Kitasatospora cheerisanensis sp. nov., a new species of the genus Kitasatospora that produces an antifungal agent.
An actinomycete, strain YC75T, which produced bafilomycin-like antifungal compounds, was identified as a member of the genus Kitasatospora on the basis of morphological and chemotaxonomic characteristics. The strain produced the aerial and fragmenting vegetative mycelia consisting of straight chains of 20 or more smooth-surfaced spores. Submerged spores were formed in tryptic soy broth. No soluble pigments were formed. Whole-cell hydrolysates contained glucose and mannose, but not galactose. The 16S rDNA sequence of YC75T was compared with those of the other representative kitasatosporae and streptomycetes. Strain YC75T formed a significant monophyletic clade with Kitasatospora phosalacinea. The levels of DNA relatedness between strain YC75T and representatives of the genus Kitasatospora ranged from 16 to 59% including K. phosalacinea (28 and 40%). It is clear from polyphasic evidence that the isolate should be classified as Kitasatospora cheerisanensis sp. nov., whose type strain is YC75T (= KCTC 2395T). The presence of galactose in whole-cell hydrolysates may not be a stable chemical marker for the genus Kitasatospora. (+info)
Sequence analysis of domains III and IV of the 23S rRNA gene of verticillate streptomycetes.
Domains III and IV of the 23S rRNA gene of 25 strains of verticillate streptomycetes were sequenced. None of the sequences was identical to any other, with regions of variability being restricted to parts of helices 54 and 64. No relationships were detected between the similarities in the sequence and the assignment to phenetic clusters as defined by the numerical taxonomy studies. Limited agreement was also found between similarity of the sequences and DNA-DNA homology values. However, species (> 70% DNA-DNA homology values)-specific diagnostic oligonucleotides generally could be defined, except for Streptomyces baldaccii. Therefore the determination of the 23S rRNA sequence may be of greater value for fingerprinting individual strains than for taxonomic or identification purposes. (+info)
Overproduction of microbial transglutaminase in Escherichia coli, in vitro refolding, and characterization of the refolded form.
(MTG) The Streptoverticillium transglutaminase gene, synthesized previously for yeast expression, was modified and resynthesized for overexpression in E. coli. A high-level expression plasmid, pUCTRPMTG-02(+), was constructed. Furthermore, to eliminate the N-terminal methionine, pUCTRPMTGD2 was constructed. Cultivation of E. coli transformed with pUCTRPMTG02(+) or pUCTRPMTGD2 yielded a large amount of MTG (200-300 mg/liter) as insoluble inclusion bodies. The N-terminal amino acid residue of the expressed protein was methionine or serine (the second amino acid residue of the mature MTG sequence), respectively. Transformed E. coli cells were disrupted, and collected pellets of inclusion bodies were solubilized with 8 M urea. Rapid dilution treatment of solubilized MTG restored the enzymatic activity. Refolded MTG, purified by ion-exchange chromatography, which had an N-terminal methionine or serine residue, showed activity equivalent to that of native MTG. These results indicated that recombinant MTG could be produced efficiently in E. coli. (+info)
Description of two novel species of the genus Kitasatospora Omura et al. 1982, Kitasatospora cineracea sp. nov. and Kitasatospora niigatensis sp. nov.
Five actinomycete strains, SK-3255T, SK-3406T, SK-3412, SK-3421 and OM-5023, were isolated using a novobiocin-containing agar medium from soil samples. These strains produced long spore chains on aerial mycelium and contained LL- and meso-diaminopimelic acids (DAPs) in the cell wall. On the basis of morphological and chemotaxonomic characteristics and phylogenetic analysis, these five strains were classified into the genus Kitasatospora. DNA-DNA hybridization and comparison of physiological characteristics revealed that strains SK-3255T and SK-3406T differed from known species. Strains SK-3406T, SK-3412 and SK-3421 were regarded as the same species. Strain OM-5023 was identified as Kitasatospora griseola. Therefore, two novel species are proposed, Kitasatospora cineracea sp. nov. and Kitasatospora niigatensis sp. nov., with the type strains K. cineracea SK-3255T (= IFO 16452T = JCM 10915T = NRRL B-23134T) and K. niigatensis SK-3406T (= IFO 16453T = JCM 10916T = NRRL B-24135T). (+info)
Purification and some properties of an inhibitor for a yeast pleiotropic drug resistant pump from Kitasatospora sp. E-420.
A strain producing an inhibitor for pleiotropic drug resistant 5 (Pdr5) was isolated using our original screening system in yeast. The strain was classified and named as Kitasatospora sp. E-420. The purified inhibitor (molecular weight = 1,193 by FAB-MS) inhibited a Pdr5-mediated efflux of cycloheximide and cerulenin. The intracellular accumulation of a fluorescent dye, rhodamine 123, by the inhibitor was also confirmed. Some physicochemical data suggested that the Pdr5-specific inhibitor was different from an immunosuppressant, FK506, reported as the only inhibitor of Pdr5 in vivo. (+info)
Prodiginine (prodigiosin-like) pigments from Streptoverticillium rubrireticuli, an organism that causes pink staining of polyvinyl chloride.
Red pigments were extracted from Streptoverticillium rubrireticuli strain 100-19, an organism frequently incriminated in pink staining of polyvinyl chloride. These pigments were identified as undecylprodiginine and butylcycloheptylprodiginine. (+info)
Secretion of active-form Streptoverticillium mobaraense transglutaminase by Corynebacterium glutamicum: processing of the pro-transglutaminase by a cosecreted subtilisin-Like protease from Streptomyces albogriseolus.
The transglutaminase secreted by Streptoverticillium mobaraense is a useful enzyme in the food industry. A fragment of transglutaminase was secreted by Corynebacterium glutamicum when it was coupled on a plasmid to the promoter and signal peptide of a cell surface protein from C. glutamicum. We analyzed the signal peptide and the pro-domain of the transglutaminase gene and found that the signal peptide consists of 31 amino acid residues and the pro-domain consists of 45 residues. When the pro-domain of the transglutaminase was used, the pro-transglutaminase was secreted efficiently by C. glutamicum but had no enzymatic activity. However, when the plasmid carrying the S. mobaraense transglutaminase also encoded SAM-P45, a subtilisin-like serine protease derived from Streptomyces albogriseolus, the peptide bond to the C side of 41-Ser of the pro-transglutaminase was hydrolyzed, and the pro-transglutaminase was converted to an active form. Our findings suggest that C. glutamicum has potential as a host for industrial-scale protein production. (+info)
A new IL-1 receptor inhibitor 139A: fermentation, isolation, physico-chemical properties and structure.
Interleukin (IL)-1 is known to be a cytokine which plays a major role in pathological conditions like septic shock, inflammation and auto-immune disease, hence, methods that reduce the activity of IL-1 have an impact on clinical medicine. Inhibiting the binding of IL-1 to IL-1 receptors is one of the methods. A new inhibitor of IL-1 receptor, 139A, was isolated from the fermentation broth of Streptomyces sp.139. It was extracted from the broth filtrate, purified by Diaion HP-20, cation exchange resin and DEAE Sephadex A-25. 139A was identified as polysaccharide, its structure was elucidated on the basis of spectral analysis, the immobilized ligand IL-1 receptor binding assay (IL-ILRBA) proved 139A can competitively inhibits the binding of IL-1 to IL-1 receptors. (+info)