Inoue Kengo, Widada Jaka, Nakai Seiichiro, Endoh Takayuki, Urata Masaaki, Ashikawa Yuji, Shintani Masaki, Saiki Yuko, Yoshida Takako, Habe Hiroshi, Omori Toshio, Nojiri Hideaki
Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Biosci Biotechnol Biochem. 2004 Jul;68(7):1467-80. doi: 10.1271/bbb.68.1467.
Southern hybridization analysis of the genomes from the newly-isolated 10 carbazole (CAR)-utilizing bacteria revealed that 8 of the isolates carried gene clusters homologous to the CAR-catabolic car operon of Pseudomonas resinovorans strain CA10. Sequencing analysis showed that two car operons and the neighboring regions of Pseudomonas sp. strain K23 are nearly identical to that of strain CA10. In contrast to strains CA10 and K23, carEF genes did not exist downstream of the car gene cluster of Janthinobacterium sp. strain J3. In the car gene clusters, strains CA10, K23 and J3 have Rieske-type ferredoxin as a component of carbazole dioxygenase, although Sphingomonas sp. strain KA1 possesses a putidaredoxin-type ferredoxin. We confirmed that this putidaredoxin-type ferredoxin CarAc can function as an electron mediator to CarAa of strain KA1. In the upstream regions of the carJ3 and carKA1 gene clusters, ORFs whose deduced amino acid sequences showed homology to GntR-family transcriptional regulators were identified.
对新分离出的10株咔唑(CAR)利用菌的基因组进行Southern杂交分析表明,其中8株携带与树脂糖假单胞菌CA10菌株的CAR分解代谢car操纵子同源的基因簇。测序分析表明,假单胞菌属K23菌株的两个car操纵子及其邻近区域与CA10菌株几乎相同。与CA10和K23菌株不同,詹氏菌属J3菌株的car基因簇下游不存在carEF基因。在car基因簇中,CA10、K23和J3菌株具有Rieske型铁氧化还原蛋白作为咔唑双加氧酶的一个组分,尽管鞘氨醇单胞菌属KA1菌株拥有一个恶臭假单胞菌型铁氧化还原蛋白。我们证实,这种恶臭假单胞菌型铁氧化还原蛋白CarAc可以作为KA1菌株CarAa的电子介质。在carJ3和carKA1基因簇的上游区域,鉴定出了其推导氨基酸序列与GntR家族转录调节因子具有同源性的开放阅读框。