Z. Řeháček

Effect of Tween 80 on alkaloid-producing cultures ofClaviceps paspali

Addition of Tween 80 to submerged cultures ofClaviceps paspali (Stevens and Hall) growing in a simple defined medium increased biomass formation and caused a temporary change in alkaloid synthesis intensity. The Tween-supplemented culture reached maximal alkaloid yields four days earlier than the control. The shift of alkaloid production was associated with a shift of organic acids and amino acids in the cell-pool. Thus the maximal formation of alkaloids was characterized by a decrease in the amount of citric acid, by the disappearance of succinic, fumaric and oxaloacetic acids and by increased accumulation of methionine, cysteine, alanine and histidine. The slow alkaloid synthesis was accompanied by a relatively high content of citric and succinic acids in the cell-pool and by the absence of methionine. The data are consistent with the hypothesis that ergot alkaloids participate in the regulation of the metabolism of cultures.

Taxonomic characteristic of the strain ETH 7437 producing granaticin

The streptomycete strain ETH 7437, producing granaticin, was characterized according to present taxonomic standards and compared with the strainsStreptomyces olivaceus (Waksman) Waksman and Henrici,Streptomyces coelicolor (Müller) andStreptomyces violaceruber (Waksman & Curtis). The strain ETH 7437 formed gray aerial mycelium, straight and flexuos sporophores, even single hooks and open spirals. Normally the amount of spores in chains was over 50, the surface of spores being smooth. The strain utilizedd-glucose,d-arabinose,d-xylose andd-fructose. The strain was capable of forming antibiotic pigment stably. For the strain ETH 7437 the termStreptomyces granaticolor was proposed.

Preparation of inoculum from sclerotia of the ascomyceteClaviceps purpurea

Slices of pleetenchymatous tissues of the purified sclerotia are used for preparation of saprophytic cultures of Claviceps purpurea producing ergot alkaloids (Kirchhoff, 1929; GrSger & Tyler, 1963). This procedure is often accompanied by contamination which decreases the number of the obtained cultures available for inoculation purposes. In this paper we wish to describe a procedure according to which it is possible to prepare more of the inoculation material and at the same time to reduce the hazard of contamination considerably. Sclerotia of the strain Claviceps purpurea (Fr.) Tul. were used in our work. For the disinfection of their surface following procedures were employed: 1) Soaking of sclerotia in water followed by 25 min submersion in a sublimate solution (0.1%) with an admixture of tween (0.1%); 2) plunging of sclerotia (30 min) into a solution of famosept containing tween (0.1%); 3) mechanical cleaning with scalpel, dipping into a 50% solution of propanol (2 min) and a 4% formaldehyde solution (2 min) (GrSger & Tyler, 1963). The efficiency of the disinfection procedures was examined after a 7 day incubation of the sclerotia washed in water on malt agar (8 ~ Bal.) at 28 ~ C. The best results have been shown by procedure 3 yielding a massive myeelial growth proving tha t the germinating ability of the cultures remained unaffected. For the preparation of the inoculum the disinr

Some relations between the morphology and composition of the mycelium ofstreptomyces erythreus and the biosynthesis of erythromycin

SummaryStreptomyces erythreus formed two types of mycelium hyphae under submerged conditions in a complex medium: Inoculum spores produced long, little branched hyphae, from which thick hyphae grew at the beginning of the rise in production activity. The synthetic activity of the mycelium changed concurrently with the intensity of growth of the microorganism. The mycelium was most productive during the phase of early autolysis.With increasing growth intensity and production activity, the content of sugar components of mycelium cell walls decreas ed. In completely hydrolyzed media of different ages the concentration of alanine, glutamic acid and glycine changed concurrently with the growth and production activity of the actinomycete.α-aminobutyric acid was detected only in mycelium hydrolyzates from the beginning of the sharp decrease in growth intensity of the micro-organism.AbstractStreptomyces erythreus формируется двумя типамимицелия hyphae под во дуусловия в сложной ср еде: Inoculum спор производств а давно, мало разветвле нныхhyphae, из которого выро с густой hyphae на начало рас ти в сфере производства деятельности. Синтетич еской активностимицел ий изменилось одноврем енно сИнтенсивность ро ста микроорганизмаМиц елий был самым продукти вным в течениераннем эт апе autolysis.По мере усиления и нтенсивности и роста производственной деят ельности, содержание са харамицелий компонент ов клеточной стенки decreas изд. В средствах массов ой информации полность ю hydrolyzed разных возрастов к онцентрацииаланин, glutamic кислота и глицин измени лосьаланин, glutamic кислота и глицин изменилосьодн овременно с ростом про изводства иДеятельно сть actinomycete. га-aminobutyric кислота была обнаружена только в мицелий hydrolyzates с начала от резкого снижения тем пов роста интенсивности от микроорганизмов.

Incidence and possibilities of screening antifungal non-polyene antibiotics in Actinomycetes.

The author studied 114 strains of actinomycetes recovered from different geographical localities with different vegetation. Under submerse cultivation conditions 57% of the strains displayed antibiotic activity—48% antibacterial, 56% antifungal. As distinct from intracellular antibiotics, production of extracellular antibiotics was manifestly influenced by the composition of the fermentation medium. The commonest antifungal antibiotics were polyenes and non-polyenes, in the ratio 2∶1 Twenty strains produced non-polyene antifungal antibiotics, 14 of which corresponded to substances described in the literature. Only three strains produced antifungal non-polyenes singly; the others simultaneously formed basic antibacterial antibiotics of the type of streptomycin, streptothricin and neomycin or (also) polyene antibiotics, as a third component. Non-polyenes and polyenes did not occur together in the mycelium only or in fermentation fluid filtrate only. No correlation was found between the formation of non-polyenes and the place of origin of the strain. The possibility of screening new antibiotics is discussed.AbstractИсследовали 114 штаммов актиномицетов, вЫделеннЫх на географически различнЫх местах с разнообразнЫм растительнЫм покровом. В условиях глубинной культивации 57% штаммов оказались антибиотически эффективнЫми: 48% из них—против бактерий и 56%— против грибков. У внеклеточнЫх антибиотиков, в отличие от внутриклеточнЫх, заметно проявлялось влияние состава ферментационной средЫ. ПротивогрибнЫе антибиотики бЫли представленЫ преимущественно полиенами и неполиенами (в соотношении 2∶1) Группу неполиеновЫх антибиотиков образовали 20 штаммов, в том числе 14 из них соответствовали веществам, описаннЫм в литературе. Только 3 штамма образовали отдельнЫе антифунгальнЫе неполиенЫ, остальнЫе же одновременно продуцировали основнЫе антибактерийнЫе антибиотики типа стрептомицина, стрептотрицина и неомицина, или же одновременно еще третий компонент смеси—полиеновЫе антибиотики. Только в мицелии или в фильтрате фермен-тационной жидкости неполиенЫ не встречались одновременно с полиенами. Не наблюдалось корреляции менду образованием неполиенов и географическим распространением штамма.—Обсуждаются возможности скрининга новЫх антибиотиков.

Biogenesis of peptide antibiotics

A simple synthetic medium with glycine,l-tryptophan anddl-alanine was developed for study of antimycin A biogenesis. The strain produced 550 mg of mycelium dry weight and 200 μg of antimycin A per 100 ml of this medium. Intense biosynthesis of antibiotic was initiated in early fermentation and optimum yields were achieved after an additional 96 h incubation. No absolute relationship between growth rate and antimycin A production intensity was found. Biogenesis of antimycin A via tryptophan and formylkynurenine is suggested and similarity in early biogenesis of both antimycin A and actinomycin is considered.

Characterization and comparative studies ofStreptomyces sp. 246 producing a cytostatic antibiotic

The authors submit the results of taxonomic comparative studies of the strainStreptomyces sp. 246, which produces a polypeptide type cytostatic antibiotic. Strain 246 is characterized by tufts of straight sporophores of the “Rectus-Flexibilis” type, smooth spores arranged in chains (over 10 spores in a chain), yellow aerial and substrate mycelium, a negative test for melanin synthesis, utilization of glucose, arabinose, xylose, mannitol, fructose and rhamnose and inability to grow on sucrose, inositol, raffinose and cellulose. The taxonomic characters ofStreptomyces sp. 246 are identical with those of the strainStreptomyces chrysomallus JA 1449-1 and differ manifestly from those ofStreptomyces antibioticus strains (producing actinomycins, antimycin A and oleandomycin), fromStreptomyces cinereoruber ETH 7451 (producing rhodomycin) and from the strainStreptomyces sp. 4127 (producing actinomycin D).

Electrochromatography of amino acid mixtures in fermentation liquors

The substitution of the phenolic chromatography dimension, by electrophoresis at 500 to 1000 volts, in a veronal buffer, pH 8.6, μ 0.02 followed by triple rechromatography in a completely miscible butanol, acetic acid, water solvent system, leads to clear, easily reproducible electrochromatography patterns. A reliable economical chromatography cabinet is described. The technique allows the analysis of many samples conveniently, and some important characteristics of unidentifiable components can be readily predicted.AbstractПрименение электрофореза при 500–1000 вольт в вероналовом буферном растворе при pH 8,6, μ 0,02 вместо проявления в феноле и последующая тройная рехроматография во вполне смешивающейся системе бутанол—уксусная кислота—вода (4∶1∶1) дают отчетливые и легко воспроизводимые результаты. Описано экономное хроматографическое устройство. Метод делает возможным надежный анализ большого количества образцов, причем нетрудно предсказать некоторые важные характеристики неидентифицированных веществ.

Cytochrome system ofStreptomyces antibioticus producing antimycin A

Relatively little is known about the pathway of hydrogen (electron) transfer to molecular oxygen in the genus Strepto~nyces. Information concerning even the existence of cytochromes in these microorganisms is meagre (Sato, 1940; Heim, Silver & Birk, 1957; Inoue, 1958). This investigation was undertaken to seek the cytochrome system of Streptomyces antibioticus NRRL 2838, the producer of antimycin A which has been found to block the electron transport chain specifically between cytochrome b and c (Chance

Quantitative determination of ristocetin by a microbiological diffusion method

SummaryThe basic factors influencing the microbiological diffusion titration of ristocetin were studied. The results were used as a basis for the elaboration of directions for the routine determination of this antibiotic in fermentation liquids, usingBacillus subtilis as the test microorganism and spontin as the standard. The basic conditions for obtaining sharply defined inhibition zones and reproducible results are as follows:(1)Titration medium without the addition of glucose; pH after sterilization 6.5;(2)Density of the inoculum corresponding to 3.2×107 to 5×107 spores/ml;(3)Preculture (60 min. at 20°C) of the titration medium, together with the specimens, so as to reduce the unfavourable effect of diffusion of the antibiotic on the quality of the inhibition zones;(4)Lowering of the sodium chloride, potassium chloride and copper sulphate concentration in the analysed specimens by diluting them to the maximum degree possible.AbstractИсследовались основные факторы, влияющие на микробиологическую диффузную титрацию ристоцетина. На основе полученных результатов была разработана инструкция для рутинных определений этого антибиотика в ферментационных средах с применением Bacillus subtilis в качестве тест-микроба и спонтина в качестве стандарта. Основными условиями для получения отчетливых зон ингибиции и воспроизводимых результатов являются:(1)Титрационная среда без прибавления глюкозы с pH 6,5 после стерилизации.(2)Густота используемго инокулума, отвечающая 3,2×107–5×107 спор в 1 мл.(3)Предварительная культивация (60 мин. при 20° C) титрационной панели с нанесенными на нее образцами с целью ограничения неблагоприятного влияния диффузии антибиотика на качество зон угнетения.(4)Понижение концентрации хлористого натрия, хлористого калия и сернокислой меди в анаизируемых образдах путем максимального возможного разведения образцоб.