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The Studies on Strain-improving and Product Isolation for Nosiheptide Production

Author: LouShuangFeng
Tutor: HeJianYong
School: Shenyang Pharmaceutical University
Course: Microbial and Biochemical Pharmacy
Keywords: nosiheptide strain-improving rational screening isolation and purification biosynthesis
CLC: TQ464.7
Type: Master's thesis
Year: 2005
Downloads: 35
Quote: 2
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Abstract


In this paper, the strain-improving, the extraction and purification as well as the biosynthesis and metabolism regulation of nosiheptide were studied by using the nosiheptide producer-Streptomyces actuosus A01 as a starting strain.First, the strain D-45, which was a high-yield and stable strain with a productivity of 2100 ug/ml, was obtained by a procedure of natural screening. Investigation of different mutagen and different dosage of each kind of mutagen lead to the confirmation of a suitable mutagenesis condition - UV 90s. Based on the investigation of the correlation between solid fermentation and flask liquid fermentation, an agar-column solid fermentation method was adopted, replacing the flask liquid fermentation. As a result, the screening efficiency was improved greatly.Secondly, the screening models of tolerance to cysteine、Methionine、methylamine and nosiheptide were established. Results showed that the probability of obtaining high-yield strain increased obviously by screening for strains which were tolerance to cysteine or methylamine. As a result, the high-yield strain C1-11 with a fermentation titer of 2583 ug/ml and strain S4-20 with a fermentation titer of 2499ug/ml were obtained. By adopting a combined screening models, a stable high-yield strain MSC1-72 with a fermentation titer of 2961ug/ml ,which was tolerant to cysteine、Methionine、methylamine simultaneously, was obtained.In addition, an isolation and purification procedure of nosiheptide from fermentation broth was established in this paper. a macroporous absorption resin MN-200 was used first time in the isolation of nosiheptide and a suitable technical condition of crystallization was determined after investigation of a variety of conditions of crystallization. The total recovery rate of nosiheptide from fermentation broth was 53%, and the purity of product was increased greatly with a titer 852ug / mg. Besides, the process of preparation of nosiheptide premix was established in laboratory and the quality of the premix sample was detected and analyzed according the veterinary pharmacopeia.Finally, the possible biosynthesis pathway of serine and cysteine, which are the precursor for nosiheptide biosynthesis, in Streptomyces actuosus was investigated. experiment results showed that serine, the precursor of nosiheptide biosynthesis, may come from the 3-phosphoglyceric acid produced by EMP pathway.

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CLC: > Industrial Technology > Chemical Industry > Pharmaceutical chemical industry > Drug production of biological products > Amino acids,peptides,proteins
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