Accelerated Tetramethylpyrazine Production from Glucose using Diammonium Phosphate

2,3,5,6-Tetramethylpyrazine, or TMP, is a popular food flavor additive which also has a potential in dietotherapy functions, such as in cardiovascular and cerebrovascular health. However, current production methods have low yields and efficiency. The China University of Petroleum researchers led by Zijun Xiao have found a temperature-controlled preparation method for TMP with improved yield and efficiency. 

First, glucose is biosynthesized by Bacillus subtilis CICC 10211 for 29 hours in a fermentor at 37°C, pH 7.0. The speed of the stirring during biosynthesis is at 500 rpm with 1.0 vvm of airflow. The biosynthesis will yield action, the precursor of TMP. After biosynthesis, acetoin is transferred to another reactor and mixed with diammonium phosphate at 95°C for 2.5 hours. A temporary by-product was formed but decomposed quickly under higher temperature. The method also received consistent results when tested using other strains of bacteria. Crystallized TMP with high purity (99.9%) was harvested by filtration.

The method can be expanded for other acetoin-producing strains and it gives high product yield and production efficiency for TMP production from renewable materials.