Study On Metabolomics Of Recombinant Bacillus Subtilis RH44

In this paper, the metabonomics of Bacillus subtilis RH44 were investigated primarily.Through comparison of mass graphics , the boil ethanol extract methods was confirmed; the comparison of separation results, the optimal mobile phases and separate gradient were confined: A :1mM HCOONH4 , B: 80% methanol (contain 1mM HCOONH4); gradient was 0～10min A:98 %- 40%,10～20 min A:40 %- 24%。The metabolites contrast under different nitrogen sources and culture time were studied, and also the comparison of Bacillus subtilis 168 and Bacillus subtilis RH44 strains. According to the fore-study, the best proper nitrogen of Bacillus subtilis RH44 was NaNO3 and NH4NO3, but some papers used (NH4)2SO4 as nitrogen source. To investigate the effects of nitrogen source on metabolites, when use NaNO3 and NH4NO3, NADH and NADPH content were lower than (NH4)2SO4; Comparing of different culture times :when 12h, the strains was in logarithm growth phase,and the synthesize rate of riboflavin was high, so the metabolites content of PP pate was low;when 24h, the strains was in stable phase, the synthesize rate of riboflavin was low, the metabolites content accumulated;Comparing of different strains:the metabolize pate of B. subtilis 168 does not produce riboflavin, so the metabolites content of PP pate was low, and the metabolites content of TCA and EMP pates was high; B. subtilis RH44 produces riboflavin, the metabolites content of PP pate was higher.