Biosynthesis of fusaruside based on recombinant Pichia pastoris

Subjects: Biology >> Bioengineering submitted time 2019-03-05 Cooperative journals: 《中国生物工程杂志》

田园 李艳玲

Abstract： Objective: To construct a fusaruside-producing strain of Pichia pastoris, and to solve the source of the natural small molecule immunosuppressant fusaruside. METHODS: The genes related to the synthesis of fusaruside, the 3-position desaturase (Δ3(E)-SD) and 10-position desaturase (Δ10(E)-SD) genes, were amplified from Fusarium graminearum PH-1. ; And the co-expression vector of the two genes was constructed by the 2A peptide strategy, and transformed into Pichia GS115 for inducible expression of the two enzymes; after the induced Pichia cells were treated with methanol and dichloromethane, the efficient Liquid chromatography-mass spectrometry (HPLC-MS) was used to detect the product changes. Results: The 3-desaturase and 10-desaturase were successfully co-expressed in Pichia pastoris. SDS-PAGE showed that the molecular weight of the 3-desaturase was about 48 kDa, and the molecular weight of the 10-desaturase was about 65 kDa; HPLC- MS showed that recombinant yeast could produce fusaruside. Conclusion: Compared with the fusaruside native strain Fusarium, the yeast has a shorter fermentation time and higher yield, which lays a foundation for the further development and application of fusaruside.

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