Discovery Of Novel Bioactive Compounds Of Two Medicinal Fungi And Biotransformation Of Traditional Chinese Herbs

Medicinal fungi are widely distributed and rich in China,it has a wide variety of secondary metabolites,the secondary metabolites have the effects on anti-tumor,enhancing mmunity,anti infection,microbial secondary metabolites play an important role in the field of medicine and important resource for drug.Medicinal fungi as an important part of traditional Chinese medicine,fully taping the secondary metabolites of medicinal fungi can be used as a clinical drug to provide medicinal ingredients more pilot or direct.Medicinal fungi under laboratory conditions,many biological synthesis gene cluster of silence in silence.Inonotus sanghuang and Eurotium cristatum are important medicinal fungus,two kinds of medicinal fungi is worthy of further development and utilization.In the artificial culture of I.sanghuang,isolation and identification of secondary metabolites produced by the to ten,while in the wild state to isolation and identification of 50 kinds of I.sanghuang;E.cristatum in the state only identified more than ten kinds of metabolites of artificial rice cultivation and gene group sequencing analysis showed that E.cristatum 39 secondary metabolite biosynthesis gene cluster,suggesting that most of the biosynthetic gene cluster in the artificial culture condition is in a state of silence,no expression or expression is very low.Therefore,the urgent need for artificial cultivation of two species of medicinal fungi silent gene activation state,thus to obtain more metabolites.1.To study the effect of co-cultivation on the production of secondary metabolites of E.cristatum and I.sanghuang,and to analyze the single culture and co-cultivation fermentation of E.cristatum with I.sanghuang by high performance liquid chromatography(HPLC)differences in analysis.The results howed that two new absorption peaks appeared in the chromatographic samples of E.cristatum and I.sanghuang co-cultivation in the retention time 6.5-7.0 min,surmisedthat both co-cultivation produced novel bioactive compounds.2.Tot discusses the influence of fungal elicitor on I.sanghuang.A strategy that added fungal elicitor to I.sanghuang was performed shined with blue light during growth,and compared with the growing of the I.sanghuang without fungal elicitor and light irradiation,single E.cristatum elicitor,then analysed the differences of HPLC fingerprint of ethyl acetate extractions after ethanol extraction of fermentation,the results show that the I.sanghuang with E.cristatum elicitor appear 3 new absorption peaks during growing at Rt=19.584 min,38.142 min,45.771 min,but thefermentation of E.cristatum or single I.sanghuang didn’t show these peaks.It illustrates that adding fungal elicitor and blue light irradiation to the growing process of I.sanghuang can induce them produce novel secondary metabolites.Moreover,with the stimulation of elicitor and blue light,some gene clusters which are silent in normal condition were activated,and then played a positive control role to increase the metabolite of I.sanghuang.Add in the training process of I.sanghuang elicitor and blue light irradiation to its function so that they produce novel secondary metabolites,further infer I.sanghuang in elicitor and blue light stimulation,some of the I.sanghuang in normal condition silent gene cluster expression may be activated,which played a positive role,the increase of metabolite I.sanghuang.3.In our project,we generated about 39.6 Gb bases in total on Illumina HiSeq sequencing platform.After assembling all samples together and filtering the abundance,we got 42,266 Unigenes,the total length,average length,N50,and GC content of Unigenes are 83,891,727 bp,1,984 bp,3,678 bp and 49.88% respectively.And then annotate Unigenes by aligning with 7 functional databases,finally,31,474(NR:74.47%),7,789(NT:18.43%),20,462(Swissprot:48.41%),21,231(KOG:50.23%),23,067(KEGG:54.58%),8,871(GO:20.99%),and 22,653(InterPro:53.60%)Unigenes are annotated.For functional annotation results,we detected 54,420 CDS by Transdecoder.We also detected 4,516 SSR distributed on 3,280 Unigenes.It was found from the data that the metabolism of I.sanghuang cells was more active under the stimulation of fungal elicitor and blue light.These data will lay a foundation for the synthesis of some unknown metabolites of mulberry yellow stimulated by fungal elicitor and blue light.4.This part mainly discusses I.sanghuang and E.cristatum transformation of Chinese herbal medicines.The substrates with Rice-Lonicerae japonicae Flos in solid-state fermentation,then the fermentation products were extracted by the aid of ultrasonic with 95% ethanolic respectively.The differences of finger-prints and the changes of active contents in Lonicerae japonicae Flos pre and post-fermentation were analyzed by HPLC,and compared the antimicrobial activity of 95% ethanol extracts by double plate punching method.The result was indicated that chlorogenic acid and rutin in Lonicerae japonicae Flos by solid-state fermentation with Aspergillus sp.were respectively reduced by 20.14 mg/g and 1.73 mg/g.While luteoloside was increased by 4.76 mg/g.Meanwhile,not only the compounds of a and b were remarkably increased,but also the HPLC chromatograms were arised the newabsorption peak after 17.5 min of retention time;The chlorogenic acid and rutin in Honeysuckle Leaves were respectively reduced by 11.74 mg/g and 0.99 mg/g.While luteoloside was increased by 5.07 mg/g;In addition,the ability of 95% ethanolic extract anti-bacterial activity was stronger than the unfermented and control group.E.cristatum and I.sanghuang can grow and ferment in different medical materials,the use of it to transforming of Pueraria lobata 、Astragalus membranaceus,not only improve the amount of total flavonoids of medicinal materials,improve the content of some compounds,can make the resistance matrix to produce new substances.