红曲黄酒传统酿造用曲中的微生物菌群及挥发性风味组分分析
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摘要
采用固相微萃取-气相色谱-质谱联用技术对红曲和白曲中的挥发性风味组分进行分析鉴定,并利用主成分分析和正交偏最小二乘法判别分析模型确定不同酒曲中的特征香气成分,同时运用MiSeq高通量测序技术解析红曲和白曲中的微生物菌群结构。结果表明:从红曲和白曲中共检测出70种挥发性成分,且红曲和白曲的挥发性风味组分存在显著差异;红曲中的优势微生物包括芽孢杆菌属、魏斯氏菌属、片球菌属、黑曲霉、紫红曲霉和黄曲霉等,而白曲中的优势微生物包括泛生菌属、肠杆菌属、魏斯氏菌属、酿酒酵母、少根根霉和印度毛霉等。研究结果为阐明红曲黄酒微生物的产香机理和提升红曲黄酒风味品质提供一定的理论依据。
Headspace solid phase microextraction coupled with gas chromatography (HS-SPME-GC-MS) was applied to identify the volatile flavor components of two traditional fermentation starters (Baiqu and Hongqu) for Hongqu glutinous rice wine. The characteristic aroma components were determined by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Also, MiSeq high throughput sequencing technology was used to analyze the structure of microbial flora in the two fermentation starters. Results showed that a total of 70 volatile components were detected in the fermentation starters, and that there was a significant difference in the volatile profiles of Hongqu and Baiqu.The dominant microorganisms in Hongqu were Bacillus sp., Weasella sp., Staphylococcus sp., Aspergillus niger, A. oryzae and A. flavus, while the dominant microorganisms in Baiqu were Pantoea sp., Enterobacter sp., Weasella sp., Saccharomyces cerevisiae, Rhizopus arrhizus and Mucor indicus. The research results provide a theoretical basis for elucidating the mechanism of aroma production and for improving the flavor quality of Hongqu glutinous rice wine.
Headspace solid phase microextraction coupled with gas chromatography (HS-SPME-GC-MS) was applied to identify the volatile flavor components of two traditional fermentation starters (Baiqu and Hongqu) for Hongqu glutinous rice wine. The characteristic aroma components were determined by principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Also, MiSeq high throughput sequencing technology was used to analyze the structure of microbial flora in the two fermentation starters. Results showed that a total of 70 volatile components were detected in the fermentation starters, and that there was a significant difference in the volatile profiles of Hongqu and Baiqu.The dominant microorganisms in Hongqu were Bacillus sp., Weasella sp., Staphylococcus sp., Aspergillus niger, A. oryzae and A. flavus, while the dominant microorganisms in Baiqu were Pantoea sp., Enterobacter sp., Weasella sp., Saccharomyces cerevisiae, Rhizopus arrhizus and Mucor indicus. The research results provide a theoretical basis for elucidating the mechanism of aroma production and for improving the flavor quality of Hongqu glutinous rice wine.
引文
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[31]余培斌.改善绍兴黄酒麦曲品质的初步研究[D].无锡:江南大学,2013:15-33.
[2]TIAN Y,HUANG J,XIE T,et al.Oenological characteristics,amino acids and volatile profiles of Hongqu rice wines during pottery storage:effects of high hydrostatic pressure processing[J].Food Chemistry,2016,203:456-464.DOI:10.1016/j.foodchem.2016.02.116.
[3]薛景波.黄酒接种生麦曲微生物群落结构及分离培养微生物的产酶、产香性质分析[D].?无锡:江南大学,2016:1-7.
[4]LV X,JIA R,LI Y,et al.Characterization of the dominant bacterial communities of traditional fermentation starters for Hong Qu glutinous rice wine by means of MALDI-TOF mass spectrometry fingerprinting,16S rRNA gene sequencing and species-specific PCRs[J].Food Control,2016,67:292-302.DOI:10.1016/j.foodcont.2016.03.005.
[5]左楠楠.红曲黄酒酿造关键技术的研究[D].杭州:浙江师范大学,2013:1-4.
[6]吕旭聪,翁星,黄若兰,等.红曲黄酒酿造用曲及传统酿造过程中酵母菌的多样性研究[J].中国食品学报,2012,12(1):182-190.DOI:10.3969/j.issn.1009-7848.2012.01.028
[7]LV X,HUANG X,ZHANG W,et al.Yeast diversity of traditional alcohol fermentation starters for Hong Qu glutinous rice wine brewing,revealed by culture-dependent and culture-independent methods[J].Food Control,2013,34(1):183-190.DOI:10.1016/j.foodcont.2013.04.020.
[8]余建平.红曲的研究现状及应用[J].科技信息,2010(32):538-539.DOI:10.3969/j.issn.1001-9960.2010.32.467.
[9]江坤文.传统小曲生产的特点[J].现代食品科技,1989(2):43.
[10]董洋,刘亚琼,刘姗姗.杂粮红曲酒高级醇及其挥发性风味物质研究[J].酿酒科技,2017(3):55-59.DOI:10.13746/j.njkj.2016378.
[11]GHOSH A,BHADURY P.Insights into bacterioplankton community structure from Sundarbans mangrove ecoregion using Sanger and Illumina MiSeq sequencing approaches:a comparative analysis[J].Genomics Data,2017,11:39-42.DOI:10.1016/j.gdata.2016.11.017.
[12]LIU M,TANG Y,ZHAO K,et al.Determination of the fungal community of pit mud in fermentation cellars for Chinese strong-flavor liquor,using DGGE and Illumina MiSeq sequencing[J].Food Research International,2017,91:80-87.DOI:10.1016/j.foodres.2016.11.037.
[13]ROESCH L F,FULTHORPE R R,RIVA A,et al.Pyrosequencing enumerates and contrasts soil microbial diversity[J].ISME Journal,2007,1(4):283-290.DOI:10.1038/ismej.2007.53.
[14]ZHENG Q,LIN B,WANG Y,et al.Proteomic and high-throughput analysis of protein expression and microbial diversity of microbes from 30-and 300-year pit muds of Chinese Luzhou-flavor liquor[J].Food Research International,2015,75:305-314.DOI:10.1016/j.foodres.2015.06.029.
[15]KUCZYNSKI J,STOMBAUGH J,WALTERS W A,et al.Using QⅡME to analyze 16S rRNA gene sequences from microbial communities[J].Current Protocols in Bioinformatics,2012,27(1):1-20.DOI:10.1002/0471250953.bi1007s36.
[16]王鹏,吴群,徐岩.中国白酒发酵过程中的核心微生物群及其与环境因子的关系[J].微生物学报,2018(1):142-153.DOI:10.13343/j.cnki.wsxb.20170060.
[17]李红蕾,冯涛,荣志伟.黄酒酿制过程中风味物质变化规律的研究[J].食品工业,2011,32(12):69-72.
[18]宋伟,胡婉君,徐宗季,等.湖南省主要早籼稻品种挥发性物质种类及含量[J].中国农业科学,2017,50(2):348-361.DOI:10.3864/j.issn.0578-1752.2017.02.013.
[19]陈丙友,韩英,张鑫,等.酒醅温度调控对清香型白酒发酵过程的影响[J].食品与发酵工业,2016,42(6):44-49.DOI:10.13995/j.cnki.11-1802/ts.201606008.
[20]WU X,LI Y,WANG Q,et al.Effects of berberine and pomegranate seed oil on plasma phospholipid metabolites associated with risks of type 2 diabetes mellitus by U-HPLC Q-TOF-MS[J].Journal of Chomatography B Analytical Technologies in the Biomedical&L i f e S c i e n c e s,2 0 1 5,1 0 0 7(1 4):11 0-1 2 0.D O I:1 0.1 0 1 6/j.foodres.2016.11.037.
[21]WORLEY B,PPWERS R.Multivariate analysis in metabolomics[J].Current Metabolomics,2013,1(1):92-107.DOI:10.2174/2213235X11301010092.
[22]QI Y,PI Z,LIU S,et al.A metabonomic study of adjuvant-induced arthritis in rats using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry[J].Molecular BioSystems,2014,10(10):2617-2625.DOI:10.1039/c4mb00131a.
[23]李思涛,黄小玲,吴时光,等.极低出生体重早产儿尿代谢组学研究[J].中华儿科杂志,2017,55(6):434-438.DOI:10.3760/cma.j.issn.0578-1310.2017.06.008.
[24]郭敏,黄永光,邱树毅,等.高通量测序在酱香白酒微生态多样性研究中的应用[J].中国酿造,2017,36(5):146-151.DOI:10.11882/j.issn.0254-5071.2017.05.031.
[25]刘芸雅.绍兴黄酒发酵中微生物群落结构及其对风味物质影响研究[D].无锡:江南大学,2015:20-23.
[26]MARTENS H,DAWOUD E,VERACHTERT H.Synthesis of aroma compounds by wort enterobacteria during the first stage of lambic fermentation[J].Journal of the Institute of Brewing,1992,98(5):421-425.DOI:10.1002/j.2050-0416.1992.tb01126.x.
[27]林晓庆,黄志清,刘志彬,等.米根霉液态发酵产糖化酶工艺条件研究[J].酿酒科技,2014(9):9-13.DOI:10.13746/j.njkj.2014.0166.
[28]宋克伟,胡建华,魏金旺,等.米根霉在牛栏山基酒生产中的应用[J].酿酒科技,2015(2):73-75.DOI:10.13746/j.njkj.2014333.
[29]赵伟,赵兵涛,次仁潘多,等.酿酒酵母代谢有机酸对乙醇发酵的影响[J].可再生能源,2017,35(7):971-977.DOI:10.13941/j.cnki.21-1469/tk.2017.07.004.
[30]LV X,CAI Q,KE X,et al.Characterization of fungal community and dynamics during the traditional brewing of Wuyi Hong Qu glutinous rice wine by means of multiple culture-independent methods[J].Food Control,2015,54:231-239.DOI:10.1016/j.foodcont.2015.01.046.
[31]余培斌.改善绍兴黄酒麦曲品质的初步研究[D].无锡:江南大学,2013:15-33.