Screeing Of High Yield Strain Of Tetramethylpyraszine From Chinese Jiuqu And Optimizating Of Its Fermentation Conditions

Tetramethylpyrazine(TTMP)also Known as 1 igustiazine,which is an alkaloid extracted from Chinese herbal medicine Ligusticum chuanxiong Hort and mainly used in the field of medicine and perfume industry.TTMP is not only an important liquor flavor-related compound but also in possession of special pharmacological functions benefical to people’s health.In this study,a TTMP producing strain Bacillus subtilis R19 was screened from Jiang-flavour Daqu of xiaoerbulake liquor.To get a high TTMP producing strain,the hereditary stabile mutant T451,was obtained by Atmospheric and Room Temperature Plasma(ARTP)treatment of Bacillus subtilis R19,Studies on T451 was carried out.The main contents and results as follows:(1)The production of TTMP of R19 was 267.25 mg/L,Preliminary optimization was carried out and 2.51 g/L TTMP was obtained under the optimized medium composition(100 g/L maltose,40 g/L soybean cake powder,15 g/L yeast extract,30 g/L diammonia phosphate)and cultivation condition(inoculation amount 6%,initial pH 6.5,200 r/min).(2)The hereditary stabile mutant T451 was obtained by Atmospheric and Room Temperature Plasma(ARTP)treatment of Bacillus subtilis R19,The production of TTMP was 1.19 g/L.(3)The differences between mutant T451 and Bacillus subtilis R19 was compared,such as colonial morphology and strain growth curve.(4)Optimization and fermentation control was carried out and 1.61 g/L TTMP was obtained under the optimized medium composition.Response surface methodology(RSM)was applied to optimize the culture condition.2.28 g/L TTMP was obtained under the optimized condition.2.34 g/L TTMP was obtained by applying the feeding strategy,3.52 g/L TTMP was obtained by applying the pH control strategy.(5)TTMP was prepared from mutant T451 under solid-state fermentation.On the basis of the result of one-factor-time experiments,Response surface methodology(RSM)was applied to optimize the culture condition.4.13 g/kg TTMP was obtained under the optimized condition.(6)T451 was co-cultured with Yeast,Lactobacillus or Geotrichum candidum.Yeast and Geotrichum candidum was found a positive effect to increase the TTMP.