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首页> 外文期刊>Applied Microbiology and Biotechnology >Identification and characterization of a novel (-)-vibo-quercitol 1-dehydrogenase from Burkholderia terrae suitable for production of (-)-vibo-quercitol from 2-deoxy-scyllo-inosose
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Identification and characterization of a novel (-)-vibo-quercitol 1-dehydrogenase from Burkholderia terrae suitable for production of (-)-vibo-quercitol from 2-deoxy-scyllo-inosose

机译：来自Burkholderia Terrae的新型（ - ） - vibo- Quercitol 1-脱氢酶的鉴定与表征，适用于2-脱氧 - Scyllo-inososose的（ - ） - vibo-槲皮素的生产

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摘要

(-)-vibo-Quercitol is a deoxyinositol (1L-1,2,4/3,5-cyclohexanepentol) that naturally occurs in oak species, honeydew honey, wines aged in oak barrels, and Gymnema sylvestre and is a potential intermediate for pharmaceuticals. We found that (-)-vibo-quercitol is stereoselectively synthesized from 2-deoxy-scyllo-inosose by the reductive reaction of a novel (-)-vibo-quercitol 1-dehydrogenase found in the proteomes of Burkholderia, Pseudomonas, and Arthrobacter. Among them, Burkholderia terrae sp. AKC-020 (40-1) produced a (-)-vibo-quercitol 1-dehydrogenase appropriate for synthesizing (-)-vibo-quercitol with a high diastereomeric excess. The enzyme was strongly induced in Bu. terrae cells when quercitol or 2-deoxy-scyllo-inosose was used as carbon source in the culture medium. The enzyme is NAD(H)-dependent and shows highly specific activity for (-)-vibo-quercitol and myo-inositol among the substrates tested. The enzyme gene (qudh) was obtained by PCR using degenerate primers based on the N-terminal and internal amino acid sequences of the purified enzyme, followed by thermal asymmetric interlaced PCR. The qudh gene showed homology with inositol 2-dehydrogenase (sharing 49.5% amino acid identity with IdhA from Sinorhizobium meliloti 1021). We successfully produced several recombinant (-)-vibo-quercitol 1-dehydrogenases and related enzymes identified by genome database mining using an Escherichia coli expression system. This revealed that scyllo-inositol dehydrogenase (IolX) in Bacillus subtilis can catalyze the reduction of 2-deoxy-scyllo-inosose to yield scyllo-quercitol, a stereoisomer of (-)-vibo-quercitol. Thus, we successfully identified two enzymes to produce both stereoisomers of deoxyinositols that are rare in nature.
机译：（ - ） - Vibo-Quercitol是一种脱氧肌醇（1L-1,2,4 / 3,5-环己己糖醇），天然发生在橡木种类，蜜露蜂蜜，橡木桶中的葡萄酒，以及Gymnema Sylvestre，是一个潜在的中间体药品。我们发现（ - ） - vibo- Quercitol通过在Burkowneria，假蛋白酶蛋白质蛋白质和关节杆菌的蛋白质组中发现的新（ - ） - vibo- Quercitol 1-脱氢酶的还原反应来由2-脱氧 - Scyllo-inosose合成。其中，Burkholderia Terrae SP。 AKC-020（40-1）制备了适于合成（ - ） - vibo-蜡的1-脱氢酶，其具有高度非映异构体过量。酶被强烈诱导在Bu。当蜡或2-脱氧 - Scyllo-Inososose用作培养基中的碳源时，Terrae细胞。酶是NAD（h） - 依赖性，并在测试的基材中对（ - ） - vibo- Quercitol和肌肌醇的高度比活性表示。通过PCR使用基于N-末端和纯化的酶的内氨基酸序列的脱底引物获得酶基因（QUDH），然后是热不对称的相互扫描PCR。 QuDH基因显示与肌醇2-脱氢酶的同源性（与Sinorhizobium Meliloti 1021的IDHA共用49.5％氨基酸同一性）。我们通过大肠杆菌表达系统成功地生产了几种重组（ - ） - vibo- Quercol 1-脱氢酶和由基因组数据库挖掘鉴定的相关酶。这揭示了枯草芽孢杆菌中的氏菌肌醇脱氢酶（IOLX）可以催化2-脱氧 - Scyllo-oisosose的还原，得到Scyllo- Quercitol，（ - ） - vibo- Quercitol的立体异构体。因此，我们成功地确定了两种酶，以产生脱氧肌醇的两个立体异构体，所述脱氧肌醇本质上很少。

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