酒醅中精氨酸利用菌株的分离筛选及其对浓香型白酒中瓜氨酸积累的影响

Characterization of arginine utilization strains from fermented grains and evaluation of their contribution to citrulline accumulation in Chinese Luzhou-flavor spirits

【目的】分析浓香型白酒酒醅发酵过程中氨基甲酸乙酯前体物质瓜氨酸含量显著增加的原因,确定酒醅中能够利用精氨酸并积累瓜氨酸的微生物,为解析白酒中氨基甲酸乙酯的形成机制提供研究基础和理论依据。【方法】采用高通量筛选技术,从浓香型白酒酒醅中分离具有高精氨酸利用能力和高瓜氨酸积累特性的菌株,并通过基因型和表现型验证以及模拟窖内发酵验证它们对瓜氨酸积累的贡献。【结果】共筛选获得20株具有高精氨酸利用能力的菌株,其中Lactococcus garvieae LD3,Bacillus amyloliquefaciens BG5,Pediococcus acidilactici PH7和Staphylococcus pasteuri SH11具有较高的瓜氨酸生成能力,并可使酒醅中瓜氨酸含量显著增加。【结论】筛选获得的4类微生物均能够通过ADI途径代谢积累瓜氨酸,是导致酒醅瓜氨酸含量增加的原因。

[Objective] This study aimed to characterize microorganisms responsible for accumulation of ethyl carbamate precursor citrulline in fermented grains during Luzhou-flavor spirits fermentation process, to provide theoretical basis for clarifying mechanisms of ethyl carbamate formation. [Methods] High-throughput strain screening approach was used to isolate strains with high arginine utilization and high citrulline accumulation capability. Essential genes that comprise the arginine deiminase pathway of these isolates were verified and a modified Chinese liquor fermentation process was conducted by adding these strains. [Results] A total of 20 strains with high arginine utilizing ability were obtained. Among them, Lactococcus garvieae LD3, Bacillus amyloliquefaciens BGS, Pediococcus acidilactici PH7 and Staphylococcus pasteuri SH11 exhibited high efficacy to produce citrulline from arginine. These strains could also increase citrulline in fermented grains. [Conclusion] The accumulation of citrulline in fermented grains was confirmed to be corresponding to arginine utilization by Lactococcus garvieae, Bacillus amyloliquefaciens, Pediococcus acidilactici and Staphylococcus pasteuri through the arginine deiminase pathway.