Effect of the seeding culture media on the biosynthesis of eremomycin

Relationship of eremomycin biosynthesis to the quantity and quality of the inoculate (age, aeration rate, number of subcultures) was studied. The seed medium composition for cultivating the inoculate was defined. A procedure for cultivation of the seed material for biosynthesis of eremomycin providing an increase in the antibiotic yield by 24 per cent was developed.     

Nutrient media for the isolation and accumulation of Listeria

Dried culture media for the isolation and accumulation of Listeria from pathological material and foodstuffs have been developed. The media are suitable for use in bacteriological and sanitary-hygienic practice. The optimum nutrient base has been selected: dried broth (from sprat hydrolysate), produced by the Research and Manufacturing Amalgamation "Culture Media". The optimum concentrations of ingredients, stimulating the growth of Listeria and inhibiting the growth of associated microbes, have been experimentally established. The samples of died accumulation and isolation culture media ensuring the growth of L. monocytogenes, diluted 10(-6), after 24-hour incubation at 37 degrees C have been obtained. The possibility of using these media for the bacteriological diagnosis of listeriosis in pregnant women has been demonstrated.     

Enzyme immunoassay for the determination of the glycopeptide antibiotic eremomycin

An indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed using rabbit polyclonal antibodies against the eremomycin-glucose oxidase conjugated antigen. This technique allows the glycopeptide antibiotic eremomycin to be determined both in aqueous solutions (with a sensitivity as high as 0.1 ng/ml) and in blood plasma. The cross-reactivity of the antibodies with vancomycin was 0.4% of that for eremomycin, while teicoplanin was almost not recognized. Experiments with blood plasma samples diluted 1: 10 showed that the assay was linear over the concentration range 1–30 ng/ml and that the variation coefficient did not exceed 10%. The high sensitivity and selectivity of this test make it suitable for pharmacokinetic studies and drug monitoring analysis.     

Determination of the bacterial contamination of nutrient media for the biosynthesis of antibiotics

Contamination of separate components of nutrient media used in biosynthesis of antibiotics was tested. It was shown that corn steep liquor, corn and soybean meals were the most contaminated components. The results provided determination of the nutrient media contamination levels by the additive factor with respect to the percentage of the components. The safe period of 2-3 hours for the storage of the nutrient media was determined. After that period the media contamination levels markedly increased.     

Nutrient optimization for cellulase biosynthesis by a newly isolatedCellulomonas sp.

Summary A cellulolytic bacterium has been isolated from soil and identified as aCellulomonas sp. Optimization of media components and environmental factors led to a 17-fold increase in cellulase activity within 40 h. The trace elements Zn²⁺, Co²⁺ and Cu²⁺ caused severe inhibition, which could be reduced by the addition of EDTA. The enzymes endoglucanase, exoglucanase and xylanase were always present irrespective of the carbon source used.

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Resumen Se ha aíslado e identificado comoCellulomonas sp. una bacteria cellulolítica del suelo. Mediante la optimización de los componentes del medio de crecimiento y de los factores ambientales se consiguió un incremento de 17 veces de la actividad cellulásica en 40 horas. Los elementos traza Zn²⁺, Co²⁺, y Cu²⁺ causaron una severa inhibición que pudo reducirse gracias a la adición de EDTA. Independientemente de la fuente de carbono utilizada los enzimas endoglucanasa, exoglucanasa y xilanasa se encontraron siempre presentes en el medio.

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Résumé Une bactérie cellulolytique a été isolée à partir du sol et identifiée commeCellulomonas sp. L'activité cellulase en 40 h a été accrue 17 fois par optimisation des constituants du milieu et des conditions de culture. Des traces de Zn²⁺, Co²⁺, et Cu²⁺ déterminent une sévère inhibition, qui peut être réduite par l'addition d'EDTA. Les enzymes endoglucanase, exoglucanase et xylanase sont toujours présentes, quelque soit la source de carbone utilisée.

     

The structure of antibiotic eremomycin B

A new biologically active component, antibiotic eremomycin B, was isolated from the culture liquid of Amycolatopsis orientalis subsp. eremomycini, the producing strain for antibiotic eremomycin. Its structure was established by NMR spectroscopy and mass spectrometry. Eremomycin B was shown to differ from eremomycin by the presence of an N-carboxymethyl substituent in the disaccharide eremosamine fragment.     

Structure of eremosamine--an amino sugar from the antibiotic eremomycin

A new amino sugar named eremosamine was isolated from hydrolysate of eremomycin, an antibiotic belonging to the group of polycyclic glycopeptides. Crystalline derivatives of the amino sugar i. e. methyleremosaminide and methyl-N,O-acetyleremosaminide (alpha- and beta-anomers) were prepared. The data on PMR study and optic properties of the compounds showed that eremosamine had the structure of 2,3,6-tridesoxy-3-amino-3-C-methyl-L-arabinohexose.     

Modification of eremomycin by amine groups

Possible modification of eremomycin, a novel glycopeptide antibiotic by the amine groups with acylating agents such as Ac2O/MeOH and CH3(CH2)7COCl/Et3N and alkylating agents such as CH3CHO, NaBH and NaBH3CN was studied. N-Acetyl, N,N'-diacetyl. N-pelargoil, N-ethyl, N,N'-diethyl and N,N',N"-triethyl derivatives of eremomycin were prepared. Their structure was asserted and the order of the substitute introduction was determined. The antimicrobial activity against Bacillus subtilis and Staphylococcus aureus was assayed. It was found that with introduction of the ethyl substitutes to the eremomycin molecule the antibiotic activity lowered insignificantly whereas the acylation resulted in its decreasing by 1-2 orders.     

Experimental study of histamine-liberating properties of eremomycin

Eremomycin, a new glycopeptide antibiotic showed the same as ristomycin, polymyxins B and M and kanamycin histamine liberating properties. Liberation of endogenic histamine induced destructive lesions on the mucosa of the gastrointestinal tract. The most pronounced lesions were observed after intraperitoneal administration of the antibiotic. When eremomycin was administered intravenously or subcutaneously the affection of the gastrointestinal tract epithelium was less pronounced. After subplantar administration of the antibiotics pad edema in albino rats was observed. The most pronounced edema was after the use of ristomycin and the polymyxins. After the use of eremomycin and kanamycin it was at the average 2 times less pronounced. Preliminary administration of dimedrol decreased intensity of the pad edema induced by the antibiotics.     

Nutrient requirements of lactococci in defined growth media

Many attempts have been made for the last six decades to design defined media for species of the lactococcus group. The general outcome of the studies suggests that this group is heterogeneous with respect to specific requirements for nutrients. Lactococcal species are limited in various metabolic pathways. Early attempts to trace the required nutrients were not always successful because of the poor quality of analysis and the presence of impurities in the medium components. Received: 15 January 1999 / Received revision: 6 April 1999 / Accepted: 9 April 1999     

Nutrient profiling reveals potent inducers of trichothecene biosynthesis in Fusarium graminearum

Fusarium head blight is one of the most important diseases of wheat worldwide due to crop losses and the contamination of grains with trichothecene mycotoxins. The biosynthesis of trichothecenes by Fusarium spp. is highest during infection, but relatively low levels are produced from saprophytic growth in axenic culture. A strain of Fusarium graminearum was constructed where the promoter from the TRI5 trichothecene biosynthesis gene was fused to GFP. Using this strain in large-scale nutrient profiling, a variety of amines were identified that significantly induce TRI5 expression. Analysis of trichothecene levels in the culture filtrates revealed accumulation of the toxin to over 1000 ppm in response to these inducers, levels either greater than or equivalent to those observed during infection. From this work, we propose that products of the arginine-polyamine biosynthetic pathway in plants may play a role in the induction of trichothecene biosynthesis during infection.     

Nutrient regulation of epothilone biosynthesis in heterologous and native production strains

Fermentation media with different initial concentrations of ammonium and phosphate salts were used to study the inhibitory effects of those ions on growth and production of epothilone in Sorangium cellulosum and Myxococcus xanthus. The native epothilone producer, S. cellulosum was more sensitive to ammonium and phosphate than the heterologous producer, M. xanthus. An ammonium concentration of 12 mM reduced epothilone titers by 90% in S. cellulosum but by only 40% in M. xanthus. When 5 mM phosphate was added to the medium, production in both strains was 60% lower. Higher phosphate concentrations had little additional effect on M. xanthus titers, but epothilone production with 17 mM extra-cellular phosphate in S. cellulosum was 95% lower than in the control condition. The effect of iron supplementation to the fermentation medium was also investigated. Both strains showed best production with 20 microM iron added to the medium.