Propane monooxygenase and NAD+-dependent secondary alcohol dehydrogenase in propane metabolism by Gordonia sp strain TY-5
Propane monooxygenase and NAD+-dependent secondary alcohol dehydrogenase in propane metabolism by Gordonia sp strain TY-5
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DOI：
10.1128/jb.185.24.7120-7128.2003
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发表时间：
2003-12-01
影响因子：
3.2
通讯作者：
Kato, N
中科院分区：
文献类型：
--
作者：
Kotani, T;Yamamoto, T;Kato, N
一种新的分离物，戈登氏菌菌株TY-5，能够在丙烷和具有C-13至C-22碳链的正烷烃上生长作为唯一的碳源。在全细胞反应中，检测到显著的丙烷氧化为2-丙醇。一个基因簇命名prmABCD，它编码一个假定的双核含铁多组分单加氧酶，包括大和小的羟化酶，NADH依赖性受体氧化还原酶，和耦合蛋白的组成部分，进行了克隆和测序。prmB被破坏的突变体（prmB：：Kan（r））失去了在丙烷上生长的能力，并且北方印迹分析显示在丙烷上生长期间诱导了prm基因的多顺反子转录。这些结果表明prmABCD基因产物在细菌的丙烷氧化中发挥着重要作用。在prm基因的下游，鉴定了编码NAD（+）依赖性仲醇脱氢酶的开放阅读框（adh 1），并纯化和表征了蛋白质。北方印迹分析结果和一个破坏突变体（adh 1：：Kan（r））的生长特性表明Adh 1在丙烷代谢中起主要作用。另外两种NAD（+）依赖性仲醇脱氢酶（Adh 2和Adh 3）也被发现参与2-丙醇氧化。基于这些结果，我们得出结论，戈登氏菌菌株TY-5氧化丙烷通过单加氧酶介导的亚末端氧化2-丙醇。
A new isolate, Gordonia sp. strain TY-5, is capable of growth on propane and n-alkanes with C-13 to C-22 carbon chains as the sole source of carbon. In whole-cell reactions, significant propane oxidation to 2-propanol was detected. A gene cluster designated prmABCD, which encodes the components of a putative dinuclear-iron-containing multicomponent monooxygenase, including the large and small subunits of the hydroxylase, an NADH-dependent acceptor oxidoreductase, and a coupling protein, was cloned and sequenced. A mutant with prmB disrupted (prmB::Kan(r)) lost the ability to grow on propane, and Northern blot analysis revealed that polycistronic transcription of the prm genes was induced during its growth on propane. These results indicate that the prmABCD gene products play an essential role in propane oxidation by the bacterium. Downstream of the prm genes, an open reading frame (adh1) encoding an NAD(+)-dependent secondary alcohol dehydrogenase was identified, and the protein was purified and characterized. The Northern blot analysis results and growth properties of a disrupted mutant (adh1::Kan(r)) indicate that Adh1 plays a major role in propane metabolism. Two additional NAD(+)-dependent secondary alcohol dehydrogenases (Adh2 and Adh3) were also found to be involved in 2-propanol oxidation. On the basis of these results, we conclude that Gordonia sp. strain TY-5 oxidizes propane by monooxygenase-mediated subterminal oxidation via 2-propanol.