干酪乳杆菌LC2W原生质体的制备与再生

Protoplast Preparation and Regeneration of Lactobacillus casei LC2W

以干酪乳杆菌LC2W为出发菌株,对其进行原生质体制备与再生。通过单因素试验研究菌龄、溶菌酶质量浓度、酶解时间及酶解温度对干酪乳杆菌LC2W原生质体制备的影响,并对制备条件进行正交设计优化。结果表明:干酪乳杆菌LC2W原生质体制备的最佳条件为:菌龄8h、酶质量浓度10mg/mL、酶解时间75min、酶解温度42℃,在此条件下制备原生质体并进行再生实验,原生质体形成率达到97.15%,再生率可达到31.25%。研究结果可为L.casei LC2W食品级外源表达系统的构建提供初步的基础。

The protoplast of Lactobacillus casei LC2W was prepared and regenerated in this study.The effects of cell age,lysozyme concentration,enzymolysis time and enzymolysis temperature on protoplast preparation were investigated according to the single factor tests.The conditions for protoplast preparation of L.casei LC2W were optimized by orthogonal tests.The results revealed that the optimal parameters for protoplast preparation of L.casei LC2W were cell age of 8 h,lysozyme concentration of 10 mg/mL,enzymolysis time of 75 min and enzymolysis temperature of 42 ℃.Under these optimal conditions,the protoplast formation rate reached up to 97.15% and the regeneration rate of L.casei LC2W was 31.25%.