The Active Metabolites Of Trichoderma Harzianum From Wild Edible Mushroom Based On One Strain Many Compounds Strategy

Fungi can produce many novel and biologically active secondary metabolites,and the secondary metabolites of many fungi has been used as a leading drug for the treatment of various diseases.The wild edible mushroom in Yunnan Province was a delicious food.At the same time,it contain a variety of active substances,such as anti-tumor and anti-oxidation substances.The research on nutrition and active ingredients of mushroom was received more attention.However,there were few studies on the secondary metabolites of its endophytic fungi.Therefore,the thesis focused on the study of secondary metabolites of Trichoderma harzianum in the wild edible mushroom Tricholoma matsutake in Yunnan Province based on one strain many compounds（OSMAC）.We hoped to find more active compounds with novel structures.This paper was mainly divided into four chapters:In the first chapter,the research progress of active ingredient in edible wild mushroom was summarized;the research progress on secondary metabolites of Trichoderma harzianum was summarized;the research status of the secondary metabolites of microorganisms under the one strain many compounds（OSMAC）strategy was summarized.In the second chapter,101 strains of endophytic fungi isolated from wild edible mushroom Tricholoma matsutake in Yunnan Province were studied.After the fermentation of the 101 strains,the fermentation broth was extracted.12 strains with good chemical diversities were screened according to TLC color.The target strain SM-6with good chemical diversities and stable metabolism was selected by rescreening 12 strains with the same method.SM-6 was identified as Trichoderma harzianum by ITS sequencing.In the third chapter,the fungus Trichoderma harzianum was fermented with solid yam medium.Six compounds were separated from the extract of the ferments of Trichoderma harzianum,including three new compounds : triharzianin A（1）,triharzianin B（2）,triharzianin C（3）and three known compounds :15-hydroxyacorenone（4）,trichoacorenols B（5）,harzianone（6）.Compounds 1-6 were tested for antifungal activity in vitro and the feeding attraction activity of young silkworms.Compounds 4-6 showed inhibitory effect against Aspergillus fumigatus and Trichoderma sp.,with the MIC value between 32-64 μg/m L.Compound 5 and the extract on blank yam medium showed feeding attraction activity with the attraction rate above 90%.The attraction rate of compounds 1,3 was around 80%,and the weight changes of silkworms were-3.9% and-1.0%.Compound 1 was tested for α-glucosidase,angiotensin-converting enzyme（ACE）,acetylcholinesterase inhibitory activity,cytotoxic activity against five tumors,and anticoagulant activity.Compound 1 did not show any activities.In the fourth chapter,the secondary metabolites of Trichoderma harzianum cultured by solid peanut was studied.11 compounds were isolated and identified from the crude extraction,including one new compound: triharzin A（5）and ten known compounds: 4-hydroxybenzaldehyde（1）,4-hydroxybenzoic acid（2）,p-Hydroxybenzoic acid methyl ester（3）,1H-indole-3-carboxaldehyde（4）,cyclonerodiol B（6）,（3S,5R,6S,9R）-3,6-dihydroxy-5,6-dihydro-β-ionol（7）,cycloerodiol（8）,ergosterol endoperoxide（9）,ergosterol（10）,β-sitosterol（11）.The vitro antibacterial activity of compounds 1-8 was tested for Aspergillus fumigatus and Trichoderma sp.,with the MICs value of compounds 5-8 128-256 μg/m L,and the MIC value of other compounds above 256 μg/m L.Compared with the positive control nystatin,the antifungal activity was not obvious.Simultaneously we tested the feeding attractant activity of compounds1,3-8,the crude extracts,and the extract of blank peanut medium.The attraction rate of compounds 1,3,6 exceeded 87%.Compund 5 did not show any activity,and the weight change of silkworm was-3.9%.