State Key Laboratory of Food Science and Technology, Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China.
Bioprocess Biosyst Eng. 2010 Oct;33(8):953-9. doi: 10.1007/s00449-010-0419-5. Epub 2010 Mar 21.
To examine the effects of ammonium salts on tetramethylpyrazine (TTMP) production by Bacillus subtilis CCTCC M 208157, different ammonium salts were tested, and diammonium phosphate (DAP) was found to have a predominant effect on stimulating TTMP synthesis. The DAP requirements for TTMP production were then investigated, experimental results showed that higher concentrations of DAP favored TTMP production, while both the ammonium and phosphate ions exhibited inhibitory effects on the cell growth and precursor 3-hydroxy-2-butanone accumulation. Based on the results above, a DAP feeding strategy was 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% compared to that of the batch TTMP fermentation, respectively. To our knowledge, these results, i.e., TTMP yields in flask or fermenter fermentations, were new records on TTMP fermentation by B. subtilis.
为了考察铵盐对枯草芽孢杆菌 CCTCC M 208157 生产四甲基吡嗪（TTMP）的影响，测试了不同的铵盐，发现磷酸二铵（DAP）对刺激 TTMP 合成有显著影响。然后研究了 DAP 对 TTMP 生产的需求，实验结果表明，较高浓度的 DAP 有利于 TTMP 的生产，而铵离子和磷酸根离子对细胞生长和前体 3-羟基-2-丁酮的积累均表现出抑制作用。基于上述结果，开发并在进一步的实验中验证了 DAP 补料策略。通过应用所提出的补料分批策略，在摇瓶和发酵罐实验中，TTMP 的最大浓度分别达到 7.46 和 7.34 g/L，与分批 TTMP 发酵相比，分别提高了 55.1%和 29.0%。据我们所知，这些结果，即在摇瓶或发酵罐发酵中 TTMP 的产率，是枯草芽孢杆菌发酵生产 TTMP 的新纪录。
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