D-gluconate dehydratase from Alcaligenes.
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Biomedical subjects
Publications and source records attributed to K Kersters.
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Improved methods for the identification and grouping of bacteria by polyacrylamide gel electrophoresis of soluble proteins are described. Electrophoretic protein patterns were obtained in rigorously standardized comditions. The results were much more reproducible than any described previously. Some of the factors affecting reproducibility were; growth conditions, time and speed of centrifugation of extracts, and conditions of gel electrophoresis. Protein patterns were compared by computing correlation coefficients from normalized densitometric tracings and clustering the strains by the unweighted average pair group method. As model systems, both Agrobacterium and Zymomonas were used because of differences in the sharpness of the peaks. The methodwas applied to 42 Agrobacterium strains. The agreement with the results of clustering by either phenotypic tests or DNA:DNA hybridization was excellent. Computerized comparisons of electrophoretic protein patterns can be a fast, easy and powerful tool for classification and identification of bacteria.
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The species Escherichia adecarboxylata was examined for DNA relatedness to the "Erwinia herbicola-Enterobacter agglomerans" complex and to other members of the family Enterobacteriaceae. DNA-DNA hybridizations (nitrocellulose filter method) showed that strains received as E. adecarboxylata were highly related to each other (73-100% homology). Three strains of E. agglomerans and one strain of E. herbicola showed, respectively, 77, 96, 97 and 92% relatedness with the labelled DNA of E. adecarboxylata. Two groups (E2 and E3) of "atypical coliforms" previously described by Gavini et al. (1983) showed high reassociation values (76-79% and 80-89%, respectively) with E. adecarboxylata. Most of these strains produced similar or nearly identical protein electrophoregrams. All these strains were therefore classified in E. adecarboxylata. This taxon yielded hybridization values lower than 53% with the previously described phenetic or genetic groups belonging to or related to the "herbicola-agglomerans" complex and values lower than 64% with 56 other species of the Enterobacteriaceae. It was concluded that E. adecarboxylata is a species different from E. agglomerans and the other species of the family Enterobacteriaceae. A new definition of the species E. adecarboxylata is presented.
A collection of 31 strains received as Agrobacterium tumefaciens and A. radiobacter was subjected to detailed phenotypic and genomic studies. These strains were recovered from plants, soil, water and clinical specimens. Type strains of A. tumefaciens, A. radiobacter, A. rhizogenes and A. rubi were also included. The strains were tested for their ability to use 169 organic compounds as sources of carbon and energy. In addition, 11 conventional characters were studied for each strain. Relatedness among the strains was assessed by determining the extent of reassociation in heterologous DNA preparations. S1 nuclease and diethylaminoethyl-cellulose filters were used to separate reassociated from non-reassociated nucleotide sequences, and to determine the thermal stability of related nucleotide sequences. The resultant data revealed the following points: regardless of their phytopathogenic effects, 3-ketolactose-producing A. tumefaciens and A. radiobacter strains group into one species; this species contains 9 taxa which can be differentiated from each other by phenotypic and genomic characters; clinical isolates did not induce tumours on plants and clustered in three taxa of this species; the clinical and ecological significance of these organisms is not known; the present classification of the genus Agrobacterium is based on phytopathogenicity and does not reflect the phylogenetic relationships amongst these bacteria; as proposed previously by several workers, the genus Agrobacterium should be divided into 3 species on the basis of phenotypic and genomic characteristics. Different aspects of the classification and nomenclature of Agrobacterium are discussed.