Chemically Induced Marine Fungi And Enzymatic Hydrolyzed Sea Cucumber Derived Anti-alzheimer's Disease And Other Biological Active Components

In the long-term research,researchers were keen to obtain natural products from nature,most of which can be used as leading compounds for the treatment of diseases or have potential medicinal value.As the birthplace of life,the ocean had produced a rich variety of species and rich resources due to its unique environment,so it has broad prospects for development.Marine microorganisims have the great potential to produce new bioactive substances due to their special living environment and they are relatively easy to use sustainably.Screening of medicinal active substances from marine microorganisms has become a hot research topic.In recent years,researchers have isolated a large number of secondary metabolites from marine fungi,and some compounds have shown good anti-Alzheimer's disease related activity.This study was based on the hypoxia hypothesis,the oxidative stress hypothesis,the inflammatory hypothesis,etc.in the pathogenesis of Alzheimer's disease.The activities?acetylcholinesterase inhibitory activity,antioxidant activity?was screened for laboratory-purified strains,resulting three target strains,Aspergillus terreus C23-3,Penicillium corylophilum TBG1-17,and Hypocrea lixii DLEN2008010.Using the"OSMAC"strategy combined with 6 different concentrations?1?mol/L,10?mol/L,100?mol/L,1 mmol/L,10 mmol/L?of inducers CuCl₂,sodium butyrate,procaine,ZnCl₂,SAHA,and 5-azaC,the strains were chemically induced in four mediums of PSB,malt extract,brown rice and soybean to activate the silencing gene cluster,which resulted in the diversity and yield of secondary metabolites of fungi.In further,using double biological activities?anti-oxidation and acetylcholinesterase inhibition?as the basis of activity tracking,TLC fingerprint and HPLC fingerprint,10 better induction groups were selected for further small-scale fermentation.Finally,C23-3-Soybean-5-azaC-10 mmol/L induction group was subjected to a large amount of fermentation.19 compounds were isolated therefrom,and 8 compounds were structurally identified,compounds 1,2,3,4,6,7 and 8 were known compounds,and compound 9 was a new compound 6,7-dimethoxyl-psoralenol,compounds?1,4,6,7,8,9?were the same series of isoflavone compounds.All compands were tested for their anti-acetylcholinesterase?AChE?activity,scavenging DPPH free radicals activity,larvicidal activity and anti-microbials activity.Compound 2?butyrolactone I?showed moderate DPPH free radical scavenging activity.Compand 1 showed moderate AChE inhibitory activity.Compand 1,2,5,7,8 exhibited potent larvicidality against brine shrimp.Compand 2,4,7 showed broad-spectrum anti-pathogenic activity.Compounds 5,6 showed good antifungal activity.The structures and activities of other compounds are also being further analyzed.The marine fungus Penicillium corylophilum TBG1-17 was fermented in a large amount of sea salt potato medium,and seven compounds were isolated therefrom.One compound structure was identified and its activity was determined,which had weak antioxidant activity,It also showed good activities against Candida albicans and weak resistance to C.albicans and MRSA.The enzymatic hydrolysate of sea cucumber was extracted with organic solvents of different polarities,and the active components were tracked by acetylcholinesterase inhibition self-development.Silica gel column chromatography,Sephadex LH-20 gel column and HPLC were used to obtained components with higher acetylcholinesterase inhibitory activity.In addition,MTT method was used to investigate the effect of total fat-soluble components of sea cucumber enzymatic hydrolysate on the growth of microglial cells?BV-2 cells?.LPS-induced cells were used to establish a model of cellular inflammation;and the effects on the levels of NO and IL-6 in LPS-induced BV-2 cells were detected by Griess method and ELISA method.The results showed that the TLC spots with acetylcholinesterase inhibitory activity showed no significant UV and fluorescence absorption,and the inhibition rate of the most active component C4-2-E was 91.04%at the concentration of 0.25?g/?L.The TLF of sea cucumber had no significant inhibitory effect on BV-2 cells within a certain concentration range,and could significantly inhibit the secretion of NO and IL-6 produced by LPS-induced BV-2 cells.Therefore,the TLF of sea cucumber enzymatic hydrolysate may have certain effects of anti-neuroinflammation and Alzheimer's disease,and the chemical structure of specific active substances needs further analysis and identification.The research in this paper showed that the"OSMAC"strategy can effectively activate the silencing gene of marine fungus by epigenetic modification,which provides an effective way for the development of anti-AD active substances;sea cucumber extract has better acetylcholinesterase inhibitory activity and its TLF have the potential of neuroprotection,both of them have good research value in the research and development of anti-AD drugs.