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Both organisms contained at least four alkane monooxygenase gene homologs (<jats:italic>alkB1<\/jats:italic>,<jats:italic>alkB2<\/jats:italic>,<jats:italic>alkB3<\/jats:italic>, and<jats:italic>alkB4<\/jats:italic>). In both strains, the<jats:italic>alkB1<\/jats:italic>and<jats:italic>alkB2<\/jats:italic>homologs were part of<jats:italic>alk<\/jats:italic>gene clusters, each encoding two rubredoxins (<jats:italic>rubA1<\/jats:italic>and<jats:italic>rubA2<\/jats:italic>;<jats:italic>rubA3<\/jats:italic>and<jats:italic>rubA4<\/jats:italic>), a putative TetR transcriptional regulatory protein (<jats:italic>alkU1<\/jats:italic>;<jats:italic>alkU2<\/jats:italic>), and, in the<jats:italic>alkB1<\/jats:italic>cluster, a rubredoxin reductase (<jats:italic>rubB<\/jats:italic>). The<jats:italic>alkB3<\/jats:italic>and<jats:italic>alkB4<\/jats:italic>homologs were found as separate genes which were not part of<jats:italic>alk<\/jats:italic>gene clusters. Functional heterologous expression of some of the rhodococcal<jats:italic>alk<\/jats:italic>genes (<jats:italic>alkB2<\/jats:italic>,<jats:italic>rubA2<\/jats:italic>, and<jats:italic>rubA4<\/jats:italic>[NRRL B-16531];<jats:italic>alkB2<\/jats:italic>and<jats:italic>rubB<\/jats:italic>[Q15]) was achieved in<jats:italic>Escherichia coli<\/jats:italic>and<jats:italic>Pseudomonas<\/jats:italic>expression systems.<jats:italic>Pseudomonas<\/jats:italic>recombinants containing rhodococcal<jats:italic>alkB2<\/jats:italic>were able to mineralize and grow on C<jats:sub>12<\/jats:sub>to C<jats:sub>16<\/jats:sub><jats:italic>n<\/jats:italic>-alkanes. All rhodococcal alkane monooxygenases possessed the highly conserved eight-histidine motif, including two apparent alkane monooxygenase signature motifs (LQRH[S\/A]DHH and NYXEHYG[L\/M]), and the six hydrophobic membrane-spanning regions found in all alkane monooxygenases related to the<jats:italic>Pseudomonas putida<\/jats:italic>GPo1 alkane monooxygenase. The presence of multiple alkane hydroxylases in the two rhodococcal strains is reminiscent of other multiple-degradative-enzyme systems reported in<jats:italic>Rhodococcus<\/jats:italic>.<\/jats:p>","DOI":"10.1128\/aem.68.12.5933-5942.2002","type":"journal-article","created":{"date-parts":[[2002,11,26]],"date-time":"2002-11-26T00:04:48Z","timestamp":1038269088000},"page":"5933-5942","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":208,"title":["Gene Cloning and Characterization of Multiple Alkane Hydroxylase Systems in<i>Rhodococcus<\/i>Strains Q15 and NRRL B-16531"],"prefix":"10.1128","volume":"68","author":[{"given":"L. G.","family":"Whyte","sequence":"first","affiliation":[{"name":"Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec, Canada H4P 2R2"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"T. H. 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