Novel fungal phenylpyruvate reductase belongs to d-isomer-specific 2-hydroxyacid dehydrogenase family
Novel fungal phenylpyruvate reductase belongs to d-isomer-specific 2-hydroxyacid dehydrogenase family
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DOI：
10.1016/j.bbapap.2011.05.024
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发表时间：
2011-12-01
影响因子：
3.2
通讯作者：
Takaya, Naoki
中科院分区：
文献类型：
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作者：
Fujii, Taiki;Shimizu, Motoyuki;Takaya, Naoki
我们发现了一株产苯乳酸的真菌菌株荧光威克汉姆菌TK1，并从真菌无细胞提取物中纯化了苯丙酮酸还原酶(PPR)。PPR同时使用NADPH和NADH作为辅助因子，更倾向于前者。酶反应和真菌培养产生光学活性的D-聚乳酸。克隆了PPR(PPRA)基因，并在大肠杆菌中进行了表达。天然和重组PPR的纯化制剂均以羟基苯丙酮酸、乙醛和羟基丙酮酸为底物，而不是丙酮酸、草酰乙酸酯或苯甲酰甲酸酯。预测的PPR蛋白与D-异构体特异性2-羟基酸脱氢酶家族中的蛋白具有序列相似性。系统发育分析表明，预测的PPR蛋白与真菌预测的蛋白一起构成了一组新的乙氧基酸/羟基丙酮酸还原酶。该真菌有效地将苯丙氨酸和苯丙酮酸转化为D-聚乳酸。这些化合物上调了PPRA的转录，表明它在真菌的苯丙氨酸代谢中发挥了作用。(C)2011爱思唯尔B.V.保留所有权利。
We discovered the phenyllactate (PLA)-producing fungal strain Wickerhamia fluorescens TK1 and purified phenylpyruvate reductase (PPR) from fungal cell-free extracts. The PPR used both NADPH and NADH as cofactors with more preference for the former. The enzyme reaction as well as the fungal culture produced optically active D-PLA. The gene for the PPR (pprA) was cloned and expressed in Escherichia coli cells. Purified preparations of both native and recombinant PPR used hydroxyphenylpyruvate, glyoxylate and hydroxypyruvate as substrates but not pyruvate, oxaloacetate or benzoylformate. The predicted PPR protein had sequence similarity to proteins in the D-isomer-specific 2-hydroxyacid dehydrogenase family. Phylogenetic analyses indicated that the predicted PPR protein together with fungal predicted proteins constitutes a novel group of glyoxylate/hydroxypyruvate reductases. The fungus efficiently converted phenylalanine and phenylpyruvate to D-PLA. These compounds up-regulated the transcription of pprA, suggesting that it plays a role in fungal phenylalanine metabolism. (C) 2011 Elsevier B.V. All rights reserved.