| The secondary metabolites produced by actinomycetes are rich in variety and diverse in activity,and are widely used in drug development.In recent years,with the continuous increase of drug-resistant strains,scientific researchers continue to expand the scope of screening of active strains and try new screening methods,placing the hope of overcoming drug-resistant strains on the discovery of new compounds.New species of actinomycetes contain new genes and new metabolic pathways,and are the main research materials for exploring and discovering new metabolites.In this study,ten new species of actinomycetes were used as research materials.Based on genome information analysis,biological activity detection and metabolite spectroscopy analysis,strains were screened and metabolites were explored.Complete genome sequencing and splicing of ten new species of actinomycetes,and bioinformatics analysis.Through Gene Mark prediction,genome non-coding RNA prediction,genome protein coding gene function annotation and anti SMASH online analysis,the results show that the actinomycete genome is a Scaffold sequence,which can be connected into a circular genome.For the functional annotation of the protein-coding genes of the genome,the GO database shows that the strains are mainly related to the biological process of the protein-coding gene types and numbers;the KEGG metabolic pathway shows that carbohydrate metabolism and amino acid metabolism are the main components;COG annotations indicate that the encoded protein genes are mainly involved The basic functions of cells.Through Anti SMASH online analysis,it was found that 10 strains contained abundant secondary metabolite synthesis gene clusters.By comparing the information of biosynthetic gene clusters,strains with high metabolic potential were screened out.It was further screened by antibacterial activity detection and product NMR HSQC-TOCSY spectroscopy.Through comparative analysis,the strain Streptomyces lasuensis TRM68005 contains more secondary metabolite NMR gene clusters and strong antibacterial activity than other strains,and the HSQC-TOCSY spectrum has a secondary metabolite signal area.Therefore,the strain TRM 68005 is judged It has a strong potential for the synthesis of secondary metabolites.The selected new species of Streptomyces TRM 68005 was fermented and prepared by silica gel column chromatography,gel column chromatography and high performance liquid chromatography.A total of 5 monomer compounds were separated.Through 1H NMR and 13 C NMR and other nuclear magnetic resonance spectrum data,combined with the comparison and analysis of the spectrum data in the microspectral database and literature,five compounds were identified as:2-Methyl butyl propyl phthalate(1),(E)-6-Decenyl Acetate(2),methyl 11-hydroxyhexadecanoate(3),ntetradecanoic acid(4),di-2-ethylhexyl phthalate(DEHP)(5).The suspected new species of Streptomyces TRM 64462 found in the previous whole-genome sequence analysis was systematically classified and identified through morphological characteristics,genome information,chemical classification characteristics,and physiological and biochemical characteristics.It was identified that the strain represented a new species of Streptomyces.And confirmed the systematic taxonomy status of this strain.According to the isolation source of strain TRM 64462,it was named Streptomyces alhagi.In this study,10 new species of actinomycetes were analyzed by bioinformatics,which improved the information of new species and revealed the molecular level functions of the strains.And through the combination of nuclear magnetic resonance spectroscopy analysis and traditional biological activity screening strategies,comprehensive information from all aspects,explore the potential and research value of strains to synthesize metabolites,and provide guidance for the subsequent excavation of secondary metabolites of strains.It also shows the diversification of mining methods for secondary metabolites. |
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