高产橙花叔醇的酵母细胞工厂创建

Construction of cell factories for high production of nerolidol in Saccharomyces cerevisiae

橙花叔醇为萜类精油成分,具有抗菌、抗肿瘤和抗氧化等多种生物学活性。为了实现在酿酒酵母中异源生产橙花叔醇,该研究首先将猕猴桃Actinidia chinensis来源的橙花叔醇合酶(NES)基因通过密码子优化、人工合成后转入底盘菌株FPP-001中获得工程菌株NES-001,其橙花叔醇产量达到2.71 mg·L^(-1)。在进一步工作中,橙花叔醇合酶的N端通过连接肽GGGS融合法尼烯合酶(FPS)后得到橙花叔醇产量显著提高的工程菌NES-002,其产量是NES-001的59.80倍,达到162.07 mg·L^(-1)。最终,通过恢复NES-002中色氨酸生物合成缺陷基因TRP1得到工程菌NES-003,该菌在高密度发酵体系中橙花叔醇产量能达到1 711.53 mg·L^(-1)。该研究为微生物发酵法生产橙花叔醇等倍半萜化合物提供了基础。

Nerolidol is an important constituent of terpenoid essential oil and has excellent anti-tumor, anti-bacterial, and anti-oxi- dative properties. For realizing heterogenous production of nerolidol, our research firstly integrated the codon-optimized Actiniclia sinen- sis nerolidol synthase gene （NES） into the terpenoid chassis strain FPP-001, and obtained NES-001 that could produce 2. 71 mg· L-1 nerolidol. Then, the N-terminal of the NES was fused with FPS by linker peptide GGGS. With this strategy, nerolidol production im- proved by 59.80-fold, reaching 162.07 mg· L-1. Finally, by introduction of auxotrophic marker TRP1 in NES-002, the resulting strain NES-003 could produce 1 711.53 mg· L-1 by high cell density fermentation method. This study provides the basis for the fer- mentative production of nerolidol and other sesquiterpenoids.