食品与发酵工业 >

2022 , Vol. 48 >Issue 11: 24 - 29

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.029692

研究报告

黄水芽胞杆菌抗逆性潜力挖掘及乙醇耐受机制分析

  • 程坤 ,
  • 刘蕊 ,
  • 郑佳 ,
  • 翟磊 ,
  • 姚粟
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  • 1(中国食品发酵工业研究院有限公司,中国工业微生物菌种保藏管理中心,北京,100015)
    2(宜宾五粮液股份有限公司,四川 宜宾,644000)
第一作者:博士,高级工程师(姚粟教授级高级工程师为通信作者,E-mail:milly@china-cicc.org)

收稿日期: 2021-10-16

  修回日期: 2021-11-03

  网络出版日期: 2022-06-23

基金资助

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(2019JJ008)

收起

Stress resistance potential mining of Bacillus aquiflavi 3H-10 and the mechanism of its ethanol tolerance

  • CHENG Kun ,
  • LIU Rui ,
  • ZHENG Jia ,
  • ZHAI Lei ,
  • YAO Su
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  • 1(China National Research Institute of Food and Fermentation Industries Co.Ltd., China Center of Industrial Culture Collection, Beijing 100015, China)
    2(Wuliangye Yibin Co.Ltd., Yibin 644000, China)

Received date: 2021-10-16

  Revised date: 2021-11-03

  Online published: 2022-06-23

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摘要

黄水芽胞杆菌(Bacillus aquiflavi)3H-10分离自浓香型白酒的黄水,是我国四川省宜宾地区首株酿酒微生物新种菌株,具有较好的产蛋白酶活力且菌种安全性风险较低,在白酒发酵过程中具有潜在应用价值。为了进一步挖掘该菌株的抗逆性潜力,从乙醇、酸、碱、NaCl和温度等方面对菌株开展特性研究,并从基因层面探究其抗逆性潜在机制。结果表明,菌株可在4%乙醇环境生长,对乙醇具有一定耐受性,推测菌株对乙醇的耐受性是其与环境适应性进化的结果。基因分析发现,菌株含有数量较多的热休克蛋白编码基因,使菌株在乙醇胁迫环境中能够产生应激反应,减弱乙醇对细胞的毒害作用;菌株基因组中含有完整的乙醇代谢途径,可将乙醇依次氧化生成乙醛和乙酸,后者进入柠檬酸循环,最终生成二氧化碳和水。因此,丰富的热休克蛋白与完整的乙醇代谢途径是菌株耐受乙醇的主要原因。

关键词： 黄水芽胞杆菌; 抗逆性; 乙醇耐受; 热休克蛋白; 代谢途径

本文引用格式

程坤 , 刘蕊 , 郑佳 , 翟磊 , 姚粟 . 黄水芽胞杆菌抗逆性潜力挖掘及乙醇耐受机制分析[J]. 食品与发酵工业, 2022 , 48(11) : 24 -29 . DOI: 10.13995/j.cnki.11-1802/ts.029692

Abstract

Bacillus aquiflavi 3H-10, is a novel strain isolated from yellow water of strong-flavor Baijiu in Yibin, Sichuan province, China. It showed good protease-producing activity with low safety risks as a potential strain in Baijiu fermentation. In order to explore the stress resistance potential of the strain B. aquiflavi 3H-10, the tolerance to ethanol, acid, alkali, NaCl and temperature and the mechanism of stress resistance was analyzed. The results showed that the strain 3H-10 was tolerant to 4% ethanol, which might resulted from adaptive evolution in environments. Genetic analysis demonstrated that there were multiple heat shock protein-encoding genes in the genome of strain 3H-10, which allowed stress responses to ethanol and weaken the toxic effect of ethanol on cells. Furthermore, the strain 3H-10 harbored genes responsible for a complete ethanol metabolic pathway, which can enable ethanol oxidization to acetaldehyde and acetic acid. Acetic acid enters the citric acid cycle and finally transforms to CO2 and water. Therefore, abundant heat shock proteins and the complete ethanol metabolic pathway were primary reasons for ethanol tolerance in strain 3H-10. This study provides theoretical basis for the practical application of B. aquiflavi 3H-10 in the fermentation of strong-flavor Baijiu.

Key words： Bacillus aquiflavi; stress resistance; ethanol tolerance; heat shock protein; metabolic pathway

参考文献

[1] XU Y Q, SUN B G, FAN G S, et al.The brewing process and microbial diversity of strong flavour Chinese spirits:A review[J].Journal of the Institute of Brewing, 2017, 123(1):5-12.
[2] 晋湘宜, 蔡开云, 陈萍, 等.中国白酒酿酒微生物研究概述[J].酿酒, 2020, 47(5):16-21.
JIN X Y, CAI K Y, CHEN P, et al.Overview of the research on brewing microorganisms of Chinese liquor[J].Liquor Making, 2020, 47(5):16-21.
[3] 胡玉婕, 朱秀玲, 丁延芹, 等.芽孢杆菌的耐盐促生机制研究进展[J].生物技术通报, 2020, 36(9):64-74.
HU Y J, ZHU X L, DING Y Q, et al.Research progress on salt tolerance and growth-promoting mechanism of Bacillus[J].Biotechnology Bulletin, 2020, 36(9):64-74.
[4] 李长福, 吴影, 周子吕, 等.碳源对凝结芽孢杆菌耐酸特性的影响及其机制研究[J].食品与发酵工业, 2019, 45(24):16-21.
LI C F, WU Y, ZHOU Z L, et al.Effect and mechanisms of carbon sources on acid tolerance of Bacillus coagulans[J].Food and Fermentation Industries, 2019, 45(24):16-21.
[5] 郭辉祥, 余东, 龙远兵, 等.发酵期间多粮浓香型白酒窖内黄水成分的变化趋势[J].中国酿造, 2020, 39(1):82-87.
GUO H X, YU D, LONG Y B, et al.Change trend of Huangshui components in multiple-grains strong-flavor Baijiu fermentation pit during the fermentation period[J].China Brewing, 2020, 39(1):82-87.
[6] XIE J Y, CHENG K, ZHAO D, et al.Bacillus aquiflavi sp.nov., isolated from yellow water of strongly flavored Chinese Baijiu[J].International Journal of Systematic and Evolutionary Microbiology, 2020, 70(5):3 406-3 412.
[7] KANEHISA M, GOTO S.KEGG:Kyoto encyclopedia of genes and genomes[J].Nucleic Acids Research, 2000, 28(1):27-30.
[8] 郭宴君, 樊璐璐, 何永吉, 等.芽孢杆菌菌株TYF-B5-5的产乙醇特性及其全基因组测序分析[J].山西农业科学, 2020, 48(1):35-39;44.
GUO Y J, FAN L L, HE Y J, et al.Ethanol production optimization and whole genome sequencing analysis of Bacillus sp.TYF-B5-5[J].Journal of Shanxi Agricultural Sciences, 2020, 48(1):35-39;44.
[9] 饶冉. 极端环境微生物的适应机理及应用[J].安徽农业科学, 2012, 40(27):13 512-13 515.
RAO R.The adaptive mechanism and application of extremophilic microorganisms[J].Journal of Anhui Agricultural Sciences, 2012, 40(27):13 512-13 515.
[10] 陈星宇, 马信, 孙长龙, 等.嗜盐微生物的研究进展[J].盐科学与化工, 2019, 48(2):1-4.
CHEN X Y, MA X, SUN C L, et al.The research progress of halophilic microorganisms[J].Journal of Salt Science and Chemical Industry, 2019, 48(2):1-4.
[11] MIN S, KIM H G, LEE B, et al.Protein transfer learning improves identification of heat shock protein families[J].PLoS One, 2021, 16(5):e0251865.
[12] 匡素芳, 彭仁.耐有机溶剂微生物的耐受机制、改造策略和应用[J].生物技术, 2017, 27(6):607-612.
KUANG S F, PENG R.Tolerance mechanism, improvement strategy and application of organic-solvent-tolerant microorganisms[J].Biotechnology, 2017, 27(6):607-612.
[13] MIAO Y J, XIONG G T, LI R Y, et al.Transcriptome profiling of Issatchenkia orientalis under ethanol stress[J].AMB Express, 2018, 8(1):39.
[14] 李伟丽, 秦琦, 李良, 等.热休克蛋白对枯草芽孢杆菌抗逆性和乙醇产量的影响[J].安徽农业科学, 2008, 36(21):8 963-8 965.
LI W L, QIN Q, LI L, et al.The study of using hsp in genetic engineered Bacillus subtilis to increase its resistance to adversity and its production of ethanol[J].Journal of Anhui Agricultural Sciences, 2008, 36(21):8 963-8 965.
[15] 鞠建松, 马宁, 赵冉冉, 等.假坚强芽胞杆菌中乙醇降解相关酶的克隆、表达及酶学特性[J].微生物学报, 2013, 53(4):363-371.
JU J S, MA N, ZHAO R R, et al.Cloning, expression and characterization of alcohol dehydrogenase and aldehyde dehydrogenase from Bacillus pseudofirmus OF4[J].Acta Microbiologica Sinica, 2013, 53(4):363-371.
[16] 何迎粉, 何荣荣, 刘敦华, 等.海藻糖与酿酒酵母乙醇耐受性相关性的研究进展[J].中国酿造, 2020, 39(11):1-4.
HE Y F, HE R R, LIU D H, et al.Correlation between trehalose and ethanol tolerance in Saccharomyces cerevisiae[J].China Brewing, 2020, 39(11):1-4.
[17] PEREIRA M D, ELEUTHERIO E C, PANEK A D.Acquisition of tolerance against oxidative damage in Saccharomyces cerevisiae[J].BMC Microbiology, 2001, 1:11.
[18] SEKINE T, KAWAGUCHI A, HAMANO Y, et al.Desensitization of feedback inhibition of the Saccharomyces cerevisiae γ-glutamyl kinase enhances proline accumulation and freezing tolerance[J].Applied and Environmental Microbiology, 2007, 73(12):4 011-4 019.
[19] HIRASAWA T, YOSHIKAWA K, NAKAKURA Y, et al.Identification of target genes conferring ethanol stress tolerance to Saccharomyces cerevisiae based on DNA microarray data analysis[J].Journal of Biotechnology, 2007, 131(1):34-44.
[20] OKOCHI M, KURIMOTO M, SHIMIZU K, et al.Increase of organic solvent tolerance by overexpression of manXYZ in Escherichia coli[J].Applied Microbiology and Biotechnology, 2007, 73(6):1 394-1 399.
[21] AONO R.Improvement of organic solvent tolerance level of Escherichia coli by overexpression of stress-responsive genes[J].Extremophiles:Life Under Extreme Conditions, 1998, 2(3):239-248.
[22] RAMOS J L, DUQUE E, GALLEGOS M T, et al.Mechanisms of solvent tolerance in Gram-negative bacteria[J].Annual Review of Microbiology, 2002, 56(1):743-768.
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