为筛选适合樱桃果酒发酵的优良酿酒酵母,从樱桃及土壤中采集样本,经2,3,5-氯化三苯基四氮唑(2,3,5-triphenyltetrazolium chloride,TTC)显色法初筛,果汁发酵法复筛,结合在不同条件下的菌株发酵耐受性实验,筛选出具有较强产乙醇能力和良好耐受性的酵母,然后对目的菌株进行分子生物学鉴定,并运用顶空固相微萃取-气相色谱-质谱联用(headspace solid phase micro-extraction combine with gas chromatography mass spectrometry,HS-SPME/GC-MS)与香气活度值(odor activity value, OAV)分析果酒香气成分。结果表明:编号为J35的酵母菌,在果汁含量为35%、糖度25°Bx条件下产乙醇量达到12.57%vol;其在糖度45°Bx、pH 2.0、酒精度18%vol、SO2质量浓度250 mg/L的条件下具有较好产气性能;经菌株形态和rDNA的相似性比对后,J35鉴定为酿酒酵母;其发酵的樱桃果酒为深红色,口感柔和、香气馥郁,含有辛酸乙酯、癸酸乙酯、异戊醇、苯乙醇、苯甲醛、辛酸、β-香茅醇等樱桃酒典型香气成分。
The aim of this study was to screen out Saccharomyces cerevisiae which was suitable for the fermentation of cherry wine from cherries and soils. The colour method, 2,3,5-triphenyltetrazolium chloride (TTC), and the fruit juice fermentation method were used to screen and rescreen these samples. Combined with the fermentation tolerance test under different conditions, the yeast that has strong ethanol production ability and good tolerance was screened out. And then molecular identification of the target strain was conducted. Headspace solid phase micro-extraction combined with gas chromatography mass spectrometry (HS-SPME/GC-MS) and odor activity value (OAV) were applied to analyze aroma components in cherry fruit wine. The results showed that the yeast J35 can produced ethanol by 12.57%vol under conditions that 35% juice content and 25°Bx sugar content. Furthermore, yeast J35 showed good gas production performance under conditions that sugar content was 45°Bx, pH 2.0, alcohol content 18%vol and SO2 content 250 mg/L. After the morphology and rDNA analysis, J35 was characterized as Saccharomyces cerevisiae. Cherry fruit wine fermented by J35 were dark red, soft taste, rich aroma, and contained ethyl octanoate, hydrazine typical aroma components of cherry wine including ethyl acetate, isoamyl alcohol, phenylethyl alcohol, benzaldehyde, octanoic acid, and β-citronellol.
[1] 李志友,李启要.樱桃果酒发酵条件的优化[J].贵州农业科学,2016,44(3):152-155.
[2] 陈杨.樱桃的贮藏保鲜与加工[J].新农村,2018(6):35-36.
[3] KIRAKOSYAN A,SEYMOUR E M,LLANES D E U,et al.Chemical profile and antioxidant capacities of tart cherry products[J].Food Chemistry,2009,115(1):20-25.
[4] 姜东琪,张云舒,武晓玮,等.酿酒酵母对樱桃酒挥发性组分及感官品质的影响[J].中国酿造,2019,38(8):94-99.
[5] 牛云蔚.樱桃酒的特征风味及品质调控研究[D].无锡:江南大学,2012.
[6] 范兆军.响应面优化樱桃果酒发酵条件的研究[J].农产品加工,2015(9):22-25.
[7] 秦伟帅,姜凯凯,孙玉霞,等. SPME-GC3MS联用测定“红灯”樱桃酒中香气成分[J].食品工业,2016,37(9):296-298.
[8] 于海燕,张妮,许春华.樱桃酒发酵过程中的主要香气成分及其变化[J].食品与发酵工业,2011,37(10):159-163.
[9] 许程,王晶,孙艳艳,等.樱桃酒专用酵母的筛选鉴定[J].食品科技,2017,42(11):14-18.
[10] SUN S Y,GONG H S,JIANG X M,et al.Selected non-Saccharomyces wine yeasts in controlled multistarter fermentations with Saccharomyces cerevisiae on alcoholic fermentation behaviour and wine aroma of cherry wines[J].Food Microbiology,2014, 44:15-23.
[11] 李凯,王金晶,李永仙,等.红芯火龙果酒酿酒酵母的筛选及鉴定[J].食品与发酵工业,2019,45(5):115-120.
[12] 刘慧.现代食品微生物学[M].北京:中国轻工业出版社,2011.
[13] 李凯,王金晶,李永仙,等.红心火龙果果酒特征香气分析[J].食品与发酵工业,2019,45(13):217-223.
[14] 左勇,刘利平,鞠帅,等.无花果果酒酵母的筛选及发酵性能研究[J].现代食品科技,2013,29(6):1 293-1 296.
[15] 胡志平,柯锋,叶静萱,等.三种香型白酒蒸馏前后总酯的差异性分析[J].酿酒科技,2019,296(2):81-84.
[16] 中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会.GB/T 15038—2006,葡萄酒、果酒通用分析方法[S].北京:中国标准出版社,2006.
[17] 贾瑞楠,单万祥,李永仙,等.橙酒酵母的分离、筛选及鉴定[J].食品与发酵工业,2018,44(7):122-127.
[18] NY/T 1508—2017,绿色食品果酒[S].北京:中国标准出版社,2017.
[19] SUN S Y,GONG H S,LIU W L,et al.Application and validation of autochthonous Lactobacillus plantarum starter cultures for controlled malolactic fermentation and its influence on the aromatic profile of cherry wines[J].Food Microbiology,2016,55(5):16-24.
[20] 王春,许德富,税梁扬,等.泸型白酒主要物质组分在蒸馏过程中的变化[J].酿酒科技,2009,183(9):61-64.
[21] 杜晶,余培斌,苏琪,等.杨梅果酒酿造酵母的筛选、鉴定及耐受性研究[J].食品与发酵工业,2015,41(9):34-39.
[22] GB 2760—2014,食品添加剂使用标准[S]北京:中国标准出版社,2014.
[23] 范文来,徐岩.酒类风味化学[M].北京:中国轻工业出版社,2014.
[24] SUN S Y,JIANG W G,ZHAO Y P.Characterization of the aroma-active compounds in five sweet cherry cultivars grown in Yantai (China)[J]. Flavour and Fragrance Journal, 2010, 25(4):206-213.
[25] 张婷,陈小伟,张沙沙,等.不同酒曲发酵生产木薯酒香气成分GC-MS分析[J].食品研究与开发,2019,40(2):125-131.
[26] SUN S Y,CHE C Y,SUN T F,et al.Evaluation of sequential inoculation of Saccharomyces cerevisiae and Oenococcus oeni strains on the chemical and aromatic profiles of cherry wines[J].Food Chemistry,2013,138(4):2 233-2 241.
[27] LAURE R,CLARK A C,SCHMIDTKE L M,et al.A robust method for quantification of volatile compounds within and between vintages using headspace-solid-phase micro-extraction coupled with GC-MS- Application on Semillon wines[J].Analytica Chimica Acta,2010,660(1-2):149-157.
[28] 肖作兵,周璇,牛云蔚,等.GC-O结合OAV分析樱桃酒的特征香气成分[J].中国食品学报,2017,17(8):246-254.
[29] FRAILE P,JULIN GARRIDO,CARMEN ANCN. Influence of a Saccharomyces cerevisiae Selected Strain in the Volatile Composition of Rose Wines. Evolution during Fermentation[J]. Journal of Agricultural and Food Chemistry, 2000, 48(5):1 789-1 798.
[30] HAUSCH B J,LORJAROENPHON Y,CADWALLADER K R.Flavor Chemistry of Lemon-Lime Carbonated Beverages[J]. Journal of Agricultural and Food Chemistry,2015,63(1):112-119.
[31] 李华敏,孙舒扬,黄萍萍,等.戴尔有孢圆酵母WA19与酿酒酵母F33混合发酵在樱桃酒酿造中的应用[J].食品科学,2018, 39(24):123-130.
[32] 肖作兵.樱桃酒特征香气成分研究[D].上海:上海应用技术学院,2016.
[33] 陶永胜.贺兰山东麓霞多丽葡萄酒的香气成分活性及其指纹特征研究[J].宁夏林业, 2019(3): 55-61.
