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芝麻香型白酒发酵过程中乳酸菌多样性及其演替规律

邢敏钰1 杜海1 徐岩1

(1.工业生物技术教育部重点实验室江南大学生物工程学院酿酒科学与酶技术中心, 江苏无锡 214122)

【摘要】【背景】乳酸菌是白酒发酵过程中一类非常重要的微生物, 其种类及动态变化对于白酒品质具有重要影响。然而, 目前对于芝麻香型白酒发酵过程中乳酸菌群落结构及其演替规律的认识并不全面。【目的】揭示芝麻香型白酒发酵过程中乳酸菌的多样性及菌群的演替规律, 为更好地探索白酒酿造机理和控制白酒品质提供生物学依据。【方法】利用高通量测序技术对芝麻香型白酒发酵过程中乳酸菌菌群演替进行跟踪分析, 同时采用实时荧光定量PCR对发酵过程中乳酸菌的生物量进行定量分析。【结果】高通量测序结果显示, 芝麻香型白酒发酵过程涉及5个属的乳酸菌:魏斯氏菌属 (Weissella) 、片球菌属 (Pediococcus) 、乳杆菌属 (Lactobacillus) 、明串珠菌属 (Leuconostoc) 和乳球菌属 (Lactococcus) , 共计43种乳酸菌。其中, 在发酵过程中平均相对丰度大于0.5%的乳酸菌有10种, 分别是类肠膜魏斯氏菌 (Weissella paramesenteroides) 、食窦魏斯氏菌 (Weissella cibaria) 、融合魏斯氏菌 (Weissella confusa) 、戊糖片球菌 (Pediococcus pentosaceus) 、假肠膜明串珠菌 (Leuconostoc pseudomesenteroides) 、发酵乳杆菌 (Lactobacillus fermentum) 、植物乳杆菌 (Lactobacillus plantarum) 、副干酪乳杆菌 (Lactobacillus paracasei) 、耐酸乳杆菌 (Lactobacillus acetotolerans) 和Lactobacillus sp.。在堆积发酵过程中, Weissella属占细菌总量的50%以上, 其次是Pediococcus属和Lactobacillus属, 而Leuconostoc属和Lactococcus属相对较少。在窖池发酵过程中Lactobacillus属的乳酸菌逐渐成为优势细菌, 尤其是Lactobacillus sp.在窖池发酵中后期相对丰度达到80%以上。实时荧光定量PCR结果显示, 在堆积发酵和窖池发酵前期乳酸菌总量变化不大;从窖池发酵5 d开始, 乳酸菌总量迅速上升, 30 d时达到最大值。【结论】对白酒发酵过程中乳酸菌种类及动态变化的研究有助于探究白酒酿造过程中乳酸菌功能, 进而解析白酒酿造机理, 最终达到控制白酒品质的目的。

【关键词】 乳酸菌;多样性;演替规律;

【DOI】

【基金资助】 国家自然科学基金 (31501469, 31530055) ; 江苏省自然科学基金 (BK20150143) ; 中央高校基本科研业务费专项资金 (JUSRP11537) ; 江苏省现代工业发酵协同创新中心行业发展项目; 中国白酒“3C”计划;

Diversity and succession of lactic acid bacteria during sesame-flavor liquor fermentation

XING Min-Yu1 DU Hai1 XU Yan1

(1.Key Laboratory of Industrial Biotechnology, Ministry of Education; Center for Brewing Science and Enzyme Technology, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China 214122)

【Abstract】[Background] Lactic acid bacteria are important in liquor fermentation, and the diversity and succession of lactic acid bacteria have important influence on liquor quality. However, the structure and succession of lactic acid bacterial communities are not clear during sesame-flavor liquor fermentation. [Objective] We studied the diversity and succession of lactic acid bacterial communities during sesame-flavor liquor fermentation for better process control and product quality. [Methods] High-throughput sequencing was used to analyze lactic acid bacterial communities during sesame-flavor liquor fermentation. Biomass of lactic acid bacteria was quantified by real-time qPCR. [Results] Lactic acid bacteria in sesame-flavor liquor fermentation included Weissella, Pediococcus, Lactobacillus, Leuconostoc, and Lactococcus, classified to 43 species. Ten species of lactic acid bacteria were observed with relative abundance higher than 0.5%, including Weissella paramesenteroides, Weissella cibaria, Weissella confuse, Pediococcus pentosaceus, Leuconostoc pseudomesenteroides, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus paracasei, Lactobacillus acetotolerans and Lactobacillus sp. During heap-fermentation, Weissella was more than half of the total bacteria, followed by Pediococcus and Lactobacillus. Leuconostoc and Lactococcus were present in low numbers. During pit-fermentation, Lactobacillus became the predominant bacteria, and the relative abundance of Lactobacillus sp. was up to 80% at the mid-to-late stage of pit-fermentation. During heap-fermentation and the early stage of pit-fermentation, the biomass of lactic acid bacteria changed little. After 5 days of pit-fermentation, the biomass of lactic acid bacteria increased fast, and reached the maximum on day 30. [Conclusion] The knowledge of diversity and succession of lactic acid bacteria during sesame-flavor liquor fermentation will help understand the roles of lactic acid bacteria in liquor production with better process control and product quality.

【Keywords】 Lactic acid bacteria; Diversity; Succession;

【DOI】

【Funds】 National Natural Science Foundation of China (31501469, 31530055); Natural Science Foundation of Jiangsu Province, China (BK20150143); Fundamental Research Funds for the Central Universities in China (JUSRP11537); Collaborative Innovation Center of Modern Fermentation Industry in Jiangsu Province, China; The “3C” Plan of Chinese Liquor;

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    References

    [1] Liu SN, Han Y, Zhou ZJ. Lactic acid bacteria in traditional fermented Chinese foods [J]. Food Research International, 2011, 44 (3): 643-651

    [2] Wu ZF, Zhuang BW, Weng PF, et al. Fermentation quality characteristics and flavor formation changes during the process of pickled wax gourd in eastern Zhejiang [J]. International Journal of Food Properties, 2016, 19 (2): 409-419

    [3] le Lay C, Coton E, le Blay G, et al. Identification and quantification of antifungal compounds produced by lactic acid bacteria and propionibacteria [J]. International Journal of Food Microbiology, 2016, 239: 79-85

    [4] Gerez CL, Torres MJ, de Valdez GF, et al. Control of spoilage fungi by lactic acid bacteria [J]. Biological Control, 2013, 64 (3): 231-237

    [5] Magnusson J, Ström K, Roos S, et al. Broad and complex antifungal activity among environmental isolates of lactic acid bacteria [J]. FEMS Microbiology Letters, 2003, 219 (1): 129-135

    [6] Dalié DKD, Deschamps AM, Richard-Forget F. Lactic acid bacteria-potential for control of mould growth and mycotoxins: a review [J]. Food Control, 2010, 21 (4): 370-380

    [7] Wu Q, Chen LQ, Xu Y. Yeast community associated with the solid state fermentation of traditional Chinese Maotai-flavor liquor [J]. International Journal of Food Microbiology, 2013, 166 (2): 323-330

    [8] Wu LL, Wang HY, Xu Y, et al. Differences of lactic acid bacteria community between soy sauce aroma style and light aroma style liquor fermentation [J]. Microbiology China, 2013, 40 (12): 2182-2188 (in Chinese)

    [9] Qi YM. Investigation on sesame-flavor liquor production [J]. Liquor-making Science & Technology, 2009 (8): 140-142 (in Chinese)

    [10] Liu MM, Wang JG, Sun PP, et al. Application of bran starter in the production of sesame-flavor liquor [J]. Liquor-making Science & Technology, 2013 (3): 69-70, 74 (in Chinese)

    [11] Zhang B. Production and application of distiller’s yeast used especially for the production of sesame-flavor liquor [J]. Liquor Making, 2012, 39 (6): 38-41 (in Chinese)

    [12] Wu Q, Ling J, Xu Y. Starter culture selection for making Chinese sesame-flavored liquor based on microbial metabolic activity in mixed-culture fermentation [J]. Applied and Environmental Microbiology, 2014, 80 (14): 4450-4459

    [13] Xu ZJ. Analysis of brewing microbiology diversity of the sesame-favor liquor [D]. Hangzhou: Master’s Thesis of Zhejiang Sci-Tech University, 2012 (in Chinese)

    [14] Wang HY, Zhang XJ, Zhao LP, et al. Analysis and comparison of the bacterial community in fermented grains during the fermentation for two different styles of Chinese liquor [J]. Journal of Industrial Microbiology & Biotechnology, 2008, 35 (6): 603-609

    [15] de Pasquale I, di Cagno R, Buchin S, et al. Microbial ecology dynamics reveal a succession in the core microbiota involved in the ripening of pasta filata Caciocavallo Pugliese cheese [J]. Applied and Environmental Microbiology, 2014, 80 (19): 6243-6255

    [16] Michel E, Monfort C, Deffrasnes M, et al. Characterization of relative abundance of lactic acid bacteria species in French organic sourdough by cultural, qPCR and MiSeq high-throughput sequencing methods [J]. International Journal of Food Microbiology, 2016, 239: 35-43

    [17] Zhang XL, Tian XQ, Ma LY, et al. Biodiversity of the symbiotic bacteria associated with toxic marine dinoflagellate Alexandrium tamarense [J]. Journal of Biosciences and Medicines, 2015, 3 (6): 23-28

    [18] Zhang Y. Community structures and functions of lactic acid bacteria during Maotai-flavor liquor fermentation [D]. Wuxi: Master’s Thesis of Jiangnan University, 2015 (in Chinese)

    [19] Wang HY, Zhang XJ, Xu Y, et al. Analysis of the bacterial community in fermented grains of strong aroma style and roasted sesame aroma style liquor [J]. Liquor-Making Science & Technology, 2008 (2): 86-89, 91 (in Chinese)

    [20] Toh H, Morita H, Tsuji H, et al. Complete genome sequence of Lactobacillus acetotolerans RIB 9124 (NBRC 13120) isolated from putrefied (hiochi) Japanese sake [J]. Journal of Biotechnology, 2015, 214: 214-215

    [21] Basso TO, Gomes FS, Lopes ML, et al. Homo- and hetero-fermentative lactobacilli differently affect sugarcane-based fuel ethanol fermentation [J]. Antonie Van Leeuwenhoek, 2014, 105 (1): 169-177

    [22] Blandino A, Al-Aseeri ME, Pandiella SS, et al. Cereal-based fermented foods and beverages [J]. Food Research International, 2003, 36 (6): 527-543

    [23] Sudun, Wulijideligen, Arakawa K, et al. Interaction between lactic acid bacteria and yeasts in airag, an alcoholic fermented milk [J]. Animal Science Journal, 2013, 84 (1): 66-74

    [24] Lee ME, Jang JY, Lee JH, et al. Starter cultures for kimchi fermentation [J]. Journal of Microbiology and Biotechnology, 2015, 25 (5): 559-568

    [25] Jung JY, Lee SH, Jeon CO. Kimchi microflora: history, current status, and perspectives for industrial kimchi production [J]. Applied Microbiology and Biotechnology, 2014, 98 (6): 2385-2393

    [26] Zhu KL. Research on screening, high cell density cultivation and mixture of lactic acid bacteria for pickle fermentation [D]. Wuxi: Master’s Thesis of Jiangnan University, 2014 (in Chinese)

This Article

ISSN:0253-2654

CN: 11-1996/Q

Vol 45, No. 01, Pages 19-28

January 2018

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Article Outline

Abstract

  • 1 Materials and methods
  • 2 Results and analyses
  • 3 Conclusions
  • References