Feedback inhibition of the prephenate dehydratase from Saccharomyces cerevisiae and its mutation in huangjiu (Chinese rice wine) yeast
Feedback inhibition of the prephenate dehydratase from Saccharomyces cerevisiae and its mutation in huangjiu (Chinese rice wine) yeast
复制标题
酿酒酵母预苯酸脱水酶的反馈抑制及其在黄酒酵母中的突变
DOI：
10.1016/j.lwt.2020.110040
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发表时间：
2020-11-01
影响因子：
6
通讯作者：
Mao, Jian
中科院分区：
文献类型：
--
作者：
Liu, Shuangping;Yang, Qilin;Mao, Jian
2-苯乙醇是一种常见于发酵食品中的芳香化合物，是黄酒（中国米酒）中含量最多的芳香化合物之一。在黄酒中发现的2-苯乙醇是由酿酒酵母HJ01产生的，它比酿酒酵母W303型菌株产生的2-苯乙醇多得多。除了预苯酸脱水酶的行为（PDT; EC 4.2.1.51）外，酿酒酵母对2-苯乙醇生物合成的反馈调节已经被仔细研究过。在S. cerevisiae HJ01中发现了PDT突变（I161K、L239P和Q250H），基于结构分析表明存在保守的L-Phe结合位点。结果表明，l -苯丙氨酸对酿酒葡萄球菌HJ01和W303的PDT有抑制作用。PDTHJ01的比活性是PDTW303的1.27倍。催化效率（kcat/Km, 58.9 ~ 51.7 L/(mmol)）。s))和热稳定性，但对反馈抑制的敏感性较低（K-i, 3.97 ~ 3.35 mM）。因此，酿酒酵母HJ01的PDT突变导致黄酒中发现的2-苯乙醇浓度较高。
2-Phenylethanol, an aromatic compound often found in fermented foods, is among the most abundant aromatic compounds in huangjiu (Chinese rice wine). The 2-phenylethanol found in huangjiu is produced by Saccharomyces cerevisiae HJ01, which produces much more 2-phenylethanol than the type strain, S. cerevisiae W303. Feedback regulation of 2-phenylethanol biosynthesis in S. cerevisiae has been carefully studied, except for the behavior of prephenate dehydratase (PDT; EC 4.2.1.51). Mutation (I161K, L239P and Q250H) in PDT were found in S. cerevisiae HJ01, and a structure-based analysis suggested the presence of conserved L-Phe binding sites. The purified PDT from S. cerevisiae strains HJ01 and W303 were found to be inhibited by L-Phe. The specific activity of the PDTHJ01 was 1.27-fold greater than that of the PDTW303. It also showed greater substrate affinity (Km, 11.1-15.9 mM), catalytic efficiency (kcat/Km, 58.9 to 51.7 L/(mmol.s)), and thermal stability, but was less susceptible to feedback inhibition (K-i, 3.97 to 3.35 mM). Thus, mutations in the PDT from S. cerevisiae HJ01 contribute to the higher 2-phenylethanol concentration found in huangjiu.