毕赤酵母中外源表达脂肪酰-CoA还原酶生产脂肪醇

Production of fatty alcohol in Komagataella pastoris by heterologous expression of fatty acyl-CoA reductase

摘要:

【目的】通过在毕赤酵母Komagataella pastoris GS115中外源表达来源于霍霍巴[Simmondsia chinensis (Jojoba)]的脂肪酰-CoA还原酶Jojoba FAR，利用微生物发酵生产脂肪醇。【方法】以质粒pRL105为模板PCR扩增获得霍霍巴脂肪酰-CoA还原酶的编码基因，以pGAPZαA为载体构建重组表达质粒pGAP-far，并通过电转化法转入K. pastoris GS115，筛选转化子并发酵，气相色谱-质谱联用检测发酵产物。【结果】构建了毕赤酵母重组菌株pGAPZ-far-GS115，通过摇瓶发酵检测到脂肪醇的合成。随后在7 L规模的发酵罐上发酵验证，得到脂肪醇产量为134.74 mg/L，产率为1.22 mg/(L·h)。【结论】实现了脂肪醇在毕赤酵母中的生物合成，为工业上利用毕赤酵母生产脂肪醇奠定了一定基础。

关键词:脂肪酰-CoA还原酶，毕赤酵母，生物发酵，脂肪醇

Abstract:

[Objective] We selected Simmondsia chinensis (Jojoba) fatty acyl-CoA reductase Jojoba FAR for heterologous expression in Komagataella pastoris strain GS115 to produce fatty alcohol by fermentation. [Methods] Gene encoding this fatty acyl-CoA reductase was amplified from plasmid pRL105 and cloned into the plasmid pGAPZαA; the recombinant expression plasmid was transformed into K. pastoris GS115 by electrotransformation. We detected fatty alcohol from positive transformant by GC-MS. [Results] We constructed a recombinant K. pastoris GS115 strain pGAPZ-far-GS115 that produced fatty alcohols in flask fermentation. Afterwards, a fed-batch fermentation was used for further verification. And the engineered strain produced 134.74 mg/L fatty alcohols with a productivity of 1.22 mg/(L·h). [Conclusion] Our findings prove the possibility to produce fatty alcohol in K. pastoris GS115, provide the potential for industrial production of fatty alcohol by K. pastoris.

Key words:Fatty acyl-CoA reductase, Komagataella pastoris, Fermentation, Fatty alcohol