Cell protein profiles of submerged cultures of Streptomyces aureofaciens cultivated in the absence or presence of 12 microM benzyl thiocyanate (BT) were analyzed by one-dimensional SDS polyacrylamide gel electrophoresis. Substantial increase in the intensity of the 13, 35, 37, 60, and 100 kDa protein bands was observed in cultures treated with BT. Similar increase in the 35, 37, and 60 kDa bands was found in a mutant blocked in the last chlortetracycline biosynthesis step. Effect of BT on the solid medium-grown cultures was also observed, with a more intensive substrate mycelium pigmentation and alteration in the spore size and shape as the most characteristic features. Earlier studies of BT effect involving those on the stimulation of chlortetracycline biosynthesis are summarized and a possible signal-transducing mechanism is discussed from the point of view of adaptation of S. aureofaciens to the uncoupling of oxidative phosphorylation.

• MeSH
• Bacterial Proteins metabolism   MeSH
• Microscopy, Electron MeSH
• Streptomyces aureofaciens drug effects   metabolism   ultrastructure   MeSH
• Tetracyclines biosynthesis   MeSH
• Thiocyanates pharmacology   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Bacterial Proteins MeSH
• benzyl thiocyanate MeSH Browser
• Tetracyclines MeSH
• Thiocyanates MeSH

The localization of anhydrotetracycline oxygenase and glucose-6-phosphate dehydrogenase (EC 1.1.1.49) was studied by determining the enzyme activities in subcellular fractions obtained by differential centrifugation of the mycelia of Streptomyces aureofaciens after lysozyme treatment. Glucose-6-phosphate dehydrogenase was a typical cytoplasmic enzyme both in the low- and high-production strain. Anhydrotetracycline oxygenase was found in the membrane fraction of the low-production strain. In the high-production strain, it was detected in several fractions, the highest activity being found in cytoplasm. The presence of 10 microM benzyl thiocyanate in the culture medium significantly changed the distribution of the latter enzyme in both strains. The redistribution of the enzymes is discussed with respect to tetracycline over-production.

• MeSH
• Cell Fractionation methods   MeSH
• Glucosephosphate Dehydrogenase metabolism   MeSH
• Magnesium pharmacology   MeSH
• Kinetics MeSH
• Oxygenases metabolism   MeSH
• Streptomyces aureofaciens drug effects   enzymology   MeSH
• Thiocyanates pharmacology   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• anhydrotetracycline oxygenase MeSH Browser
• benzyl thiocyanate MeSH Browser
• Glucosephosphate Dehydrogenase MeSH
• Magnesium MeSH
• Oxygenases MeSH
• Thiocyanates MeSH

Streptomyces aureofaciens glucosidizes 1,2,4-trihydroxy-9,10-anthraquinone (purpurin) added to the cultivation medium to yield the corresponding 2-beta-D-glucoside. The identity of the glucoside was demonstrated by comparing its physico-chemical properties with data of an authentic sample prepared synthetically. A further chemical glucosidation of the acetylated 2-beta-D-glucoside gives rise to 2-(hepta-O-acetyl-beta-gentiobiosyl)-4-(tetra-O-acetyl-beta-D-gluc opyranosyl) purpurin. All the derivatives are immunoactive.

• MeSH
• Anthraquinones * MeSH
• Chemical Phenomena MeSH
• Chemistry MeSH
• Fermentation MeSH
• Glucosides biosynthesis   MeSH
• Mass Spectrometry MeSH
• Lectins immunology   metabolism   MeSH
• Magnetic Resonance Spectroscopy MeSH
• Spectrophotometry, Infrared MeSH
• Streptomyces aureofaciens metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Anthraquinones * MeSH
• Glucosides MeSH
• Lectins MeSH
• purpurin anthraquinone MeSH Browser

ATP diphosphohydrolase activity and inorganic pyrophosphatase reached two maxima during cultivation of the low- and high-producing variant of Streptomyces aureofaciens under conditions of phosphate limitation, i.e. after 30 and 70 h of cultivation. Increased levels of inorganic phosphate in a medium inhibitory to biosynthesis of chlortetracycline markedly decreased the levels of both enzymes. The ATP diphosphohydrolase activity was detected both in the supernatant and membrane fractions of the cell-free preparation of the mycelium.

• MeSH
• Apyrase metabolism   MeSH
• Chlortetracycline biosynthesis   MeSH
• Phosphoric Monoester Hydrolases metabolism   MeSH
• Phosphates pharmacology   MeSH
• Culture Media MeSH
• Membranes enzymology   MeSH
• Pyrophosphatases metabolism   MeSH
• Streptomyces enzymology   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Apyrase MeSH
• Chlortetracycline MeSH
• Phosphoric Monoester Hydrolases MeSH
• Phosphates MeSH
• Culture Media MeSH
• Pyrophosphatases MeSH

A case is made out for regarding secondary metabolism as part of normal cell growth, related to its interactions with the environment. Secondary metabolism is widespread, especially in fungi and actinomycetes, and is not to be regarded as confined to the production of antibiotics and other special substances. It is part of the normal maturation process. Examples are given of the influence of secondary metabolism in ecological systems. It is also shown that cell productivity can be related to age structure. Secondary metabolism is thus linked with growth, although in many cases this may not be obvious in laboratory work. Initiation of production will arise from the system which regulates growth and differentiation. These processes are little understood at present, but it is clear that the factors involved differ in different instances and that they involve a very great variety of biochemical and physiological processes.

• MeSH
• Actinomycetales metabolism   MeSH
• Bacteria growth & development   metabolism   MeSH
• Ecology MeSH
• Fungi growth & development   metabolism   MeSH
• Penicillins biosynthesis   MeSH
• Plants metabolism   MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• Penicillins MeSH

The rate of protein synthesis in Streptomyces aureofaciens, measured by incorporation of U-14C-L-leucine into cells, fluctuated during the production phase in the range of 10-15% of the values determined in the phase of intensive growth. Tetracycline partially inhibited the protein synthesis during the growth phase only. The proteins synthesized between the 6th and 18th hour of growth, were 75% degraded by the 48th hour. The DNA synthesis, measured by means of incorporation of 2-14C-thymine into the mycelium, occurred predominantly during the first 24 h of cultivation. Similarly, DNA synthesized between the 6th and 12th hour of cultivation was degraded by 75% after 48 h. The turnover of culture proteins is thus caused largely by degradation of old cells and growth of new ones which are more resistant to tetracycline. The activity of alanine aminotransferase and aspartate aminotransferase increase substantially towards the end of fermentation.

• MeSH
• Alanine Transaminase metabolism   MeSH
• Aspartate Aminotransferases metabolism   MeSH
• Bacterial Proteins metabolism   MeSH
• Chloramphenicol pharmacology   MeSH
• DNA, Bacterial metabolism   MeSH
• Leucine metabolism   MeSH
• Streptomyces aureofaciens metabolism   MeSH
• Tetracyclines pharmacology   MeSH
• Thymine metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Alanine Transaminase MeSH
• Aspartate Aminotransferases MeSH
• Bacterial Proteins MeSH
• Chloramphenicol MeSH
• DNA, Bacterial MeSH
• Leucine MeSH
• Tetracyclines MeSH
• Thymine MeSH

• MeSH
• Anti-Bacterial Agents biosynthesis   MeSH
• Bacteria growth & development   metabolism   MeSH
• Bacterial Proteins metabolism   MeSH
• RNA, Bacterial metabolism   MeSH
• DNA, Bacterial metabolism   MeSH
• DNA metabolism   MeSH
• Fungi growth & development   metabolism   MeSH
• RNA metabolism   MeSH
• Spores, Bacterial physiology   MeSH
• Streptomyces growth & development   metabolism   MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• Anti-Bacterial Agents MeSH
• Bacterial Proteins MeSH
• RNA, Bacterial MeSH
• DNA, Bacterial MeSH
• DNA MeSH
• RNA MeSH

The relationship was studied between the energy metabolism of the actinomycete Streptomyces aureofaciens and the biosynthesis of chlorotetracycline by this organism. The energy charge values in a culture of low-production strain were almost identical with those of a production variant but the total sum of adenylates was about 10 times higher. In the stationary growth phase both strains evinced a drop in energy charge values followed by a rise to the original level. An increase in the concentration of inorganic phosphate in fermentation medium caused a suppression of antibiotic formation in the lowproduction strain and further rise in the total adenylate level. The expression of the energy charge in Streptomyces aureofaciens acquires a complex character owing to the participation, apart from the adenylate system, of high-molecular polyphosphates as energy donors and the probable lack of a regulating mechanism such as the adenylate kinase reaction.

• MeSH
• Adenine Nucleotides metabolism   MeSH
• Adenosine Diphosphate metabolism   MeSH
• Adenosine Monophosphate metabolism   MeSH
• Adenosine Triphosphate metabolism   MeSH
• Chlortetracycline biosynthesis   MeSH
• Energy Metabolism MeSH
• Phosphates metabolism   MeSH
• Genetic Variation MeSH
• Streptomyces aureofaciens metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Adenine Nucleotides MeSH
• Adenosine Diphosphate MeSH
• Adenosine Monophosphate MeSH
• Adenosine Triphosphate MeSH
• Chlortetracycline MeSH
• Phosphates MeSH

The activity of ATP-glucokinase and of polyphosphate glucokinase was examined during growth of the actinomycete Streptomyces aureofaciens 8425 under conditions of intense chlortetracycline (CTC) synthesis. ATP-glucokinase was active in the strain only during the logarithmic phase of culture growth; the activity of polyphosphate glucokinase appears only at the end of the logarithmic phase of growth and rises in parallel with the rate of CTC biosynthesis in the stationary phase. During the rise of activity of polyphosphate glucokinase and of CTC biosynthesis the cells accumulate sugar phosphates, mainly glucose-6-phosphate. It appears that the biosynthesis of CTC in Streptomyces aureofaciens takes place at the expense of glycolysis, using up the high-energy phosphate of high-molecular polyphosphates.

• MeSH
• Adenosine Triphosphate metabolism   MeSH
• Cell-Free System MeSH
• Chlortetracycline biosynthesis   MeSH
• Sugar Phosphates biosynthesis   MeSH
• Phosphates metabolism   MeSH
• Phosphotransferases metabolism   MeSH
• Glucokinase metabolism   MeSH
• Streptomyces aureofaciens enzymology   growth & development   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Adenosine Triphosphate MeSH
• Chlortetracycline MeSH
• Sugar Phosphates MeSH
• Phosphates MeSH
• Phosphotransferases MeSH
• Glucokinase MeSH

• MeSH
• Acetates metabolism   MeSH
• Alanine metabolism   MeSH
• Glutamates metabolism   MeSH
• Glutarates metabolism   MeSH
• Bicarbonates metabolism   MeSH
• Colorimetry MeSH
• Malonates metabolism   MeSH
• Propionates metabolism   MeSH
• Pyruvates metabolism   MeSH
• Carbon Radioisotopes MeSH
• Streptomyces aureofaciens metabolism   MeSH
• Succinates metabolism   MeSH
• Tetracycline biosynthesis   MeSH
• Uracil metabolism   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Acetates MeSH
• Alanine MeSH
• Glutamates MeSH
• Glutarates MeSH
• Bicarbonates MeSH
• Malonates MeSH
• Propionates MeSH
• Pyruvates MeSH
• Carbon Radioisotopes MeSH
• Succinates MeSH
• Tetracycline MeSH
• Uracil MeSH