Soluble di-iron monooxygenases (SDIMOs) are key enzymes in the bacterial oxidation of hydrocarbons, and have applications in environmental and industrial biotechnology. SDIMOs from pure cultures are unlikely to represent the total diversity of this enzyme family, so we used polymerase chain reaction to survey the diversity of SDIMO alpha subunit genes in environmental samples, ethene enrichments and ethene-degrading bacterial isolates. From 178 cloned amplicons, 98 restriction fragment length polymorphism types were seen, from which 75 representative SDIMO sequences were obtained; 45 from environmental samples, 25 from enrichments and seven from isolates. The sequences were diverse, including genes similar to ethene (etnC), propene (amoC, pmoC), propane (prmA) and butane (bmoX) monooxygenases, in addition to many novel sequences comprising a new SDIMO group (group 6). Environmental samples showed the highest diversity, with strong representation of group 6 SDIMOs and prmA-like genes. Ethene stimulation of samples resulted in increased frequencies of group 4 SDIMOs (etnC-like). Four ethene-utilizing Mycobacterium isolates (NBB1-NBB4) from enrichments all contained etnC; one isolate (NBB4) also contained three additional SDIMO genes (bmoX-like, amoC-like and group 6). The primers, database, clone libraries and strains reported here provide a resource for future bioremediation and biocatalysis studies, with particular relevance for chlorinated alkene and alkane compounds.

译文

:可溶性二铁单加氧酶(SDIMOs)是烃类细菌氧化中的关键酶,并已应用于环境和工业生物技术中。来自纯培养物的SDIMO不太可能代表该酶家族的总多样性,因此我们使用聚合酶链反应来调查环境样品,乙烯富集和乙烯降解细菌分离物中SDIMOα亚基基因的多样性。从178个克隆的扩增子中,观察到98个限制性片段长度多态性类型,从中获得了75个代表性的SDIMO序列。来自环境样品的45个,来自浓缩物的25个和来自分离株的7个。这些序列是多种多样的,除了包括新的SDIMO组(第6组)的许多新颖序列外,还包括与乙烯(etnC),丙烯(amoC,pmoC),丙烷(prmA)和丁烷(bmoX)单加氧酶相似的基因。环境样品显示出最高的多样性,强烈代表了第6组SDIMOs和prmA样基因。乙烯对样品的刺激导致第4组SDIMO(etnC样)的频率增加。来自浓缩物中的四个利用乙烯的分枝杆菌分离株(NBB1-NBB4)均包含etnC;一个分离株(NBB4)还包含三个其他SDIMO基因(bmoX样,amoc样和第6组)。此处报道的引物,数据库,克隆文库和菌株为将来的生物修复和生物催化研究提供了资源,尤其与氯化烯烃和烷烃化合物有关。

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