RESUMO
To examine what causes increased viscosity in culture broth in Streptomyces fradiae culture, various natural nitrogen sources were investigated. Extracellular protease activity increased with culture time and decomposed the natural nitrogen source into amino acids. In the case of gluten meal, after a culture time of 5 d, concentrations of glutamic acid and aspartic acid had increased to 600 and 200 mg/L, respectively, which were about 3- and 2-fold as high as levels in cultures under similar conditions using Pharmamedia. For various amino acids tested, the addition of glutamic acid or aspartic acid mixture to the culture medium raised the apparent viscosity to its highest demonstrated value, 260 mPa.s after 5 d of culture, which was 3-fold higher than without amino acids. Consumption of the decomposed glutamic acid and aspartic acid was dependent on the activities of glutamate dehydrogenase and aspartate aminotransferase, respectively. When ammonium ion was used as the nitrogen source, cell concentration reached 1.75 g/L measured as an intracellular nucleic acid concentration, which was about 2.3-fold higher than that with any other natural nitrogen source. However, apparent viscosity was only 75 mPa.s, a value one-third that of the amino acid mixture, and 70% of the pellets were bigger than 1.2 x 10(4) microm(2). In the case of gluten meal or the amino acid mixture, pellets bigger than 1.2 x 10(4) microm(2) comprised only 8%. This demonstrates that consumption of some amino acids affected the formation of filamentous morphology, which caused an increase in the apparent viscosity of the culture broth, and the apparent viscosity was not caused by the mycelial concentration but the mycelial morphology.
Assuntos
Nitrogênio/metabolismo , Streptomyces/citologia , Aminoácido Oxirredutases/metabolismo , Aminoácidos/metabolismo , Ácido Aspártico/farmacologia , Meios de Cultura , Endopeptidases/metabolismo , Glutamato Desidrogenase/metabolismo , Ácido Glutâmico/farmacologia , ViscosidadeRESUMO
Corn starch and soybean oil are suitable carbon sources for the production of tetracycline by Streptomyces aureofacience CG-1. However, it could not produce more than 6 g/l of tetracycline even if initial corn starch concentration was increased to more than 100 g/l. It was confirmed by shaking flask experiments that the k(L)a in a mixture of 2% soybean oil in water was four folds compared with that without soybean oil. With the addition of soybean oil to the starch medium in a shaking flask, tetracycline production was significantly improved. By scaling-up to a 5.5-l airlift bioreactor from 500-ml Erlenmeyer flask, more than 10 g/l of tetracycline was produced with the addition of 60 g/l of soybean oil to the medium containing 100 g/l of corn starch. The dissolved oxygen level in the airlift bioreactor containing soybean oil was higher than that without soybean oil. This suggests that soybean oil is not only a suitable carbon source but is also a surface-active agent which may accelerate the oxygen transfer. This may lead to the possibility of the enhanced production of tetracycline at a low cost in airlift bioreactor.
RESUMO
To investigate why more tylosin was produced when Streptomyces fradiae T1558 was cultured in a rapeseed oil medium than in a glucose or starch medium, we measured the activity of methylmalonyl-CoA carboxyltransferase (EC 2.1.3.1) and intracellular propionic acid. The activity of the enzyme, which catalyzes the formation of the precursor of tylosin, protylonolide, was 0.19 U/mg protein in 5 days of culture in rapeseed oil medium, which was 2.5- and 1.3-fold that with the glucose or starch medium, respectively. The intracellular propionic acid concentration was 1.2 g/g of dry weight, which was 4.3- and 2.1-fold that with the glucose or starch medium, respectively. The addition of propionic acid increased tylosin production in batch culture: when 0.2 g/l (final concentration) propionic acid was added to the glucose medium, 3.8 g/l tylosin was produced in 10 days of culture, 4.7-fold the amount without propionic acid. These findings suggest that in glucose medium, intracellular propionic acid is a limiting factor because of the low activity of methylmalonyl-CoA carboxyltransferase of the tylosin biosynthesis pathway.