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HCCB10124 preliminary study of antifungal activity

Author: WangXueJun
Tutor: ChenDaiJie
School: Shanghai Normal University
Course: Microbiology
Keywords: HCCB10124 Antifungal activity Identification of bacteria 16SrDNA sequence Optimization of fermentation Separation and purification
CLC: R96
Type: Master's thesis
Year: 2008
Downloads: 58
Quote: 0
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Abstract


In this study, a production of Shanghai the to benefit Biopharmaceutical R \u0026 D center screened broad spectrum of highly active anti-fungal product HCCB10124 strains , antibacterial spectrum measured through the paper - sensitive plate method , found that more than 20 species of fungi , some of which are plant and animal pathogens, significant antagonism and inhibition of Staphylococcus aureus and Sarcina lutea ; different concentrations and mycelium extract diluted , antifungal activity was measured and found high activity . 16SrDNA sequence analysis , this strain Streptomyces Streptomyces albireticuli having a sequence homology of 99% . Prior to the separation and purification of Streptomyces HCCB10124 preliminary optimization of the fermentation conditions and the medium components , and in order to obtain a higher yield of active metabolites . The optimized medium basic components : 2% glucose , 2% soluble starch , 4% drug medium , 0.04% ammonium molybdate , 0.4% of calcium carbonate . Detecting the product of the biological activity in the mycelium extract and fermentation supernatants on the temperature, pH stability; The method of using the the Czech eight solvent system paper chromatography conducted research and found that the product on the biological activity on the temperature, pH value than stable. The preliminary results of the study as a foundation for further separation and extraction . By pH precipitation , organic solvent extraction and the resin adsorption, the silica gel column chromatography , medium pressure chromatography chromatography experiments to determine the workflow for the first fermentation supernatant was adjusted to pH 6 centrifugal not removed solids is removed , and a large macroporous resin adsorption acetone extract of the supernatant and the mycelium , removing the dye, different concentrations of ethanol elution, the eluate was concentrated and subjected to silica gel column chromatography with acetone : methanol = 10:1,5:1 subparagraph 3:1,2:1,1:1,100 % methanol elution, sufficiently remove the pigments, the initial separation of the active material . Finally, by medium pressure chromatography experiments , to remove part of the impurities , to achieve the sample enrichment and purification purposes . In view of the the Streptomyces HCCB10124 produced antifungal substance with broad-spectrum , high activity , high stability, etc. , and therefore has the potential , it is important to value and theoretical significance . Its production of antimicrobial active substance is expected to develop as a can be widely applied to the fungal disease biocontrol or human and animal fungal diseases of the antifungal antibiotics.

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CLC: > Medicine, health > Pharmacy > Pharmacology
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