西藏传统青稞酒的生产菌株选育及生产技术研究
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摘要
西藏青稞酒是藏族人民的最重要的传统饮品,至今已有1300多年的历史。本文通过对从藏曲中筛选出的优良菌株再进行一系列物理和化学的诱变选育,以期获得更优良的菌株,并以此制备纯种曲进行青稞酒酿造,并对其酿造工艺条件进行了优化试验研究。研究结果表明:
(1)糖化菌种的选育。对从藏曲中筛选的18株优良糖化菌中,结合糖化透明圈初筛和麸皮培养基复筛,得到ZM6用于诱变选育的出发菌株,并对其采用了紫外线和LiCl复合诱变、DES诱变及自然分离,再结合糖化透明圈初筛和麸皮培养基复筛,最终选育得到一株糖化力高(糖化力比ZM6提高了185%,液化力提高了48.8%)的优良突变株ZM6.1.2。其遗传性状稳定,生长发酵快,菌丝密集粗短。
(2)酵母菌种的选育。对从藏曲中筛选的3株优良酵母菌,通过发酵速度、酵母耐酒精、死亡温度、最适发酵温度以及最适发酵pH值的比较,得到ZY2用于诱变选育的出发菌株,对其分别采用乙醇热冲击和细胞紫外诱变,结合高温驯养、TTC平板显色,最终选育到一株耐38℃高温的酵母突变株ZY2.1.3.2。其遗传性状稳定,生长均一快速,生理耐性好,发酵力高(产酒达到了7.3%)。
(2)制纯种曲工艺条件的研究。将经过诱变选育的糖化菌ZM6.1.2和酵母菌ZY2.1.3.2,通过单因素分析,得出制曲工艺:①菌悬液的准备:于活化好的菌种斜面加入5mL无菌水,刮下孢子或菌体细胞,打散备用;②将过40目的青稞粉和麸皮粉按4:1的比例混合,在100℃烘箱中干燥过夜(17h);③将准备好的混合粉加入16%的霉菌菌悬液、4%的酵母菌菌悬液及30%的水混匀,30℃下培养24h,在45℃烘箱中干燥3h～5h,即得纯种曲。
(3)青稞酒发酵条件工艺的研究。分别就青稞蒸煮时间、加曲量、投料品温,进行分析得到青稞酒发酵优化工艺:取适量青稞,洗净,加适量水煮约60min,凉至品温26℃,添加煮前青稞干重0.40%的曲,拌匀加入瓷盆,加盖。瓷盆外加盖如棉絮等可保温的材料,于28-30℃发酵72h。发酵完毕后,按1.0g(酒醅重):1.5mL加入20℃凉开水,浸泡4h,两层纱布过滤,滤液即为青稞酒。实验证明,经诱变后的菌种制成曲发酵的青稞酒,酒体风格上保持了传统青稞酒的清雅、醇和的特点,风味好,适于工业化生产质量稳定的西藏青稞酒。
With a long history of more than 1300 years, Highland barley wine has served as an indispensable drink for Tibetans.In this thesis, selecting superior stains from the superior tranditional Tibet starters, making pure culture starters with the selected-superior stains and fermenting Highland barley wine with the pure culture starters are studied , in order to improve the quality of tranditional Tibet wine.
(1) To breed strains .Comparing 18 superior diastatic strains which were isolated from Tibet starters, mould ZM6 have been selected for further studies.Then,ZM6 was mutagenized by UV,UV-LiCl and DES, combined natural selection and transparent-loop in plate, and we obtained one mutant ZM6.1.1 which had strong diastatic power (higher 185% than ZM6); We gained one yeast mutant YS2.1.3.2 which came from YS2 that was controlled by ethanol and heat impacting, UV radiating on its cell and protoplast, combine high temperature domesticating and TTC coloration in plate .Mutant YS2.1.3.2 had advantages of stable transmissibility ,fast-synchronous growth velocity, better physiological tolerance, strong fermenting power (alcoholicity was 7.3%).
(2) To make culture stater. Via single-analysis test, we obtained the optimal processes of making the Tibei starter :①Preparation of inoculun suspensions :Inoculum suspensions were made by adding 5 mL of sterile physiological salt solution (0.85% w/v NaCl) onto each pure culture slant. The biomass was gently scraped off the agar by means of an inoculation loop;②The ratio of mixed flours is 80g( highland barley flours):20g (wheat bran flours),drying at 100 for overnight (about 17h );③Mixed flours,16% mould suspension,4% yeast suspension ,30% water ,cultulating at 30℃for 24h,then drying at 45℃for 3-5h.
(3)To control sound fermenting process. Cooking highland barley ,the dosage of starter and intitial temperature were studied ,and we arrived at the optimal fermenting process of highland barley wine in Tibet :starters were mixed with cooked highland barley whose water-absorbed ratio is 150%~155%.The ratio of starters to mass is 0.4g(dry production starter ):100g(dry highland barley).A fungal solid-state fermentation was undertaking in a dish which have been wrapped with quilt to keep the initial temperature . After 72h, the fermented mass was mixed with added water at the ratio of 100g to 150 mL for 4h, and then filtrated with cloth.
(1)糖化菌种的选育。对从藏曲中筛选的18株优良糖化菌中,结合糖化透明圈初筛和麸皮培养基复筛,得到ZM6用于诱变选育的出发菌株,并对其采用了紫外线和LiCl复合诱变、DES诱变及自然分离,再结合糖化透明圈初筛和麸皮培养基复筛,最终选育得到一株糖化力高(糖化力比ZM6提高了185%,液化力提高了48.8%)的优良突变株ZM6.1.2。其遗传性状稳定,生长发酵快,菌丝密集粗短。
(2)酵母菌种的选育。对从藏曲中筛选的3株优良酵母菌,通过发酵速度、酵母耐酒精、死亡温度、最适发酵温度以及最适发酵pH值的比较,得到ZY2用于诱变选育的出发菌株,对其分别采用乙醇热冲击和细胞紫外诱变,结合高温驯养、TTC平板显色,最终选育到一株耐38℃高温的酵母突变株ZY2.1.3.2。其遗传性状稳定,生长均一快速,生理耐性好,发酵力高(产酒达到了7.3%)。
(2)制纯种曲工艺条件的研究。将经过诱变选育的糖化菌ZM6.1.2和酵母菌ZY2.1.3.2,通过单因素分析,得出制曲工艺:①菌悬液的准备:于活化好的菌种斜面加入5mL无菌水,刮下孢子或菌体细胞,打散备用;②将过40目的青稞粉和麸皮粉按4:1的比例混合,在100℃烘箱中干燥过夜(17h);③将准备好的混合粉加入16%的霉菌菌悬液、4%的酵母菌菌悬液及30%的水混匀,30℃下培养24h,在45℃烘箱中干燥3h～5h,即得纯种曲。
(3)青稞酒发酵条件工艺的研究。分别就青稞蒸煮时间、加曲量、投料品温,进行分析得到青稞酒发酵优化工艺:取适量青稞,洗净,加适量水煮约60min,凉至品温26℃,添加煮前青稞干重0.40%的曲,拌匀加入瓷盆,加盖。瓷盆外加盖如棉絮等可保温的材料,于28-30℃发酵72h。发酵完毕后,按1.0g(酒醅重):1.5mL加入20℃凉开水,浸泡4h,两层纱布过滤,滤液即为青稞酒。实验证明,经诱变后的菌种制成曲发酵的青稞酒,酒体风格上保持了传统青稞酒的清雅、醇和的特点,风味好,适于工业化生产质量稳定的西藏青稞酒。
With a long history of more than 1300 years, Highland barley wine has served as an indispensable drink for Tibetans.In this thesis, selecting superior stains from the superior tranditional Tibet starters, making pure culture starters with the selected-superior stains and fermenting Highland barley wine with the pure culture starters are studied , in order to improve the quality of tranditional Tibet wine.
(1) To breed strains .Comparing 18 superior diastatic strains which were isolated from Tibet starters, mould ZM6 have been selected for further studies.Then,ZM6 was mutagenized by UV,UV-LiCl and DES, combined natural selection and transparent-loop in plate, and we obtained one mutant ZM6.1.1 which had strong diastatic power (higher 185% than ZM6); We gained one yeast mutant YS2.1.3.2 which came from YS2 that was controlled by ethanol and heat impacting, UV radiating on its cell and protoplast, combine high temperature domesticating and TTC coloration in plate .Mutant YS2.1.3.2 had advantages of stable transmissibility ,fast-synchronous growth velocity, better physiological tolerance, strong fermenting power (alcoholicity was 7.3%).
(2) To make culture stater. Via single-analysis test, we obtained the optimal processes of making the Tibei starter :①Preparation of inoculun suspensions :Inoculum suspensions were made by adding 5 mL of sterile physiological salt solution (0.85% w/v NaCl) onto each pure culture slant. The biomass was gently scraped off the agar by means of an inoculation loop;②The ratio of mixed flours is 80g( highland barley flours):20g (wheat bran flours),drying at 100 for overnight (about 17h );③Mixed flours,16% mould suspension,4% yeast suspension ,30% water ,cultulating at 30℃for 24h,then drying at 45℃for 3-5h.
(3)To control sound fermenting process. Cooking highland barley ,the dosage of starter and intitial temperature were studied ,and we arrived at the optimal fermenting process of highland barley wine in Tibet :starters were mixed with cooked highland barley whose water-absorbed ratio is 150%~155%.The ratio of starters to mass is 0.4g(dry production starter ):100g(dry highland barley).A fungal solid-state fermentation was undertaking in a dish which have been wrapped with quilt to keep the initial temperature . After 72h, the fermented mass was mixed with added water at the ratio of 100g to 150 mL for 4h, and then filtrated with cloth.
引文
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