食品与发酵工业 >

2022 , Vol. 48 >Issue 19: 50 - 57

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.029928

研究报告

一株耐酸乳杆菌Lactobacillus acetotolerans FBKL1.0204的筛选鉴定及耐酸、产酸特性

  • 赵皓静 ,
  • 冯婧煕 ,
  • 王晓丹 ,
  • 周鸿翔 ,
  • 罗小叶 ,
  • 邱树毅 ,
  • 班世栋
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  • 1(贵州大学 酿酒与食品工程学院,贵州 贵阳,550025)
    2(贵州省发酵工程与生物制药重点实验室(贵州大学),贵州 贵阳,550025)
    3(中国农业大学 生物学院,北京,100193)
第一作者：硕士研究生(班世栋讲师为通信作者,E-mail:sdban@gzu.edu.cn)

收稿日期: 2021-11-08

  修回日期: 2022-03-07

  网络出版日期: 2022-11-01

基金资助

贵州省科技计划项目(黔科合支撑[2021]一般106);贵州省科技厅人才项目(黔科合平台人才[2018]5251);中国轻工业浓香型白酒固态发酵重点实验室开放基金项目(2019JJ014);贵州大学引进人才科研项目(贵大人基合字(2019)03);薏仁米酒生产工艺关键技术的研究与开发项目(GZDX20190925)

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Screening, identification and characterization of acid tolerant Lactobacillus acetotolerans FBKL1.0204

  • ZHAO Haojing ,
  • FENG Jingxi ,
  • WANG Xiaodan ,
  • ZHOU Hongxiang ,
  • LUO Xiaoye ,
  • QIU Shuyi ,
  • BAN Shidong
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  • 1(School of Liquor and Food Engineering, Guizhou University, Guiyang 550025, China)
    2(Key Laboratory of Fermentation Engineering and Bio-Pharmacy of Guizhou Province, Guizhou University, Guiyang 550025, China)
    3(College of Biological Sciences, China Agricultural University, Beijing 100193, China)

Received date: 2021-11-08

  Revised date: 2022-03-07

  Online published: 2022-11-01

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摘要

耐酸乳杆菌是浓香型白酒窖池发酵中后期酒醅中的优势菌,此时酒醅酸性较强。采用传统分离纯化法分离酒醅中细菌,经形态学特征和生理生化特性分析,并结合16S rDNA基因序列同源性分析和管家基因pheS和rpoA序列同源性分析及耐酸性筛选,分离鉴定出1株耐酸乳杆菌(Lactobacillus acetotolerans)FBKL1.0204。采用比浊法、电极电位法和高效液相色谱法测定菌株在不同酸胁迫下的生物量、pH和有机酸的变化。耐酸特性研究表明,在乳酸、乙酸和盐酸酸化的酸性发酵环境中,FBKL1.0204能耐受pH 4的酸性环境,极端生存pH值为3;产酸特性研究表明,FBKL1.0204为异型乳酸发酵,主要代谢生成乳酸和乙酸,最终使发酵液pH值降至3.5左右,在乙酸调节pH值为6的MRS培养基中,其代谢生成乳酸和乙酸含量最高,分别为(28.32±0.54)g/L和(2.73±0.52)g/L。由此可见,FBKL1.0204具有优良的耐酸和产酸特性,有利于其在白酒酿造过程中生长繁殖并丰富白酒风味。

关键词： 浓香型白酒; 酒醅; 耐酸乳杆菌; 分离; 鉴定; 耐酸特性; 产酸特性

本文引用格式

赵皓静 , 冯婧煕 , 王晓丹 , 周鸿翔 , 罗小叶 , 邱树毅 , 班世栋 . 一株耐酸乳杆菌Lactobacillus acetotolerans FBKL1.0204的筛选鉴定及耐酸、产酸特性[J]. 食品与发酵工业, 2022 , 48(19) : 50 -57 . DOI: 10.13995/j.cnki.11-1802/ts.029928

Abstract

Lactobacillus acetotolerans is the dominant bacteria in the middle and late fermentation in the pits of Luzhou-flavor Baijiu, which produces excessive acids and makes strong acidity of fermented grains. A strain FKL1.0204 in fermented grains was isolated and identified by morphological characteristics, physiological and biochemical characteristics, 16S rDNA gene sequence analysis, pheS and rpoA gene sequence analysis and acid tolerance screening. It was identified as Lactobacillus acetotolerans. Strain FKL1.0204 could withstand the acidic environment of pH 4, and the extreme survival pH was 3. The study of acid production characteristics showed that it was hetero-lactic acid fermentation, which mainly metabolized lactic acid and acetic acid, and finally lowers the pH of fermentation liquid to about 3.5. The contents of lactic acid and acetic acid were the highest in the MRS medium under pH 6 (modified by acetic acid), (28.32±0.54) g/L and (2.73±0.52) g/L respectively.

Key words： Luzhou-flavor Baijiu; fermented grains; Lactobacillus acetotolerans; screening; identification; acid resistance; acid production

参考文献

[1] 徐军. 浓香型枝江白酒香味成分的分析研究[D]. 武汉: 华中农业大学, 2019.
XU J. Study on aroma components of strong-aroma Zhijiang Baijiu[D]. Wuhan: Huazhong Agricultural University, 2019.
[2] HU X L, DU H, XU Y. Identification and quantification of the caproic acid-producing bacterium Clostridium kluyveri in the fermentation of pit mud used for Chinese strong-aroma type liquor production[J]. International Journal of Food Microbiology, 2015, 214:116-122.
[3] ZHENG J, WU C, HUANNG J, et al. Spatial distribution of bacterial communities and related biochemical properties in Luzhou-flavor liquor-fermented grains[J]. Journal of Food Science, 2014, 79(12): M2 491-M2 498.
[4] DI MARTINO C, TESTA B, LETIZIA F, et al. Effect of exogenous proline on the ethanolic tolerance and malolactic performance of Oenococcus oeni[J]. Journal of Food Science and Technology, 2020, 57(11): 3 973-3 979.
[5] ZOU W, ZHAO C Q, LUO H B. Diversity and function of microbial community in Chinese strong-flavor Baijiu ecosystem: A review[J]. Frontiers in Microbiology, 2018, 9:671.
[6] WANG X S, DU H, XU Y. Source tracking of prokaryotic communities in fermented grain of Chinese strong-flavor liquor[J]. International Journal of Food Microbiology, 2017, 244:27-35.
[7] SUN W N, XIAO H Z, PENG Q, et al. Analysis of bacterial diversity of Chinese Luzhou-flavor liquor brewed in different seasons by Illumina Miseq sequencing[J]. Annals of Microbiology, 2016, 66(3):1 293-1 301.
[8] 刘森, 李林光, 李可, 等. 中国浓香型白酒窖池窖泥中原核微生物群落的空间异质性[J]. 食品科学, 2013, 34(21):221-226.
LIU S, LI L G, LI K, et al. Spatial heterogeneity of prokaryotic microbial communities in Luzhou-flavor liquor pit mud[J]. Food Science, 2013, 34(21):221-226.
[9] WANG H Y, ZHANG X J, ZHAO L P, 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 and Biotechnology, 2008, 35(6):603-609.
[10] LI X R, MA E B, YAN L Z, et al. Bacterial and fungal diversity in the traditional Chinese liquor fermentation process[J]. International Journal of Food Microbiology, 2011, 146(1):31-37.
[11] ENTANI E, MASAI H, SUZUKI K I. Lactobacillus acetotolerans, a new species from fermented vinegar broth[J]. International Journal of Systematic Bacteriology, 1986, 36(4):544-549.
[12] 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.
[13] YANG X P, TENG K L, ZHANG J, et al. Transcriptome responses of Lactobacillus acetotolerans F28 to a short and long term ethanol stress[J]. Scientific Reports, 2017, 7:2650.
[14] LIU J Y, DENG Y, PETERS B M, et al. Transcriptomic analysis on the formation of the viable putative non-culturable state of beer-spoilage Lactobacillus acetotolerans[J]. Scientific Reports, 2016, 6:36753.
[15] LI H, HE R Y, XIONG X M, et al. Dynamic diversification of bacterial functional groups in the Baiyunbian liquor stacking fermentation process[J]. Annals of Microbiology, 2016, 66(3):1 229-1 237.
[16] 刘文容. 黄酒陈酿过程中酸败乳酸菌的分离鉴定及其特性研究[D]. 无锡:江南大学, 2017.
LIU W R. Studies on the isolation, identification and characteristics of spoilage lactic acid bacteria in storage of Chinese rice wine[D]. Wuxi: Jiangnan University, 2017.
[17] 布坎南·吉本斯. 伯杰氏细菌鉴定手册[M]. 第九版.北京: 科学出版社, 1984.
BUCHANAN GIBBONS. Berger’s Bacteria Identification Handbook[M]. 9th edition. Beijing: Science Press, 1984.
[18] 袁峥, 赵瑞香, 牛生洋, 等. 酸胁迫下嗜酸乳杆菌菌体形态的扫描电镜观察[J]. 食品工业科技, 2012, 33(24):199-201.
YUAN Z, ZHAO R X, NIU S Y, et al. Observation of the mycelia morphology of Lactobacillus acidophilus with scanning electron microscope under acid stress[J]. Science and Technology of Food Industry, 2012, 33(24):199-201.
[19] 陈笑语, 张力元, 纪海玉, 等. 渤海虾酱中益消细菌的分离和鉴定[J]. 中国食品学报, 2020, 20(3):258-265.
CHEN X Y, ZHANG L Y, JI H Y, et al. Isolation and identification of beneficial bacteria in Bohai shrimp paste[J]. Journal of Chinese Institute of Food Science and Technology, 2020, 20(3):258-265.
[20] ENDO A, TANIZAWA Y, ARITA M. Isolation and Identification of Lactic Acid Bacteria from Environmental Samples[M]. New York: Springer New York, 2019.
[21] 韩之皓, 郭帅, 黄天, 等. 复合益生菌在乳酸菌饮料中的发酵特性和功能性质研究[J]. 中国食品学报, 2020, 20(11):129-138.
HAN Z H, GUO S, HUANG T, et al. Studies on fermentation characteristics and functional properties of compound probiotics in lactic acid bacteria beverage[J]. Journal of Chinese Institute of Food Science and Technology, 2020, 20(11):129-138.
[22] 王芳, 廖勤俭, 安明哲, 等. 高效液相色谱同时检测白酒中四大酸的方法[J]. 酿酒科技, 2018(5):114-116.
WANG F, LIAO Q J, AN M Z, et al. HPLC for detecting the content of four main acids in Baijiu simultaneously[J]. Liquor-Making Science & Technology, 2018(5):114-116.
[23] TANAKA K, TAJIRI S, SAWADA R, et al. Acid-toleracnt lactic acid bacterium isolated from rice vinegar[J].Impact Journals, 2015, 10(3): 2 347-4 580.
[24] 翟磊, 程宵宵, 苏姣姣, 等. 一株食醋污染菌CICC 10774的鉴定及其生长代谢特性[J]. 微生物学通报, 2016, 43(7):1 524-1 531.
ZHAI L, CHENG X X, SU J J, et al. Identification and characterization of strain CICC 10774 causing vinegar spoilage[J]. Microbiology China, 2016, 43(7):1 524-1 531.
[25] 柴丽娟, 钱玮, 钟小忠, 等. 浓香型白酒发酵过程中窖内古菌群落分布特征[J]. 生物工程学报, 2020, 36(12):2 635-2 643.
CHAI L J, QIAN W, ZHONG X Z, et al. Distribution of archaeal community in the mud pit during strong-flavor Baijiu fermentation[J]. Chinese Journal of Biotechnology, 2020, 36(12):2 635-2 643.
[26] DE ANGELIS M, GOBBETTI M. Environmental stress responses in Lactobacillus: A review[J]. Proteomics, 2004, 4(1):106-122.
[27] CARPENTER C E, BROADBENT J R. External concentration of organic acid anions and pH: Key independentvariables for studying how organic acids inhibit growth of bacteria in mildly acidic foods[J]. Journal of Food Science,2009, 74(1): R12-R15.
[28] 张明阳, 张娟, 刘龙, 等. 精氨酸代谢途径抗酸关键基因对乳酸乳球菌Lactococcus lactis NZ9000胁迫抗性的影响[J]. 微生物学通报, 2017,44(2): 314-324.
ZHANG M Y, ZHANG J, LIU L, et al. Influence of key acid-resistant genes in arginine metabolism on stress tolerance in Lactococcus lactis NZ9000[J]. Microbiology China, 2017, 44(2): 314-324.
[29] 孟掉琴, 吴霞, 岳田利, 等. 混菌发酵苹果浊汁的益生菌筛选及其发酵动力学模型构建[J]. 食品科学, 2019, 40(12):153-159.
MENG D Q, WU X, YUE T L, et al. Screening of probiotics for mixed culture fermentation of cloudy apple juice and construction of kinetic models[J]. Food Science, 2019, 40(12):153-159.
[30] 黄桂东. Lactobacillus brevis NCL912的耐酸特性及其酸胁迫下差异表达蛋白的研究[D]. 南昌: 南昌大学, 2011.
HUANG G D. Acid tolerance of lactobaillus brevis NCL912 and its differentially expressed protein under acid strese[D]. Nanchang: Nanchang University, 2011.
[31] 杨帆. 酱香型白酒中乳酸代谢机理及调控策略的研究[D]. 无锡: 江南大学, 2020.
YANG F. Lactic acid metabolic mechanism investigation and control strategy development during Chinese Jiang-flavor liquor making[D]. Wuxi: Jiangnan University, 2020.
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