低高级醇啤酒酵母的选育及应用
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摘要
高级醇是啤酒中主要的风味物质,适量的高级醇能赋予啤酒丰满的口感,增加酒体的协调性,过量则会给啤酒带来异杂味,使人头晕、头痛,并引起人体某些疾病。降低啤酒中高级醇的含量是现代啤酒企业所追求的主要目标之一,菌种改良和发酵条件是控制高级醇的主要方法,淡雅、清香、低高级醇、绿色健康的啤酒必将是21世纪的发展方向。
本试验从实验室收藏的啤酒酵母菌株出发,利用紫外线和硫酸二乙酯对出发酵母菌株进行复合诱变处理,根据酵母菌高级醇的合成代谢与乳酸代谢途径之间的关系,采用乳酸培养基、碳酸钙培养基和TTC上层培养基三重筛选获得优良菌株。同时采用响应面分析方法对发酵条件进行优化,得到最优发酵参数。利用具有全夹套制冷,C.I.P自动清洗系统的1000 L不锈钢发酵设备,进行中试啤酒发酵。
出发菌株经过紫外线-DES(硫酸二乙酯)复合诱变处理后,高级醇含量降低了24.34%。响应面分析方法优化得到最佳发酵条件：发酵温度10.91℃,氨基酸含量为172.83mg/L,接种量为3.44%,高级醇含量为70.8 mg/L。
中试12°P啤酒发酵结果显示高级醇含量降低明显,总高级醇产量70.67mg/L,低于市售国内外知名大企业啤酒高级醇含量的平均值,达到了国际优质啤酒的标准。对发酵啤酒其他成分进行分析检测显示,酒精度为4%、双乙酰0.071mg/L、总酯29.1mg/L、总醛18.51mg/L、总酸3.15ml/L各项指标均达到国家标准,酒液淡黄、清澈透明、富有光泽、酒质柔和、浓香醇厚、有明显酒花香和麦芽香,并且具有啤酒特有的爽口苦味和杀口力。
Higher alcohols is the main flavor substance in the beer, adequate higher alcohols can give beer the full taste and coordination body,the beer will have different and miscellaneous flavor if higher alcohols content is excess and make people headache and vertigo and cause some human diseases. Reducing the content of higher alcohols in beer is one of main objectives that the modern beer enterprise pursued.The main method of control higher alcohols are strain improvement and conditions of fermentation, Quietly elegant, light fragrance, low content of higher alcohols, green and healthy of beer will be the development direction of 21st century'.
According to the relation that between constructive metabolism of higher alcohols and lactic acid metabolic pathway,the good strain was got by compound mutation of UV-DES with triplication screening of lactic acid, calcium carbonate and TTC upper stratum medium of the preserved strain. The fermentation conditions were optimized by using reponse surface analytical method and the optimization fermentation parameters were got.The pilot production of beer fermentation was made by using 1000L stainless steel fermentation equipment with refrigeration of whole jacket, C.I.P automatic cleaning system.
The content of higher alcohols of the strains was reduced by 24.34% after compound mutation of UV-DES. The fermentation conditions were got by using reponse surface analytical method, the optimization fermentation parameters are: fermentation temperature 10.91℃, content of amino acids 172.83mg/L, inoculum size 3.44% and the content of higher alcohols is 70.8 mg/L.
The whole content of higher alcohols of the pilot beer is 70.67mg/L and lower than the average value of higher alcohols content of the beer made in well-known domestic and overseas big enterprise. The pilot beer quality come up to advanced world standards. The fermentation parameters of pilot beer are:alcoholicity 4%、biacetyl 0.071mg/L、total ester 29.1mg/L、total aldehyde 18.51mg/L、total acid 3.15ml/L, all indexes meet the national standards,the beer is yellowish,clean and clear,lustrous,body soft and thick aroma and has obviously fragrance of hop and malt and typical refreshing bitter taste and stimulation power.
本试验从实验室收藏的啤酒酵母菌株出发,利用紫外线和硫酸二乙酯对出发酵母菌株进行复合诱变处理,根据酵母菌高级醇的合成代谢与乳酸代谢途径之间的关系,采用乳酸培养基、碳酸钙培养基和TTC上层培养基三重筛选获得优良菌株。同时采用响应面分析方法对发酵条件进行优化,得到最优发酵参数。利用具有全夹套制冷,C.I.P自动清洗系统的1000 L不锈钢发酵设备,进行中试啤酒发酵。
出发菌株经过紫外线-DES(硫酸二乙酯)复合诱变处理后,高级醇含量降低了24.34%。响应面分析方法优化得到最佳发酵条件：发酵温度10.91℃,氨基酸含量为172.83mg/L,接种量为3.44%,高级醇含量为70.8 mg/L。
中试12°P啤酒发酵结果显示高级醇含量降低明显,总高级醇产量70.67mg/L,低于市售国内外知名大企业啤酒高级醇含量的平均值,达到了国际优质啤酒的标准。对发酵啤酒其他成分进行分析检测显示,酒精度为4%、双乙酰0.071mg/L、总酯29.1mg/L、总醛18.51mg/L、总酸3.15ml/L各项指标均达到国家标准,酒液淡黄、清澈透明、富有光泽、酒质柔和、浓香醇厚、有明显酒花香和麦芽香,并且具有啤酒特有的爽口苦味和杀口力。
Higher alcohols is the main flavor substance in the beer, adequate higher alcohols can give beer the full taste and coordination body,the beer will have different and miscellaneous flavor if higher alcohols content is excess and make people headache and vertigo and cause some human diseases. Reducing the content of higher alcohols in beer is one of main objectives that the modern beer enterprise pursued.The main method of control higher alcohols are strain improvement and conditions of fermentation, Quietly elegant, light fragrance, low content of higher alcohols, green and healthy of beer will be the development direction of 21st century'.
According to the relation that between constructive metabolism of higher alcohols and lactic acid metabolic pathway,the good strain was got by compound mutation of UV-DES with triplication screening of lactic acid, calcium carbonate and TTC upper stratum medium of the preserved strain. The fermentation conditions were optimized by using reponse surface analytical method and the optimization fermentation parameters were got.The pilot production of beer fermentation was made by using 1000L stainless steel fermentation equipment with refrigeration of whole jacket, C.I.P automatic cleaning system.
The content of higher alcohols of the strains was reduced by 24.34% after compound mutation of UV-DES. The fermentation conditions were got by using reponse surface analytical method, the optimization fermentation parameters are: fermentation temperature 10.91℃, content of amino acids 172.83mg/L, inoculum size 3.44% and the content of higher alcohols is 70.8 mg/L.
The whole content of higher alcohols of the pilot beer is 70.67mg/L and lower than the average value of higher alcohols content of the beer made in well-known domestic and overseas big enterprise. The pilot beer quality come up to advanced world standards. The fermentation parameters of pilot beer are:alcoholicity 4%、biacetyl 0.071mg/L、total ester 29.1mg/L、total aldehyde 18.51mg/L、total acid 3.15ml/L, all indexes meet the national standards,the beer is yellowish,clean and clear,lustrous,body soft and thick aroma and has obviously fragrance of hop and malt and typical refreshing bitter taste and stimulation power.
引文
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