| TTMP were easily detected in different Chinese-liquor such as stronger flavor,Mao-flavor,and Zhima-flavor,but very poor in light fragrant Chinese liquor.Many research reported that TTMP is not only one kind of aroma,but also has health function such as promoting micro-circulation,improving immunity.As one of the main style of Chinese-liquor,light fragrant Chinese liquor should to improve its flavor,enhance its qulity further,meanwhile increase its health factor.In this study,Bacillus were isolated from Daqu for light fragrant Chinese liquor brewing.Then the strains selected for higher yield of tetramethylpyazine(TTMP)were identified by molecular biological meth od and analyzed in evolutionary aspect.Furthermore,the optimal conditions of producing TTMP were optimized and the initial application experiment was implemented.The mainly results were as follows:Total 15 strains of Bacillus were isolated from Daqu for light fragrnat Chinese liquor brewing.Their micro-morphology and clone characterize were observed.Three strains with higher TTMP yield were selected by gas chromatorgraphy,and identified as two Bacillus subtilis and one Bacillus flexus,and numbered as B76,B83 and B64,respectively,by molecular method.The synthesis conditions of strain B83 for TTMP were optimized,the results indicated that: the best raw materials was the combinatio of rice and sorghum,and the ratio of rice and sorghum was 8:2,the fermentation time was 5 days,the heat treatment time and temperatue was 4 h and 90 ℃,respectively.Under these conditions,the most TTMP concentration run up to 1600ug/ml.In order to lay the foundation for the practical application of B83,this study designed two usage methods in laboratory.One was the distillate of the culture of B83 under the optimal conditions were distilled with the fermented sorghum once more.The other one was solid culture of B83 were added into the brewing system with Daqu.The TTMP of new Chinese liquor were detected.The resulte showed that TTMP concentration of new Chinese liquor increased with the increasing of the quantity of distillate of liquid B83 culture and the solid culture of B83.When the added with 10%,the TTMP of new Chinese liquor reached up to 25ug/ml-30ug/ml.At last,Bacilluls B83 were used in brewing workshop.When added as solid culture at 1% with Daqu,the fermented sorghum had not changes in major indexes such as moistue,starch and sugar,and even the new Chinese liuqor had not changes in gas chromatogram skeleton components concentration.The new Chinese liquor kept the major characterization of light fragrant Chiese liquor and had stronger soft feel by professional critic.And the TTMP of new CHinese liquor increased to 5.3 mg/ml,an increase of nearly 3 times than the control. |
