酒酒球菌液氮超低温保存  

Storage of Oenococcus oeni in liquid nitrogen

作　　者：杜立业[1,2] 王华[1,2] 金刚[1,2] 李翠霞[3] 李华[1,2] 

机构地区：[1]西北农林科技大学葡萄酒学院,杨凌712100 [2]陕西省葡萄与葡萄酒工程技术研究中心,杨凌712100 [3]西北农林科技大学生命科学学院,杨凌712100

出　　处：《微生物学报》2011年第9期1263-1269,共7页Acta Microbiologica Sinica

基　　金：农业部"948"项目(2009-Z29)~~

摘　　要：【目地】为安全、长期的保藏酒酒球菌,本文研究了菌体生长时间、冷冻方法、解冻温度、菌密度以及保护剂等对酒酒球菌细胞冷冻存活率的影响,找到最优液氮超低温保存方法。【方法】采用平板计数法测定冷冻存活率。【结果】实验结果表明酒酒球菌的最佳保存方法为:首先在稳定期前期离心收集菌体;其次加入保护剂(20 g/L酵母浸提物,40V/V甘油,20 g/L蔗糖,30 g/L谷氨酸钠)稀释菌体,使菌密度为109CFU/mL;然后直接投入液氮冷冻;最后在37℃温水浴中迅速解冻。保存6个月后,其中21株酒酒球菌的冷冻存活率达到99%以上。【结论】初步研究表明酵母浸提物,甘油,蔗糖,谷氨酸钠复合保护剂对酒酒球菌的保护效果较好,液氮超低温保存可用于酒酒球菌的长期保存。[Objective]To develop cryopreservation for Oenococcus oeni.[Methods] We investigated the influence of the age and concentration of the strain in the protective medium,rate of freezing,temperature of thawing and the protective medium on the number of viable cells after cryopreservation by colony counting.[Results] We got the highest number of viable cells after cryopreservation by collecting cells in the early stationary growth phase,keeping their concentration in the protective medium at 109 CFU/mL,transferring cells into liquid nitrogen directly and thawing in a water bath at 37℃.The ratio of viable cells was above 99% for 21 of the Oenococcus oeni after storing in liquid nitrogen for 6 months.[Conclusion] We The storage of Oenococcus oeni in liquid nitrogen is convenient for strains preserved in culture collections.

关 键 词：酒酒球菌 液氮超低温保存 冷冻存活率 

分 类 号：TS261.13[轻工技术与工程—发酵工程]