...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >Functional Characterization of an NADPH Dependent 2-Alkyl-3-ketoalkanoic Acid Reductase Involved in Olefin Biosynthesis in Stenotrophomonas maltophilia
【24h】

Functional Characterization of an NADPH Dependent 2-Alkyl-3-ketoalkanoic Acid Reductase Involved in Olefin Biosynthesis in Stenotrophomonas maltophilia

机译：在斯托科托洛尼亚酸芽孢杆菌中烯烃生物合成中涉及烯烃生物合成的NADPH依赖性2-烷基-3-酮烷酸还原酶的功能表征

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

OleD is shown to play a key reductive role in the generation of alkenes (olefins) from acyl thioesters in Stenotrophomonas maltophilia. The gene coding for OleD clusters with three other genes, oleABC, and all appear to be transcribed in the same direction as an operon in various olefin producing bacteria. In this study, a series of substrates varying in chain length and stereochemistry were synthesized and used to elucidate the functional role and substrate specificity of OleD. We demonstrated that OleD, which is an NADP(H) dependent reductase, is a homodimer which catalyzes the reversible stereospecific reduction of 2-alkyl-3-ketoalkanoic acids. Maximal catalytic efficiency was observed with syn-2-decyl-3-hydroxytetradecanoic acid, with a k_(cat)/K_m 5- and 8-fold higher than for syn-2-octy1-3-hydroxydodecanoic acid and syn-2-hexyl-3-hydroxydecanoic acid, respectively. OleD activity was not observed with syn-2-butyl-3-hydroxyoctanoic acid and compounds lacking a 2-alkyl group such as 3-ketodecanoic and 3-hydroxydecanoic acids, suggesting the necessity of the 2-alkyl chain for enzyme recognition and catalysis. Using diastereomeric pairs of substrates and 4 enantiopure isomers of 2-hexyl-3-hydroxydecanoic acid of known stereochemistry, OleD was shown to have a marked stereochemical preference for the (2R,3S)-isomer. Finally, experiments involving OleA and OleD demonstrate the first 3 steps and stereochemical course in olefin formation from acyl thioesters; condensation to form a 2-alkyl-3-ketoacyl thioester, subsequent thioester hydrolysis, and ketone reduction.
机译：显示OLED在斯内特科洛尼亚酸辣椒胺酰胺中的酰基硫代酯的产生中发挥关键还原作用。编码具有三种其他基因的OLED簇的基因，ob，并且似乎在与各种烯烃产生细菌中的操纵子相同的方向转录。在本研究中，合成了一系列在链长和立体化学中变化的基质，并用于阐明OLED的功能作用和底物特异性。我们证明，作为NADP（H）依赖性还原酶的OLED是催化2-烷基-3-酮烷烃的可逆立体分子减少的同型二聚体。用SYN-2-癸基-3-羟基二甲酸观察到最大催化效率，k_（猫）/ k_m 5-和8倍高于SYN-2-octy1-3-羟基二癸酸和同步2-己基-3-羟基二癸酸分别。未观察到OLED活性，用SYN-2-丁基-3-羟基乙酸和缺少2烷基如3-酮二烷和3-羟基二癸酸的化合物，表明酶识别和催化的2-烷基链的必要性。使用已知立体化学的2-己基-3-羟二癸酸的二羟基甲酸的非对映异构体对，显示出对（2R，3S） - 溶剂的标记立体化学偏好。最后，涉及Olea和OLED的实验证明了烯烃形成的前3个步骤和立体化学过程从酰基硫代酯中形成;缩合形成2-烷基-3-酮烷基酯，随后的硫酯水解和酮还原。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号