Assuntos
Infecções por Actinomycetales/diagnóstico , Transplante de Medula Óssea/efeitos adversos , Transplante de Medula Óssea/imunologia , Bactéria Gordonia/isolamento & purificação , Hospedeiro Imunocomprometido , Sepse/diagnóstico , Infecções por Actinomycetales/tratamento farmacológico , Adulto , Antibacterianos , Sequência de Bases , DNA Bacteriano/análise , Quimioterapia Combinada/administração & dosagem , Feminino , Seguimentos , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Leucemia Mielogênica Crônica BCR-ABL Positiva/cirurgia , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Medição de Risco , Sepse/tratamento farmacológico , Índice de Gravidade de Doença , Transplante Homólogo/efeitos adversos , Resultado do TratamentoRESUMO
Two cis-1,4-polyisoprene (isoprene rubber) degrading bacteria, strains VH2 and Y2K, were identified as strains of the species Gordonia polyisoprenivorans belonging to the Corynebacterineae, a suborder of the order Actinomycetales. Both showed characteristic growth and degradation of isoprene rubber as described previously for the type strain of G. polyisoprenivorans Kd2 (DSM 44302(T)). For strain VH2 the chemotaxonomic properties were investigated, and DNA-DNA hybridization experiments with the type strain revealed the affiliation to the species G. polyisoprenivorans. The comparison of the 16S rDNA sequences, and especially hyper variable regions of these, led to the classification of strain Y2K to the same species. At present, the species G. polyisoprenivorans comprises three different isolates which share the ability to degrade isoprene rubber potently but which were obtained from different geographic regions.
Assuntos
Actinomycetales/classificação , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Borracha/metabolismo , Actinomycetales/química , Actinomycetales/genética , Sequência de Bases , Cromatografia Líquida de Alta Pressão , Ácidos Graxos/análise , Dados de Sequência Molecular , RNA Ribossômico 16S/análise , Alinhamento de Sequência , Especificidade da Espécie , Vitamina K 2/análiseRESUMO
A gene transfer system for Rhodococcus opacus PD630 based on electroporation was established and optimized employing the Escherichia coli-Rhodococcus shuttle vectors pNC9501 and pNC9503 as well as the E. coli-Corynebacterium glutamicum shuttle vector pJC1 as suitable cloning vectors for R. opacus PD630, resulting in transformation efficiencies up to 1.5 x 10(5) CFUs/microgram plasmid DNA. Applying the optimized electroporation protocol to the pNC9501-derivatives pAK68 and pAK71 harboring the entire PHB synthesis operon from Ralstonia eutropha and the PHA synthase gene phaC1 from Pseudomonas aeruginosa, respectively, recombinant PHA biosynthesis was established in R. opacus PD630 and the TAG-negative mutant ROM34. Plasmid pAK68 enabled synthesis and accumulation of poly(3HB) in R. opacus PD630 and ROM34 during cultivation under storage conditions from 1% (w/v) gluconate, of poly(3HB-co-3HV) from 0.2% (w/v) propionate and of poly(3HV) from 0.1% (w/v) valerate. Under storage conditions, recombinant strains of PD630 and ROM34 harboring pAK71 were able to synthesize and accumulate PHA of the medium chain length hydroxyalkanoic acids 3HHx, 3HO, 3HD and 3HDD from 0.1% (w/v) hexadecane or octadecane and a copolyester composed of 3HHp, 3HN and 3HUD from 0.1% (w/v) pentadecane or heptadecane. In the recombinant strains of PD630 and ROM34, the thiostrepton-induced overexpression of a 20 kDa protein was observed with its N-terminus exhibiting a homology of 60% identical amino acids to TipA from Streptomyces lividans.
