Butane metabolism in Pseudomonas butanovora

丁香假单胞菌中的丁烷代谢

• 批准号: 7056745
• 负责人: DANIEL J ARP
• 金额: $ 24.23万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7056745
• 关键词: Pseudomonas; bacterial genetics; butanols; electron spin resonance spectroscopy; electron transport; enzyme substrate; gene mutation; genetic mapping; genetic transduction; microorganism metabolism; oxidoreductase; oxygenases; protein purification; site directed mutagenesis

DESCRIPTION (provided by applicant): Gaseous and liquid alkanes can serve as bacterial growth substrates. Alkane monooxygenases initiate the metabolism of alkanes by catalyzing the oxidation of the alkanes to alcohols. While metabolism of methane and liquid alkanes (e.g., octane) is well studied, much less is known about the oxidation of short-chain, gaseous alkanes. The applicant's research has focused on BMOs as representatives of this understudied group of alkane utilizers and on the complete pathway of butane metabolism. Previous results with three butane-utilizing bacteria indicated a surprising level of diversity among the enzymes that harvest butane as well as those involved in the subsequent metabolism of the alcohols. The investigators now propose to focus on just one of these bacteria, Pseudomonas butanovora. The specific aims of the project are: 1. Characterize the diiron containing BMO, which is similar to soluble MMO, but does not oxidize methane. 2. The applicants discovered two clusters of genes, each coding for a butanol dehydrogenase and a putative butyraldehyde dehydrogenase, which use parallel pathways of electron transport. The applicant now proposes to identify the components of these parallel pathways. 3. Previous studies indicated independent regulation of the production of BMO, butanol dehydrogenases and butyraldehyde dehydrogenases by butane and intermediates in the pathway. The investigators will examine the molecular basis of the regulation of induction of the butane pathway in P. butanovora. The proposed experiments may provide unique insights to the basis of substrate specificity in the broad class of diiron monooxygenases that oxidize nonpolar molecules. Studies of parallel pathways will be relevant to other metabolic pathways where toxic metabolites are necessarily produced as part of the metabolism of a growth supporting substrate.

描述（由申请人提供）：气态和液态烷烃可作为细菌生长基质。烷烃单加氧酶通过催化烷烃氧化成醇来启动烷烃的代谢。 而甲烷和液体烷烃的代谢（例如，辛烷）的氧化研究得很好，但对短链气态烷烃的氧化知之甚少。 申请人的研究集中在BMO作为这一未充分研究的烷烃利用者群体的代表以及丁烷代谢的完整途径上。 以前的结果与三个丁烷利用细菌表明一个令人惊讶的水平的多样性之间的酶收获丁烷以及参与随后的代谢的醇。 研究人员现在建议只关注其中一种细菌，即丁酸假单胞菌。 该项目的具体目标是：1。表征含二铁的BMO，其类似于可溶性MMO，但不氧化甲烷。 2.申请人发现了两个基因簇，每个基因簇编码丁醇脱氢酶和推定的丁醛脱氢酶，它们使用平行的电子传递途径。 申请人现在提出鉴定这些平行途径的组分。 3.以前的研究表明，丁烷和中间产物对BMO、丁醇脱氢酶和丁醛脱氢酶的产生具有独立的调节作用。 研究人员将研究P. butanovora中丁烷途径诱导调节的分子基础。 所提出的实验可能提供独特的见解的基础上的底物特异性在广泛的类二铁单加氧酶，氧化非极性分子。 平行途径的研究将与其他代谢途径相关，在这些代谢途径中，作为生长支持基质代谢的一部分，必然会产生有毒代谢物。

项目成果

Roles for the two 1-butanol dehydrogenases of Pseudomonas butanovora in butane and 1-butanol metabolism.

丁香假单胞菌的两种 1-丁醇脱氢酶在丁烷和 1-丁醇代谢中的作用。

• DOI: 10.1128/jb.184.16.4343-4350.2002
• 发表时间: 2002
• 期刊: Journal of bacteriology
• 影响因子: 3.2
• 作者: Vangnai,AlisaS;Sayavedra-Soto,LuisA;Arp,DanielJ
• 通讯作者: Arp,DanielJ

Propionate inactivation of butane monooxygenase activity in 'Pseudomonas butanovora': biochemical and physiological implications.

“丁酸假单胞菌”中丁烷单加氧酶活性的丙酸灭活：生化和生理学影响。

• DOI: 10.1099/mic.0.2007/008441-0
• 发表时间: 2007
• 期刊: Microbiology (Reading, England)
• 作者: Doughty,DM;Halsey,KH;Vieville,CJ;Sayavedra-Soto,LA;Arp,DJ;Bottomley,PJ
• 通讯作者: Bottomley,PJ

Diversity in butane monooxygenases among butane-grown bacteria.

丁烷生长的细菌中丁烷单加氧酶的多样性。

• DOI: 10.1128/aem.65.10.4586-4593.1999
• 发表时间: 1999
• 期刊: Applied and environmental microbiology
• 影响因子: 4.4
• 作者: Hamamura,N;Storfa,RT;Semprini,L;Arp,DJ
• 通讯作者: Arp,DJ

Evidence for modified mechanisms of chloroethene oxidation in Pseudomonas butanovora mutants containing single amino acid substitutions in the hydroxylase alpha-subunit of butane monooxygenase.

丁烷单加氧酶羟化酶 α 亚基中含有单个氨基酸取代的丁酸假单胞菌突变体中氯乙烯氧化机制改变的证据。

• DOI: 10.1128/jb.00189-07
• 发表时间: 2007
• 期刊: Journal of bacteriology
• 影响因子: 3.2
• 作者: Halsey,KimberlyH;Doughty,DavidM;Sayavedra-Soto,LuisA;Bottomley,PeterJ;Arp,DanielJ
• 通讯作者: Arp,DanielJ

Molecular analysis of the soluble butane monooxygenase from 'Pseudomonas butanovora'.

• DOI: 10.1099/00221287-148-11-3617
• 发表时间: 2002-11
• 期刊: Microbiology
• 影响因子: 1.5
• 作者: M. Sluis;L. Sayavedra-Soto;D. Arp