老白干酒曲中一株高产β-苯乙醇酵母菌的分离、鉴定及其产香特性研究
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摘要
采用平板涂布法从老白干香型酒曲中筛选获得一株高产β-苯乙醇的酵母,命名为F11601;通过菌落形态、细胞结构和分子生物学方法进行鉴定为季也蒙毕赤酵母。该菌株具有较好的β-苯乙醇耐受性和NaCl耐受性,能在p H 1~10范围内生长,属于耐高温酵母。通过单因素试验优化获得该菌合成β-苯乙醇的最佳条件,在该条件下β-苯乙醇合成量达到1.66 g/L。利用豆芽汁培养基研究了其发酵产香特性,在豆芽汁培养基中能产近20种挥发性风味物质。本文报道了大曲来源的季也蒙毕赤酵母产β-苯乙醇研究,为其在老白干香型白酒中应用提供理论基础。
A yeast designated as F11601 with high-yield for β-phenylethanol was isolated and purified from Laobaigan-flavor Daqu by traditional microbial separation, then the yeast was identified as types of Meyerozyma guilliermondii, by the morphological and physiological characteristics and its 26 S rDNA D1/D2 sequence. Studies on the tolerance showed that strain F11601 presented high tolerance to β-phenylethanol and NaCl. It can grow in the pH within the scope of 1 to 10, and it is a thermophilic yeast. By the experiments of single factor, the optimal conditions for β-phenylethanol production were obtained. Under the condition, the yeast F11601 yielded β-phenylethanol of 1.66 g/L. It was tested for aroma production in bean sprouts medium, and there were nearly twenty kinds of volatile components present in the fermentation of medium. This is the study about β-phenylethanol production of Meyerozyma guilliermondii from Daqu, and it provides theoretical basis for its application in Laobaigan-flavor Baijiu.
A yeast designated as F11601 with high-yield for β-phenylethanol was isolated and purified from Laobaigan-flavor Daqu by traditional microbial separation, then the yeast was identified as types of Meyerozyma guilliermondii, by the morphological and physiological characteristics and its 26 S rDNA D1/D2 sequence. Studies on the tolerance showed that strain F11601 presented high tolerance to β-phenylethanol and NaCl. It can grow in the pH within the scope of 1 to 10, and it is a thermophilic yeast. By the experiments of single factor, the optimal conditions for β-phenylethanol production were obtained. Under the condition, the yeast F11601 yielded β-phenylethanol of 1.66 g/L. It was tested for aroma production in bean sprouts medium, and there were nearly twenty kinds of volatile components present in the fermentation of medium. This is the study about β-phenylethanol production of Meyerozyma guilliermondii from Daqu, and it provides theoretical basis for its application in Laobaigan-flavor Baijiu.
引文
[1]丁云连,范文来,徐岩,等.老白干香型白酒香气成分分析[J].酿酒, 2008(4):109-113.
[2]朱会霞,张煜行,程宗志,等.衡水老白干酒发酵主要风味物质变化规律研究[J].酿酒科技, 2015(2):36-39.
[3]吕浩.老白干香型酒感官品评特色[J].酿酒, 2009(1):43-44.
[4] HAIYAN F, WENLAI F, YAN X. Charac terization of key odorants in Chinese chixiang aroma-type liquor by gas chromatography-olfactometry, quantitative measurements, aroma recombination, and omission studies[J]. Journal of Agricultural and Food Chemistry, 2015, 63(14):3660-3668.
[5] SHUANG C, DONG W, YAN X. Characterization of odor-active compounds in sweet-type Chinese rice wine by aroma extract dilution analysis with special emphasis on sotolon[J]. Journal of Agricultural and Food Chemistry, 2013, 61(40):9712-9718.
[6] IZZREEN N Q M N, PETERSEN M A, HANSEN A S. Volatile compounds in crumb of whole-meal wheat bread fermented with different yeast levels and fermentation temperatures[J]. Cereal Chemistry,2016, 93(2):209-216.
[7] FENG Y, CAI Y, SUN-WATERHOUSE D, et al.Approaches of aroma extraction dilution analysis(AEDA)for headspace solid phase microextraction and gas chromatography-olfactometry(HS-SPME-GCO):Altering sample amount, diluting the sample or adjusting split ratio?[J]. Food Chemistry, 2015,187:44-52.
[8] DE-LA-FUENTE-BLANCO A, SAENZ-NAVAJAS M, FERREIRA V. On the effects of higher alcohols on red wine aroma[J]. Food Chemistry, 2016, 210:107-114.
[9]燕雯,张正茂,刘拉平.顶空固相微萃取-气质联用分析小麦馒头制作过程中的挥发性成分变化[J].食品科学, 2012(12):254-258.
[10]陈双,罗涛,徐岩,等.我国黄酒酵母和酿酒原料对黄酒中β-苯乙醇含量的影响[J].中国酿造, 2009(4):23-26.
[11]姚粟,于学健,白飞荣,等.中国传统发酵食品用微生物菌种名单的研究[J].食品与发酵工业, 2017,43(9):238-258.
[12]蒋慧,吴玉新,庄靓,等.融合魏斯氏菌和异常威克汉姆酵母混菌发酵荞麦酸面团馒头的香气物质特征[J].食品工业科技, 2018, 39(15):234-241.
[13] OOI B G, WANAMAKER L E, MARKUSZEWSKI B M, et al. Genetic and enological analysis of selected Saccharomyces cerevisiae strains for wine production[J]. International Journal of Food Science and Technology, 2008, 43(6):1111-1120.
[14] PATETKO A, SILINS R, SCHERBAKA R, et al.2-Phenylethanol production by Kluyveromyces marxianus on glucose and lactose substrates[J]. Journal of Biotechnology, 2016, 231S:S62-S63.
[15]明红梅,周健,陈蒙恩,等.大曲中异常威克汉姆酵母发酵产苯乙醇的条件优化[J].湖北农业科学,2015(14):3492-3496.
[16]巴尼特,佩恩,亚罗.酵母菌的特征与鉴定手册[M].青岛:青岛海洋大学出版社, 1991:17-32.
[17]黄筱萍,刘兰,熊大维,等.一株高产2-苯乙醇酵母菌的筛选及鉴定[J].食品与生物技术学报, 2016,35(5):531-536.
[18]曲佳乐,闵伟红,王巧玲,等.一株产木糖醇菌株的生理生化及分子生物学鉴定[J].食品科学, 2011,32(11):171-175.
[19]连鑫,杨锡洪,解万翠,等.中国传统虾酱中产香酵母的分离鉴定及其耐盐性分析[J].现代食品科技,2014, 30(7):92-97.
[20]秦晓杰,许皎姣,索娜,等.草莓灰霉病拮抗酵母菌的筛选鉴定及对采后草莓品质的影响[J].食品工业科技, 2012, 33(21):330-335.
[21]张馨予,侯红漫.一株产胆固醇氧化酶酵母菌的鉴定及发酵条件优化[J].食品工业科技, 2011, 32(6):234-237.
[22]高云慨,张荣意,钟利文,等. 1株新分离拮抗酵母菌株对芒果炭疽病生防效果及其分类鉴定[J].热带生物学报, 2015, 6(1):47-52.
[23] STARK D, ZALA D, MUNCH T, et al. Inhibition aspects of the bioconversion of L-phenylalanine to2-phenylethanol by Saccharomyces cerevisiae[J]. Enzyme and Microbial Technology, 2003, 32(2):212-223.
[24]张强,郭元,韩德明.酿酒酵母乙醇耐受性的研究进展[J].化工进展, 2014, 33(1):187-192.
[25]贺家明,赵祥颖,陈苏萍.酵母耐乙醇性状及提高其耐性的途径[J].酿酒科技, 2000(2):84.
[26]沈萍.微生物学实验[M].北京:高等教育出版社,2006:106-108.
[27]娜日苏,苏亚拉图,高凤芹.耐高温耐酸耐酒精酵母的筛选与鉴定[J].草业科学, 2013, 30(10):1625-1632.
[28] ZHENG X, YAN Z, NOUT M J R, et al. Characterization of the microbial community in different types of Daqu samples as revealed by 16S rRNA and 26S rRNA gene clone libraries[J]. World Journal of Microbiology&Biotechnology, 2015, 31(1):199-208.
[29]黄建华,张煜行,张希君,等.不同入池时间对衡水老白干酒醅发酵质量的影响[J].酿酒科技, 2007(2):51-54.
[30]梁慧,马海霞,李来好.腊鱼产香酵母菌的筛选及其发酵产香特性初步研究[J].食品工业科技, 2011,32(12):213-217.
[31]陈金丽,郭阳,薛洁,等.优良野生葡萄酒酵母的筛选及性能评价[J].食品与发酵工业, 2012, 38(5):106-111.
[32]张志民.衡水老白干香型的初步研究[J].酿酒, 1998(2):14-17.
[33]霍丽娜,王运霄,李达.老白干香型原酒的微量成分与风格特点分析[J].酿酒科技, 2015(10):44-46.
[34]周海洋,王士敏,于金侠,等.一种快速测量白酒中棕榈酸乙酯的方法[J].酿酒科技, 2014(10):120-122.
[35]廖勤俭,练顺才,李杨华,等.高效液相色谱四级杆飞行时间质谱联用技术检测白酒中高级脂肪酸[J].酿酒科技, 2013(10):97-99.
[36]杨霄,崔志峰.酵母生物转化生产2-苯乙醇的研究进展[J].应用与环境生物学报, 2006, 12(1):140-144.
[37]吴荣荣,张志民,张煜行,等.老白干大曲中酿酒酵母y1发酵特性的研究[J].中国酿造, 2011, 30(12):104-106.
[2]朱会霞,张煜行,程宗志,等.衡水老白干酒发酵主要风味物质变化规律研究[J].酿酒科技, 2015(2):36-39.
[3]吕浩.老白干香型酒感官品评特色[J].酿酒, 2009(1):43-44.
[4] HAIYAN F, WENLAI F, YAN X. Charac terization of key odorants in Chinese chixiang aroma-type liquor by gas chromatography-olfactometry, quantitative measurements, aroma recombination, and omission studies[J]. Journal of Agricultural and Food Chemistry, 2015, 63(14):3660-3668.
[5] SHUANG C, DONG W, YAN X. Characterization of odor-active compounds in sweet-type Chinese rice wine by aroma extract dilution analysis with special emphasis on sotolon[J]. Journal of Agricultural and Food Chemistry, 2013, 61(40):9712-9718.
[6] IZZREEN N Q M N, PETERSEN M A, HANSEN A S. Volatile compounds in crumb of whole-meal wheat bread fermented with different yeast levels and fermentation temperatures[J]. Cereal Chemistry,2016, 93(2):209-216.
[7] FENG Y, CAI Y, SUN-WATERHOUSE D, et al.Approaches of aroma extraction dilution analysis(AEDA)for headspace solid phase microextraction and gas chromatography-olfactometry(HS-SPME-GCO):Altering sample amount, diluting the sample or adjusting split ratio?[J]. Food Chemistry, 2015,187:44-52.
[8] DE-LA-FUENTE-BLANCO A, SAENZ-NAVAJAS M, FERREIRA V. On the effects of higher alcohols on red wine aroma[J]. Food Chemistry, 2016, 210:107-114.
[9]燕雯,张正茂,刘拉平.顶空固相微萃取-气质联用分析小麦馒头制作过程中的挥发性成分变化[J].食品科学, 2012(12):254-258.
[10]陈双,罗涛,徐岩,等.我国黄酒酵母和酿酒原料对黄酒中β-苯乙醇含量的影响[J].中国酿造, 2009(4):23-26.
[11]姚粟,于学健,白飞荣,等.中国传统发酵食品用微生物菌种名单的研究[J].食品与发酵工业, 2017,43(9):238-258.
[12]蒋慧,吴玉新,庄靓,等.融合魏斯氏菌和异常威克汉姆酵母混菌发酵荞麦酸面团馒头的香气物质特征[J].食品工业科技, 2018, 39(15):234-241.
[13] OOI B G, WANAMAKER L E, MARKUSZEWSKI B M, et al. Genetic and enological analysis of selected Saccharomyces cerevisiae strains for wine production[J]. International Journal of Food Science and Technology, 2008, 43(6):1111-1120.
[14] PATETKO A, SILINS R, SCHERBAKA R, et al.2-Phenylethanol production by Kluyveromyces marxianus on glucose and lactose substrates[J]. Journal of Biotechnology, 2016, 231S:S62-S63.
[15]明红梅,周健,陈蒙恩,等.大曲中异常威克汉姆酵母发酵产苯乙醇的条件优化[J].湖北农业科学,2015(14):3492-3496.
[16]巴尼特,佩恩,亚罗.酵母菌的特征与鉴定手册[M].青岛:青岛海洋大学出版社, 1991:17-32.
[17]黄筱萍,刘兰,熊大维,等.一株高产2-苯乙醇酵母菌的筛选及鉴定[J].食品与生物技术学报, 2016,35(5):531-536.
[18]曲佳乐,闵伟红,王巧玲,等.一株产木糖醇菌株的生理生化及分子生物学鉴定[J].食品科学, 2011,32(11):171-175.
[19]连鑫,杨锡洪,解万翠,等.中国传统虾酱中产香酵母的分离鉴定及其耐盐性分析[J].现代食品科技,2014, 30(7):92-97.
[20]秦晓杰,许皎姣,索娜,等.草莓灰霉病拮抗酵母菌的筛选鉴定及对采后草莓品质的影响[J].食品工业科技, 2012, 33(21):330-335.
[21]张馨予,侯红漫.一株产胆固醇氧化酶酵母菌的鉴定及发酵条件优化[J].食品工业科技, 2011, 32(6):234-237.
[22]高云慨,张荣意,钟利文,等. 1株新分离拮抗酵母菌株对芒果炭疽病生防效果及其分类鉴定[J].热带生物学报, 2015, 6(1):47-52.
[23] STARK D, ZALA D, MUNCH T, et al. Inhibition aspects of the bioconversion of L-phenylalanine to2-phenylethanol by Saccharomyces cerevisiae[J]. Enzyme and Microbial Technology, 2003, 32(2):212-223.
[24]张强,郭元,韩德明.酿酒酵母乙醇耐受性的研究进展[J].化工进展, 2014, 33(1):187-192.
[25]贺家明,赵祥颖,陈苏萍.酵母耐乙醇性状及提高其耐性的途径[J].酿酒科技, 2000(2):84.
[26]沈萍.微生物学实验[M].北京:高等教育出版社,2006:106-108.
[27]娜日苏,苏亚拉图,高凤芹.耐高温耐酸耐酒精酵母的筛选与鉴定[J].草业科学, 2013, 30(10):1625-1632.
[28] ZHENG X, YAN Z, NOUT M J R, et al. Characterization of the microbial community in different types of Daqu samples as revealed by 16S rRNA and 26S rRNA gene clone libraries[J]. World Journal of Microbiology&Biotechnology, 2015, 31(1):199-208.
[29]黄建华,张煜行,张希君,等.不同入池时间对衡水老白干酒醅发酵质量的影响[J].酿酒科技, 2007(2):51-54.
[30]梁慧,马海霞,李来好.腊鱼产香酵母菌的筛选及其发酵产香特性初步研究[J].食品工业科技, 2011,32(12):213-217.
[31]陈金丽,郭阳,薛洁,等.优良野生葡萄酒酵母的筛选及性能评价[J].食品与发酵工业, 2012, 38(5):106-111.
[32]张志民.衡水老白干香型的初步研究[J].酿酒, 1998(2):14-17.
[33]霍丽娜,王运霄,李达.老白干香型原酒的微量成分与风格特点分析[J].酿酒科技, 2015(10):44-46.
[34]周海洋,王士敏,于金侠,等.一种快速测量白酒中棕榈酸乙酯的方法[J].酿酒科技, 2014(10):120-122.
[35]廖勤俭,练顺才,李杨华,等.高效液相色谱四级杆飞行时间质谱联用技术检测白酒中高级脂肪酸[J].酿酒科技, 2013(10):97-99.
[36]杨霄,崔志峰.酵母生物转化生产2-苯乙醇的研究进展[J].应用与环境生物学报, 2006, 12(1):140-144.
[37]吴荣荣,张志民,张煜行,等.老白干大曲中酿酒酵母y1发酵特性的研究[J].中国酿造, 2011, 30(12):104-106.