Showing papers in "The Journal of Antibiotics in 1979"

Thienamycin, a new beta-lactam antibiotic. I. Discovery, taxonomy, isolation and physical properties.

Abstract: A new β-lactam antibiotic, named thienamycin, was discovered in culture broths of Streptomyces MA4297. The producing organism, subsequently determined to be a hitherto unrecognized species, is designated Streptomyces cattleya (NRRL 8057). The antibiotic was isolated by adsorption on Dowex 50, passage through Dowex 1, further chromatography on Dowex 50 and Bio-Gel P2, and final purification and desalting on XAD-2. Thienamycin is zwitterionic, has the elemental composition C11H16N2O4S (M. W.=272.18) and possesses a distinctive UV absorption (λmax=297 nm, e=7, 900). Its β-lactam is unusually sensitive to hydrolysis above pH 8 and to reaction with nucleophiles such as hydroxylamine, cysteine and, to a lesser degree, the primary amine of the antibiotic itself. The latter reaction results in accelerated inactivation at high antibiotic concentrations.

Herbimycin, a new antibiotic produced by a strain of Streptomyces.

Abstract: Herbimycin, a new antibiotic, was isolated from the fermentation broth of Streptomyces hygroscopicus strain No. AM-3672, a soil isolate. The molecular formula of herbimycin was determined to be C30H42N2O9. Herbimycin was found to have potent herbicidal activity against most mono- and di-cotyledonous plants, especially against Cyperus microiria STEUD. However, Oryza sativa showed strong resistance to herbimycin.

Rapamycin (AY-22,989), a new antifungal antibiotic. IV. Mechanism of action.

Abstract: Rapamycin, an antifungal antibiotic produced by Streptomyces hygroscopicus showed a strong candicidal activity, which could not be reversed by sterols. It had no effect on efflux of K+, Pi or U.V. absorbing materials and cell permeability of Candida albicans. Thus, in its action it differs from the polyenes. Mechanism of action of rapamycin appears to be different from many known antifungal agents. In C. albicans, rapamycin at the minimum growth inhibitory concentration inhibited: 1) phosphate incorporation into nucleic acids, 2) acetate incorporation into lipids and 3) substrate respiration of amino acids. The effect on amino acid metabolism was expressed as inhibition of oxidative deamination. At low concentrations rapamycin caused degradation of P32-labeled intracellular macromolecules. Inhibition of threonine incorporation into cell wall and leucine incorporation into cellular protein was observed at relatively higher concentrations of rapamycin. The antibiotic showed no effect on cell-free protein synthesizing systems of Escherichia coli, rat liver and C. albicans and in the mitochondrial enzyme systems. Whether the lethal action of rapamycin on C. albicans is primarily due to one of the above effects or is the result of combined effect on some of these biosynthetic parameters remains to be established.

Antibiotics from basidiomycetes. IX. Oudemansin, an antifungal antibiotic from Oudemansiella mucida (Schrader ex Fr.) Hoehnel (Agaricales).

Abstract: From mycelial cultures of Oudemansiella mucida a crystalline optically active antibiotic, oudemansin (2), has been isolated; its structure is closely related to strobilurin A (1). The relative configuration of oudemansin have been determined by X-ray analysis. The antibiotic exhibits strong antifungal properties and inhibits respiration in fungi, cells of the ascitic form of EHRLICH carcinoma, and rat liver mitochondria.

Holomycin and an antibiotic (MM 19290) related to tunicamycin, metabolites of Streptomyces clavuligerus.

Abstract: Streptomyces clavuligerus produces penicillin N, several cephalosporins and the beta-lactamase inhibitor clavulanic acid. The detection, isolation and properties of further metabolites of this culture, MM 21801 and MM 19290, are described. MM 21801 was identified as the antibiotic holomycin. MM 19290 was shown to be related to tunicamycin, an antibiotic complex obtained from cultures of Streptomyces lysosuperificus.

Antitumor anthracycline antibiotics, aclacinomycin A and analogues. I. Taxonomy, production, isolation and physicochemical properties.

Abstract: Aclacinomycin A and B, two major components of a new antitumor antibiotic complex, and their 19 analogues were produced by a culture of strain No. MA144-M1, which was identified as Streptomyces galilaeus. They were isolated by chelation with copper ion and silicic acid chromatography, and characterized by physicochemical methods in the anthracycline group of antibiotics.

Olivanic acids, a family of beta-lactam antibiotics with beta-lactamase inhibitory properties produced by Streptomyces species. I. Detection, properties and fermentation studies.

Abstract: The screening of soil actinomyces for beta-lactamase inhibitors is described. Using a plate test a number of strains of Streptomyces were found to produce beta-lactamase inhibitory activity designated olivanic acid complex. Factors affecting the production of this complex by Streptomyces olivaceus ATCC 21379 are reported. The complex showed antibacterial activity and also inhibited a number of different types of beta-lactamase in a progressive manner. Certain ampicillin-resistant bacteria are rendered sensitive to ampicillin in the presence of olivanic acid complex at a concentration which alone did not inhibit growth.

Pamamycin: a new antibiotic and stimulator of aerial mycelia formation

Abstract: Pamamycin is a new antibiotic isolated from Streptomyces alboniger ATCC 12461. The antibiotic is active in vitro against Gram-positive bacteria, Neurospora, and Mycobacteria. The compound also acts as a streptomycete differentiation effector. It stimulates aerial mycelia formation in the producing organism. The new antibiotic of elemental composition C36H63NO7 is completely different from puromycin, also produced by this strain. The present communication deals with the isolation, properties, and preliminary characterization of pamamycin.

Antitumor anthracycline antibiotics, aclacinomycin a and analogues. II. Structural determination.

Abstract: The structures of aclacinomycins A and B and 19 analogues were determined by a combination of chemical conversions and degradations and spectral interpretations.

Studies on Actinomycetales producing antibiotics only on agar culture. I. Screening, taxonomy and morphology-productivity relationship of Streptomyces halstedii, strain SF-1993.

Abstract: Soil Actinomycetales that produce antibiotics during growth on agar but not in submerged culture were searched for and about 25 strains were obtained One of these, Streptomyces halstedii, strain SF-1993, which produces an antibiotic designated SF-1993 was studied taxonomically and morphologically The antibiotic-productivity of strain SF-1993 was correlated with mycelial morphology The vegetative mycelium was filamentous in antibiotic-producing agar cultures, but fragmented in non-producing submerged cultures By maintaining submerged cells filamentous, production of antibiotic in the submerged fermentations was accomplished Filamentation was maintained by the use of diluted media or non-fragmented mutant substrains

Antibiotics from basidiomycetes. VII. Crinipellin, a new antibiotic from the basidiomycetous fungus Crinipellis stipitaria (Fr.) Pat.

Abstract: A crystalline antibiotic, which we have named crinipellin, was isolated from submerged cultures of the basidiomycete Crinipellis stipitaria, strain No. 7612. High resolution mass spectrometry yielded the formula C22H28O5. The antibiotic is most active against Gram-positive bacteria, although yeasts and filamentous fungi are affected to a lesser extent. Crinipellin exhibits high in vitro inhibitory activity against the ascitic form of EHRLICH carcinoma. The incorporation of precursors of DNA-, RNA-, and protein syntheses in EHRLICH carcinoma (and in Bacillus brevis) cells was completely inhibited at 5(10) microgram/ml. In Bacillus brevis the inhibition of the incorporation of uridine was found to be due to an interference by crinipellin with the transport of the precursor into the cells.

Equisetin, an antibiotic from Fusarium equiseti NRRL 5537, identified as a derivative of N-methyl-2,4-pyrollidone

Abstract: Sir: Various species of Fusarium produce toxins that are implicated in mycotoxicoses in several areas around the world. During a survey of this genus, we found that one species, Fusarium equiseti (CORDA) SACCARDO, produced an antibiotic in yields of 5 g/kg when grown on corn grit medium at room temperature (20-.24°C). Production, isolation and biological activity of the antibiotic, assigned the trivial name equisetin, have been reported'). Equisetin is active against several strains of Gram-positive bacteria-Bacillus subtilis, Mycobacterium phlei and Staphylococcus aureus-and the Gram-negative bacteria Neisseria perflava, at concentrations of 0.5-. 4.01tg/ml of growth substrate; however, it did not inhibit other Gram-negative bacteria tested nor fungi. The LD50 in mice is 63 mg/kg body weight. Chemical and spectroscopic evidence indicate equisetin to be an N-methyl tetramic acid (1methyl-3-acyl-5-hydroxymethyl-2,4-dione) as shown in Fig. 1. The proposed structure is unique among metabolites insofar as it contains a hydroxymethyl group at position C5 which can form an intramolecular hydrogen bond bridge with the acyl carbonyl group of the /j-triketone system. Elemental analysis and high resolution mass spectroscopy of equisetin (m.p. 65 -. 66°C), as well as microanalysis of its copper salt and tetrahydro derivative gave a molecular composition of C22H3,NO,. The UV spectrum of equisetin with characteristic pH-dependent shifts, its IR spectrum (Table 1), formation of its copper salt and positive FeCI3 and TiCl3 tests are similar to those given by the acyl tetramic acid, tenuazonic acid'-' and the N-methyl tetramic acid, decahydroerythroskyrine3 ) . Presence of a hydroxymethyl group at C5 and an N-methyl lactam in the /3-triketone structure was indicated by NMR, mass spectroscopy and chemical transformations. The 170.0453 fragFig. 1.

Antifungal activity upon Saccharomyces cerevisiae of iturin A, mycosubtilin, bacillomycin L and of their derivatives; inhibition of this antifungal activity by lipid antagonists.

Abstract: The antifungal activity of three antibiotics of the iturin group: iturin A, mycosubtilin, bacillomycin L and of eleven methylated and acetylated derivatives of these antibiotics was tested upon Saccharomyces cerevisiae. The lowest MIC values were found for natural antibiotics. The substitution of polar groups diminished the antifungal activity. Various lipids, sterols, fatty acids, fatty acid methyl esters and phospholipids were tested as inhibitors of the antifungal activity of iturin A, mycosubtilin and bacillomycin L. Cholesterol was the strongest inhibitor upon the three antibiotics; ergosterol, oleic acid and cis-vaccenic acid were less potent inhibitors. Among phospholipids, phosphatidyl choline inhibited bacillomycin L and iturin A while diphosphatidyl glycerol inhibited bacillomycin L and mycosubtilin. The inhibitory effect appeared to be dependent on the nature of both the hydrophilic group and the fatty acid part of phospholipids.

Studies on auromomycin.

Abstract: A new antitumor antibiotic, named auromomycin, was isolated from the culture broth of Streptomyces macromomyceticus, a macromomycin-producing strain. The antibiotic was recovered from the culture filtrate by salting out with ammonium sulfate and further purified by successive application of ion-exchange chromatography on Amberlite IRA-93 (Cl form) and DEAE-Sephadex (OH form), Gel filtration on Sephadex G-50 and hydrophobic chromatography on Octyl-Sepharose CL-4B. The antibiotic is an acidic polypeptide with a molecular weight of 12, 500 and an isoelectric point of pH 5.4 and consists of 16 different amino acids. It has characteristic absorption maxima at 273 nm and 357 nm in the ultraviolet spectrum and two minima at 280 nm and 350 nm in the optical rotatory dispersion spectrum. Auromomycin exhibits antibacterial activity not only against Gram-positive bacteria, but also Gram-negative bacteria. Antitumor activities of auromomycin were revealed against EHRLICH ascites carcinoma, ascites sarcoma 180, L1210 leukemia and LEWIS lung carcinoma.Auromomycin was found to be converted into macromomycin by adsorption chromatography on Amberlite XAD.

Olivanic acids, a family of beta-lactam antibiotics with beta-lactamase inhibitory properties produced by Streptomyces species. II. Isolation and characterisation of the olivanic acids MM 4550, MM 13902 and MM 17880 from Streptomyces olivaceus.

Abstract: The olivanic acids MM 4550, MM 13902 and MM 17880 are members of a new family of β-lactam antibiotics. An isolation and purification process utilising ion-pair extraction and ion-exchange chromatography is described and the metabolites are characterized by physico-chemical and biological properties.

Studies of Metallobleomycins by Electronic Spectroscopy,Electron Spin Resonance Spectroscopy, and Potentiometric Titration

Abstract: The 1:1 bleomycin-A2-Cu(II) complex shows an absorption maximum at 595 nm (epsilon 120), circular dichroism extrema at 555 nm (delta epsilon + 1.21) and 665 nm (-0.61), and electron spin resonance (ESR) signal with g = 2.211, g = 2.055, and A = 178 x 10(-4) cm-1. The formation constant (log K = 12.630) and deprotonation constant (PKc = 3.585) of the 1:1 bleomycin-Cu(II) complex were determined by computer analysis of potentiometric data. The results of potentiometric titration also indicate that the stability of bleomycin-metal complexes is in the order Fe(II) less than Co(II) less than Ni(II) less than Cu(II) greater than Zn(II) and that these divalent metal complexes have a similar coordination environment. The bleomycin-Cu(II) complex has substantially a square-pyramidal configuration in which the secondary amine nitrogen, pyrimidine(N-1) ring nitrogen, deprotonated peptide nitrogen of histidine residue, and histidine imidazole(N-1) nitrogen coordinate to Cu(II) as planar ligand donors, and the alpha-amino nitrogen as axial donor. The specific Cu(II)-binding site of bleomycin has been compared with that of human serum albumin.

DNA strand scission induced by adriamycin and aclacinomycin A.

Abstract: The binding of adriamycin and aclacinomycin A with PM2 DNA, and the consequent cleavage of DNA have been demonstrated by agarose gel electrophoresis, using an ethidium bromide assay. Adriamycin was observed to induce a single strand scission of DNA in the presence of a reducing agent, but aclacinomycin A caused much less degree of DNA breaks. The DNA cleavage was enhanced by Cue2+ and Fe2+, but not significantly by Ni2+, Zn2+, Mg2+ and Ca2+, suggesting that reduction and auto-oxidation of the quinone moiety and H2O2 production participate in the DNA-cutting effect. The DNA degradation was dependent upon concentrations of the anthracyclines and CuCl2. The degree of DNA cleavage at 0.04 mM adriamycin was similar to that at 0.4 mM aclacinomycin A in the presence of 1 mM NADPH and 0.4 mm CuCl2. DNA was degraded to small fragments at 0.4 mM adriamycin and 0.2 mM CuCl2. The anthracycline-induced DNA cleavage was stimulated by H2O2, but partially inhibited by potassium iodide, superoxide dismutase, catalase and nitrogen gas atmosphere. The results suggested that both free radical of anthracycline quinones and hydroxyl radical directly react with DNA strands.

Biosynthetic relationships among the secalonic acids. Isolation of emodin, endocrocin and secalonic acids from Pyrenochaeta terrestris and Aspergillus aculeatus.

Abstract: Cynodontin, emodin, endocrocin and secalonic acids A, E and G have been isolated from five strains of Pyrenochaeta terrestis. Aspergillus aculeatus produces emodin, endocrocin and secalonic acids B, D and F. No cynodontin was detected. Isolation of emodin in small amounts supports previous evidence that it is an intermediate in secalonic acid biosynthesis. Isolation of cynodontin and endocrocin, which are co-produced with secalonic acids in other organisms, suggests that these compounds are formed by a common branching pathway. A natural isolate of P. terrestris contained variant strains which produced different relative amounts of secalonic acids A, E and G. From the combinations of secalonic acids produced in organisms so far examined it is concluded that precursor tetrahydroxanthone units or formed in pairs differing in stereo-chemistry only at C-5 or at the trans-invariant C-6 : C-10a positions. A possible biosynthetic pathway is discussed.

Four further antibiotics related to olivanic acid produced by Streptomyces olivaceus: fermentation, isolation, characterisation and biosynthetic studies.

Abstract: Four β-lactam antibiotics with β-lactamase inhibitory activity MM 22380, MM 22381, MM 22382 and MM 22383 containing the carbapenem nucleus have been isolated from aculture of Streptomyces olivaceus ATCC 31365. Fermentation conditions for their productionand methods for their isolation are described. Evidence for a biosynthetic link between thesecompounds and the previously described olivanic acid derivatives MM 4550, MM 13902 andMM 17880 is presented.

Isolation and characterization of antibiotic X-14547A, a novel monocarboxylic acid ionophore produced by Streptomyces antibioticus NRRL 8167.

Abstract: A novel carboxylic acid ionophore, antibiotic X-14547A, closely related to the polyether antibiotics has been isolated along with four other metabolites from fermented cultures of a new strain of Streptomyces antibioticus The structure, determined by X-ray analysis of the R(+)-1-amino-1-(4-bromophenyl)-ethane salt contained pyrrole carbonyl and trans-butadienyl chromophores in addition to the unusual tetrahydroindane bicyclic ring system A second novel metabolite was identified as 3-ethyl-1,3-dihydro-3-methoxy-2H-indol-2-one

Biological studies on the degradation products of 3-[(s)-1'-phenylethylamino]propylaminobleomycin: a novel analog (pepleomycin)

Abstract: Pepleomycin (PEP), 3-[(S)-1'-phenylethylamino]propylaminobleomycin has potent activity and is less pulmonary toxic than bleomycin (BLM). Biological activity and toxicity of the following degradation products of PEP have been studied in detail: the product of carbamoyl migration (ISO), the product of decarbamylation (DC), the product of ring closure of the side chain on the pyrimidine moiety (RC), the depyruvamide product (DP) and the product of an enzymatic inactivation (DA). These degradation products showed much lower activity than PEP in vitro: antimicrobial and anti-HeLa activities, inhibition of DNA synthesis in AH66 cells and the DNA strand cleavage. Acute toxicity and pulmonary toxicity were tested in mice. Results indicated much lower acute toxicity corresponding to the decreased in vitro activity when compared to PEP. DP and RC did not cause lung fibrosis in mice, while ISO and DC showed 1/2.6 and 1/5.7 degree of pulmonary toxicity, respectively, in comparison with PEP.

A new antitumor antibiotic, bactobolin produced by Pseudomonas.

Abstract: Sir: A new chlorine-containing antibiotic, bactobolin, has been found in the culture broth of Pseudoinonas BMG13-A7. Bactobolin inhibits the growth of Gram-positive and Gram-negative bacteria, and experimental animal tumors, and is structurally related to actinobolin produced by Streptomyices griseoviridis var. atrofaciens1,2). Psendonionas BMG 13-A7 was cultured at 27°C for 24 hours on a rotatory shaker (180 rpm) in a 500-m1 Erlenmeyer flask which contained 110 ml of a medium consisting of maltose 1.5 %, yeast extract 0.3%, NZ-amine (A type) 1.0% and NaCl 0.3%. (pH 7.4). This culture (110 ml) was inoculated into 15 liters of the same medium in a 30-liter jar fermentor and the fermentation was carried out at 27°C under agitation at 250 rpm and aeration at 15 liters/minute. The 40hour fermented broths in two fermentors were combined and centrifuged at 10,000 rpm to yield a supernatant (27 liters, 52 μg/ml). Concentrations of bactobolin were determined by the cylinder plate method against Escherichia coli K-12 using crystalline bactobolin (1,000μg/ml) as an assay standard. The antibiotic in the supernatant was adsorbed on a column of Amberlite XAD-2 resin (1,500 nil) and eluted with 50% aqueous methanol. The active eluate (4,270 ml) was concentrated to dryness and a crude powder (13.4 g, 71μrg/mg) was obtained. The crude powder in water (20 ml) was purified by column chromatography on Amberlite CG-50 (Type I, 70 % NH4+, 600 ml) developed with water to give an yellowish powder (1.7 g, 489μg/mg). The powder was further purified by repeated chromatography, yielding a colorless powder (940 mg, 719 μg/mg) of bactobolin. Crystallization of the powder from a mixture of water (1.5 ml) and ethanol (0.5 ml) gave colorless needles of bactobolin (604 mg, 1,000 μg/mg). Bactobolin (1) shows mp 196~197°C (decomp.), [a]24D -26.7° (c 1.0, water), pKa' 7.3 and 8.4. Anal. calcd. for C14H20N2O6Cl2: C 43.88, H 5.26, N 7.31, 0 25.05. Found: C 43.57, H 5.22, N 7.33, 0 24.99. The molecular formula is derived by FD-MS of 1 (m/e 382, M+) and by high-resolution MS of monoN-acetylbactobolin (2) (m/e 424.0789, calcd. mol. wt. for C16H22N2O7Cl2: 424.0801). UV max.: 276 nm (e 10,000) in water, 262 nm (L: 11,200) in 0.1 N HCl and 287 nm (e 17,000) in 0.1 N NaOH. The IR spectrum is shown in Fig. 1. The PMR chemical shifts are represented in Table 1. The CMR spectra (D2O) shows signals of 14 carbons (184.7s, 173.9s, 167.6s, 85.3s, 84.3s, 75.9d, 72.6d, 70.1d, 50.0d, 47.7d, 42.0d, 40.3t, 19.lq and 19.0q). The antibiotic 1 gives positive ninhydrin, RYDON-SMITH and 2,4dinitrophenylhydrazine reactions, and is soluble in water and methanol. By high-voltage paper electrophoresis with 3,500 V for 15 minutes in formic acid acetic acid water (1: 3 : 36, v/v), the antibiotic 1 moves to the cathode with Rm (relative mobility to alanine) 0.68. Rf values of thin-layer chromatography on Silica gel G (Merck, Art. 5715) developed with ethyl acetate -

Synthesis of a tight-binding, multisubstrate analog inhibitor of gentamicin acetyltransferase I.

Abstract: Gentamicin acetyltransferase I will catalyze acyl transfer from chloroacetylcoenzyme A to form 3-N-chloroacetylgentamicin. This product can be linked to coenzyme A to form a multisubstrate analog by nucleophilic displacement of the chlorine by the sulfur of coenzyme A. The analog can be purified by selective binding to cationic and anionic ion exchange resins. Kinetic analysis of a time-dependent onset and reversal of inhibition of gentamicin acetyltransferase I by the purified multisubstrate analog yields an inhibition constant of 5-20×10-10M. The inhibitor does not potentiate antibiotic activity against resistant Escherichia coli. Nevertheless, the effectiveness of the tight-binding between the enzyme and the multisubstrate analog demonstrates that inhibitors of resistance can be designed and prepared by specific enzymatic synthesis.

Neoxaline, a new alkaloid produced by Aspergillus japonicus. Production, isolation and properties.

Abstract: A new alkaloid named neoxaline has been isolated from culture broth of Aspergillus japonicug Fg-551 by solvent extraction and silica gel chromatography. The compound does not possess antimicrobial activities, but weakly stimulates the central nervous system. The molecular formula of neoxaline has been determined as C23H25N5O4 on the basis of elemental analysis and its mass spectrometry.

The comparative .BETA.-lactamase resistance and inhibitory activity of 1-oxa cephalosporin, cefoxitin and cefotaxime.

Abstract: The β-lactamase stability and inhibitory activity of 1-oxa cephalosporin, (6R, 7R)-7-[[carboxy(4-hydroxyphenyl)acetyl]amino]-7-methoxy-3-[[(1-methyl-1H-tetrazol-5-yl)thio]methyl]-8-oxo-5-oxa-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid, was investigated and compared to that of cefoxitin and cefotaxime. There was no detectable β-lactamase hydrolysis of 1-oxa cephalosporin, cefotaxime and cefoxitin when incubated with β-lactamases of plasmid or chromosomal origin which were primarily cephalosporinases or enzymes which hydrolyzed both penicillins and cephalosporins. The β-lactamase inhibitory activity of 1-oxa cephalosporinwas comparable to that of cefoxitin and cefotaxime. At equal molar concentration of substrate and inhibitor, cefoxitin, cefotaxime and 1-oxa cephalosporin effectively inhibited cephalosporinase hydrolysis of cephaloridine. Cefoxitin and cefotaxime were more effective inhibitors than the 1-oxa cephalosporin against a Providencia enzyme, whereas cefotaxime and 1-oxa cephalosporin were more effective inhibitors of a Citrobactercephalosporinase.