ABSTRACT
Genes for catechol 1,2- and 2,3-dioxygenases were cloned. These enzymes hold important positions in the ortho and meta pathways of the metabolism of aromatic carbons by microbial associations that consume the following volatile organic compounds in pilot minireactors: toluene, styrene, ethyl benzene, o-xylene, m-xylene, and naphthalene. Genes of both pathways were found in an association consuming m-xylene; only genes of the ortho pathway were found in associations consuming o-xylene, styrene, and ethyl benzene, and only genes of the meta pathway were found in associations consuming naphthalene and toluene. Genes of the ortho pathway (C120) cloned from associations consuming o-xylene and ethyl benzene were similar to corresponding genes located on the pND6 plasmid of Pseudomonas putida. Genes of the ortho pathway from associations consuming o-xylene and m-xylene were similar to chromosomal genes of P. putida. Genes of the meta pathway (C230) from associations consuming toluene and naphthalene were similar to corresponding genes formerly found in plasmids pWWO and pTOL.
Subject(s)
Bacteria/enzymology , Dioxygenases/genetics , Genes, Bacterial/genetics , Hydrocarbons, Aromatic/metabolism , Bacteria/genetics , Catechol 1,2-Dioxygenase , Catechol 2,3-Dioxygenase , Cloning, Molecular , Dioxygenases/classification , Phylogeny , Plasmids/genetics , Pseudomonas putida/enzymology , Pseudomonas putida/geneticsABSTRACT
A range of species of four mixed bacterial cultures was studied by molecular systematics methods with the use of 16S rRNA genes. The cultures had been developed for application in minireactors, to degrade volatile organic compounds (VOCs): ethyl benzene, m-xylene, styrene, and o-xylene. A sample of 30 plasmid rDNA clones was obtained for each of the mixed cultures. The clones were analyzed by RFLP according to two restriction sites. Major variants of the 16S-rDNA sequences, corresponding to the most abundant species, were determined for each association. Sequencing of four clones of predominant 16S-rDNAs showed that the culture consuming ethyl benzene was dominated by Pseudomonas fluorescens; o-xylene, by Achromobacter xylosoxydans; styrene, by Pseudomonas veronii; and m-xylene, by Delftia acidovorans. Minor components of all four cultures were generally similar. They included species of the genera Sphingobacter, Rhizobium, Mesorhizobium, Pedobacter, and Paenibacillus. Sampling sequencing of genes for 16S rRNA cloned from total genomic DNA allowed quantitative determination of the composition of actual bacterial associations consuming VOCs in minireactors.
Subject(s)
Bacteria/metabolism , DNA, Ribosomal/genetics , Organic Chemicals/metabolism , RNA, Ribosomal, 16S/genetics , Bacteria/classification , Bacteria/genetics , Base Sequence , DNA Primers , Polymorphism, Restriction Fragment Length , VolatilizationABSTRACT
The dependence of toluene elimination capacity on its load was obtained in five small-scale reactors filled with glass beads carrying biocatalyst cells. With increase in operation time the calculated maximal elimination capacity was shown to increase along with biomass density in the biocatalyst bed. Fivefold increase in trickling intensity did not affect the reactor performance. A simplified mathematical model for evaluation of minimal required biocatalyst bed volume at certain loading was developed based on experimental dependence of elimination capacity vs. loading.