一种用于枯草杆菌发酵生产四甲基吡嗪的补料策略

A Feeding Strategy for Tetramethylpyrazine Production by Bacillus subtilis based on the Stimulating Effects of Ammonium Phosphate

考察了铵盐对Bacillus subtilis CCTCC M 208157发酵产TTMP的影响,发现磷酸铵盐(DAP)对发酵体系中TTMP的合成有显著促进作用。进一步考察了TTMP合成对DAP的需求,结果表明,高浓度的DAP有利于TTMP的合成,但高浓度的铵根离子对细胞的生长和前体(3-羟基丁酮,乙偶姻)有抑制作用。基于以上结果,建立了一种铵盐A策略,并应用于摇瓶发酵和7.5-l发酵罐培养,TTMP浓度达7.46 g/L和7.34 g/L,与分批发酵相比分别提高了55.1%和29.0%。

In order to investigate the effects of ammonium salts on tetramethylpyrazine （TTMP） production by Bacillus subtilis CCTCC M 208157, different ammonium salts concentration was tested, and diammonium phosphate （DAP） was found to have a predominant effect on stimulating TTMP synthesis. Then DAP requirements for TTMP production was further investigated, the experimental results showed that higher concentration of DAP favored TTMP production, while both ammonium and phosphate ions exhibited inhibitory effects on the cell growth and precursor 3-hydroxy-2-butanone （HB） accumulation. Based on the above results, a DAP feeding strategy had been developed and verified in further experiments. By applying the proposed fed-batch strategy, the maximum TTMP concentrations reached 7.46 and 7.34 g/L in flask and fermenter experiments, increased by 55.1% and 29.0 % respectively compared to that of the batch TTMP fermentation.