The Separation And Purification Of The Active Substance Of Spongeous Bacteria And Its Structural Analysis

In recent decades,land microbial resources have been developed and utilized in large quantities.Due to the high-pressure,high-salt,and oligotrophic marine environments,the active substances extracted from marine organisms have stronger and more biological activity.The separation of different compounds from Marine life means that there is a promising natural product.Through the activity screening(including herbicidal,insecticidal,antibacterial,and anti-tumor),it was found that the fermentation broth of the sponge endophyte NH1403 had good herbicidal activity against barnyard grass and purslane.It has good anti-tumor activity on human hepatoma cells(HepG2).In this experiment,the antitumor activity was tracked and the strain was isolated to produce an antitumor active metabolite.The analysis and preparation were carried out by extraction,thin layer chromatography and high performance liquid chromatography(HPLC)analysis.The three active substances,namely A1,B1 and B2,were found to have antitumor activity.Because the fermentation broth had better herbicidal activity,it was further verified whether the three compounds had herbicidal activity test.It was found that compound A1 and B2 had inhibitory activity on the germination of barnyard grass seeds and purslane seeds.Due to the small amount of compounds B1 and B2,it is not enough to carry out structural analysis,only the compound A1 was subjected to structural analysis.By high resolution mass spectrometry analysis,the relative molecular mass of A1 compound was 402.The pure product was analyzed by 1H-NMR,13C-NMR,DEPT90,DEPT135,1H-1HCOSY,HMBC,HSQC NMR and the molecular formula was C24H38O5,and its structure was worked out.A quantitative analysis method for the determination of the content of compound A1 during fermentation and isolation and purification was established by HPLC.Through the observation of the morphological characteristics of the NH1403 strain,the phenomenon of physiological and biochemical experiments and 16SrDNA gene sequence of tests and the establishment of the further growth of trees,determine the NH 1403 strains belong to bacillus genus.