Formation of cyclosporins by Tolypocladium inflatum.

Submerged cultures of the low-production strain of Tolypocladium inflatum DSM 63544 formed a mixture of cyclosporins (CS) consisting of CS-A, CS-B, "CS-3" and "CS-4". Glucose, sucrose and maltose were highly favored for biomass production but provided a different physiological state necessary for CS biosynthesis. Not only the magnitude of CS production but also the proportion of individual components of the CS mixture were affected by the C source. Intensive CS synthesis was in correlation with the formation of CS-3. Lower yields of CS were accompanied by an increased proportion of CS-A in the CS complex. The best specific production of CS was achieved on the glucose medium, the highest yield of CS-A on the maltose medium. There was no remarkable relationship between the biomass formation and the intensity of CS synthesis.

Substrate regulation of elymoclavine formation by some saccharides.

Regulation of the production of clavine alkaloids, especially elymoclavine, by sucrose, maltose, and mixtures of these saccharides was studied in submerged cultures of strains Claviceps purpurea 129/35 and Claviceps sp. SD-58. The data were statistically processed on an EC 1040 computer. Fermentation medium containing sucrose (80 g/l) in combination with glucose (20 g/l) was the best for elymoclavine formation. Retarded release of glucose from maltose increased the formation of elymoclavine and suppressed the synthesis of undesirable extracellular glucans. Carbon source can affect both the total amount of produced alkaloids and the relative proportion of individual clavines in the alkaloid mixture.

[Morphological differentiation of cells in submerged cultures of Claviceps purpurea (Fr.) Tul., producing alkaloids]. / Morfologicheskaia differentsiatsiia kletok v pogruzhennykh kul'turakh Claviceps purpurea (Fr.) Tul., obrazuiushchikh alkaloidy

Differentiation of the cells of the submerged culture of Claviceps purpurea (Fr.) Tul. was studied by electron microscopy. Two types of oviform cell were found: (1) the conidia which had one nucleus and vacuolized cytoplasm and were not involved in the production of alkaloids; (2) the chlamydospores with two nuclei, homogeneous cytoplasm, and high content in lipids. The chlamydospores, like the cells of sclerotia, were found to produce alkoloids.

Production of alkaloids and differentiation in a submerged culture of Claviceps purpurea (Fr.) Tul.

The submerged culture of the Claviceps purpurea strain studied was polymorphous. In the process of alkaloid synthesis, cytodifferentiation preceded biochemical differentiation. The onset of the alkaloid phase was characterized by: predominance of chlamydospores in the culture, the presence of vegetative cells with reduced or arrested proliferation, maximum acetylCoA carboxylase activity, the maximum amount of total fatty acids, an over-average cell pool tryptophan level and minimum tryptophan synthetase activity. Intracellular ricinoleic acid was an indicator of differentiation of the culture towards alkaloid formation and also of alkaloid synthesis. The cytodifferentiation period in the initial phases of fermentation, when the cell has several alternative possibilities of development, is regarded as the most sensitive sector of the regulatory mechanisms of alkaloid formation.

Correlation of certain alterations in metabolic activity with alkaloid production by submerged Claviceps.

Alkaloid biosynthesis in Claviceps paspali MG-6 was favored by unbalanced growth. A positive correlation between the rate of protein turnover and alkaloid formation was noted. The pattern of the orthophosphate content in the mycelium resembled that in the ripening sclerotia of the parasitic strains. Alkaloids were revealed as potentially effective in energy metabolism. Reduced adenosine triphosphate (ATP) utilization and an increase in the ATP pool were found to be favorable for alkaloid production. Acetyl-coenzyme A carboxylase activity and the level of cell lipids were directly related to the intensity of alkaloid biosynthesis. An inverse relationship was observed between the activities of the tricarboxylic acid and glycoxylate cycles and the rate of alkaloid formation.

Respiratory chain of antimycin A-producing Streptomyces antibioticus.

An active respiratory chain system was demonstrated in sonically treated mycelium of Streptomyces antibioticus, the producer of antimycin A. The respiratory electron transfer from substrate to oxygen proceeded successively through flavoprotein(s), b-, c-, and a-type cytochromes, and terminated with the cyanide-sensitive cytochrome oxidase. The cytochrome composition of the culture was not affected by the age of the mycelium, the intensity of antimycin A production, or differences in the media. Slater factor, coenzyme Q, and vitamin K were not interposed as hydrogen carriers in the respiratory chain between flavoproteins and cytochromes. The oxidation of reduced nicotinamide adenine dinucleotide and succinate was unaffected by antimycin A. Evidence is presented in support of the absence of the antimycin A-sensitive site from the electron transport system of S. antibioticus.