Haigler B E, Pettigrew C A, Spain J C
Air Force Civil Engineering Support Agency, Tyndall Air Force Base, Florida 32403-6001.
Appl Environ Microbiol. 1992 Jul;58(7):2237-44. doi: 10.1128/aem.58.7.2237-2244.1992.
Pseudomonas sp. strain JS150 was isolated as a nonencapsulated variant of Pseudomonas sp. strain JS1 that contains the genes for the degradative pathways of a wide range of substituted aromatic compounds. Pseudomonas sp. strain JS150 grew on phenol, ethylbenzene, toluene, benzene, naphthalene, benzoate, p-hydroxybenzoate, salicylate, chlorobenzene, and several 1,4-dihalogenated benzenes. We designed experiments to determine the conditions required for induction of the individual pathways and to determine whether multiple substrates could be biodegraded simultaneously. Oxygen consumption studies with whole cells and enzyme assays with cell extracts showed that the enzymes of the meta, ortho, and modified ortho cleavage pathways can be induced in strain JS150. Strain JS150 contains a nonspecific toluene dioxygenase with a substrate range similar to that found in strains of Pseudomonas putida. The presence of the dioxygenase along with multiple pathways for metabolism of substituted catechols allows facile extension of the growth range by spontaneous mutation and degradation of mixtures of substituted benzenes and phenols. Chlorobenzene-grown cells of strain JS150 degraded mixtures of chlorobenzene, benzene, toluene, naphthalene, trichloroethylene, and 1,2- and 1,4-dichlorobenzenes in continuous culture. Under similar conditions, phenol-grown cells degraded a mixture of phenol, 2-chloro-, 3-chloro, and 2,5-dichlorophenol and 2-methyl- and 3-methylphenol. These results indicate that induction of appropriate biodegradative pathways in strain JS150 permits the biodegradation of complex mixtures of aromatic compounds.
假单胞菌属菌株JS150是作为假单胞菌属菌株JS1的非包膜变体分离得到的,JS1含有多种取代芳香化合物降解途径的基因。假单胞菌属菌株JS150能在苯酚、乙苯、甲苯、苯、萘、苯甲酸盐、对羟基苯甲酸盐、水杨酸盐、氯苯以及几种1,4 - 二卤代苯上生长。我们设计了实验来确定诱导各个途径所需的条件,并确定多种底物是否能同时被生物降解。对全细胞的氧气消耗研究以及对细胞提取物的酶活性测定表明,间位、邻位和修饰邻位裂解途径的酶在菌株JS150中均可被诱导。菌株JS150含有一种非特异性甲苯双加氧酶,其底物范围与恶臭假单胞菌菌株中发现的类似。双加氧酶的存在以及取代儿茶酚的多种代谢途径使得通过自发突变和降解取代苯和苯酚的混合物能够轻松扩展生长范围。菌株JS150的氯苯培养细胞在连续培养中能降解氯苯、苯、甲苯、萘、三氯乙烯以及1,2 - 二氯苯和1,4 - 二氯苯的混合物。在类似条件下,苯酚培养细胞能降解苯酚、2 - 氯苯酚、3 - 氯苯酚、2,5 - 二氯苯酚以及2 - 甲基苯酚和3 - 甲基苯酚的混合物。这些结果表明,在菌株JS150中诱导适当的生物降解途径可实现芳香化合物复杂混合物的生物降解。