Difference of microbial community stressed in artificial pit muds for <i>Luzhou</i>
-flavour liquor brewing revealed by multiphase culture-independent technology

Zhang, Liqiang; Zhou, Rongqing; Niu, Meican; Zheng, Jia; Wu, Chongde

doi:10.1111/jam.12943

Cited by 33 publications

(30 citation statements)

References 41 publications

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“…The new pit was a cellar in which a batch of fermentations were performed just after construction with artificial pit mud. The eubacterial genera detected in new PM were similar to the results reported in our previous study [22]. The eubacteria detected in new PM also appeared in the corresponding ZP samples, with the exception of Bacillus, which was detected in daqu [28].…”

Section: Discussionsupporting

confidence: 89%

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Characterization of Interphase Microbial Community in Luzhou-Flavored Liquor Manufacturing Pits of Various Ages by Polyphasic Detection Methods

Huang

Liu³

et al. 2017

Journal of Microbiology and Biotechnology

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It is vital to understand the changing characteristics of interphase microbial communities and interspecies synergism during the fermentation of Chinese liquors. In this study, microbial communities in the three indispensable phases (pit mud, zaopei, and huangshui) of Luzhou-flavored liquor manufacturing pits and their shifts during cellars use were first investigated by polyphasic culture-independent approaches. The archaeal and eubacterial communities in the three phases were quantitatively assessed by combined phospholipid ether lipids/phospholipid fatty acid analysis and fluorescence in situ hybridization. In addition, qualitative information regarding the microbial community was analyzed by PCR-denaturing gradient gel electrophoresis. Results suggested that the interphase microbial community profiles were quite different, and the proportions of specific microbial groups evolved gradually. Anaerobic bacteria and gram-positive bacteria were dominant and their numbers were higher in pit mud (10 cells/g) than in huangshui (10 cells/ml) and zaopei (10cells/g). Hydrogenotrophic methanogenic archaea were the dominant archaea, and their proportions were virtually unchanged in pit mud (around 65%), whereas they first increased and then decreased in zaopei (59%-82%-47%) and increased with pit age in huangshui (82%-92%). Interactions between microbial communities, especially between eubacteria and methanogens, played a key role in the formation of favorable niches for liquor fermentation. Furthermore, daqu (an essential saccharifying and fermentative agent) and metabolic regulation parameters greatly affected the microbial community.

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Section: Discussionsupporting

confidence: 89%

“…Whole genomic DNA was extracted according to methods by Zhou et al [21]. A nested PCR was employed, and all PCR primers used in this study were derived from a previous study [22]. Before DGGE analysis, PCR products were examined by electrophoresis on 1% agarose gels.…”

Section: Characterization Of the Microbial Community Based On Pcr-dggmentioning

confidence: 99%

Characterization of Interphase Microbial Community in Luzhou-Flavored Liquor Manufacturing Pits of Various Ages by Polyphasic Detection Methods

Huang

Liu³

et al. 2017

Journal of Microbiology and Biotechnology

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“…However, the genera that increased included also facultatively anaerobic bacteria, for example Weissella (27). To our knowledge, Fonticella and Hazenella were detected for the first time in SFB production, suggesting that the APM microbial communities and their succession are more complex than previously known (12,16,17).…”

Section: Discussionmentioning

confidence: 62%

“…This APM can be used to line the fermentation cellars, and high-quality SFB can be obtained from the first three production batches in only 1 year (14,15), showing that a beneficial microbial community structure for SFB production can be successfully created in a short brewing period. Microbial communities in APM have been studied using a range of cultivation and molecular methods (12,16,17). However, our knowledge of the molecular mechanisms of APM microbial dynamics and associated functional variation and of the role of this variation in high-quality SFB production remains limited.…”

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confidence: 99%

Structural and Functional Changes in Prokaryotic Communities in Artificial Pit Mud during Chinese Baijiu Production

et al. 2020

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The Chinese alcoholic beverage strong-flavor baijiu (SFB) gets its characteristic flavor during fermentation in cellars lined with pit mud. Microbes in the pit mud produce key precursors of flavor esters. The maturation time of natural pit mud of over 20 years has promoted attempts to produce artificial pit mud (APM) with a shorter maturation time. However, knowledge about the molecular basis of APM microbial dynamics and associated functional variation during SFB brewing is limited, and the role of this variability in high-quality SFB production remains poorly understood. We studied APM maturation in new cellars until the fourth brewing batch using 16S rRNA gene amplicon sequencing, quantitative PCR, metaproteomics, and metabolomics techniques. A total of 36 prokaryotic classes and 195 genera were detected. Bacilli and Clostridia dominated consistently, and the relative abundance of Bacilli decreased along with the APM maturation. Even though both amplicon sequencing and quantitative PCR showed increased abundance of Clostridia, the levels of most of the Clostridium proteins were similar in both the first- and fourth-batch APM samples. Six genera correlated with eight or more major flavor compounds in SFB samples. Functional prediction suggested that the prokaryotic communities in the fourth-batch APM samples were actively engaged in organic acid metabolism, and the detected higher concentrations of proteins and metabolites in the corresponding metabolic pathways supported the prediction. This multi-omics approach captured changes in the abundances of specific microbial species, proteins, and metabolites during APM maturation, which are of great significance for the optimization of APM culture technique. IMPORTANCE Strong-flavor baijiu (SFB) accounts for more than 70% of all Chinese liquor production. In the Chinese baijiu brewing industry, artificial pit mud (APM) has been widely used since the 1960s to construct fermentation cellars for production of high-quality SFB. To gain insights at the systems level into the mechanisms driving APM prokaryotic taxonomic and functional dynamics and into how this variation is connected with high-quality SFB production, we performed the first combined metagenomic, metaproteomic, and metabolomic analyses of this brewing microecosystem. Together, the multi-omics approach enabled us to develop a more complete picture of the changing metabolic processes occurring in APM microbial communities during high-quality SFB production, which will be helpful for further optimization of APM culture technique and improvement of SFB quality.

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“…It has been used to characterize the archaeal community in complex environments [18,19]. In our previous researches, the microbial communities in PM were characterized by FISH using various order-specific oligonucleotide probes [20][21][22][23][24]. The composition of four orders methanogenicarchaeal (methanosarcinales, methanobacteriales, methanomicrobiales and methanococcales) in various aged PM were also estimated quantitatively [25].…”

Section: Introductionmentioning

confidence: 99%

Design, Optimization and Verification of 16S rRNA Oligonucleotide Probes of Fluorescence in-situ Hybridization for Targeting Clostridium spp. and Clostridium kluyveri

Hu¹,

Huang

et al. 2018

Journal of Microbiology and Biotechnology

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Difference of microbial community stressed in artificial pit muds for Luzhou -flavour liquor brewing revealed by multiphase culture-independent technology

Abstract: APM cultivation technology was necessary to promote enriching functional liquor-brewing microbes into APMs. These results may facilitate understanding the microbial succession during APMs manufacture.

Cited by 33 publications

References 41 publications

Characterization of Interphase Microbial Community in Luzhou-Flavored Liquor Manufacturing Pits of Various Ages by Polyphasic Detection Methods

Characterization of Interphase Microbial Community in Luzhou-Flavored Liquor Manufacturing Pits of Various Ages by Polyphasic Detection Methods

Structural and Functional Changes in Prokaryotic Communities in Artificial Pit Mud during Chinese Baijiu Production

Design, Optimization and Verification of 16S rRNA Oligonucleotide Probes of Fluorescence in-situ Hybridization for Targeting Clostridium spp. and Clostridium kluyveri

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