为了提高短双歧杆菌的冻干存活率和工业化生产效率,首先系统测定短双歧杆菌在不同分子质量糖(醇)类及蛋白类保护剂中的冻干存活率,其次研究不同分子质量糖类复配及其与蛋白类复配的保护效果,再次测定复合保护剂添加其他小分子物质对冻干保护存活率的影响,最后优化冻干前菌泥与保护剂的比例与浓度。结果表明,三糖及以下的小分子糖对短双歧杆菌的冻干保护效果较好,棉子糖和山梨糖醇保护效果最好,2者复配进一步提高冻干存活率,达55%左右。复配蛋白质及抗坏血酸、谷胱甘肽、甜菜碱、微量元素等小分子不会提高对菌体的冻干保护效果,但蛋白质的适量添加可提高菌粉的疏松度。菌泥与保护剂以干物质质量比1∶1.2混合,样品干物质总质量分数为25%时,短双歧杆菌的冻干存活率接近80%,活菌数达(1.61±0.27)×1012 CFU/g。
In order to improve the freeze-drying survival rate and industrial production efficiency of Bifidobacterium breve cell mass, the lyophilized survival rate of B. breve in different molecular weight of saccharide(alcohol) and proteins was systematically determined and the ratio and concentration of the wet bacterial mass and protectant were optimized. The results showed that raffinose and sorbitol had the best protective effect and their combination further increased the survival rate of lyophilization to about 55%. Addition of protein or small molecules such as ascorbic acid, glutathione, betaine and trace elements did not improve the lyophilization protection effect. Nevertheless, the optimal amount of protein improved the porosity of bacterial powder. When the wet bacterial mass mixed with the protective agents were at ratio of 1∶1.2 and the total dry matter fraction was 25%, the survival rate of B. breve was reached to 80% and the viable cells were up to (1.61±0.27)×1012 CFU/g.
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