Physiological and molecular genetic analyses of vinyl chloride and ethene biodegradation in Nocardioides sp strain JS614
Physiological and molecular genetic analyses of vinyl chloride and ethene biodegradation in Nocardioides sp strain JS614
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DOI：
10.1007/s00203-004-0749-2
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发表时间：
2005-02-01
影响因子：
2.8
通讯作者：
Gossett, JM
中科院分区：
文献类型：
--
作者：
Mattes, TE;Coleman, NV;Gossett, JM
诺卡氏菌属菌株JS614利用氯乙烯和乙烯作为碳源和能源。JS614在自然衰减和乙烯生物地球化学循环中具有重要作用，可用于生物修复、生物催化和代谢工程，但需要对菌株JS614中氯乙烯和乙烯同化的生理和遗传学基础有一个基本的了解。在JS614无细胞提取液中检测到烯烃单加氧酶(AkMO)活性和环氧烷：辅酶M转移酶(EaCoMT)活性。脉冲场凝胶电泳法检测到JS614中含有290kb的pNoc614。固化实验和聚合酶链式反应表明，pNoc614编码氯乙烯/乙烯降解基因。从一个克隆中回收了JS614氯乙烯/乙烯分解代谢基因及其侧翼DNA(34.8kb)。在一个可能的操纵子中发现了AkMO和EaCoMT基因，该操纵子包括辅酶A转移酶、酰基辅酶A合成酶、脱氢酶和还原酶基因。与该基因簇相邻的是一个差异转录的基因簇，它编码了可能的辅酶M生物合成酶。逆转录-聚合酶链式反应证实了氯乙烯和乙烯诱导的几个基因的性质。在Fosid克隆上也发现了编码可能的质粒结合、整合和分割功能的基因。
Nocardioides sp. strain JS614 utilizes vinyl chloride and ethene as carbon and energy sources. JS614 could be influential in natural attenuation and biogeochemical ethene cycling, and useful for bioremediation, biocatalysis and metabolic engineering, but a fundamental understanding of the physiological and genetic basis of vinyl chloride and ethene assimilation in strain JS614 is required. Alkene monooxygenase (AkMO) activity was demonstrated in whole-cell assays and epoxyalkane:coenzyme M transferase (EaCoMT) activity was detected in JS614 cell-free extracts. Pulsed-field gel electrophoresis revealed a 290-kb plasmid (pNoc614) in JS614. Curing experiments and PCR indicated that pNoc614 encodes vinyl chloride/ethene-degradation genes. JS614 vinyl chloride/ethene catabolic genes and flanking DNA (34.8 kb) were retrieved from a fosmid clone. AkMO and EaCoMT genes were found in a putative operon that included CoA transferase, acyl-CoA synthetase, dehydrogenase, and reductase genes. Adjacent to this gene cluster was a divergently transcribed gene cluster that encoded possible coenzyme M biosynthesis enzymes. Reverse transcription-PCR demonstrated the vinyl chloride- and ethene-inducible nature of several genes. Genes encoding possible plasmid conjugation, integration, and partitioning functions were also discovered on the fosmid clone.