单亲灭活德氏乳杆菌和乳酸乳球菌原生质体融合条件优化  

Optimization of Conditions for Protoplast Fusion between Single Inactivated Lactobacillus delbrueckii and Lactococcus lactis

作　　者：李丽[1] 房杰[1] 黄洁洁[1] 付瑞燕[1] 

机构地区：[1]安徽农业大学茶与食品科技学院,安徽合肥230036

出　　处：《食品科学》2012年第5期193-198,共6页Food Science

基　　金：国家"973"计划项目(2007CB707803);安徽农业大学稳定和引进人才科研资助项目

摘　　要：利用单亲灭活原生质体技术对德氏乳杆菌FQ菌株和乳酸乳球菌FL菌株的原生质体进行融合,考察原生质体制备、再生和融合条件的影响因素。结果表明:制备德氏乳杆菌FQ菌株原生质体最适条件为温度37℃,在含有10μg/mL变溶菌素和1mg/mL溶菌酶溶液中超声处理90min。在此条件下,原生质体再生率可达6.36%。乳酸乳球菌FL菌株添加1mg/mL甘氨酸处理后,用10mg/mL溶菌酶37℃恒温酶解90min,原生质体形成率可达99.97%。65℃处理乳酸乳球菌FL菌株原生质体120min,原生质体灭活率可达96.89%。融合实验结果表明,在PEG6000 400g/L(含0.02mol/L MgCl2和0.01mol/L CaCl2)、融合时间5min、融合温度20℃、pH6.5的条件下促融,德氏乳杆菌FQ菌株和乳酸乳球菌FL菌株原生质体的融合率可达2.72×10-6。Single inactivated protoplast fusion technique was used for the fusion of Lactobacillus delbrueckii FQ with Lactococcus lactis FL. The protoplast formation, regeneration and fusion conditions were examined in this study. Results showed that the optimal enzymolysis conditions for L. delbrueckii FQ were ultrasound treatment for 90 rain at 37℃ in the presence of 10 μ g/mL mutanolysin and 1 mg/mL lysozyme. Under these conditions, the protoplast regeneration rate reached 6.36%. The protoplast formation rate of L lactis FL was up to 99.97% after 90 min of treatment with 10 mg/mL lysozyme in the presence of 1 mg/mL glycine. L lactis FL protoplasts showed an inactivated rate of 96.89% after 120 rnin of exposure to 65℃. The fusion rate between Lactobacillus delbrueckii FQ and Lactococcus lactis FL reached 2.72 × 10^-6 after 5 rain of infusion at pH 6.5 in the presence of 400 g/L PEG 6000 （included 0.02 mol/L MgCh and 0.01 mol/L CaCl2）.

关 键 词：单亲灭活 德氏乳杆菌 乳酸乳球菌 原生质体融合 

分 类 号：Q813.2[生物学—生物工程] Q939.117