优良刺葡萄酒酵母的选育及应用研究
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摘要
为了获得一株发酵性能优良、性质稳定的刺葡萄酒酵母,以指导生产实践,酿造优质刺葡萄酒。本课题首先从带皮渣的刺葡萄汁发酵液中筛选出一株较优酵母。然后对其生长、耐受及发酵特性进行研究。最后通过正交试验建立了科学合理的刺葡萄酒发酵工艺。主要研究结果如下：
1.设置刺葡萄汁发酵试验,确定以起酵速度、发酵气味、凝聚性能、产酒精度、感官评定为初筛指标,筛选出编号分别为Y6、Y24、Y101、Y102、Y121的5株酵母。以安琪葡萄酒活性干酵母为对照,用各纯种酵母酿造刺葡萄酒,以酿造酒的理化品质和感官评定为指标,复筛出酿造刺葡萄酒的优良酵母菌株Y102。通过观察酵母Y102在WL营养琼脂培养基上的菌落特征及子囊孢子、假丝孢子、掷孢子的生长情况,结合酵母菌鉴定手册及Cavazza关于WL营养琼脂培养基鉴定酵母的理论,初步判定酵母Y102为酿酒酵母属(Saccharomyces cerevisiae)酵母。
2.测定酵母Y102生长曲线,了解其生长周期,确定20h为其对数生长期的中期。通过L9(33)正交试验对酵母Y102生长条件进行优化,确定其最佳生长条件为：pH5.0,温度30℃,转速200r/min。OD560值为0.798,菌密度为1.2×108个/mL。设计耐受性试验,测定出酵母Y102的各项耐性指标：耐酒精量为18%vol；耐高温为40℃；耐高糖为60%；耐高酸为pH1.8；耐二氧化硫为200mg/L；耐KCl为2.0mol/L;耐饥饿为18天。对比安琪葡萄酒活性干酵母,酵母Y102耐受性能较好。设计发酵试验,测定出酵母Y102的发酵特性：对于含糖20%、pH3.5的刺葡萄果浆,酵母Y102在接种7小时后开始发酵,120h后达到高峰,8天后主发酵结束。与安琪葡萄酒活性干酵母比较,酵母Y102的起酵时间更快,并且发酵速率也更快,缩短了发酵周期,节省了发酵成本。在24℃条件下,发酵结束后,酵母Y102发酵产酒精11.5%vol,残糖2.1g/L。对比安琪葡萄酒活性干酵母,酵母Y102发酵性能更好,适合酿造刺葡萄酒。
3.以发酵周期,酒精度、残糖、总酸,感官评定分数为综合指标,对初始糖度、初始pH值、发酵温度、酵母接种量进行单因素酿酒试验。通过对最终刺葡萄成品酒的综合评价,得出酵母Y102酿造刺葡萄酒的最佳初始糖度为22%；最佳初始pH值为3.5；最佳发酵温度为24℃；最佳酵母接种量为4%。以初始pH3.0、pH3.5、pH4.0;初始糖度20%、22%、24%；发酵温度22℃、24℃、26℃为因素水平,以感官评定为指标,采用L9(33)正交试验对刺葡萄酒发酵工艺条件进行优化。确定最佳发酵组合为：pH3.5,初始糖度20%,发酵温度22℃,感官评定分数为94。对该工艺条件进行验证试验,感官评定分数为95分,与理论值相比差异不显著。各项理化指标均达到GB15037-2006的基本要求。
In order to obtain a thorn wine yeast who has good and stable fermentation performance. To guide the production of brewing quality thorn wine.The first, an optimal yeast was selected from grape juice fermentation.Then study the growth, tolerance and fermentation characteristics.Finally,established a scientific and rational orthogonal thorn wine fermentation.
1.Seted thorn juice fermentation tests.Established fermentation rate,fermentation odor, pool performance, alcohol production and sensory evaluation as indicators,selected 5 strains of yeast, Y6、Y24、Y101、Y102、Y121. Angel active dry yeast was the control, brewing thorn wine with the pure yeast, the wine sensory evaluation and physical-chemical quality was indicators. Rescreening a good thorn wine yeast strain Y102.Oberving the colony characteristics of Y102 in WL nutrient agar medium,and growth of Ascospores, Candida spores, spore throwing. According to the yeast identification manual and identificational theory of WL nutrient agar medium of Cavazza, preliminary determination Y102 is the Saccharomyces cerevisiae yeast.
2. Determination of the growth curve of Y102, Understanding of its growth cycle, determined its mid-logarithmic phase was 20h.Optimal growth condition of Y102 with L9 (33) Orthogonal. To determine the optimum growth conditions: pH5.0,temperature 30℃, speed 200r/min. OD560 value of 0.798, the bacterial density of 1.2×108a/mL. Design tolerance test, to determine the endurance of Y102:amount of resistance to alcohol 18% vol; high temperature is 40℃; resistant to high glucose 60%; resistance to acid pH1.8; resistance to sulfur dioxide for the 200mg/L; resistant to KC1 was 2.0mol/L; resistance to starvation for 18 days. Comparison Angel wine active dry yeast, Y102 has better tolerated performance. Design fermentation tests, determination of the fermentation characteristics of Y102:after 7 hours,the fermentationg was started in the grape berries of Sugar 20%, pH3.5. Reached a peak after 120h,8 days after the end of the main fermentation. Comparison Angel wine active dry yeast, Y102 has the faster starting fermentation time and fermentation rate, shorten the fermentation period, saving the cost of fermentation. In the conditions at 24℃, after the end of fermentation,Y102 producted 11.5% vol ethanol. Residual sugar 2.1g/L.Y102 has better fermentation performance for brewing wine thorn.
3.The fermentation period, alcohol, residual sugar, total acid, sensory evaluation scores for the composite indicator. Testing the initial sugar content, initial pH, fermentation temperature, yeast inoculation.By evaluating the final grape wine, obtained the best initial sugar content of 22%; optimal initial pH 3.5; optimum fermentation temperature was 24℃; the best yeast inoculum of 4%. The initial pH3.0, pH3.5, pH4.0; initial sugar content of 20%,22%,24%; fermentation temperature 22℃,24℃,26℃for the factor level.The sensory evaluation as indicators, optimization of fermentation conditions. Determine the best combination of fermentation:pH3.5, 20% of the initial sugar content, fermentation temperature 22℃, sensory evaluation scores for the 94. Test the verify of the conditions, the sensory evaluation score of 95 points, was not significant compared with the theoretical value. The physical-chemical indicators meet the basic requirements of GB15037-2006.
1.设置刺葡萄汁发酵试验,确定以起酵速度、发酵气味、凝聚性能、产酒精度、感官评定为初筛指标,筛选出编号分别为Y6、Y24、Y101、Y102、Y121的5株酵母。以安琪葡萄酒活性干酵母为对照,用各纯种酵母酿造刺葡萄酒,以酿造酒的理化品质和感官评定为指标,复筛出酿造刺葡萄酒的优良酵母菌株Y102。通过观察酵母Y102在WL营养琼脂培养基上的菌落特征及子囊孢子、假丝孢子、掷孢子的生长情况,结合酵母菌鉴定手册及Cavazza关于WL营养琼脂培养基鉴定酵母的理论,初步判定酵母Y102为酿酒酵母属(Saccharomyces cerevisiae)酵母。
2.测定酵母Y102生长曲线,了解其生长周期,确定20h为其对数生长期的中期。通过L9(33)正交试验对酵母Y102生长条件进行优化,确定其最佳生长条件为：pH5.0,温度30℃,转速200r/min。OD560值为0.798,菌密度为1.2×108个/mL。设计耐受性试验,测定出酵母Y102的各项耐性指标：耐酒精量为18%vol；耐高温为40℃；耐高糖为60%；耐高酸为pH1.8；耐二氧化硫为200mg/L；耐KCl为2.0mol/L;耐饥饿为18天。对比安琪葡萄酒活性干酵母,酵母Y102耐受性能较好。设计发酵试验,测定出酵母Y102的发酵特性：对于含糖20%、pH3.5的刺葡萄果浆,酵母Y102在接种7小时后开始发酵,120h后达到高峰,8天后主发酵结束。与安琪葡萄酒活性干酵母比较,酵母Y102的起酵时间更快,并且发酵速率也更快,缩短了发酵周期,节省了发酵成本。在24℃条件下,发酵结束后,酵母Y102发酵产酒精11.5%vol,残糖2.1g/L。对比安琪葡萄酒活性干酵母,酵母Y102发酵性能更好,适合酿造刺葡萄酒。
3.以发酵周期,酒精度、残糖、总酸,感官评定分数为综合指标,对初始糖度、初始pH值、发酵温度、酵母接种量进行单因素酿酒试验。通过对最终刺葡萄成品酒的综合评价,得出酵母Y102酿造刺葡萄酒的最佳初始糖度为22%；最佳初始pH值为3.5；最佳发酵温度为24℃；最佳酵母接种量为4%。以初始pH3.0、pH3.5、pH4.0;初始糖度20%、22%、24%；发酵温度22℃、24℃、26℃为因素水平,以感官评定为指标,采用L9(33)正交试验对刺葡萄酒发酵工艺条件进行优化。确定最佳发酵组合为：pH3.5,初始糖度20%,发酵温度22℃,感官评定分数为94。对该工艺条件进行验证试验,感官评定分数为95分,与理论值相比差异不显著。各项理化指标均达到GB15037-2006的基本要求。
In order to obtain a thorn wine yeast who has good and stable fermentation performance. To guide the production of brewing quality thorn wine.The first, an optimal yeast was selected from grape juice fermentation.Then study the growth, tolerance and fermentation characteristics.Finally,established a scientific and rational orthogonal thorn wine fermentation.
1.Seted thorn juice fermentation tests.Established fermentation rate,fermentation odor, pool performance, alcohol production and sensory evaluation as indicators,selected 5 strains of yeast, Y6、Y24、Y101、Y102、Y121. Angel active dry yeast was the control, brewing thorn wine with the pure yeast, the wine sensory evaluation and physical-chemical quality was indicators. Rescreening a good thorn wine yeast strain Y102.Oberving the colony characteristics of Y102 in WL nutrient agar medium,and growth of Ascospores, Candida spores, spore throwing. According to the yeast identification manual and identificational theory of WL nutrient agar medium of Cavazza, preliminary determination Y102 is the Saccharomyces cerevisiae yeast.
2. Determination of the growth curve of Y102, Understanding of its growth cycle, determined its mid-logarithmic phase was 20h.Optimal growth condition of Y102 with L9 (33) Orthogonal. To determine the optimum growth conditions: pH5.0,temperature 30℃, speed 200r/min. OD560 value of 0.798, the bacterial density of 1.2×108a/mL. Design tolerance test, to determine the endurance of Y102:amount of resistance to alcohol 18% vol; high temperature is 40℃; resistant to high glucose 60%; resistance to acid pH1.8; resistance to sulfur dioxide for the 200mg/L; resistant to KC1 was 2.0mol/L; resistance to starvation for 18 days. Comparison Angel wine active dry yeast, Y102 has better tolerated performance. Design fermentation tests, determination of the fermentation characteristics of Y102:after 7 hours,the fermentationg was started in the grape berries of Sugar 20%, pH3.5. Reached a peak after 120h,8 days after the end of the main fermentation. Comparison Angel wine active dry yeast, Y102 has the faster starting fermentation time and fermentation rate, shorten the fermentation period, saving the cost of fermentation. In the conditions at 24℃, after the end of fermentation,Y102 producted 11.5% vol ethanol. Residual sugar 2.1g/L.Y102 has better fermentation performance for brewing wine thorn.
3.The fermentation period, alcohol, residual sugar, total acid, sensory evaluation scores for the composite indicator. Testing the initial sugar content, initial pH, fermentation temperature, yeast inoculation.By evaluating the final grape wine, obtained the best initial sugar content of 22%; optimal initial pH 3.5; optimum fermentation temperature was 24℃; the best yeast inoculum of 4%. The initial pH3.0, pH3.5, pH4.0; initial sugar content of 20%,22%,24%; fermentation temperature 22℃,24℃,26℃for the factor level.The sensory evaluation as indicators, optimization of fermentation conditions. Determine the best combination of fermentation:pH3.5, 20% of the initial sugar content, fermentation temperature 22℃, sensory evaluation scores for the 94. Test the verify of the conditions, the sensory evaluation score of 95 points, was not significant compared with the theoretical value. The physical-chemical indicators meet the basic requirements of GB15037-2006.
引文
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