1,3-poly(glycerol phosphate) chains in the cell wall of Streptomyces rutgersensis var. castelarense VKM Ac-238.

The cell wall of Streptomyces rutgersensis var. castelarense contains structurally different chains of 1,3-type glycerol teichoic acid. Part of the molecules consisting of 20-25 monomers, carry on every third glycerol phosphate unit (at C-2) alpha-glucosamine residues, only half of which are N-acetylated. There are chains with O-lysine groups, and free nonsubstituted ones. The chain structure has been ascertained by chemical analysis and 13C- and 1H-NMR spectroscopy.

Separation and determination of polyether carboxylic antibiotics from Streptomyces hygroscopicus NRRL B 1865 by thin-layer chromatography with flame ionization detection.

Thin-layer chromatography coupled with flame ionization detection was used to develop a method to separate and to determine simultaneously three polyether carboxylic ionophore antibiotics (abierixin, nigericin and grisorixin) produced by Streptomyces hygroscopicus NRRL B 1865. Various proportions of chloroform, methanol and formic acid (or acetic acid as a substitute for formic acid) were used in the developing solvent to determine changes in RF values of the antibiotics and to allow conditions for maximum resolution to be obtained. Development on Chromarods SII with chloroform-methanol-formic acid (97:4:0.6, v/v/v) gave satisfactory and reliable separations of the three polyether antibiotics. Under these conditions, the internal standard methyl desoxycholate was found to be suitable for their simultaneous determination in the lipid extracts of Streptomyces hygroscopicus NRRL B 1865.

Structure and synthesis of an immunoactive lipopeptide, WS1279, of microbial origin.

The structure of WS1279, isolated from Streptomyces sp. as an immunoactive lipopeptide, has been deduced on the basis of chemical and physical evidence as S-[2,3-bis(palmitoyloxy)propyl]-N alpha-palmitoyl-Cys-Asn-Ser-Gly-Gly-Ser- OH. This was confirmed by synthesis.

Purification and primary structure of C-1027-AG, a selective antagonist of antitumor antibiotic C-1027, from Streptomyces globisporus.

C-1027-AG, a selective antagonist of antitumor antibiotic C-1027, was isolated by column chromatography on DEAE-cellulose, butyl-Toyopearl and Sephadex G-50 from a culture filtrate of Streptomyces globisporus. The amino acid sequence of purified C-1027-AG was determined with a protein sequencer on the basis of fragment peptides obtained by enzymatic hydrolysis with lysylendopeptidase, V8 protease, endopeptidase AspN and chymotrypsin, after performic acid oxidation. C-1027-AG is shown to consist of a single polypeptide chain cross-linked by two disulfide bonds, and to contain a total of 110 amino acid residues with alanine and glycine as its amino- and carboxyl-termini, respectively; its molecular weight was calculated to be 10,500 daltons. The primary structure of C-1027-AG is indicated to be identical to the protein moiety of C-1027, and is highly homologous to the sequences of antitumor proteins obtained from other Streptomyces species.

The origin of the DNA induced circular dichroism of CC-1065 and analogs.

The calf thymus DNA (CT-DNA) and poly(dI-dC).poly(dI-dC) binding properties of the natural antitumor antibiotic CC-1065 and selected analogs of CC-1065 were studied by circular dichroism (CD) and absorbance methods. The results indicate that the intense long wavelength DNA-induced CD band of these molecules originates from a chiral electronic transition which is delocalized over the whole molecule. Both the covalently bound species (N-3 adenine adduct) and the reversibly bound species exhibit the characteristic spectral behavior of an inherently dissymmetric chromophore when these agents bind within the minor groove of B-form DNA. This mechanism of optical activity accounts for why CC-1065 shows a weak CD in buffer but a very intense induced CD at long wavelength when bound to DNA, why the intensity of the induced CD of CC-1065 analogs depends upon how many fused ring systems the analog contains, and why covalently bound analogs having the mirror image configuration of the natural configuration also exhibit an intense positive induced CD band at long wavelength.

Artritis de rodilla por Streptomyces Somaliensis: un caso / Arthritis of knee for streptomyces somaliensis: a case

Se presenta un caso de artritis de rodilla por Streptomyces somaliensis, raro agente causal de micetomas, que predomina en el continente africano. Es el segundo caso encontrado en la Argentina, con aislamiento e identificación microbiológica del agente causal y el único donde se demostró compromiso intra-articular

Tautomycin from the bacterium Streptomyces verticillatus. Another potent and specific inhibitor of protein phosphatases 1 and 2A.

Tautomycin inhibited the catalytic subunits of protein phosphatase-1 (Kiapp = 0.16 nM) more potently than protein phosphatase 2A (Kiapp = 0.4 nM), and the native forms of these enzymes in mammalian, protozoan and plant extracts were inhibited in a similar manner. Protein phosphatase 2B was inhibited 10,000-fold less potently, while two other phosphatases and six protein kinases were unaffected at 10 microM. Okadaic acid prevented the binding of tautomycin to protein phosphatase 2A, indicating a common binding site for both inhibitors. The different relative potencies of tautomycin and okadaic acid for protein phosphatases 1 and 2A suggest that parallel use of both inhibitors may help to identify physiological substrates for each enzyme.

Procedure for the purification of streptavidin by hydrophobic interaction chromatography.

A procedure is described for the purification of hydrophobic microbial proteins such as streptavidin from Streptomyces avidinii, using Benzyl-DC bead cellulose as the column material. The separation is rapid with a high loading capacity and sufficient resolution for preparative uses. Advantages are discussed especially for industrial purposes.

Three-dimensional structure of an alpha-amylase inhibitor HAIM as determined by nuclear magnetic resonance methods.

The three-dimensional structure of an alpha-amylase inhibitor, HAIM, composed of 78 amino acids, was analyzed by two-dimensional NMR techniques. Sequence-specific assignments were made for the amino acid residues from Ile-6 to Cys-72. Distance geometry analysis of the interresidue NOEs revealed that the HAIM molecule consists of two beta-sheets, as is the case in a homologous alpha-amylase inhibitor, Tendamistat, though one of its beta-strands is much shorter than that of Tendamistat. The combination of molecular modeling from Tendamistat and distance geometry analysis was confirmed to be useful for our purpose.

DNA strand scission by the novel antitumor antibiotic leinamycin.

Leinamycin is a recently discovered antitumor antibiotic with an unusual 1,3-dioxo-1,2-dithiolane structure. It preferentially inhibits the incorporation of [3H]thymidine into the acid-insoluble fraction of Bacillus subtilis. In vitro, leinamycin causes single-strand cleavage of supercoiled double-helical pBR322 DNA in the presence of thiol cofactors. Scavengers of oxygen radical did not supress the DNA-cleaving activity. Thiol-activated leinamycin binds calf thymus DNA at 4 degrees C and thermal treatment of the leinamycin-DNA adduct released a chemically modified leinamycin from the complex. The lack of cytotoxicity and DNA-cleaving activity for S-deoxyleinamycin indicates that the 1,3-dioxo-1,2-dithiolane moiety is essential for the activity of leinamycin. Thus, the primary cellular target of leinamycin appears to be DNA. It binds DNA and causes single-strand break at low concentrations, which may account for the potent antitumor activity.

Proton NMR studies of apo-neocarzinostatin from Streptomyces carzinostaticus. Sequence-specific assignment and secondary structure.

The sequence-specific resonance assignment of apo-neocarzinostatin from Streptomyces carzinostaticus was carried out from two-dimensional proton-NMR spectra. The assignments were obtained for the backbone protons of 111 of the 113 residues of the protein, missing the two C alpha H of one glycine but including 3 of the 4 prolines. The majority of side chain protons were also assigned. The secondary structure derived from the analysis of sequential connections corresponds to ten beta-strands separated by clearly identified loops and turns. Inter-strand connectivities and slowly exchanging amide protons confirm the presence of the two disulfide bridges from Cys37 to Cys47 and from Cys88 to Cys93 and indicate a global folding similar to that of the similar proteins, actinoxanthin and macromomycin, for which crystallographic data are available.

A common ancestor for human placental 17 beta-hydroxysteroid dehydrogenase, Streptomyces coelicolor actIII protein, and Drosophila melanogaster alcohol dehydrogenase.

A computer-based comparison of the amino acid sequences of human placental 17 beta-hydroxysteroid dehydrogenase and Streptomyces coelicolor actIII protein, which is important in the synthesis of the antibiotic actinorhodin, gives an alignment score 12.15 standard deviations higher than that of 1000 comparisons of randomized sequences of these proteins. The probability of getting this score by chance is 3 x 10(-34). Comparison of actIII protein with Drosophila melanogaster alcohol dehydrogenase yields a score of 10.3 standard deviations (P = 3.5 x 10(-25)). Based on these similarities, we propose that 17 beta-hydroxysteroid dehydrogenase, actIII protein, and Drosophila alcohol dehydrogenase are derived from a common ancestor.

Resorthiomycin, a novel antitumor antibiotic. I. Taxonomy, isolation and biological activity.

Resorthiomycin, a novel antitumor antibiotic, was isolated from the fermentation broth of a strain of Streptomyces collinus by ethyl acetate extraction, silica gel chromatography and HPLC. Resorthiomycin exhibited an in vitro cytotoxic activity against mouse leukemia L5178Y cells (IC50, 15.5 micrograms/ml) and also inhibited the clonogenic activity of a multidrug-resistant mutant of human hepatoma PLC/PRF/5 cells to a greater extent than that of the parental cells. On the other hand, this antibiotic does not possess any antibacterial or antifungal activity.

A new inhibitor of protein kinase C, RK-286C (4'-demethylamino-4'-hydroxystaurosporine). II. Isolation, physico-chemical properties and structure.

RK-286C, a new inhibitor of protein kinase C, has been found by the bleb-forming assay using K562 cells. It was produced by Streptomyces sp. RK-286 and purified by solvent extraction, silica gel chromatography and preparative HPLC. Spectrometric analysis revealed that the structure is 4'-demethylamino-4'-hydroxystaurosporine.

Probestin, a new inhibitor of aminopeptidase M, produced by Streptomyces azureus MH663-2F6. I. Taxonomy, production, isolation, physico-chemical properties and biological activities.

Probestin has been isolated as part of a program designed to find microorganism-produced inhibitors of aminopeptidase M from Streptomyces azureus MH663-2F6. It was purified by use of column chromatography of Amberlite XAD-4, silica gel, YMC-gel, Toyopearl HW-40, YMC D-ODS-5 (HPLC) and then isolated as colorless powders. Probestin is competitive with the substrate, and the inhibition constant (Ki) of it was 1.9 x 10(-8) M.

Probestin, a new inhibitor of aminopeptidase M, produced by Streptomyces azureus MH663-2F6. II. Structure determination of probestin.

Probestin, a new inhibitor of aminopeptidase M, has been isolated from the culture broth of Streptomyces azureus MH663-2F6. The 1H and 13C NMR studies and amino acid analysis confirmed the presence of one 3-amino-2-hydroxy-phenylbutanoic acid, leucine and two proline residues in the molecule. Stereochemistries of these amino acids were determined by HPLC analysis. The fragmentation pattern shown in the mass spectrum and the chemical analysis on probestin clarified the amino acid sequence. Thus the structure of probestin was defined as (2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl-L-leucyl-L-prolyl-L-pro line.

Preparative high-performance liquid chromatography of minor products of Streptomyces cinnamonensis.

Analytical and preparative high-performance liquid chromatography of 3 phenazines and furonaphthoquinone derivative on reversed-phase column are described. The mobile phase was methanol and water. The injected amount of the mixture was about 30 mg for a preparative chromatographic run requiring 80 min. Substances were detected directly in the column effluent by UV detection.

Novel cytocidal antibiotics, glucopiericidinols A1 and A2. Taxonomy, fermentation, isolation, structure elucidation and biological characteristics.

Two novel antibiotics, glucopiericidinols A1 (1) and A2 (2) were isolated from the cultured broth of Streptomyces sp. OM-5689. The structures of these two compounds were deduced employing spectroscopic analyses. These antibiotics showed potent cytocidal activities against HeLa S3 cells in vitro (MIC 1: 0.39 microgram/ml, 2: 0.10 microgram/ml) when the cells were exposed to the antibiotics for 3 days. Although 1 and 2 showed no activity at 1,000 micrograms/ml against various Gram-positive and Gram-negative bacteria, yeast or fungi, they did have inhibitory activity against Piricularia oryzae (MIC of 1: 125 micrograms/ml, of 2: 31 micrograms/ml).

Leinamycin, a new antitumor antibiotic from Streptomyces: producing organism, fermentation and isolation.

Leinamycin (DC 107) is newly discovered antitumor antibiotic with an unusual 1,3-dioxo-1,2-dithiolane structure. Five different producing strains were isolated from soils collected in Japan during 1985-1988 and were taxonomically assigned as Streptomyces. Fermentation studies indicate: Leinamycin was unstable in culture broth. A chemically defined medium could be designed for a preferable production. Streptomyces sp. S-140 grew on medium supplemented with Zn2+ and high porus polymer resin and accumulated 32 micrograms/ml of leinamycin. Improved isolation methods are described along with identification of mikamycin A co-produced with leinamycin by the strain S-140.