The ZbYME2 gene from the food spoilage yeast Zygosaccharomyces bailii confers not only YME2 functions in Saccharomyces cerevisiae, but also the capacity for catabolism of sorbate and benzoate, two major weak organic acid preservatives
The ZbYME2 gene from the food spoilage yeast Zygosaccharomyces bailii confers not only YME2 functions in Saccharomyces cerevisiae, but also the capacity for catabolism of sorbate and benzoate, two major weak organic acid preservatives
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DOI：
10.1046/j.1365-2958.2001.02686.x
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发表时间：
2001-11-01
影响因子：
3.6
通讯作者：
Piper, PW
中科院分区：
文献类型：
--
作者：
Mollapour, M;Piper, PW
影响食品腐败微生物对山梨酸盐和苯甲酸盐的抗性的一个因素是这些微生物是否能够催化这些防腐剂化合物的降解。几种真菌通过β-酮己二酸途径代谢苯甲酸，包括苯甲酸酯羟基化为4-羟基苯甲酸酯。酿酒酵母不能使用苯甲酸盐作为唯一的碳源，这显然是由于缺乏苯甲酸-4-羟化酶活性。然而，来自食品腐败酵母Zygosaccharomyces bailii的单个基因在S.酿酒酵母细胞，可以使后者在苯甲酸盐、山梨酸盐和苯丙氨酸上生长。虽然ZbYME 2基因是Z. bailii，其ZbYme 2 p产物与迄今为止研究的其他真菌苯甲酸-4-羟化酶几乎没有同源性，所有这些酶似乎都是微粒体细胞色素P450。相反，ZbYme 2 p与S.酿酒酵母线粒体蛋白Yme 2 p/Rna 12 p/Prp 12 p，并且当在S.在苯甲酸盐上生长的酿酒酵母主要定位于线粒体。与S.酿酒酵母，最明显的是在yme 1，yme 2细胞，可能涉及增加的内源性氧化应激的有害影响。ZbYME 2的异源表达补充了这些表型，但它也赋予了天然S.酿酒酵母Yme 2 p无法提供。S.利用苯甲酸盐。表达ZbYME 2的酿酒酵母需要功能性线粒体呼吸链，但不需要线粒体的天然Yme 1 p和Yme 2 p。
A factor influencing resistances of food spoilage microbes to sorbate and benzoate is whether these organisms are able to catalyse the degradation of these preservative compounds. Several fungi metabolize benzoic acid by the beta -ketoadipate pathway, involving the hydroxylation of benzoate to 4-hydroxybenzoate. Saccharomyces cerevisiae is unable to use benzoate as a sole carbon source, apparently through the lack of benzoate-4-hydroxylase activity. However a single gene from the food spoilage yeast Zygosaccharomyces bailii, heterologously expressed in S. cerevisiae cells, can enable growth of the latter on benzoate, sorbate and phenylalanine. Although this ZbYME2 gene is essential for benzoate utilization by Z. bailii, its ZbYme2p product has little homology to other fungal benzoate-4-hydroxylases studied to date, all of which appear to be microsomal cytochrome P450s. Instead, ZbYme2p has strong similarity to the matrix domain of the S. cerevisiae mitochondrial protein Yme2p/Rna12p/Prp12p and, when expressed as a functional fusion to green fluorescent protein in S. cerevisiae growing on benzoate, is largely localized to mitochondria. The phenotypes associated with loss of the native Yme2p from S. cerevisiae, mostly apparent in yme1,yme2 cells, may relate to increased detrimental effects of endogenous oxidative stress. Heterologous expression of ZbYME2 complements these phenotypes, yet it also confers a potential for weak acid preservative catabolism that the native S. cerevisiae Yme2p is unable to provide. Benzoate utilization by S. cerevisiae expressing ZbYME2 requires a functional mitochondrial respiratory chain, but not the native Yme1p and Yme2p of the mitochondrion.