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Biosynthesis of Fusaruside Based on Recombinant Pichia pastoris |
Yuan TIAN(),Yan-ling LI |
College of Life Science, Shandong First Medical University & Shandong Academy of Medical Sciences, Taian 271016, China |
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Abstract Objective: A strain of Pichia pastoris producing fusaruside was constructed to solve the source problem of fusaruside, a selective immunosuppressive molecule.Method: Two related biosynthetic genes coding delta 3(E)-sphingolipid desaturase [Δ3(E)-SD] and delta 10(E)-sphingolipid desaturase [Δ10(E)-SD] were amplified from fungus Fusarium graminearum PH-1. 2A peptide based strategy was used to construct the co-expression vector. Then the 2A polyprotein construct was transformed into Pichia pastoris GS115 for induction. Finally, P. pastoris cells were extracted with methanol and dichloromethane, and the extract was detected by high performance liquid chromatography mass spectrometer (HPLC-MS).Result: The two desaturases were successfully co-expressed in P. pastoris. SDS-PAGE showed that the molecular weight of Δ3(E)-SD and Δ10(E)-SD were about 48kDa and 65kDa, respectively. HPLC-MS indicated that fusaruside could be produced by the recombinant yeast.Conclusion: Compared with fusarium which producing fusaruside, the engineered yeast had shorter fermentation time and higher yield, laying a foundation for further development and application of fusaruside.
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Received: 20 December 2018
Published: 05 August 2019
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Corresponding Authors:
Yuan TIAN
E-mail: tianyuan2005hit@163.com
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