Food and Fermentation Industries >

2025 , Vol. 51 >Issue 21: 114 - 122

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.042973

Screening of high-yielding tetramethylpyrazine Bacillus and its application in Jiangxiangxing Baijiu fermentation

  • BIN Zhiqiang ,
  • LI Shuangyu ,
  • ZHANG Tianqi ,
  • LI Chenyao ,
  • LIU Yafeng ,
  • HUANG Zhiguo ,
  • SUN Jinxu ,
  • GUO Xuewu
Expand
  • 1(Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China)
    2(Key Laboratory of Brewing Biotechnology and Application of Sichuan Province, Sichuan University of Science and Engineering, Yibin 644000, China)
    3(College of Life Science, Hengshui University, Hengshui 053000, China)

Received date: 2025-04-09

  Revised date: 2025-06-12

  Online published: 2025-11-21

Fold

Abstract

To screen Bacillus with high tetramethylpyrazine (TTMP) production and explore their application in Jiangxiangxing Baijiu fermentation, 364 Bacillus were isolated from Daqu and fermented grains.Six high- TTMP -producing candidates were selected through halo formation tests, liquid-state enzyme activity assays, and bran-based solid-state fermentation evaluations.Strain DQ-B102 produced 61.42 mg/L TTMP in solid-state fermentation, significantly outperforming other strains.Identification results classified DQ-B102 as Bacillus velezensis.Simulated fermentation tests demonstrated that DQ-B102 inoculation significantly enhances TTMP yield.At 15% inoculum dosage, TTMP reached 144.18 mg/L, representing a 1.54-fold increase compared to the control group.High-throughput sequencing revealed that DQ-B102 optimized bacterial community structure by suppressing Clostridium and enriching Bacillus.Increasing the inoculum to 15% inhibited Saccharomyces but elevated Aspergillus abundance to 19.50%.GC-MS analysis detected 34 volatile flavor compounds, including 8 alcohols, 9 esters, 7 aldehydes, 4 phenols, and 6 other compounds.The 5% inoculum group exhibited the highest flavor diversity with 34 compounds.Increased inoculum concentration enhanced pyro-sauce aroma contribution.This study successfully isolated Bacillus velezensis DQ-B102, a high-yield TTMP producer.The strain significantly improved TTMP content and pyro-sauce aroma intensity in liquor through efficient bioconversion of acetoin to TTMP and strategic regulation of microbial community structure.These findings provide valuable microbial resources for quality enhancement of Jiangxiangxing Baijiu.The results establish a scientific foundation for optimizing fermentation processes and developing functional microbial agents in Baijiu production.

Key words： Jiangxiangxing Baijiu; tetramethylpyrazine; Bacillus; microbial community structure analysis; flavor substance

Cite this article

BIN Zhiqiang , LI Shuangyu , ZHANG Tianqi , LI Chenyao , LIU Yafeng , HUANG Zhiguo , SUN Jinxu , GUO Xuewu . Screening of high-yielding tetramethylpyrazine Bacillus and its application in Jiangxiangxing Baijiu fermentation[J]. Food and Fermentation Industries, 2025 , 51(21) : 114 -122 . DOI: 10.13995/j.cnki.11-1802/ts.042973

References

[1] LI J, LU J, MA Z L, et al.A green route for high-yield production of tetramethylpyrazine from non-food raw materials[J].Frontiers in Bioengineering and Biotechnology, 2022, 9:792023.
[2] LIN J G, WANG Q Q, ZHOU S M, et al.Tetramethylpyrazine:A review on its mechanisms and functions[J].Biomedicine & Pharmacotherapy, 2022, 150:113005.
[3] MÜLLER R, RAPPERT S.Pyrazines:Occurrence, formation and biodegradation[J].Applied Microbiology and Biotechnology, 2010, 85(5):1315-1320.
[4] WANG G F, SHI C G, SUN M Z, et al.Tetramethylpyrazine attenuates atherosclerosis development and protects endothelial cells from ox-LDL[J].Cardiovascular Drugs and Therapy, 2013, 27(3):199-210.
[5] ZHANG M S, GAO F, TENG F M, et al.Tetramethylpyrazine promotes the proliferation and migration of brain endothelial cells[J].Molecular Medicine Reports, 2014, 10(1):29-32.
[6] BIAN Y Y, YANG L L, SHENG W, et al.Ligustrazine induces the colorectal cancer cells apoptosis via p53-dependent mitochondrial pathway and cell cycle arrest at the G0/G1 phase[J].Annals of Palliative Medicine, 2021, 10(2):1578-1588.
[7] XIONG X M, HU Y L, YAN N F, et al.PCR-DGGE analysis of the microbial communities in three different Chinese “Baiyunbian” liquor fermentation starters[J].Journal of Microbiology and Biotechnology, 2014, 24(8):1088-1095.
[8] 王晓丹, 王婧, 朱国军, 等.酱香大曲中产四甲基吡嗪细菌的分离鉴定及其功能性研究[J].中国酿造, 2017, 36(1):55-60.
WANG X D, WANG J, ZHU G J, et al.Isolation and identification of functional bacteria with high yield of tetramethyl pyrazine from Moutai-flavor high-temperature Daqu[J].China Brewing, 2017, 36(1):55-60.
[9] 史洪丽, 余梦婷, 王世昕, 等.高温大曲中高产四甲基吡嗪芽孢杆菌的筛选及其在酒醅堆积中的应用[J].中国酿造, 2025, 44(3):59-65.
SHI H L, YU M T, WANG S X, et al.Screening of high yield tetramethylpyrazine Bacillus from high temperature Daqu and its application in the stacking of fermented grains[J].China Brewing, 2025, 44(3):59-65.
[10] 葛向阳, 李晓欢, 周庆伍, 等.浓香型白酒中高含四甲基吡嗪酿造工艺的研究[J].酿酒, 2018, 45(3):26-29.
GE X Y, LI X H, ZHOU Q W, et al.Study on brewing process of high-containing tetramethylpyrazine in Luzhou-flavor liquor[J].Liquor Making, 2018, 45(3):26-29.
[11] 吴建峰, 徐岩.白酒细菌酒曲固态培养条件下B.subtilis S12产四甲基吡嗪的合成机制[J].食品与生物技术学报, 2014, 33(1):8-15.
WU J F, XU Y.Formation mechanism of tetramethylpyrazine produced with B.subtilis S12 under the fermentation condition simulated bacterial Qu preparation used for Chinese liquor brewing[J].Journal of Food Science and Biotechnology, 2014, 33(1):8-15.
[12] 陈媛媛, 陈德茗, 李子健, 等.产乙偶姻菌株的分离鉴定及其对固态混菌发酵产物的影响[J].中国酿造, 2024, 43(3):84-89.
CHEN Y Y, CHEN D M, LI Z J, et al.Isolation and identification of acetoin-producing strain and its effect on solid-state multi-microbial fermentation products[J].China Brewing, 2024, 43(3):84-89.
[13] 阮玉磊. 小曲清香型白酒合成微生物群落的构建[D].天津:天津科技大学, 2023.
RUAN Y L.Construction of synthetic microbial community of xiaoqu qingxiang baijiu[D].Tianjin:Tianjin University of Science and Technology, 2023.
[14] 李晨瑶. 贝莱斯芽孢杆菌在酱香型白酒生产中的作用[D].天津:天津科技大学, 2023.
LI C Y.The role of Bacillus velezensis in the production of jiangxiangxing baijiu[D].Tianjin:Tianjin University of Science and Technology, 2023.
[15] 沈怡方. 白酒生产技术全书[M].北京:中国轻工业出版社, 2015.
SHEN Y F.Comprehensive Textbook on Baijiu Production Technology[M].Beijing:China Light Industry Press, 2015.
[16] 邢爽, 王亚平, 郭学武, 等.发酵条件对5种产酯酵母酒精发酵和产酯的影响[J].中国酿造, 2018, 37(2):24-28.
XING S, WANG Y P, GUO X W, et al.Effects of fermentation conditions on ethanol fermentation and ester production of five ester-producing yeasts[J].China Brewing, 2018, 37(2):24-28.
[17] 陈梦圆, 李志军, 罗爱民.高温大曲中高产四甲基吡嗪菌株的筛选及鉴定[J].酿酒科技, 2018(8):24-29.
CHEN M Y, LI Z J, LUO A M.Screening and identification of strains with high yield of tetramethylpyrazine from high-temperature daqu[J].Liquor-Making Science & Technology, 2018(8):24-29.
[18] WANG C L, LI C Y, BIN Z Q, et al.Workshop environment heterogeneity shaped the microbiome and metabolome profiles during Xiasha round of Jiangxiangxing Baijiu[J].Food Chemistry:X, 2024, 22:101264.
[19] 王婧. 酱香大曲中产吡嗪类物质芽孢杆菌的筛选及其应用研究[D].贵阳:贵州大学, 2016.
WANG J.Screening and application of pyrazine-producing Bacillus strains in Jiangxiang Daqu[D].Guiyang:Guizhou University, 2016.
[20] 姚粟, 葛媛媛, 李辉, 等.利用非培养技术研究芝麻香型白酒高温大曲的细菌群落多样性[J].食品与发酵工业, 2012, 38(6):1-6.
YAO S, GE Y Y, LI H, et al.Analysis on bacterial communities in high temperature Daqu of sesame flavor liquor through culture-free approach[J].Food and Fermentation Industries, 2012, 38(6):1-6.
[21] 王光路, 张帆, 杨旭, 等.贾湖原香型白酒高温堆积过程原核微生物的消长规律研究[J].中国酿造, 2020, 39(11):30-35.
WANG G L, ZHANG F, YANG X, et al.Growth and decline of prokaryotic microorganisms during high temperature stacking fermentation of the Jiahu original flavor Baijiu[J].China Brewing, 2020, 39(11):30-35.
[22] 梁晨, 杜海, 徐岩.大曲贮存过程中原核微生物群落结构及风味成分演替规律[J].微生物学通报, 2017, 44(2):384-393.
LIANG C, DU H, XU Y.The succession of procaryotic microbial community and the flavor components in the storage process of Daqu[J].Microbiology China, 2017, 44(2):384-393.
[23] 蒲领平, 邓杰, 卫春会, 等.贝莱斯芽孢杆菌对小曲酒酿造过程中酵母菌生长及风味物质代谢的影响[J].中国酿造, 2023, 42(2):53-57.
PU L P, DENG J, WEI C H, et al.Effects of Bacillus velesiensis on the growth and flavor metabolism of yeast during Xiaoqu Baijiu brewing[J].China Brewing, 2023, 42(2):53-57.
[24] 赵龙, 周池虹伶, 赵谋明, 等.解淀粉芽孢杆菌SWJS22和米曲霉混合制曲在酱油发酵中的应用[J].食品科学, 2017, 38(22):125-130.
ZHAO L, ZHOU C H L, ZHAO M M, et al.Application of mixed cultures of Bacillus amyloliquefaciens SWJS22 and Aspergillus oryzae in soy sauce fermentation[J].Food Science, 2017, 38(22):125-130.
[25] ZHAO M, SU X Q, NIAN B, et al.Integrated meta-omics approaches to understand the microbiome of spontaneous fermentation of traditional Chinese Pu-erh tea[J].mSystems, 2019, 4(6):e00680-19.
[26] 马宁. 酱香型习酒特征香气成分鉴定及香气协同作用研究[D].上海:上海应用技术大学, 2021.
MA N.Study on key aroma compounds and perceptual interaction among odorants in soy sauce aroma type Xi Baijiu[D].Shanghai:Shanghai Institute of Technology,2021.
[27] 刘发洋, 李璐, 游奇, 等.基于OAV分析多粮浓香型调味酒陈酿过程中风味物质的变化[J].中国酿造, 2023, 42(5):237-242.
LIU F Y, LI L, YOU Q, et al.Changes of flavor substances of multi-grain strong-flavor flavouring liquor during the aging process based on OAV[J].China Brewing, 2023, 42(5):237-242.
[28] 聂士昊, 张志伟, 汪俊卿, 等.芽孢杆菌加强菌对酱香型白酒酒醅微生物群落及蛋白酶活力的影响[J].中国酿造, 2022, 41(8):137-142.
NIE S H, ZHANG Z W, WANG J Q, et al.Effect of addition of Bacillus on microbial communities and protease activity of fermented grains of sauce-flavor Baijiu[J].China Brewing, 2022, 41(8):137-142.
Options
Outlines

/

〈 〉

Powered by Beijing Magtech Co. Ltd,.