Studies On The Production Of Actinomycin D By Streptomyces Mutabilis TRM 45540

TRM45540 strain come from Lop Nur in Xinjiang and it isolated by saline soil.This strain have variety of antibacterial activity and it identified by16 s r DNA variable as Streptomyces.mutabilis,and the main produce actinomycin D.Actinomycin D can antitumor,antiviral,antibacterial,anticancer,antituberculosis and it effect of cyclic ester peptide antibiotics.Actinomycin D phenoxazine one chromophore can be inserted between the DNA guanine and cytosine base pairs,side-chain cyclic pentapeptide which can be placed in the minor groove of the DNA double helix to promote a combination of stability,inhibition of DNA-dependent RNA polymerase vitality,blocking m RNA synthesis,interfere with the transcription process cells.Actinomycin D is the most prominent member of this family and has found clinical application as a chemotherapeutic agent,particularly in the treatment of infantile kidney tumors.In order to increase the Streptomyces.Mutabilis TRM45540 produce more actinomycin D,we use the optimization of fermentation conditions,mutation breeding strain and substrate on the levels of the three angles to explore secondary on the metabolism rules.The fermentation conditions were optimized for Streptomyces mutabilis TRM45540 to produce actinomycin D.By single factor experiment,Plackett-Burman test,the steepest ascending experiment as well as central composite design and response surface method.Results The optimized fermentation conditions were as follows: oatmeal 30.0 g/L,soybean powder 15.0 g/L,sodium chloride 19.0 g/L,potassium hydrogen phosphate 1.5 g/L,magnesium sulfate heptahydrate 2.5 g/L,calcium carbonate 1.0 g/L;rotating speed 180 r/min,initial p H value 7.0,broth content 150 m L/500 m L,inoculums size 4%,culture temperature 28?,and fermentation period 7 d.Under these optimum conditions,the yield of actinomycin D was obtained up to 679.2mg/L.Conclusion It was increased by 6.5 times compared with the original value before optimization.It takes the advantages of high yield,low impurities,cheap materials and short fermentation time for Streptomyces mutabilis TRM 45540 to produce actinomycin D.Selection of high producing strain of actinomycin D.Research on Streptomyces mutabilis TRM45540 of mutation breeding,we found the strain is easy to degradation and suitable for use UV mutagenesis,with the UV mutagenesis conditions of high positive mutation frequency,and we received UV-45540-1 and UV-45540-2 two stable high yield strain.By adding proline derivative in the culture medium TRM45540,followed by separation of raw metabolites identified except with actinomycin D,but also we found hydroxyproline be integrated into the structure of actinomycin D in the formation of discharge actinomycin A₁?single ring hydroxyproline-actinomycin D?.Comparison of these two compounds against Escherichia coli,Staphylococcus aureus Staphylococcus epidermidis,Salmonella typhii,Shigella sp,Verticillium dahliae,Fusarium oxysporum f sp.Vasinfectum more dynamic and antimicrobial activity of plant pathogens,the results showed that the antibacterial activity of actinomycin D,actinomycin slightly higher than A₁.