为筛选用于葡萄酒发酵的可降解氨基甲酸乙酯(ethyl carbamate, EC)酵母菌,对酿酒葡萄、干化葡萄、酒醅进行筛选,获得7株对EC有较强降解能力的菌株,经过发酵性能、耐乙醇以及耐二氧化硫实验,发现2株菌发酵性能较好。在此基础上,对这2株菌所发酵的葡萄酒进行EC含量、理化指标、香气组分的比较及电子鼻分析,并以商业酵母XR作为对照。筛选出目的菌株,经分子鉴定后,确定该菌株为酿酒酵母,并命名为NXU12-2。研究所获得的菌株是具有氨基甲酸乙酯降解能力的酿酒酵母,其发酵的葡萄酒品质较优。该研究为葡萄酒中EC的调控提供了优质的菌种资源,具有重要意义。
In order to screen the yeast which can degrade ethyl carbamate (EC) used in wine fermentation, multiple rounds of selection with wine grapes, dried grapes, and fermented grains were adopted in this paper and seven strains with strong capacity of degrading EC were obtained. The results of fermentation performance, ethanol resistance and sulfur dioxide resistance showed that two strains had better fermentation performance. Then the contents of EC, physicochemical indexes, aroma components and electronic nose analysis of the wines fermented by the two strains were compared, and the commercial yeast XR was used as the control. After the molecular identification of the target strain, it was identified as Saccharomyces cerevisiae and was named as NXU12-2. In this study, a strain of S. cerevisiae with the ability to degrade EC was selected, and it fermented wines are of good quality. This research would provide high quality strain resources which can control EC in wine.
[1] 顾国贤. 酿造酒工艺学[M].第二版.北京:中国轻工业出版社, 2008:475-463.
GU G X.Brewing Technology[M].Second edition.Beijing:China Light Industry Press, 2008:475-463.
[2] THIBON C, MARULLO P, CLAISSE O, et al.Nitrogen catabolic repression controls the release of volatile thiols by Saccharomyces cerevisiae during wine fermentation[J].FEMS Yeast Research, 2008, 8(7):1 076-1 086.
[3] BAAN R, STRAIF K, GROSSE Y, et al.Carcinogenicity of alcoholic beverages[J].The Lancet Oncology, 2007, 8(4):292-293.
[4] WEBER J V, SHARYPOV V I.Ethyl carbamate in foods and beverages:A review[J].Environmental Chemistry Letters, 2009, 7(3):233-247.
[5] 赵雅敏. 红酵母降解中国黄酒中氨基甲酸乙酯的研究[D].无锡:江南大学, 2012.
ZHAO Y M.Investigation of ethyl carbamate degradation in Chinese rice wine by Rhodotorula sp[D].Wuxi:Jiangnan University, 2012.
[6] 丁霞, 李巧玉, 刘凡, 等.降解浓香型白酒中氨基甲酸乙酯菌株的筛选及产酶特性研究[J].食品与发酵工业, 2018, 44(7):29-36.
DING X, LI Q Y, LIU F, et al.Isolation of microbial strains for degradation of ethyl carbamate in Luzhou-flavour Baijiu and characterization of corresponding enzymes[J].Food and Fermentation Industries, 2018, 44(7):29-36.
[7] 周建立, 康振, 刘庆涛, 等.重组酸性脲酶对黄酒中尿素和氨基甲酸乙酯的降解应用[J].生物工程学报, 2016, 32(1):74-83.
ZHOU J L, KANG Z, LIU Q T, et al.Degradation of urea and ethyl carbamate in Chinese Rice wine by recombinant acid urease[J].Chinese Journal of Biotechnology, 2016, 32(1):74-83.
[8] KOBASHI K, TAKEBE S, SAKAI T.Urethane-hydrolyzing enzyme from Citrobacter sp.[J].Chemical & Pharmaceutical Bulletin, 1990, 38(5):1 326-1 328.
[9] MOHAPATRA B R, BAPUJI M.Characterization of urethanase from Micrococcus species associated with the marine sponge (Spirasfrella species)[J].Letters in Applied Microbiology, 1997, 25(6):393-396.
[10] 杨广明. 酸性脲酶与氨基甲酸乙酯降解酶产生菌的筛选及酶的特性[D].无锡:江南大学, 2014.
YANG G M.The isolation of acid urease and urethanase producing strain and the enzymatic characterization[D].Wuxi:Jiangnan University, 2014.
[11] 姚晓瑞宁. 降解氨基甲酸乙酯酵母菌的筛选及特性分析[D].石河子:石河子大学, 2018.
YAO X R N.Screening and characterization of the yeast degrading ethyl carbamate[D].Shihezi:Shihezi University, 2018.
[12] 区棋铭, 黄雨锟, 朱国军, 等.气相色谱–质谱法测定涂料中游离氨基甲酸乙酯[J].化学分析计量, 2018, 27(2):36-39.
OU Q M, HUANG Y K, ZHU G J, et al.Determination of free ethyl carbamate in coating by gas chromatography–mass spectrometry[J].Chemical Analysis and Meterage, 2018, 27(2):36-39.
[13] 浩楠. 甘肃河西走廊葡萄酒产区苹乳发酵乳酸菌分离鉴定及其对葡萄酒品质的影响[D].兰州:甘肃农业大学, 2019.
HAO N.Isolation, identification and influence of wine quality of malolactic bacteria from Hexi Corridor in Gansu[D].Lanzhou:Gansu Agricultural University, 2019.
[14] 孙玮璇, 岳卓雅, 刘珍, 等.香柏木浸渍工艺酿造干红葡萄酒的感官质量分析[J].中国食品学报, 2017, 17(6):172-178.
SUN W X, YUE Z Y, LIU Z, et al.The sensory analysis of dry red wine fermented by Cupressus pieces maceration process[J].Journal of Chinese Institute of Food Science and Technology, 2017, 17(6):172-178.
[15] JIANG B, ZHANG Z W.A preliminary study of aroma composition and impact odorants of cabernet franc wines under different terrain conditions of the loess plateau region (China)[J].Molecules (Basel, Switzerland), 2018, 23(5):1096.
[16] 宋茹茹, 段卫朋, 祝霞, 等.戴尔有孢圆酵母与酿酒酵母顺序接种发酵对干红葡萄酒香气的影响[J].食品与发酵工业, 2019, 45(24):1-9.
SONG R R, DUAN W P, ZHU X, et al.Effects of sequential inoculation with Torulaspora delbrueckii and Saccharomyces cerevisiae on aroma profile of dry red wine[J].Food and Fermentation Industries, 2019, 45(24):1-9.
[17] 宫雪, 刘宁, 李二虎, 等.基于电子鼻的葡萄酒感官评价模型的构建[J].中国酿造, 2014, 33(5):67-71.
GONG X, LIU N, LI E H, et al.Model for wine sensory evaluation based on electronic nose[J].China Brewing, 2014, 33(5):67-71.
[18] 尤雅, 段长青, 燕国梁.扁平云假丝酵母与酿酒酵母混合发酵对葡萄酒乙醇含量及香气的影响[J].食品科学, 2018, 39(20):146-154.
YOU Y, DUAN C Q, YAN G L.Effects of mixed fermentation of Candida humilis and Saccharomyces cerevisiae on ethanol content and aroma of wine[J].Food Science, 2018, 39(20):146-154.
[19] LI W P, YAO H, CHEN K Q, et al.Effect of foliar application of fulvic acid antitranspirant on sugar accumulation, phenolic profiles and aroma qualities of Cabernet Sauvignon and Riesling grapes and wines[J].Food Chemistry, 2021, 351:129308.
[20] 李斌斌, 赵晓敏, 周鹤, 等.不同冷浸渍时间对干红葡萄酒酚类物质和色泽品质的影响[J].食品与发酵工业, 2019, 45(18):183-188, 194.
LI B B, ZHAO X M, ZHOU H, et al.Effects of different cold maceration time on phenolic substances and color of dry red wine[J].Food and Fermentation Industries, 2019, 45(18):183-188;194.
[21] SWIEGERS J H, BARTOWSKY E J, HENSCHKE P A, et al.Yeast and bacterial modulation of wine aroma and flavour[J].Australian Journal of Grape and Wine Research, 2005, 11(2):139-173.
[22] JOLLY N P, VARELA C, PRETORIUS I S.Not your ordinary yeast:Non-Saccharomyces yeasts in wine production uncovered[J].FEMS Yeast Research, 2014, 14(2):215-237.
