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紫外线照射后脂质生产率大幅提高的斯达氏油脂酵母突变体的分离与鉴定

Isolation and characterization of Lipomyces starkeyi mutants with greatly increased lipid productivity following UV irradiation.

作者信息

Takaku Hiroaki, Ebina Sayaka, Kasuga Kotoha, Sato Rikako, Ara Satoshi, Kazama Haruka, Matsuzawa Tomohiko, Yaoi Katsuro, Araki Hideo, Shida Yosuke, Ogasawara Wataru, Ishiya Koji, Aburatani Sachiyo, Yamazaki Harutake

机构信息

Department of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, 265-1 Higashijima, Akiha-ku, Niigata 956-8603, Japan.

Department of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, 265-1 Higashijima, Akiha-ku, Niigata 956-8603, Japan.

出版信息

J Biosci Bioeng. 2021 Jun;131(6):613-621. doi: 10.1016/j.jbiosc.2021.01.006. Epub 2021 Feb 10.

Abstract

The oleaginous yeast Lipomyces starkeyi is an intriguing lipid producer that can produce triacylglycerol (TAG), a feedstock for biodiesel production. We previously reported that the L. starkeyi mutant E15 with high levels of TAG production compared with the wild-type was efficiently obtained using Percoll density gradient centrifugation. However, considering its use for biodiesel production, it is necessary to further improve the lipid productivity of the mutant. In this study, we aimed to obtain mutants with better lipid productivity than E15, evaluate its lipid productivity, and analyze lipid synthesis-related gene expression in the wild-type and mutant strains. The mutants E15-11, E15-15, and E15-25 exhibiting higher lipid productivity than E15 were efficiently isolated from cells exposed to ultraviolet light using Percoll density gradient centrifugation. They exhibited approximately 4.5-fold higher lipid productivity than the wild-type on day 3. The obtained mutants did not exhibit significantly different fatty acid profiles than the wild-type and E15 mutant strains. E15-11, E15-15, and E15-25 exhibited higher expression of acyl-CoA synthesis- and Kennedy pathway-related genes than the wild-type and E15 mutant strains. Activation of the pentose phosphate pathway, which supplies NADPH, was also observed. These results suggested that the increased expression of acyl-CoA synthesis- and Kennedy pathway-related genes plays a vital role in lipid productivity in the oleaginous yeast L. starkeyi.

摘要

产油酵母斯达氏油脂酵母是一种有趣的脂质生产者,它能够产生三酰甘油(TAG),这是生物柴油生产的一种原料。我们之前报道过,与野生型相比,利用Percoll密度梯度离心法能高效获得具有高TAG产量的斯达氏油脂酵母突变体E15。然而,考虑到其在生物柴油生产中的应用,有必要进一步提高该突变体的脂质生产率。在本研究中,我们旨在获得脂质生产率比E15更高的突变体,评估其脂质生产率,并分析野生型和突变体菌株中脂质合成相关基因的表达情况。利用Percoll密度梯度离心法从经紫外线照射的细胞中高效分离出脂质生产率高于E15的突变体E15-11、E15-15和E15-25。在第3天,它们的脂质生产率比野生型高约4.5倍。所获得的突变体与野生型和E15突变体菌株相比,脂肪酸谱没有显著差异。E15-11、E15-15和E15-25与野生型和E15突变体菌株相比,酰基辅酶A合成和肯尼迪途径相关基因的表达更高。还观察到供应NADPH的磷酸戊糖途径的激活。这些结果表明,酰基辅酶A合成和肯尼迪途径相关基因表达的增加在产油酵母斯达氏油脂酵母的脂质生产率中起着至关重要的作用。

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