副多态Ogataea pho91-Δ突变体不能产生活性甲醇氧化酶可以通过PHO87基因失活来补偿。

IF 2.4 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Folia microbiologica Pub Date : 2024-12-27 DOI:10.1007/s12223-024-01236-2
Vasilina Farofonova, Azamat Karginov, Anton Zvonarev, Ekaterina Kulakovskaya, Michael Agaphonov, Tatiana Kulakovskaya
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引用次数: 0

摘要

甲基营养酵母(Ogataea parapolymorpha)细胞中有两个编码低亲和性磷酸转运蛋白的基因:PHO87(编码质膜转运蛋白)和PHO91(编码一种与酿酒酵母液泡膜转运蛋白同源的蛋白)。此前,我们报道了O. parapolymorpha PHO91的失活,由于MOX基因编码的甲醇氧化酶缺乏活性,干扰了甲醇的利用。在这项工作中,我们发现通过失活PHO87基因或将MOX基因的额外拷贝引入细胞中,这种缺陷被完全抑制。PHO91基因敲除仅在甲醇培养的细胞中降低了长链多磷酸盐的水平,而在葡萄糖培养的细胞中没有。这种效应甚至在具有额外的MOX拷贝的菌株中仍然存在,这挽救了突变体在甲醇上生长的能力。相比之下,PHO87基因敲除改变了甲醇和葡萄糖培养细胞中短链和长链多磷酸盐的水平。PHO91失活后对钒酸盐的抗性没有改变,而PHO87失活后对钒酸盐的抗性增加。我们的数据表明,在O. parapolymorpha中,Pho87和Pho91转运体在无机多磷酸盐代谢和对甲醇消耗的适应中具有不同的作用。
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Inability of Ogataea parapolymorpha pho91-Δ mutant to produce active methanol oxidase can be compensated by inactivation of the PHO87 gene.

Cells of the methylotrophic yeast Ogataea parapolymorpha have two genes encoding low-affinity phosphate transporters: PHO87, encoding the plasma membrane transporter, and PHO91, encoding a protein, which is homologous to the Saccharomyces cerevisiae vacuolar membrane transporter. Earlier, we reported that inactivation of PHO91 in O. parapolymorpha interferes with methanol utilization due to the lack of activity of methanol oxidase encoded by the MOX gene. In this work, we showed that this defect was completely suppressed by inactivating the PHO87 gene or introducing additional copies of the MOX gene into the cell. The PHO91 gene knockout decreased the level of long-chained polyphosphates only in methanol-grown cells, but not in glucose-grown cells. This effect remained even in the strain with extra copies of MOX, which rescues the ability of the mutant to grow on methanol. In contrast, the PHO87 gene knockout changed the levels of short-chained and long-chained polyphosphates in both methanol- and glucose-grown cells. Inactivation of PHO91 did not change vanadate resistance, while inactivation of PHO87 increased this resistance. Our data suggest that in O. parapolymorpha, Pho87 and Pho91 transporters have different roles in inorganic polyphosphate metabolism and adaptation to methanol consumption.

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来源期刊
Folia microbiologica
Folia microbiologica 工程技术-生物工程与应用微生物
CiteScore
5.80
自引率
0.00%
发文量
82
审稿时长
6-12 weeks
期刊介绍: Unlike journals which specialize ever more narrowly, Folia Microbiologica (FM) takes an open approach that spans general, soil, medical and industrial microbiology, plus some branches of immunology. This English-language journal publishes original papers, reviews and mini-reviews, short communications and book reviews. The coverage includes cutting-edge methods and promising new topics, as well as studies using established methods that exhibit promise in practical applications such as medicine, animal husbandry and more. The coverage of FM is expanding beyond Central and Eastern Europe, with a growing proportion of its contents contributed by international authors.
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