1. |
Mukherjee G,
Sen SK,
( 2006 ) Purification, characterization, and antifungal activity of chitinase from Streptomyces venezuelae P10. PMID : 16972135 : DOI : 10.1007/s00284-005-0412-4 Abstract >>
Streptomyces venezuelae P(10) could produce extracellular chitinase in a medium containing 0.6% colloidal chitin that was fermented for 96 hours at 30 degrees C. The enzyme was purified to apparent homogeneity with 80% saturation of ammonium sulfate as shown by chitin affinity chromatography and DEAE-cellulose anion-exchange chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the enzyme showed a molecular weight of 66 kDa. The chitinase was characterized, and antifungal activity was observed against phytopathogens. Also, the first 15 N-terminal amino-acid residues of the chitinase were determined. The chitin hydrolysed products were N-acetylglucosamine and N, N'-diacetylchitobiose.
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2. |
Everest GJ,
Cook AE,
Kirby BM,
Meyers PR,
( 2011 ) Evaluation of the use of recN sequence analysis in the phylogeny of the genus Amycolatopsis. PMID : 21671192 : DOI : 10.1007/s10482-011-9604-z Abstract >>
Partial recN gene sequences (>1 kb) were obtained from 35 type strains of the genus Amycolatopsis. Phylogenetic trees were constructed to determine the effectiveness of using this gene to predict taxonomic relationships within the genus. The use of recN sequence analysis as an alternative to DNA-DNA hybridization (DDH) for distinguishing closely related species was also assessed. The recN based phylogeny mostly confirmed the conventional 16S rRNA and gyrB gene-based phylogenies and thus provides further support for these phylogenetic groupings. As is the case for the gyrB gene, pairwise recN sequence similarities cannot be used to predict the DNA relatedness between type strains but the recN genetic distance can be used as a means to assess quickly whether an isolate is likely to represent a new species in the genus Amycolatopsis. A recN genetic distance of >0.04 between two Amycolatopsis strains is proposed to provide a good indication that they belong to different species (and that polyphasic taxonomic characterization of the unknown strain is worth undertaking).
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3. |
Bayer EA,
Kulik T,
Adar R,
Wilchek M,
( 1995 ) Close similarity among streptavidin-like, biotin-binding proteins from Streptomyces. PMID : 7632734 : DOI : 10.1016/0167-4781(95)00077-t Abstract >>
Two strains of Streptomyces venezuelae were found to produce high-affinity, biotin-binding proteins, termed streptavidin v1 and v2, respectively. Both proteins were isolated to purity, and their corresponding genes were cloned and sequenced. Compared to streptavidin from S. avidinii, streptavidin v1 had only a single amino acid substitution and streptavidin v2 showed 9 such differences. The substitutions were remarkably conservative, none of which affected the amino acid residues known to be important to the biotin-binding properties or to the structure of the tetrameric protein. The results also indicate that the biosynthesis of such biotin-binding proteins is not simply a curious anomaly in a single species of Streptomyces. It is suggested that the classification of S. avidinii as a unique species should be reconsidered. The occurrence of these proteins appears to be linked to the production of an unusual synergistic antibiotic complex.
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4. |
Mosher RH,
Camp DJ,
Yang K,
Brown MP,
Shaw WV,
Vining LC,
( 1995 ) Inactivation of chloramphenicol by O-phosphorylation. A novel resistance mechanism in Streptomyces venezuelae ISP5230, a chloramphenicol producer. PMID : 7592948 : DOI : 10.1074/jbc.270.45.27000 Abstract >>
Plasmid pJV4, containing a 2.4-kilobase pair insert of genomic DNA from the chloramphenicol (Cm) producer Streptomyces venezuelae ISP5230, confers resistance when introduced by transformation into the Cm-sensitive host Streptomyces lividans M252 (Mosher, R. H. Ranade, N. P., Schrempf, H., and Vining, L. C. (1990) J. Gen. Microbiol. 136, 293-301). Transformants rapidly metabolized Cm to one major product, which was isolated and purified by reversed phase chromatography. The metabolite was identified by nuclear magnetic resonance spectroscopy and mass spectrometry as 3'-O-phospho-Cm, and was shown to have negligible inhibitory activity against Cm-sensitive Micrococcus luteus. The nucleotide sequence of the S. venezuelae DNA insert in pJV4 contains an open reading frame (ORF) that encodes a polypeptide (19 kDa) with a consensus motif at its NH2 terminus corresponding to a nucleotide-binding amino acid sequence (motif A or P-loop; Walker, J. E., Saraste, M., Runswick, M. J., and Gay, N. J. (1982) EMBO J. 1, 945-951). When a recombinant vector containing this ORF as a 1.6-kilobase pair SmaI-SmaI fragment was used to transform S. lividans M252, uniformly Cm-resistant transformants were obtained. A strain of S. lividans transformed by a vector in which the ORF had been disrupted by an internal deletion yielded clones that were unable to phosphorylate Cm, and exhibited normal susceptibility to the antibiotic. The results implicate the product of the ORF from S. venezuelae as an enzymic effector of Cm resistance in the producing organism by 3'-O-phosphorylation. We suggest the trivial name chloramphenicol 3'-O-phosphotransferase for the enzyme.
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5. |
Virolle MJ,
Long CM,
Chang S,
Bibb MJ,
( 1988 ) Cloning, characterisation and regulation of an alpha-amylase gene from Streptomyces venezuelae. PMID : 3266752 : DOI : 10.1016/0378-1119(88)90166-7 Abstract >>
The alpha-amylase gene (aml) of Streptomyces venezuelae ATCC15068 was cloned in Streptomyces lividans TK24 using the plasmid vector pIJ702. Sub-cloning and exonuclease III deletion experiments localised the sequences required for alpha-amylase production to a segment of 2.05 kb. Low-resolution nuclease S1 mapping revealed a aml transcript of approx. 1.7 kb, and the extracellular form of alpha-amylase was estimated by SDS-polyacrylamide gel electrophoresis to be 59 kDa, suggesting that aml mRNA is monocistronic. The nucleotide sequence of aml was determined and high-resolution nuclease S1 mapping experiments identified transcripts that appeared to initiate at a promoter identical to that of the alpha-amylase gene of Streptomyces limosus [Long et al., J. Bacteriol. 169 (1987) 5745-5754]. Transcription of aml in S. venezuelae, and of the cloned gene in Streptomyces coelicolor A3(2), was induced by maltose and repressed by glucose. Glucose repression in S. coelicolor A3(2) depended on a functional glucose kinase gene. The predicted amino acid sequence of the extracellular enzyme was very similar (75% identity) to the alpha-amylase of S. limosus and shared with this enzyme a strong susceptibility to tendamistat, a potent inhibitor of mammalian alpha-amylases. Sequence inspection revealed a putative signal sequence of 28 amino acids that preceded the probable signal peptidase cleavage site.
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6. |
( 1998 ) Molecular characterization of a novel subtilisin inhibitor protein produced by Streptomyces venezuelae CBS762.70. PMID : 9773272 : Abstract >>
We report here on the isolation and identification of a gene coding for a novel subtilisin inhibitor (VSI) isolated from Streptomyces venezuelae CBS762.70. The vsi gene was isolated on a 5-kb chromosomal PvuII fragment as identified by DNA sequencing and inhibitor activity testing of the gene product. Primer extension studies revealed that the mRNA transcriptional start point was situated at -37 and -36 relatively to the ATG start codon assuming the presence of solely one promoter. Vsi promoter strength was about double of those of ermE-P1a and aph-P1, as tested with the mRNA production of the aphII gene preceded by the respective promoters. Translation of the vsi coding sequence revealed a 28 amino acids long signal peptide. The mature VSI protein consists of 118 amino acids of which 87% was verified by N-terminal amino acid sequence analysis. Compared with the already known Streptomyces proteinase inhibitors, VSI shows a relatively high amino acid identity in the conserved domains. Nevertheless, only a maximum amino acid identity of 56.1% was noticed and some highly conserved residues were substituted in VSI. As a consequence, VSI could be classified within a separate group of Streptomyces subtilisin inhibitors.
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