| Gloeostereum incarnatum S.Ito et Imai is a traditional fungi use as the medicine and food in China. Because of its clear ability as antimicrobial activity, people use it to cure some disease such as enteritis and dysenteries and so on in a long-standing tradition.The experiments use the cultivation fruitbodies and fermentation broth focus as the research objects, empHasis on systematic study and comprehensive analysis the antimicrobial activity of G. incarnatum. The specific object in this experiment is cultivation fruitbodies and strain G118 both bought and identified by Jilin Agricultural University, Fungi Research Center. Main contents and results are as follows:1. In this study, experiment using a new way which improve from traditional methods about boil to pick up the fruitbodies, acquired relatively complete fruitbodies extract. Then from the side of heat, pH and metal ions, determine the antimicrobial activity of the fruitbodies extract. Fruitbodies extract has perfect heat- Resistance and wide pH- Resistance. Low temperature as -20℃treat the fruitbodies extract 20min, the antimicrobial activity has unchanged, and high temperature as 121℃treat it 20min, the antimicrobial activity has unchanged, too; In acid and neutral pH, the antimicrobial activity of the fruitbodies extract keeps stabilize; Zn2+ ions can enhance the antibacterial activity of the fruitbodies extract evidently. Fe3+ ions can enhance the antibacterial activity also, maybe, it because that mental ions can chelate with parts of antibacterial substances, antibacterial activity has been changed. Many others mental ions have no effects about antibacterial activity. The PHysical and chemical natures of fermentation broth are approach to the fruitbodies'.2. It studies from many sides about shake flask fermentation of strains G118, and find out the best fermentation condition on antibacterial activity is: experiment use potato liquid medium (pH=7.0); Liquid volume in flask 50mL/250mL; inoculum 15% (v/v); train temperature 28℃; rotator speed 160r/min; fermentation cycle 9d.3. Experiment uses different polar organic solvents to extract the fruitbodies extract and fermentation broth. Obtained the extract liquid and raffinate liquid. Then we determine the samples, find out that the strong polar organic solvents such as ethyl acetate more easily extracted antibacterial substances from samples, and its raffinate liquid have no antibacterial activity; the weak polar organic solvents such as petroleum ether extracted the antibacterial substances from samples more difficultly; petroleum ether raffinate liquid is the only one which has antibacterial activity in the three kinds of raffinate liquids. And as ethyl ether which polar is living in the middle of other organic solvents, its antibacterial activity of extract liquid is in the middle of them too. About this experiment we found that the antibacterial substances in G. incarnatum must be a mixture with polar. In addition, there are many methods to be used in experiments to depart the G. incarnatum fruitbodies extract and fermentation broth, and found that the samples have no polysaccharide, and the ones have no protein, are show off clear antibacterial activity, but corresponding the polysaccharide liquid has no polysaccharide. The antibacterial activity of the samples treated by protease is more strongly than the one treated by sevage reagent, it maybe some antibacterial substances have lost when the samples treated by organic solvents. Sum up all the experiments before, we found that the antibacterial substances in G. incarnatum must be a small molecule mixture with polar.4. From antibacterial activity compare experiments, we found that antibacterial activity of G. incarnatum fermentation broth is stronger than fruitbodies extract clearly. The antibacterial activity of fermentation broth corresponds to 400u/mL Erythromycin and 400u/mL ChlorampHenicol, and 2.0% Benzoate (pH=4.0); the antibacterial activity of fruitbodies corresponds to 40u/mL Erythromycin and 40u/mL ChlorampHenicol, and 1.5% Benzoate (pH=4.0). Based on the studies before, G. incarnatum can be exploitated more scientifically. |
