酱香型大曲是白酒酿造的关键环节,为白酒酿造提供丰富的酶和风味物质前体。然而,过度繁殖的乳酸菌将引起大曲酸度升高并降低糖化力、发酵力和出酒率。抑菌黑曲(antibacterial black Daqu,BBQ)是一种能够抑制乳酸菌的特殊黑曲。因此,解析抑菌黑曲中抑制乳酸菌的关键代谢物将有助于降低大曲酸度。这项研究利用非靶向代谢组学分析了抑菌黑曲和黑曲的代谢物,发现抑菌黑曲中有259种显著上调的差异代谢物。香兰素、萘啶酸、2-甲基呋喃和黑色素被鉴定为潜在的抑菌物质,并通过体外纯培养乳酸菌得到了证实。抑菌黑曲中的酪氨酸代谢显著上调,该代谢通路终产物是黑色素。香兰素由苯丙氨酸代谢产生。抑菌黑曲中频繁的美拉德反应生成大量有机杂环类物质包括2-甲基呋喃和萘啶酸。综上所述,黑曲中香兰素、萘啶酸、2-甲基呋喃和黑色素的积累,使其产生抑制乳酸菌活性。该研究将有助于通过提升大曲品质来降低酱香型白酒的酸度。
Sauce-flavored Daqu is the key link of Baijiu brewing, which provides abundant enzymes and flavor precursors for Baijiu brewing. However, over-proliferated lactic acid bacteria will increase the acidity of Daqu and reduce the saccharification power, fermentation power and alcohol yield. Antibacterial black Daqu (BBQ) is a special black Daqu (BQ) that inhibits lactic acid bacteria. Therefore, analyzing the key metabolites that inhibit lactic acid bacteria in BBQ will help to reduce the acidity of Daqu. This study analyzed the metabolites of BBQ and BQ using untargeted metabolomics and found 259 differential metabolites that were significantly up-regulated in BBQ. Vanillin, nalidixic acid, 2-methylfuran, and melanin were identified as potential bacteriostatic substances and confirmed by pure culture of lactic acid bacteria in vitro. Tyrosine metabolism was significantly up-regulated in BBQ, and the end product of this metabolic pathway was melanin. Vanillin is produced by the metabolism of phenylalanine. Frequent Maillard reactions in BBQ generated a large number of organic heterocycles including 2-methylfuran and nalidixic acid. Overall, the accumulation of vanillin, nalidixic acid, 2-methylfuran, and melanin in BQ resulted in its inhibitory activity against lactic acid bacteria. This research will help to reduce the acidity of sauce-flavored Baijiu by improving the quality of Daqu.
[1] 李大和. 建国五十年来白酒生产技术的伟大成就(六)[J].酿酒, 1999,26(6):19-31.
LI D H.The great achievements of liquor production technology in the 50 years since the founding of the People's Republic of China (six)[J].Liquor Making, 1999,26(6):19-31.
[2] WU C D, QIN Z, HUANG J, et al.Characterization of microbial community in Daqu by PLFA method[J].Food Science and Technology Research, 2014, 20(1):147-154.
[3] GAN S H, YANG F, SAHU S K, et al.Deciphering the composition and functional profile of the microbial communities in Chinese Moutai liquor starters[J].Frontiers in Microbiology, 2019, 10:1 540.
[4] 唐贤华. 不同入窖酸度对浓香型白酒发酵的影响[J].酿酒, 2020, 47(3):54-57.
TANG X H.Effect of different acidity in cellar on fermented grains of Luzhou-flavor Baijiu [J].Liquor Making, 2020, 47(3):54-57.
[5] 唐贤华, 王思思, 黄睿.不同入窖糟醅在发酵过程中有机酸的变化及对酒质的影响[J].粮食与食品工业, 2020, 27(2):39-44;50.
TANG X H, WANG S S, HUANG R.Changes of organic acids in fermented grains of different pit entry grains and their effects on liquor quality[J].Cereal & Food Industry, 2020, 27(2):39-44;50.
[6] 赖登燡, 薛常有, 潘华文.入窖七因素的变化规律及相互关系的研究(四):入窖酸度[J].酿酒科技, 2011(4):43-45.
LAI D Y, XUE C Y, PAN H W.Study on the change rules of seven factors for pit entry and their correlations (IV):Pit entry acidity[J].Liquor-Making Science & Technology, 2011(4):43-45.
[7] YANG L, FAN W L,XU Y. GC×GC-TOF/MS and UPLC-Q-TOF/MS based untargeted metabolomics coupled with physicochemical properties to reveal the characteristics of different type Daqus for making soy sauce aroma and flavor type baijiu[J].LWT-Food Science and Technology, 2021, 146:111416.
[8] SONG X B, JING S, ZHU L, et al.Untargeted and targeted metabolomics strategy for the classification of strong aroma-type baijiu (liquor) according to geographical origin using comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry[J].Food Chemistry, 2020, 314:126098.
[9] DOPPLER M, KLUGER B, BUESCHL C, et al.Stable isotope-assisted evaluation of different extraction solvents for untargeted metabolomics of plants[J].International Journal of Molecular Sciences, 2016, 17(7):1 017.
[10] SMITH C A, WANT E J, O'MAILLE G, et al.XCMS:Processing mass spectrometry data for metabolite profiling using nonlinear peak alignment, matching and identification[J].Analytical Chemistry, 2006, 78(3):779-787.
[11] WIKLUND S, JOHANSSON E, SJÖSTRÖM L, et al.Visualization of GC/TOF-MS-based metabolomics data for identification of biochemically interesting compounds using OPLS class models[J].Analytical Chemistry, 2008, 80(1):115-122.
[12] TRYGG J, WOLD S.Orthogonal projections to latent structures (O-PLS)[J].Journal of Chemometrics, 2002, 16(3):119-128.
[13] YAN N, DU Y M, LIU X M, et al.A comparative UHPLC-QqQ-MS-based metabolomics approach for evaluating Chinese and North American wild rice[J].Food Chemistry, 2019, 275:618-627.
[14] GONG Y, JI P, YANG Y S, et al.Metabolic-pathway-based subtyping of triple-negative breast cancer reveals potential therapeutic targets[J].Cell Metabolism, 2021, 33(1):51-64.e9.
[15] 赵建芬, 韦寿莲, 严子军.香兰素的抑菌性及其应用的初步研究[J].微生物学杂志, 2009, 29(6):73-76.
ZHAO J F, WEI S L, YAN Z J.Microbial inhibition effect and the application of vanillin[J].Journal of Microbiology, 2009, 29(6):73-76.
[16] LAZO-LÁSCAREZ S, GUTIÉRREZ L Z, DUARTE-MARTíNEZ F, et al.Antimicrobial resistance and genetic diversity of Campylobacter spp.isolated from broiler chicken at three levels of the poultry production chain in Costa rica[J].Journal of Food Protection, 2021, 84(12):2 143-2 150.
[17] RUFIÁN-HENARES J A, MORALES F J.Functional properties of melanoidins:In vitro antioxidant, antimicrobial and antihypertensive activities[J].Food Research International, 2007, 40(8):995-1 002.
[18] RAHMANI ELIATO T, SMITH J T, TIAN Z, et al.Melanin pigments extracted from horsehair as antibacterial agents[J].Journal of Materials Chemistry B, 2021, 9(6):1 536-1 545.
[19] TRAN-LY A N, REYES C, SCHWARZE F W M R, et al.Microbial production of melanin and its various applications[J].World Journal of Microbiology & Biotechnology, 2020, 36(11):170.
[20] WANG Y R, CAI W C,WANG W P, et al.Analysis of microbial diversity and functional differences in different types of high-temperature Daqu[J].Food Science & Nutrition, 2021, 9(2):1 003-1 016.
[21] 班世栋, 王晓丹, 胡宝东, 等.酱香型大曲中水解酶系研究[J].酿酒科技, 2015(4):13-19.
BAN S D, WANG X D, HU B D, et al.Hydrolyzing enzymes in Jiangxiang Daqu[J].Liquor-Making Science & Technology, 2015(4):13-19.
