Study On Polyphasic Taxonomy And Its Secondary Metabolites Of Two New Species Of Streptomyces

In recent years,the discovery of new actinomycetes resources from conventional soil environment became more difficult,and many known actinomycetes resources are found repeatly.People began find actinomycetes resources from special circumstances or undevelopmented environment.This research is about a potential new actinomycetes species TRM 49605 separated from Lop Nur soli and a potential new actinomycetes species TRM 46399 separated from salt-removed of Alar.Based on the genetic characteristics,morphological characteristics,physiological and biochemical characteristics and chemical characteristics to the two strains were classified and identified,in determining the basis of their taxonomic status,the secondary metabolites of these two strains were isolated,and purification and structure identification,active compounds were studied the antibacterial activity,in order to obtain the potential value in use of antibiotics.1.The polyphasic taxonomy of TRM49605 and TRM46399Optimum growth temperature of strain TRM 49605 was 37 ?,optimum growth pH was pH 7-8,optimal growth salt concentration was 2%.The strain was determined to be positive for starch,catalase,milk peptone and solidification,nitrate reduction and production of H2 S,while negative for urease,oxidase,gel liquefaction and cellulose hydrolysis.The strain was found to degrade Tween 40,Tween 60 and Tween80.Whole-cell hydrolysates sugar type of strain TRM 49605 were galactose,glucose,xylose and mannose,and whole-cell amino acid type was LL-DAP(LL-diaminopimelic acid).The phosphorylation lipids type of cell membrane were PL,GL,DPG,PE,PC,PI and PIM.The major fatty acids types were(> 10%)iso C16:0(27.26%),anteiso C15:0(11.05%),C16:0(10.36%)and Summed Feature 5(11.62%).The major menaquinone type were MK-9(H6),MK-9(H4),MK-9(H8)and MK-10(H6).The G+C content of the genomic DNA of the type strain was 71.24 mol%.Based on the above charactiristics,the strain TRM 49605 T is identified as a new species of the genus Streptomyces,named Streptomyces luozhongensis sp.nov.Optimum growth temperature of strain TRM 46399 was 37 ?,growth pH range was 6-11,optimal growth NaCl concentration was 3%.TRM 46399 was determined to be positive for oxidase,hydrogen peroxide activity,lipase activity,cellulase activity,nitrate reductase activity and amylase activity,while negative for production of H2 S and protease.Whole-cell hydrolysates sugar type of TRM 46399 wereribose,xylose,mannose and galactose,and whole-cell amino acid type was LL-DAP(LL-diaminopimelic acid).The main phospholipids types were DPG,PE,PC,PI and PIM;major fatty acid types were iso-C16:0(47.84%)and iso-C14:0(13.50%).The major menaquinone type were MK-9(H6),MK-9(H8),MK-9(H10).The strain TRM 46399 showed the highest sequence similarities to Streptomyces xinghaiensis S187T(98.62%).Based on the above characteristics,the strain TRM 46399 is identified as a new species of the genus Streptomyces,named Streptomyces aqsuensis sp.nov.2.Isolation and identification of the secondary metabolites from TRM49605 and TRM46399TRM 49605 fermentation product is enriched through D101 macroporous resin,isolated and purificated by silica gel column chromatography and HPLC preparative methods,two compounds were obtained.Then using 1H NMR and 13 C NMR data analysis,combined HSQC,HMBC and other two-dimensional spectrum analysis,the structure of two compounds were identified as 3-hydroxytetrahydro-indole(H20-1)and tunicamycin VII(H20-2)It is found that tunicamycin VII has antibacterial activity to Candida albicans,Fusarium oxysporum f.sp.cucumerinum,Aspergillus flavus,Aspergillus niger,Penicillium citrinum,and it has good antibacterial activity to Staphylococcus aureus and Micrococcus luteus,but no antibacterial activity to Escherichia coli;TRM 46399 fermentation product is enriched through D101 macroporous resin,isolated and purificated by silica gel column chromatography and Sephadex LH-20,two compounds were obtained.Using 1H NMR and 13 C NMR data analysis,the structure of two compounds were identified as genistein(46399-1)and ?-adenosine(46399-2).This study suggests that there are rich actinomycetes resources in the high salinity special environment of Lop Nur desert and salt-removed of Alar.There exist new species Streptomyces in the rich actinomycetes resources,and can be used as a kind of special soil source for the discovery of new active compounds.At the same time,the classification and identification of the actinomycetes resources and the research of the secondary metabolites have promoted the discovery and utilization of the actinomycetes resources in Xinjiang,which also provided a good research material for finding of leading compounds,biocontrol strains and biological pesticide that can against the pathogenic bacteria of the local crops such as cotton pathogens in Xinjiang.