Department of Biotechnology, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
Collaborative Research Institute for Innovative Microbiology, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan.
FEMS Yeast Res. 2024 Jan 9;24. doi: 10.1093/femsyr/foae030.
The yeast Yarrowia lipolytica can assimilate n-alkane as a carbon and energy source. To elucidate the significance of phosphatidylserine (PS) in the utilization of n-alkane in Y. lipolytica, we investigated the role of the Y. lipolytica ortholog (PSS1) of Saccharomyces cerevisiae PSS1/CHO1, which encodes a PS synthase. The PSS1 deletion mutant (pss1Δ) of Y. lipolytica could not grow on minimal medium in the absence of ethanolamine and choline but grew when either ethanolamine or choline was supplied to synthesize phosphatidylethanolamine and phosphatidylcholine. The pss1Δ strain exhibited severe growth defects on media containing n-alkanes even in the presence of ethanolamine and choline. In the pss1Δ strain, the transcription of ALK1, which encodes a primary cytochrome P450 that catalyses the hydroxylation of n-alkanes in the endoplasmic reticulum, was upregulated by n-alkane as in the wild-type strain. However, the production of functional P450 was not detected, as indicated by the absence of reduced CO-difference spectra in the pss1Δ strain. PS was undetectable in the lipid extracts of the pss1Δ strain. These results underscore the critical role of PSS1 in the biosynthesis of PS, which is essential for the production of functional P450 enzymes involved in n-alkane hydroxylation in Y. lipolytica.
酵母解脂耶氏酵母可以将正烷烃作为碳源和能源同化。为了阐明磷脂酰丝氨酸(PS)在解脂耶氏酵母利用正烷烃中的意义,我们研究了酿酒酵母 PSS1/CHO1 的解脂耶氏酵母同源物(PSS1)的作用,该基因编码 PS 合酶。解脂耶氏酵母的 PSS1 缺失突变体(pss1Δ)在没有乙醇胺和胆碱的基础培养基上不能生长,但当提供乙醇胺或胆碱以合成磷脂酰乙醇胺和磷脂酰胆碱时可以生长。即使在提供乙醇胺和胆碱的情况下,pss1Δ 菌株在含有正烷烃的培养基上的生长缺陷也很严重。在 pss1Δ 菌株中,ALK1 的转录被上调,ALK1 编码一种内质网中催化正烷烃羟化的主要细胞色素 P450。然而,与野生型菌株一样,没有检测到功能性 P450 的产生,这表明 pss1Δ 菌株中没有还原 CO-差光谱。在 pss1Δ 菌株的脂质提取物中未检测到 PS。这些结果强调了 PSS1 在 PS 生物合成中的关键作用,这对于解脂耶氏酵母中参与正烷烃羟化的功能性 P450 酶的产生是必不可少的。