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不同氮源下弗氏链霉菌培养物的表观粘度对菌丝形态的依赖性。

Dependence of apparent viscosity on mycelial morphology of Streptomyces fradiae culture in various nitrogen sources.

作者信息

Choi D B, Park E Y, Okabe M

机构信息

Institute of Life Science, Chosun University, 375, Seo-Suk Dong, Dong-ku, Kwang-Ju, Korea.

出版信息

Biotechnol Prog. 2000 Jul-Aug;16(4):525-32. doi: 10.1021/bp000056x.

Abstract

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.

摘要

为研究弗氏链霉菌培养过程中导致培养液粘度增加的原因,对各种天然氮源进行了研究。胞外蛋白酶活性随培养时间增加,将天然氮源分解为氨基酸。以面筋粉为例,培养5 d后,谷氨酸和天冬氨酸浓度分别增加到600和200 mg/L,约为使用Pharmamedia在相似条件下培养时浓度的3倍和2倍。对于所测试的各种氨基酸,向培养基中添加谷氨酸或天冬氨酸混合物可使表观粘度在培养5 d后升至最高显示值260 mPa·s,比不添加氨基酸时高3倍。分解的谷氨酸和天冬氨酸的消耗分别取决于谷氨酸脱氢酶和天冬氨酸转氨酶的活性。当使用铵离子作为氮源时,以细胞内核酸浓度衡量的细胞浓度达到1.75 g/L,约为使用任何其他天然氮源时的2.3倍。然而,表观粘度仅为75 mPa·s,是氨基酸混合物粘度值的三分之一,且70%的菌球大于1.2×10⁴ 微米²。以面筋粉或氨基酸混合物为例,大于1.2×10⁴ 微米²的菌球仅占8%。这表明某些氨基酸的消耗影响了丝状形态的形成,从而导致培养液表观粘度增加,且表观粘度并非由菌丝体浓度引起,而是由菌丝体形态引起。

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