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Strain Screening and Submerged Culture Condition Study of Coriolus Versicolor Fungus Producing Anticancerous Peptide

Author: ZhanLin
Tutor: YangXiaoTong
School: Shanghai Normal University
Course: Microbiology
Keywords: Versicolor Small peptide extract Anticancer Liquid fermentation Tricine electrophoresis
CLC: S567.3
Type: Master's thesis
Year: 2006
Downloads: 45
Quote: 0
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Abstract


The subject of 13 versicolor strains submerged fermentation, to compare the different strains of the biomass is extracted with 70% ethanol extracts of the mycelium and the fermentation broth of the smaller peptide, determined by Lowry's method a small peptide extract The protein content in the material. The results showed that the overall average of 13 versicolor strain biomass 1.4746g/100ml, TV-5 strain the highest (2.0625g/100ml). Mycelium and the protein content of the extract of the fermentation broth small peptide great due to different strains differences, the mycelium small peptide extract the average protein content of 21.6%, the TV-33 content of the highest (30.4%); fermentation liquor small peptides extract an average protein content of 23.3%, TV-42 content in the highest (34.3%). MTT assay extract small peptide Molt-4 human T-lymphoid leukemia cells in vitro growth inhibition found mycelium and fermentation broth small peptide of TV-42 strains extract of Molt-4 strongest inhibition, inhibition rate reached 22.95 ± 1.00%, 20.43 ± 2.09% (200μg/ml, 72 hours). Accordingly, it is determined that the TV-42 strain was used as the target strains of the research fermentation conditions studied the fermentation time, pH and temperature on the mycelium biomass, medium amount of residual sugar, a small peptide extracted material and in vitro growth inhibitory activity of . The results show that the extract of mycelium and small peptides of the fermentation broth of Molt-4 in vitro inhibition strongest activity conditions is: The fermentation time for the sixth day (inhibition rates were 46.78 ± 3.54% 21.68 ± 2.20%); of acid conditions (inhibition rate were 49.48 ± 3.08%, 36.00 ± 0.46%); fermentation temperature of 28 ° C (inhibition rate were 36.38 ± 1.90%, 18.35 ± 3.69%). Morphological observation and flow cytometric analysis of cell cycle was found that the TV-42 small peptides mycelium and fermentation broth extract of Molt-4 inhibition and blocking cell cycle. Small peptide extract concentration of 400μg/ml, the role of Molt-4 cells after 72 hours the proportion of cells in S phase can be increased compared with the control group, S phase arrest activity. Present experiments also explored a small peptide extract Tricine electrophoretic conditions and fixed and stained to determine the optimum analysis conditions is the concentration of the separation gel is 20%, the fixative is 5% glutaraldehyde, the staining solution for 0.025% Coomassie Brilliant Blue G-250 and 10% acetic acid, bleaching solution for the bleaching solution of 10% acetic acid. Small peptide extract was determined under this condition to a molecular weight of 3496Da to 1060Da, and shows the features of the sugar binding to the peptide. This study and theoretical basis for the versicolor resources development and production applications.

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