Excavation Of Biologically Active Substances From Strain HDZK-BYSB47

Natural products from the microorganisms have great research value and development prospects.Endophyte has great potential in acquiring new natural products and developing new drugs.Especially in mining new antibacterial,anti-cancer and anti plant diseases and insect pests,it has become the focus of the discovery of new functional drugs.At present,genome mining provides new ideas and methods for the separation of new natural product from endophytes,and the discovery of target substances by genome mining has been applied to various fields.Termites have caused serious damage to: buildings,crops,books,furniture,clothing,ship,railway,paper,forest,fruit tree,dam,buried cable and so on,every year have caused huge economic losses.Biological control of termite has the characteristics of high efficiency,low toxicity,no residue and no pollution.The research group has isolated a antitermitic compound-producing endophytic bacteria HDZK-BYSB47(Bacillus subtilis)from Chamaecyparis lawsoniana,the idea and method of producing antitermitic biological agents by using the fermentation products of endophyte were proposed.Endophytic bacteria usually produce a variety of active metabolites,analyze the biosynthetic gene cluster in its genome,optimize the fermentation conditions and detect and separate the metabolites,which can help us to obtain highly active new compounds.Therefore,the strain HDZK-BYSB47 was used as the tested strain,the whole genome sequencing of the strain was carried out.To searched for bioactive products of strain HDZK-BYSB47 by combined with genome mining and bioactive screening technology.In addition,the genetic operation system of strain HDZK-BYSB47 was established to lay the foundation for subsequent genetic work of strain.In addition,the genetic operation system of strain HDZK-BYSB47 was established to lay the foundation for subsequent test of strain.The test results show that:1.A class of bioactive compounds were found in the metabolites of strain HDZK-BYSB47 fermentation.2.Combined with the biosynthetic gene cluster of HDZK-BYSB47,isolation and purification results.It was determined that the compound was Bacillaene.3.The dynamic and static coupling fermentation mode was applied in the Bacillus subtilis(HDZK-BYSB47),the production of Bacillaene was significantly improved.4.The genetic manipulation system of strain HDZK-BYSB47 was established,and the shuttle plasmid pUB110 S of Bacillus subtilis-E.coli was successfully constructed.It is the basis for further genetic research and global regulation.5.Plasmid pUB110 S is used as the carrier,the expression vector of positive regulatory factors,CodY and Spo0 A,was overexpressed in the receptor strain HDZK-BYSB47.6.The results of strain fermentation showed that when the global regulatory factor CodY was overexpressed,the target compound Bacillaene was not produced,and the corresponding new compounds were produced.When the global regulatory factor Spo0 A was overexpressed,the yield of compound Bacillaene and its structural analogues increased significantly.This study will find efficient,low toxic,environmentally friendly and economical termite biological control agent.It provides a new technology and method for more new termite biological control agents.It has a certain scientific significance and potential market application prospects.