摘要
【背景】高温堆积发酵过程是芝麻香型白酒生产过程中的一个重要环节,是芝麻香型白酒典型风格形成的关键工序。【目的】解析堆积发酵过程中原核和真核微生物群落的演替规律。【方法】分别对高温堆积酒醅样本的细菌16S rDNA V3-V4区和真菌ITS2区进行Illumina高通量测序,解析高温堆积过程中酒醅的真菌和细菌菌群结构变化规律,同时解析发酵过程中温度、水分、酸度、淀粉、还原糖的变化规律。【结果】酒醅堆积初期0-6 h,占优势的原核微生物菌属是Bacillus、Weissella和Acetobacter。堆积20 h后微生物群落结构变化显著,Weissella的比例降低,Acetobacter的比例显著增加。堆积初期,占优势的真核微生物菌属是Aspergillus、Saccharomyces和Pichia。堆积6 h后,Aspergillus比例下降,而Pichia和Saccharomyces则逐步增多,其绝对优势地位一直保持至堆积结束。酒醅微生物群落结构受翻堆操作的短暂影响(堆积20 h时),最终恢复至平衡状态。高温堆积36 h以后,酒醅中的原核和真核微生物群落均趋于稳定。还原糖数据显示堆积前期微生物糖化作用迅速,36 h后糖化趋于稳定,温度、酸度、淀粉含量参数基本平稳。【结论】酒醅高温堆积发酵过程中,原核微生物和真核微生物有其不同的变化规律。原核微生物的变化是以20 h时的翻堆为结点,翻堆前的优势菌属是Bacillus、Weissella和Acetobacter;堆积后的优势菌属是Weissella和Acetobacter,其中后者占绝对优势。与原核微生物相比,真菌微生物的结构逐步集中,到发酵结束时毕赤酵母和酵母两者占真菌总量的93%以上。芝麻香型白酒发酵过程的高温堆积环节,原核和真核微生物的菌种繁殖以及相应的菌群结构变化为稍后的高温发酵奠定了微生物基础。
[Background] High-temperature stacking is a key step among the whole sesame-flavor Baijiu fermentation process, and vital for the formation of typical flavor of sesame-flavor Baijiu. [Objective]This study aimed to analyze prokaryotic and eukaryotic microbial communities during the high-temperature stacking process. [Methods] Illumina high-throughput sequencing was performed on the bacterial 16 S rDNA V3–V4 region and the fungal ITS2 region of the high-temperature stacked Jiupei samples. The change pattern of prokaryotic and eukaryotic microbial community structure were analyzed.The changes in temperature, moisture, acidity, starch and reduced-sugar during stacking were also analyzed. [Results] At the beginning of 0–6 hours, the dominant bacteria were Bacillus, Weissella, and Acetobacter. After 20 h of stacking, the flora changed significantly. Weissella decreased and the Acetobacter increased. At the beginning, the dominant fungi were Aspergillus, Saccharomyces, and Pichia. After 6 hours, the proportion of Aspergillus dropped sharply, while Pichia and Saccharomyces gradually increased, and their absolute dominance maintained until the end of the stacking. The overall structure of the microbial community was temporarily affected by the turn-over operation(when it was piled up for 20 h), eventually returning to equilibrium. After 36 h of stacking, both prokaryotic and eukaryotic microbial community stabilized. Reduced-sugar data showed that the saccharification was rapid in the early stage, later became stable after 36 h. Temperature, acidity and starch content were also stable at this stage. [Conclusion] During the high-temperature stacking-fermentation process,fluctuation patterns of the prokaryotic and eukaryotic microbial community structure were different. The prokaryotic community changes were based on the turn-up at 20 h. The dominant species before the turn-up were Bacillus, Weissella and Acetobacte, the dominant strains after the turn-up were Weissella and Acetobacter, the latter was absolute dominant. Compared to prokaryotic community changes, the eukaryotic microbial community concentration gradually, by the end of the stacking, Pichia and Saccharomyces accounted for more than 93% of the total eukaryotic microbial. During the high-temperature stacking of the sesame-flavor Baijiu, the propagation of various prokaryotic/eukaryotic microbes and the corresponding changes of microbial communities laid the microbial basis for later high-temperature fermentation.
引文
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