【作者】 李晓霞；

【导师】 张秀红；

【作者基本信息】 山西师范大学 ， 生物学， 2016， 硕士

【摘要】 浓香、酱香、芝麻香等香型白酒都能检出一定浓度的四甲基吡嗪(TTMP),清香型白酒中TTMP则比较低。很多相关的研究表明,TTMP不仅是产香的成分,同时也具有一定的保健功能,如具有促进微循环,提高人体免疫力等多种保健功能。在人们越来越追求健康的当下,作为主要香型的清香型白酒,在改善风味,提高品质的同时,也需要增加健康因子。本研究从清香大曲中分离纯化芽孢杆菌,筛选出四甲基吡嗪产量较高的菌株,对其进行分子生物学鉴定及进化分析。进一步对四甲基吡嗪高产菌株合成TTMP条件进行优化,并完成初步的应用实验,主要结果如下:本实验从汾酒大曲中分离出15株可以产生四甲基吡嗪的芽孢杆菌。通过观察其菌落形态的一些基本特征和细胞的显微形态图对它们进行纯种确认,通过气相色谱法筛选出3株高产四甲基吡嗪的芽孢杆菌,并利用16S r DNA序列测定法进行菌种鉴定,得出三株高产四甲基吡嗪的芽孢杆菌中两株为枯草芽孢杆菌、一株为弯曲芽孢杆菌,分别编号B64,B83,B76。选取芽孢杆菌B83对其液体状态下合成TTMP的条件进行优化,利用紫外分光光度法检测发酵液中TTMP的产量,结果表明:最佳培养基为大米高粱8:2的时候四甲基吡嗪产量最高,培养时间设定为5天,热处理温度设为90℃,热处理时间为4h,铵盐量为5g,四甲基吡嗪的含量最高可达1600ug/ml。为菌株的实际应用奠定基础,本实验利用菌株B83合成TTMP条件的优化结果,接种培养芽孢杆菌B83并进行铵盐处理后,蒸馏得到串蒸增香液;同时进行芽孢杆菌B83固体培养后作为强化曲加入到酿酒体系,两种方法检测新产酒中的TTMP含量,结果发现随着串蒸增香液及芽孢杆菌固体制剂加入量的不断增加,新产酒中TTMP的含量明显增加。当加入量为10%时,新产酒中TTMP的产量高达25-30ug/ml之间。最后,将芽孢杆菌固体制剂应用于酿酒车间,发现芽孢杆菌制剂对新产酒中水分、淀粉及还原糖等主要指标影响不大,对主要色谱骨架成分也没有影响,保持了汾酒的基本特性,经专业评酒师品评认为增强了绵甜感,TTMP增加到5.3ug/ml,比对照增加了近3倍。更多还原

【Abstract】 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.更多还原