Kotani Tetsuya, Yurimoto Hiroya, Kato Nobuo, Sakai Yasuyoshi
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake, Kyoto 606-8502, Japan.
J Bacteriol. 2007 Feb;189(3):886-93. doi: 10.1128/JB.01054-06. Epub 2006 Oct 27.
In the propane-utilizing bacterium Gordonia sp. strain TY-5, propane was shown to be oxidized to 2-propanol and then further oxidized to acetone. In this study, the subsequent metabolism of acetone was studied. Acetone-induced proteins were found in extracts of cells induced by acetone, and a gene cluster designated acmAB was cloned on the basis of the N-terminal amino acid sequences of acetone-induced proteins. The acmA and acmB genes encode a Baeyer-Villiger monooxygenase (BVMO) and esterase, respectively. The BVMO encoded by acmA was purified from acetone-induced cells of Gordonia sp. strain TY-5 and characterized. The BVMO exhibited NADPH-dependent oxidation activity for linear ketones (C3 to C10) and cyclic ketones (C4 to C8). Escherichia coli expressing the acmA gene oxidized acetone to methyl acetate, and E. coli expressing the acmB gene hydrolyzed methyl acetate. Northern blot analyses revealed that polycistronic transcription of the acmAB gene cluster was induced by propane, 2-propanol, and acetone. These results indicate that the acmAB gene products play an important role in the metabolism of acetone derived from propane oxidation and clarify the propane metabolism pathway of strain TY-5 (propane --> 2-propanol --> acetone --> methyl acetate --> acetic acid + methanol). This paper provides the first evidence for BVMO-dependent acetone metabolism.
在利用丙烷的戈登氏菌属菌株TY-5中,丙烷被证明先氧化为2-丙醇,然后进一步氧化为丙酮。在本研究中,对丙酮的后续代谢进行了研究。在丙酮诱导的细胞提取物中发现了丙酮诱导蛋白,并基于丙酮诱导蛋白的N端氨基酸序列克隆了一个名为acmAB的基因簇。acm A和acm B基因分别编码一种拜耳-维利格单加氧酶(BVMO)和酯酶。从丙酮诱导的戈登氏菌属菌株TY-5细胞中纯化并表征了由acm A编码的BVMO。该BVMO对直链酮(C3至C10)和环酮(C4至C8)表现出依赖于NADPH的氧化活性。表达acm A基因的大肠杆菌将丙酮氧化为乙酸甲酯,而表达acm B基因的大肠杆菌则水解乙酸甲酯。Northern印迹分析表明,acmAB基因簇的多顺反子转录由丙烷、2-丙醇和丙酮诱导。这些结果表明,acmAB基因产物在丙烷氧化衍生的丙酮代谢中起重要作用,并阐明了菌株TY-5的丙烷代谢途径(丙烷→2-丙醇→丙酮→乙酸甲酯→乙酸+甲醇)。本文首次提供了依赖BVMO的丙酮代谢的证据。