乳酸菌氧化锆珠破壁方法的优化及适用性研究  

Optimization of zirconia beads wall-breaking method for lactic acid bacteria and study on its effect of the applicability

作　　者：潘荣荣[1,2] 王茂鹏[2] 李昌[2,3] 邸洋 荣凤君 杜寿文[2] 朱羿龙[2] 刘立明[3] 朱国强[1] 金宁一[2,3] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]军事医学科学院军事兽医研究所,省部共建吉林省人兽共患病预防与控制重点实验室,吉林长春130122 [3]温州大学病毒病研究所,浙江温州325035

出　　处：《食品工业科技》2017年第11期140-143,共4页Science and Technology of Food Industry

基　　金：国家自然科学基金(31472197);病原微生物生物安全国家重点实验室开放课题(SKLPBS1435)

摘　　要：本实验旨在优化氧化锆珠破壁方法,以期筛选出较佳的破壁条件,提高蛋白得率。实验在菌体重量、研磨buffer和研磨时间这3方面对该破壁方法进行了优化,并通过SDS-PAGE来分析其破壁效果。在优化后的条件下,同时对6株乳酸菌进行破壁处理并通过SDS-PAGE来分析其破壁效果。结果表明:在菌体重量、研磨buffer、锆珠三者的加入比例(W/V/W)为0.013 g∶300μL∶1 g的条件下,研磨15 min,提取的蛋白图谱条带清晰,丰度高;RIPA、PBS、PENP、GUS作为研磨buffer时,提取的蛋白图谱条带清晰且丰度高,而8 mol/L尿素作为研磨buffer时,图谱条带模糊且丰度低;6株乳酸菌经该方法破壁处理后,蛋白图谱条带均具有较高的丰度和清晰度。因此,对于乳酸菌的破壁,氧化锆珠破壁法具有快速、高效的优点。This study aims to optimize the zirconia beads wall-breaking method, screen out the better broken conditions for breaking lactic acid bacteria cell wall and improve the protein yield.The weight of bacteria, the grinding buffer and time were tested and selected as important aspects to optimize the wall-breaking method ,and then ,the breaking effect was analyzed and evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis（ SDS-PAGE）.Under optimized conditions, six different species of strains from lactic acid bacteria were tested for cell wall-breaking treatments with the zirconia beads method, after that,the breaking effects were analyzed and evaluated by SDS-PAGE.Results showed that, on condition that the ratio of bacteria, grinding buffer and zirconia beads （W/V/W）is 0.013 g：300 μL：1g, grinding time of 15 min, the protein pattern with clear banding,high abundance; RIPA, PBS, PENP, GUS as the grinding buffer, the protein pattern with clear banding, high abundance, while 8 mol/L urea as the grinding buffer, the protein pattern with obscure banding, low abundance. After Six strains of lactic acid bacteria were broken through this method, the protein pattern with clear banding, high abundance.Therefore, zirconia beads method is a combined rapid and efficient method for breaking lactic acid bacteria cell wall.

关 键 词：乳酸菌 氧化锆珠 优化 适用性 

分 类 号：TS201.3[轻工技术与工程—食品科学]