Tetramethylpyrazine production from edible materials by the probiotic Bacillus coagulans

2,3,5,6-Tetramethylpyrazine (TMP) has health care functions, especially for cardiovascular and cerebrovascular health. In this study, we found that Bacillus coagulans, a well-known probiotic, has the capability to produce acetoin, a precursor of TMP. The culture conditions and medium for the production of TMP by B. coagulans CICC 20138 were optimized. Then, a novel three-step process was successfully performed for the production of TMP from edible materials by B. coagulans. First, in the acetoin enrichment process, 12.61 ± 0.34 g/L acetoin was generated at 36 h. Second, in the spore enrichment process, various factors were optimized to make the bacteria produce more spores to improve the resistance to subsequent high-temperature reactions. Third, in the TMP enrichment process, the final concentration of TMP and B. coagulans spores contained in the product reached 2.54 ± 0.26 g/L and 8.81 × 10⁸ CFU/mL at 46 h, respectively. This is the first report of using a probiotic bacterium to produce TMP. Using edible materials and the probiotic strain, this work provides a novel method for the production of a TMP food additive rich in B. coagulans spores.