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在以补充葡萄糖的培养基上生长的铜绿假单胞菌142中,卤代苯甲酸分解代谢途径的诱导及邻氯苯甲酸的共代谢。

Induction of the halobenzoate catabolic pathway and cometabolism of ortho-chlorobenzoates in Pseudomonas aeruginosa 142 grown on glucose-supplemented media.

作者信息

Corbella M E, Garrido-Pertierra A, Puyet A

机构信息

Departamento de Bioquímica y Biología Molecular IV, Facultad de Veterinaria, Universidad Complutense de Madrid, Spain.

出版信息

Biodegradation. 2001;12(3):149-57. doi: 10.1023/a:1013117805732.

Abstract

The aerobic cometabolism of ortho-substituted chlorobenzoates by Pseudomonas aeruginosa strain 142 growing on glucose-supplemented medium was analyzed. The strain, which can use 2-chlorobenzoate (2-CBA) and 2,4-dichlorobenzoate (2,4-DCBA) as sole carbon and energy sources, showed high rates of 2-CBA metabolism in glucose-fed cells. In contrast, 2,4-DCBA was metabolized only after extended incubation of the full grown culture and depletion of glucose. In addition to the ortho-dehalogenation (ohb142) genes encoding the alpha and beta subunits of the oxygenase component of a 2-halobenzoate dioxygenase, strain 142 harbours a closely related ohbABCDFG gene cluster previously identified in P. aeruginosa JB2 (ohbJB2). The genes for the chlorocatechol ortho-catabolic pathway were identified and sequenced in this strain, showing a near complete identity with the clcABD operon of the pAC27 plasmid. Relative quantification of mRNA by RT-PCR shows a preferential induction of ohb142 by 2-CBA, which is abolished in glucose-grown cultures. The alternate ohbJB2 and clc genes were expressed preferentially in 2,4-DCBA grown cultures. Only ohbJB2 appears to be expressed in the presence of the carbohydrate. Detection of chlorocatechol-1,2-dioxygenase activity in 2,4-DCBA plus glucose grown cultures suggests the presence of an alternate system for the ortho-cleavage of chlorobenzoates. The recruitment of elements from two halobenzoate dioxygenase systems with different induction patterns, together with a chlorocatechol degradative pathway not repressed by carbon catabolite, may allow P. aeruginosa 142 to cometabolize haloaromatics in carbohydrate grown cultures.

摘要

分析了在补充葡萄糖的培养基上生长的铜绿假单胞菌142菌株对邻位取代氯苯甲酸酯的好氧共代谢情况。该菌株能够利用2-氯苯甲酸(2-CBA)和2,4-二氯苯甲酸(2,4-DCBA)作为唯一碳源和能源,在以葡萄糖为食的细胞中表现出较高的2-CBA代谢速率。相比之下,2,4-DCBA仅在完全生长的培养物长时间孵育且葡萄糖耗尽后才被代谢。除了编码2-卤苯甲酸双加氧酶加氧酶组分的α和β亚基的邻位脱卤(ohb142)基因外,142菌株还含有先前在铜绿假单胞菌JB2(ohbJB2)中鉴定出的密切相关的ohbABCDFG基因簇。在该菌株中鉴定并测序了氯儿茶酚邻位分解代谢途径的基因,与pAC27质粒的clcABD操纵子显示出近乎完全的同一性。通过RT-PCR对mRNA进行相对定量显示,2-CBA对ohb142有优先诱导作用,而在以葡萄糖生长的培养物中这种诱导作用消失。交替的ohbJB2和clc基因在以2,4-DCBA生长的培养物中优先表达。只有ohbJB2似乎在碳水化合物存在时表达。在2,4-DCBA加葡萄糖生长的培养物中检测到氯儿茶酚-1,2-双加氧酶活性,表明存在氯苯甲酸酯邻位裂解的替代系统。从具有不同诱导模式的两个卤苯甲酸双加氧酶系统中募集元件,以及不被碳分解代谢物抑制的氯儿茶酚降解途径,可能使铜绿假单胞菌142在碳水化合物生长的培养物中共代谢卤代芳烃。

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