紫外诱变选育高产酒精及酸的酿酒酵母
|
摘要
利用紫外辐照的方式对从葡萄表面筛选的菌株Y6进行诱变,选育出产酒精能力强、产酸能力高并且发酵性能稳定的酿酒酵母菌株。通过单因素试验选择酵母菌株距30 W紫外灯30 cm照射100 s,致死率为75.62%为最佳诱变条件。采用三苯基氯化四氮唑(TTC)法和溴甲酚绿法以及杜氏小管筛选出产酒精和产酸强的目标菌株Y_6-5,并对其遗传稳定性进行研究。结果表明,诱变菌株Y6-5与出发菌株Y6相比较,其产酒精、产酸能力分别提高了28.13%和201.41%,并且遗传稳定性良好。
The strain Y6 screened from the grape surface was mutagenized by ultraviolet irradiation, and the Saccharomyces cerevisiae strains with high alcohol production ability, high acid production ability and stable fermentation performance were screened. The optimal mutagenesis condition of yeast strain was 30 W, 30 cm, 100 s by single factor tests and the lethality rate was 75.62%. The mutant strain Y6-5 with strong alcohol and acid production was screened by triphenyltetrazolium chloride(TTC) method, bromocresol green method and Duchenne tubule. At the same time, the genetic stability of the mutant strain was studied. The results showed that compared with the original strain Y6, the alcohol and acid-producing ability of mutant strain Y6-5 was increased by 28.13% and 201.41%, respectively, and the genetic stability of the mutant strain was good.
The strain Y6 screened from the grape surface was mutagenized by ultraviolet irradiation, and the Saccharomyces cerevisiae strains with high alcohol production ability, high acid production ability and stable fermentation performance were screened. The optimal mutagenesis condition of yeast strain was 30 W, 30 cm, 100 s by single factor tests and the lethality rate was 75.62%. The mutant strain Y6-5 with strong alcohol and acid production was screened by triphenyltetrazolium chloride(TTC) method, bromocresol green method and Duchenne tubule. At the same time, the genetic stability of the mutant strain was studied. The results showed that compared with the original strain Y6, the alcohol and acid-producing ability of mutant strain Y6-5 was increased by 28.13% and 201.41%, respectively, and the genetic stability of the mutant strain was good.
引文
[1]何国庆,贾英民,丁立孝.食品微生物学[M].第2版.北京:中国农业大学出版社,2009:10-40.
[2]方佩佩,王世清,李静,等.紫外诱变法选育高产酒精及酯类酿酒酵母[J].粮油食品科技,2016(1):86-90.
[3] NIELSEN J, PRONK J T. Metabolic engineering, synthetic biology and systems biology[J]. Fems Yeast Res, 2012, 12(2):103.
[4] WANG P M, ZHENG D Q, LIU T Z, et al. The combination of glycerol metabolic engineering and drug resistance marker-aided genome shuffling to improve very-high-gravity fermentation performances of industrial Saccharomyces cerevisiae[J]. Bioresource Technol, 2012, 108(2):203-210.
[5]杨明琰,郭爱莲,沈俭,等.高产SOD酵母菌的诱变选育及发酵条件研究[J].食品科学,2005,26(10):147-150.
[6] KIM J W, CHIN Y W, PARK Y C, et al. Effects of deletion of glycerol-3-phosphate dehydrogenase and glutamate dehydrogenase genes on glycerol and ethanol metabolism in recombinant Saccharomyces cerevisiae[J].Bioproc Biosys Eng, 2012, 35(1-2):49-54.
[7]许翠,高梦祥.微生物菌种诱变筛选方法及其应用[J].长江大学学报:自然科学版,2013(12):56-60.
[8]曹礼,王晓琴,李彩霞,等.酿酒酵母菌的紫外诱变及其突变株的性能测定[J].中国酿造,2009,28(10):66-68.
[9] ZHU M, LI P, GONG X, et al. A comparison of the production of ethanol between simultaneous saccharification and fermentation and separate hydrolysis and fermentation using unpretreated cassava pulp and enzyme cocktail[J]. J Agr Chem Soc Jpn, 2012, 76(4):671-678.
[10]杨小冲,陈忠军,赵洁,等.紫外诱变选育耐酸酵母菌株及其特性研究[J].中国酿造,2017,36(10):109-113.
[11]王蕾.基于“五力模型”的新疆葡萄酒产业价值链分析[J].中国酿造,2017,36(1):196-200.
[12]周德庆.微生物学实验教程[M].第2版.北京:高等教育出版社,2006:57-85.
[13] WANG Y, ZHOU J, XIA X, et al. Probiotic potential of Lactobacillus paracasei FM-LP-4 isolated from Xinjiang camel milk yoghurt[J]. Int Dairy J, 2016, 62:28-34.
[14]陈忠军,杨小冲,赵洁,等.具高乙醇耐受力酵母菌的选育及其在猕猴桃果酒中的应用[J].食品工业科技,2018(2):141-145.
[15]边名鸿,万世旅,代梅,等.产酒酵母的筛选鉴定及其在小曲中的应用[J].中国酿造,2016,35(3):57-60.
[16]马文瑞,魏玉洁,邹弯,等.新疆葡萄酒高产酸酿酒酵母菌的筛选和发酵条件优化[J].食品与发酵工业,2017,43(7):134-140.
[17]刘洪,车振明,陈坤,等.人工接种与自然发酵泡豇豆的质地研究[J].食品工业科技,2012,33(14):111-115.
[18]罗跃中,李忠英,李继睿,等.高效酒精酵母菌选育及其特性研究[J].中国酿造,2013,32(8):71-74.
[19]郑莉烨.耐高渗啤酒酵母的选育及其发酵特性的研究[D].广州:华南理工大学,2013.
[20]熊建春.甘蔗威士忌酯香酵母菌的选育及发酵工艺研究[D].广州:华南理工大学,2010.
[21]李艾,李云凯,程磊,等.紫外诱变酿酒酵母筛选耐高温菌株[J].唐山学院学报,2013,26(3):35-37.
[22]张丽萍,程辉彩,田连生,等.植酸酶高产菌株的紫外线-空气等离子体复合诱变选育[J].中国饲料,2006(9):13-15.
[2]方佩佩,王世清,李静,等.紫外诱变法选育高产酒精及酯类酿酒酵母[J].粮油食品科技,2016(1):86-90.
[3] NIELSEN J, PRONK J T. Metabolic engineering, synthetic biology and systems biology[J]. Fems Yeast Res, 2012, 12(2):103.
[4] WANG P M, ZHENG D Q, LIU T Z, et al. The combination of glycerol metabolic engineering and drug resistance marker-aided genome shuffling to improve very-high-gravity fermentation performances of industrial Saccharomyces cerevisiae[J]. Bioresource Technol, 2012, 108(2):203-210.
[5]杨明琰,郭爱莲,沈俭,等.高产SOD酵母菌的诱变选育及发酵条件研究[J].食品科学,2005,26(10):147-150.
[6] KIM J W, CHIN Y W, PARK Y C, et al. Effects of deletion of glycerol-3-phosphate dehydrogenase and glutamate dehydrogenase genes on glycerol and ethanol metabolism in recombinant Saccharomyces cerevisiae[J].Bioproc Biosys Eng, 2012, 35(1-2):49-54.
[7]许翠,高梦祥.微生物菌种诱变筛选方法及其应用[J].长江大学学报:自然科学版,2013(12):56-60.
[8]曹礼,王晓琴,李彩霞,等.酿酒酵母菌的紫外诱变及其突变株的性能测定[J].中国酿造,2009,28(10):66-68.
[9] ZHU M, LI P, GONG X, et al. A comparison of the production of ethanol between simultaneous saccharification and fermentation and separate hydrolysis and fermentation using unpretreated cassava pulp and enzyme cocktail[J]. J Agr Chem Soc Jpn, 2012, 76(4):671-678.
[10]杨小冲,陈忠军,赵洁,等.紫外诱变选育耐酸酵母菌株及其特性研究[J].中国酿造,2017,36(10):109-113.
[11]王蕾.基于“五力模型”的新疆葡萄酒产业价值链分析[J].中国酿造,2017,36(1):196-200.
[12]周德庆.微生物学实验教程[M].第2版.北京:高等教育出版社,2006:57-85.
[13] WANG Y, ZHOU J, XIA X, et al. Probiotic potential of Lactobacillus paracasei FM-LP-4 isolated from Xinjiang camel milk yoghurt[J]. Int Dairy J, 2016, 62:28-34.
[14]陈忠军,杨小冲,赵洁,等.具高乙醇耐受力酵母菌的选育及其在猕猴桃果酒中的应用[J].食品工业科技,2018(2):141-145.
[15]边名鸿,万世旅,代梅,等.产酒酵母的筛选鉴定及其在小曲中的应用[J].中国酿造,2016,35(3):57-60.
[16]马文瑞,魏玉洁,邹弯,等.新疆葡萄酒高产酸酿酒酵母菌的筛选和发酵条件优化[J].食品与发酵工业,2017,43(7):134-140.
[17]刘洪,车振明,陈坤,等.人工接种与自然发酵泡豇豆的质地研究[J].食品工业科技,2012,33(14):111-115.
[18]罗跃中,李忠英,李继睿,等.高效酒精酵母菌选育及其特性研究[J].中国酿造,2013,32(8):71-74.
[19]郑莉烨.耐高渗啤酒酵母的选育及其发酵特性的研究[D].广州:华南理工大学,2013.
[20]熊建春.甘蔗威士忌酯香酵母菌的选育及发酵工艺研究[D].广州:华南理工大学,2010.
[21]李艾,李云凯,程磊,等.紫外诱变酿酒酵母筛选耐高温菌株[J].唐山学院学报,2013,26(3):35-37.
[22]张丽萍,程辉彩,田连生,等.植酸酶高产菌株的紫外线-空气等离子体复合诱变选育[J].中国饲料,2006(9):13-15.