │ 国际计量

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Antrodia cinnamomea is a unique resupinate basidiomycete endemic to Taiwan.  Besides the abundant triterpenoid metabolites, A. cinnamomea is known for producing antroquinonols, which were reported to have notable medicinal potential in oncology and immunology.  However, neither the biosynthetic pathway of these compounds nor the corresponding genes are currently clear.  To investigate the biosynthesis of antroquinonols in A. cinnamomea, we focused on the polyketide synthase (PKS) genes due to the similar structure of the cyclohexenone moiety of antroquinonols to the aromatic polyketide.  Four putative PKS genes, including three reducing PKSs and one non-reducing PKS, pks63787, were characterized in A. cinnamomea based on the partially deciphered genome and the constructed fosmid library.  For the first time, a gene disruption platform was established in A. cinnamomea via a protoplast-mediated transformation system.  Our study showed that the pks63787 knock-out mutant of A. cinnamomea (∆pks63787) is deficient in the biosynthesis of several aromatic metabolites which are involved in the antioxidant activity and colony morphology.  In the further study, we pointed out by phylogenetic analysis that pks63787 likely encodes an orsellinic acid synthase, whose function was double-confirmed with a complementary feeding test.  The amendment of orsellinic acid not only restores the ability of ∆pks63787 in producing its deficient pigment, benzenoids and antroquinonols, but also enhances the productivity of several antroquinonols.  These results provide direct evidence that the PKS63787 is involved in the biosynthesis of antroquinonols, and supported our hypothesis that the cyclohexenone moiety is a polyketide synthesized via the PKS63787-conducted polyketide pathway.  Along with the identification of numerous PKS63787- and orsellinic acid-mediated components, six compounds, including two benzenoids 1, 2, two antroquinonols 3, 4 and two antrocinnanoates 5, 6, were reported for the first time.  In conclusion, our study has contributed to the understanding of the PKS genes and the biosynthesis of antroquinonols in A. cinnamomea, and the adopted procedure may be conducive to genetics research focusing on natural products in A. cinnamomea and other basidiomycetes.

参考文献 ( 212 ) 〈TOP〉

Abdel-Hameed M, Bertrand RL, Piercey-Normore MD, Sorensen JL, 2016. Putative identification of the usnic acid biosynthetic gene cluster by de novo whole-genome sequencing of a lichen-forming fungus. Fungal Biology 120, 306–316.
连结：
Abe I, Morita H, 2010. Structure and function of the chalcone synthase superfamily of plant type III polyketide synthases. Natural Product Reports 27, 809–838.
连结：
Abe Y, Suzuki T, Ono C, Iwamoto K, Hosobuchi M, Yoshikawa H, 2002. Molecular cloning and characterization of an ML-236B (compactin) biosynthetic gene cluster in Penicillium citrinum. Molecular Genetics and Genomics 267, 636–646.
连结：
Adams M, Christen M, Plitzko I et al., 2010. Antiplasmodial lanostanes from the Ganoderma lucidum mushroom. Journal of Natural Products 73, 897–900.
连结：
Ahuja M, Chiang Y-M, Chang S-L et al., 2012. Illuminating the diversity of aromatic polyketide synthases in Aspergillus nidulans. Journal of the American Chemical Society 134, 8212–8221.
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引用含有DOI的文献

DOI可强化引用精确性、增强学术圈连结，并给予使用者跨平台的良好使用经验，目前在全世界已有超过五千万个对象申请DOI。 如想对DOI的使用与概念有进一步了解，请参考 （ ） 。

DOI可强化引用精确性、增强学术圈连结，并给予使用者跨平台的良好使用经验，目前在全世界已有超过五千万个对象申请DOI。如想对DOI的使用与概念有进一步了解，请参考（）。