Two distinct monooxygenases for alkane oxidation in Nocardioides sp strain CF8
Two distinct monooxygenases for alkane oxidation in Nocardioides sp strain CF8
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DOI：
10.1128/aem.67.11.4992-4998.2001
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发表时间：
2001-11-01
影响因子：
4.4
通讯作者：
Arp, DJ
中科院分区：
文献类型：
--
作者：
Hamamura, N;Yeager, CM;Arp, DJ
拟诺卡氏菌中的烷基单加氧酶从生理和遗传水平对菌株CF8进行了检测。菌株CF8可以利用链长从C-2到C-16的烷烃。丁烷培养细胞的丁烷降解被铜选择性螯合剂烯丙基硫脲强烈抑制，而正己烷、辛烷和正庚烷培养的细胞在烯丙基硫脲存在下显示出明显的丁烷降解活性。1-己炔对正丁烷和正己烷的生长有很强的抑制作用，而对正辛烷和正庚烷的生长没有影响。在丁烷、正己烷、辛烷和正庚烷培养的细胞中观察到一种特定的30 kDa的乙炔结合多肽，但在辛烷或正庚烷存在下生长的细胞中没有观察到。这些结果表明CF8菌株中存在两种单加氧酶。为扩增油假单胞菌双核铁烷基羟基酶相关基因而设计的简并引物克隆了菌株CF8的相关基因。逆转录-聚合酶链式反应和Northern印迹杂交分析表明，该基因编码的双核含铁烷烃羟基酶基因在生长在C-6以上烷烃上的细胞中表达。这些结果表明，诺卡氏菌中存在两种不同的烷烃氧化单加氧酶。菌株CF8。
Alkane monooxygenases in Nocardioides sp. strain CF8 were examined at the physiological and genetic levels. Strain CF8 can utilize alkanes ranging in chain length from C-2 to C-16. Butane degradation by butane-grown cells was strongly inhibited by allylthiourea, a copper-selective chelator, while hexane-, octane-, and decane-grown cells showed detectable butane degradation activity in the presence of allylthiourea. Growth on butane and hexane was strongly inhibited by 1-hexyne, while 1-hexyne did not affect growth on octane or decane. A specific 30-kDa acetylene-binding polypeptide was observed for butane-, hexane-, octane-, and decane-grown cells but was absent from cells grown with octane or decane in the presence of 1-hexyne. These results suggest the presence of two monooxygenases in strain CF8. Degenerate primers designed for PCR amplification of genes related to the binuclear-iron-containing alkane hydroxylase from Pseudomonas oleovorans were used to clone a related gene from strain CF8. Reverse transcription-PCR and Northern blot analysis showed that this gene encoding a binuclear-iron-containing alkane hydroxylase was expressed in cells grown on alkanes above C-6. These results indicate the presence of two distinct monooxygenases for alkane oxidation in Nocardioides sp. strain CF8.