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The Fps1p aquaglyceroporin facilitates the use of small aliphatic amides as a nitrogen source by amidase-expressing yeasts

机译:Fps1p甘油水通道蛋白可通过表达酰胺酶的酵母促进将小的脂肪族酰胺用作氮源

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摘要

Saccharomyces cerevisiae acquires a resistance to high, toxic levels of acetic acid by destabilizing Fps1p, the plasma membrane aquaglyceroporin through which this acid - in its undissociated state - enters the cell. In this study, Fps1p loss was shown to confer resistances to acetic acid, acrolein and allyl alcohol, not just in S. cerevisiae but also in the osmotolerant spoilage yeast Zygosaccharomyces rouxii. However, in Z. rouxii, the loss of Fps1p severely compromised the use of acetamide and several other small amides as sources of nitrogen, an indication that these amides enter the cells of this yeast by passive diffusion through the Fps1p pore. Saccharomyces cerevisiae cannot grow on acetamide, but was conferred with an ability to use this and other small amides as nitrogen sources by heterologous expression of a Z. rouxii ORF (ZrAMD1) with protein sequence identity to the amdS-encoded amidase of Aspergillus nidulans. This capacity of ZrAMD1-expressing S. cerevisiae to assimilate amide nitrogen was severely compromised by the loss of Fps1p. ZrAMD1 appears to encode the major amidase of Z. rouxii as a Zramd1 deletant mutant had, like the Zrfps1 deletant, lost the ability to assimilate small amides as sources of nitrogen.
机译:酿酒酵母通过破坏Fps1p(质膜水甘油糖蛋白)的稳定性而获得对高毒性水平乙酸的抗性,该酸以未解离状态进入细胞。在这项研究中,Fps1p的丢失显示出不仅对酿酒酵母而且对耐渗透性变质酵母鲁氏酵母(Zygosaccharomyces rouxii)具有乙酸,丙烯醛和烯丙醇的抗性。然而,在鲁氏沼虾中,Fps1p的丧失严重损害了乙酰胺和其他几种小酰胺作为氮源的使用,这表明这些酰胺通过Fps1p孔的被动扩散进入酵母细胞。酿酒酵母不能在乙酰胺上生长,但是通过与蛋白质构象与构巢曲霉amdS编码的酰胺酶的鲁氏梭菌ORF(ZrAMD1)异源表达,被赋予了使用该酰胺和其他小型酰胺作为氮源的能力。 Fps1p的丧失严重影响了表达ZrAMD1的酿酒酵母同化酰胺氮的能力。 ZrAMD1似乎编码鲁氏沼虾的主要酰胺酶,因为Zramd1缺失突变体与Zrfps1缺失突变体一样,失去了吸收小分子酰胺作为氮源的能力。

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