RESUMO
The Pseudomonas putida ÐÐ5 that was isolated from the slime pit of a Nizhnekamsk oil chemical factory can metabolize naphthalene via salicylate and gentisate. Catabolic genes are localized on non-conjugative IncP-7 plasmid pAK5 of about 115 kb in size. The "classical"nah-1 operon and the novel sgp-operon (salicylate-gentisate pathway) are both involved in naphthalene degradation by P. putida ÐÐ5, that was first described for Pseudomonas. The sgp-operon includes six open reading frames (ORFs) (sgpAIKGHB). The four ORFs code for the entire salicylate 5-hydroxylase - oxidoreductase component (sgpA), large and small subunits of the oxigenase component (sgpG and sgpH) and 2Fe-2S ferredoxin (sgpB). Genes for gentisate 1, 2-dioxygenase (sgpI) and fumarylpyruvate hydrolase (sgpK) are located in salicylate 5-hydroxylase genes clustering between sgpA and sgpG. The putative positive regulator for the sgp-operon (sgpR) was found upstream of the sgpA gene and oriented in the opposite direction from sgpA. The putative maleylacetoacetate isomerase gene is located apart, directly downstream from the sgp-operon. The sgp-operon organization and phylogenetic analysis of deduced amino acid sequences indicate that this operon has a mosaic structure according to the modular theory of the evolution of modern catabolic pathways.
Assuntos
Naftalenos/metabolismo , Óperon , Pseudomonas putida/enzimologia , Pseudomonas putida/genética , Sequência de Aminoácidos , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Dioxigenases/química , Dioxigenases/genética , Gentisatos/química , Gentisatos/metabolismo , Hidrolases/química , Hidrolases/genética , Oxigenases de Função Mista/química , Oxigenases de Função Mista/genética , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Oxirredutases/química , Oxirredutases/genética , Filogenia , Salicilatos/química , Salicilatos/metabolismo , cis-trans-Isomerases/química , cis-trans-Isomerases/genéticaRESUMO
Two strains (NF1 and NF3) of free-living chemoorganotrophic bacteria have been isolated from multiyear oil slime and Pedilanthus tithymaloides rhizosphere and ascribed to the genus Kaistia of the class Alphaproteobacteria on the basis of the nucleotide sequences of 16S rRNA gene and phenotypic characteristics. These strains can be assigned to ultramicrobacteria as their populations are represented by two subpopulations: (1) ultrasmall cells, on average 200-300 nm in diameter and <0.1 microm(3) in volume, of up to 60% of the total number of cells in a population, and (2) cells 400-800 nm in diameter and 0.15-0.5 microm(3) in volume, of up to 40% of the total number of cells in a population. The interaction of the isolated ultramicrobacteria strains (IUMB) with different bacterial species has been studied in cocultures grown under starvation and in complete nutrient media. It has been found that IUMB can be facultative parasites on certain species of chemoorganotrophic and phototrophic bacteria. The interaction of IUMB with prey bacteria exhibits the extracellular type of parasitism and involves establishing stable cell-cell contacts between the parasites and their prey to cause destruction of host cells.
Assuntos
Rhizobiaceae/isolamento & purificação , Rhizobiaceae/fisiologia , Microbiologia do Solo , Meios de Cultura , Bactérias Gram-Negativas/crescimento & desenvolvimento , Bactérias Gram-Positivas/crescimento & desenvolvimento , Microscopia Eletrônica , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Rhizobiaceae/genética , Rhizobiaceae/ultraestruturaRESUMO
A large collection of naphthalene-degrading fluorescent Pseudomonas strains isolated from sites contaminated with coal tar and crude oil was screened for the presence of IncP-9 plasmids. Seventeen strains were found to carry naphthalene catabolic plasmids ranging in size from 83 to 120 kb and were selected for further study. Results of molecular genotyping revealed that 15 strains were closely related to P. putida, one to P. fluorescens, and one to P. aeruginosa. All catabolic plasmids found in these strains, with the exception of pBS216, pSN11, and p8909N-1, turned out to belong to IncP-9 beta-subgroup. Plasmids pBS216, pSN11, and p8909N-1 were identified as members of IncP-9 delta-subgroup. One plasmid, pBS2, contains fused replicons of IncP-9beta and IncP-7 groups. RFLP analyses of the naphthalene catabolic plasmids revealed that organisation of the replicon correlates well with the overall plasmid structure. Comparative PCR studies with conserved oligonucleotide primers indicated that genes for key enzymes of naphthalene catabolism are highly conserved among all studied plasmids. Three bacterial strains, P. putida BS202, P. putida BS3701, and P. putida BS3790, were found to have two different salicylate hydroxylase genes one of which has no similarity to the "classic" enzyme encoded by nahG gene. Discovery of a large group of plasmid with unique nahR suggested that the regulatory loop may also represent a variable part of the pathway for catabolism of naphthalene in fluorescent Pseudomonas spp.
