Usage By The Use Of Ribosome-engineering Technology

When we screen the activity of microorganism, 95% strains don't show any activity under the available model. Many chemists pay attention to how to make good use of the non-active microorganism. My research focused on exploiting new microbial resources by the use of ribosome-egineering.A non-active strain Streptomyces roseoflavus was chosen as a parent strain and then was induced the antibiotic (chloramphenicol, streptomycin) resistant i.e. ribosome engineering technology. 270 strains of mutation were obtained, which of them showed obvious difference in the morphology and color from their parent strain. In the middle of the mutations, three strains' metabolites showed antitumor activity on mammalian cancer tsFT210 cells.A mutation Str3054 (streptomycin resistant) showed stronger cytoclasis bioactivity on mammalian cancer tsFT210 cells. It was selected to be fermented. The whole fermentation broth was separated by the means of modern chromatographic methods in a bioassay-guided separation manner. From the bioactive part, n-BuOH extract, six compounds were isolated by silica gel, Sephadex LH-20 and reverse-phase high performance liquid chromatragraphy. The structures of five compounds were elucidated mainly by use of spectroscopic methods (UV, IR, MS, 1D-NMR, 2D-NMR). They were phthalic acid diisobutyl ester, bis(2-ethylhexyl)fumarate(Z, R, R), Inosine, Uridine, cyclo-(Pro- Gly). Bis(2-ethylhexyl)fumarate(Z,R,R) was a new compound. Among them, five compounds showed the antitumor activities by the use of SRB and flow cytometry using mammalian cancer tsFT210 cells.The rpsL gene (encoding the ribosomal protein S12) in Streptomyces roseoflavus and its mutation Str3054 were cloned to test whether there is amutation in this region. But the result was negative. So it was presumed there may be a new mutation in other regions which confer the strain resistance to streptomycin.In summary, the experiment was the first report to apply the ribosome-engineering technology to obtain the active marine strains, which showed the cell cycle inhibitory activity. And then we studied its metabolites of mutation and obtained six compounds, five of which showed antitumor activities. It indicated that it is feasible to exploit new microbial resources for medicinal usage by applying the ribosome-engineering technology to non-active actinomycete in nature.