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酿酒酵母与地衣芽孢杆菌相互作用及基于蛋白组学的作用机制分析
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国家863计划项目(No. 2013AA102108,2012AA021301);国家自然科学基金项目(No. 31371822,31271921);中国博士后科学基金项目(No. 2014M550265);江苏省优势学科建设111项目(No. 111-2-06);江苏省现代工业发酵协同创新中心行业发展项目


Interactions between Saccharomyces cerevisiae and Bacillus licheniformis and interaction mechanisms based on proteomic analysis
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    摘要:

    【目的】为解析酱香型白酒发酵过程中群体微生物的酿造特征,研究酱香型白酒酿造中贡献特征风味的地衣芽孢杆菌和贡献酒精的酿酒酵母之间的相互作用。【方法】通过构建酿酒酵母纯培养及与地衣芽孢杆菌共培养发酵体系,比较不同培养体系中的生物量、乙醇产量及有机酸产量差异,并从蛋白组学角度加以分析和认识二者之间的相互作用。【结果】在共培养体系中,酿酒酵母抑制地衣芽孢杆菌生长,其自身生长不受地衣芽孢杆菌的影响,然而代谢产物却发生变化,其中乙醇及有机酸中的丙酮酸、苹果酸、乳酸、琥珀酸及酒石酸的最高产量分别高出其纯培养的11.8%、56.8%、36.3%、24.3%、48.2%及27.7%,而柠檬酸的最高产量低于其纯培养的35.1%;蛋白组分析显示,地衣芽孢杆菌诱导酿酒酵母胞内69个蛋白差异表达(>2倍),质谱鉴定出24个,主要功能为参与糖酵解过程、乙醇代谢过程、细胞壁稳定性调控及应激反应等。糖酵解和乙醇代谢途径相关蛋白对酿酒酵母混合培养条件下的代谢变化起重要作用,其余蛋白可能与微生物相互作用时的防御和适应性相关。【结论】在混合培养发酵体系中地衣芽孢杆菌能够影响酿酒酵母的乙醇及有机酸代谢,这对于白酒品质调控及微生物间相互作用都具有重要意义。蛋白组学结果为从分子层面深入认识酿酒酵母与地衣芽孢杆菌之间的相互作用提供理论基础,有利于促进酱香型白酒发酵过程中群体微生物酿造特征的解析。

    Abstract:

    [Objective] To better understand the behavior of microbial community during Chinese Maotai-flavor liquor fermentation, we studied the interaction between two functional strains, Saccharomyces cerevisiae and Bacillus licheniformis. [Methods] S. cerevisiae and B. licheniformis were tested in single and mixed culture. Biomass, ethanol and organic acid production were determined during the fermentation. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) coupled with matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was used to analyze and identify differentially expressed protein of S. cerevisiae in single and mixed culture. [Results] S. cerevisiae showed a negative effect on B. licheniformis growth and the growth of itself was barely influenced. However, ethanol, pyruvic acid, lactic acid, malic acid, tartaric acid and succinic acid produced by S. cerevisiae in mixed culture have increased by 11.8%, 56.8%, 24.3%, 36.3%, 27.7% and 48.2% compared with its single culture, respectively. Citric acid yield was reduced by 35.1%. Proteomic profile of S. cerevisiae indicated that 69 spots (differential ratio>2) were differentially expressed in mixed culture. Among them, 24 protein spots were identified, including protein related to glycolysis, ethanol fermentation, cell wall integrity, and stress response. [Conclusion] B. licheniformis affected ethanol and organic acid metabolism of S. cerevisiae. That was significant for liquor quality and microbial interactions. Proteomic analysis of the responses of S. cerevisiae to mixed culture allows a deeper understanding of interaction mechanisms between both strains and will promote the understanding of the microbial community during Chinese Maotai-flavor liquor fermentation.

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孟醒,吴群,徐岩. 酿酒酵母与地衣芽孢杆菌相互作用及基于蛋白组学的作用机制分析[J]. 微生物学通报, 2015, 42(9): 1679-1688

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  • 在线发布日期: 2015-09-02
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