高耐酸酒酒球菌常压室温等离子体诱变选育及其苹果酸-乳酸发酵研究  

Atmospheric and room temperature plasma mutagenesis and malolactic fermentation of highly acid-tolerant Oenococcus oeni

作　　者：张莉方 徐宁莉 陶瑾 胡蕾 张国强[1] ZHANG Lifang;XU Ningli;TAO Jin;HU Lei;ZHANG Guoqiang(College of Biological and Food Engineering,Anhui Polytechnic University,Wuhu 241000,Anhui,China)

机构地区：[1]安徽工程大学生物与食品工程学院,安徽芜湖241000

出　　处：《微生物学报》2023年第10期4000-4015,共16页Acta Microbiologica Sinica

基　　金：安徽省高校重大项目(2022AH04136);芜湖市科技计划(2022cg19);国家级大学生创新创业训练计划(202110363058);校级科研项目(Xjky2022091)。

摘　　要：【目的】为选育出高度耐酸性酒酒球菌(Oenococcus oeni)突变菌株,研究其胁迫耐受性能及苹果酸-乳酸发酵(malolactic fermentation,MLF)能力。【方法】以酒酒球菌SD-2a为出发菌株,通过常压室温等离子体(atmospheric and room temperature plasma,ARTP)诱变技术,筛选高耐酸性酒酒球菌突变菌株,并探究其乙醇耐受性及在模拟酒和葡萄酒条件下的MLF能力。【结果】经过ARTP诱变处理后,利用pH 3.0的胁迫传代培养和分离纯化等,获得了5株β-葡萄糖苷酶活性较好的耐酸突变菌株,且在高乙醇浓度下表现出了较好的耐乙醇性。其中突变菌株ARTP-2在模拟酒中的β-葡萄糖苷酶活性和L-苹果酸累积降解量最高,且其在葡萄酒中L-苹果酸降解速率快于出发菌株,在第18天完成MLF,发酵后的葡萄酒香气成分的含量显著高于接种SD-2a的酒样。【结论】突变菌株ARTP-2具有良好的胁迫耐受性和MLF能力,对葡萄酒的香气起到积极的作用,为进一步开发优质的MLF商业发酵剂奠定基础。[Objective]To breed highly acid-tolerant strains of Oenococcus oeni and study their stress tolerance and malolactic fermentation(MLF)performance.[Methods]The mutagenesis of O.oeni SD-2a was carried out with the atmospheric and room temperature plasma(ARTP)method.The ethanol tolerance of the mutants and the MLF performance of the mutants in simulated wine and wine were investigated.[Results]After ARTP mutagenesis,five acid-tolerant mutants with highβ-glucosidase activity were isolated by subculture under stress(pH 3.0).The mutants showed strong ethanol tolerance at high ethanol concentrations.Among them,the mutant ARTP-2 had the highestβ-glucosidase activity and L-malic acid cumulative degradation in the simulated wine.Moreover,it demonstrated faster L-malic acid degradation rate in wine than O.oeni SD-2a and completed MLF on day 18.The total concentration of aroma components in the wine fermented by ARTP-2 was significantly higher than that by O.oeni SD-2a.[Conclusion]The mutant ARTP-2 had strong stress tolerance and MLF performance and played a positive role in wine aroma.The findings laid a foundation for the further development of high-quality commercial fermenters for MLF.

关 键 词：常压室温等离子体诱变 酒酒球菌 耐酸 L-苹果酸 β-葡萄糖苷酶 香气成分 

分 类 号：TS261.1[轻工技术与工程—发酵工程]