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首页> 外文期刊>Applied Microbiology >Membrane-Associated Glucose-Methanol-Choline Oxidoreductase Family Enzymes PhcC and PhcD Are Essential for Enantioselective Catabolism of Dehydrodiconiferyl Alcohol
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Membrane-Associated Glucose-Methanol-Choline Oxidoreductase Family Enzymes PhcC and PhcD Are Essential for Enantioselective Catabolism of Dehydrodiconiferyl Alcohol

机译：膜相关的葡萄糖-甲醇-胆碱氧化还原酶家族酶PhcC和PhcD对脱氢二十二烷醇的对映选择性代谢是必不可少的

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摘要

Sphingobium sp. strain SYK-6 is able to degrade various lignin-derived biaryls, including a phenylcoumaran-type compound, dehydrodiconiferyl alcohol (DCA). In SYK-6 cells, the alcohol group of the B-ring side chain of DCA is initially oxidized to the carboxyl group to generate 3-(2-(4-hydroxy-3-methoxyphenyl)-3-(hydroxymethyl)-7-methoxy-2,3-dihydrobenzofuran-5-yl) acrylic acid (DCA-C). Next, the alcohol group of the A-ring side chain of DCA-C is oxidized to the carboxyl group, and then the resulting metabolite is catabolized through vanillin and 5-formylferulate. In this study, the genes involved in the conversion of DCA-C were identified and characterized. The DCA-C oxidation activities in SYK-6 were enhanced in the presence of flavin adenine dinucleotide and an artificial electron acceptor and were induced ca. 1.6-fold when the cells were grown with DCA. Based on these observations, SLG_09480 ( phcC ) and SLG_09500 ( phcD ), encoding glucose-methanol-choline oxidoreductase family proteins, were presumed to encode DCA-C oxidases. Analyses of phcC and phcD mutants indicated that PhcC and PhcD are essential for the conversion of (+)-DCA-C and (?)-DCA-C, respectively. When phcC and phcD were expressed in SYK-6 and Escherichia coli , the gene products were mainly observed in their membrane fractions. The membrane fractions of E. coli that expressed phcC and phcD catalyzed the specific conversion of DCA-C into the corresponding carboxyl derivatives. In the oxidation of DCA-C, PhcC and PhcD effectively utilized ubiquinone derivatives as electron acceptors. Furthermore, the transcription of a putative cytochrome c gene was significantly induced in SYK-6 grown with DCA. The DCA-C oxidation catalyzed by membrane-associated PhcC and PhcD appears to be coupled to the respiratory chain.
机译：鞘氨醇单胞菌菌株SYK-6能够降解各种木质素衍生的联芳基，包括苯基香豆素类化合物脱氢二癸二烯醇（DCA）。在SYK-6细胞中，DCA的B环侧链的醇基最初被氧化成羧基，生成3-（2-（4-羟基-3-甲氧基苯基）-3-（羟甲基）-7-甲氧基-2，3-二氢苯并呋喃-5-基）丙烯酸（DCA-C）。接下来，将DCA-C的A环侧链的醇基氧化为羧基，然后通过香兰素和5-甲酰基阿魏酸酯将所得的代谢产物催化代谢。在这项研究中，鉴定并鉴定了参与DCA-C转化的基因。在黄素腺嘌呤二核苷酸和一个人造电子受体的存在下，SYK-6中DCA-C的氧化活性增强，并在大约10分钟内被诱导。当细胞与DCA一起生长时，其毒性是其1.6倍。基于这些观察，推测编码葡萄糖-甲醇-胆碱氧化还原酶家族蛋白的SLG_09480（phcC）和SLG_09500（phcD）编码DCA-C氧化酶。对phcC和phcD突变体的分析表明，PhcC和PhcD分别对于（+）-DCA-C和（α）-DCA-C的转化是必不可少的。当在SYK-6和大肠杆菌中表达phcC和phcD时，主要在其膜级分中观察到基因产物。表达phcC和phcD的大肠杆菌膜级分催化DCA-C特定转化为相应的羧基衍生物。在DCA-C的氧化中，PhcC和PhcD有效地利用了泛醌衍生物作为电子受体。此外，推定的细胞色素c基因的转录在DCA生长的SYK-6中被显着诱导。膜相关的PhcC和PhcD催化的DCA-C氧化似乎与呼吸链相连。

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