摘要
采用脉冲强光对枯草芽孢杆菌进行诱变处理,通过耐受性测试,在抗性菌株中筛选出优良耐受菌株,并与原始菌株对比分析产酶活力及遗传稳定性,以十二烷基硫酸钠-聚丙烯酰胺凝胶电泳比较分析菌体蛋白质表达的差异。结果表明,在脉冲电压2 450 V,照射距离5 cm,脉冲40次条件下,筛选出8株抗性菌株;经初筛和复筛得到兼具高温耐受、强酸耐受和高浓度胆盐耐受性变异菌株B3和B7,产α-淀粉酶、蛋白酶、脂肪酶活力分别比原始菌株提高了67%和77%(P<0.01)、56%和71%(P<0.01)、34%和42%(P<0.05),变异菌株产酶稳定性较高(P>0.05),可遗传变异;诱变前后菌体蛋白表达出现差异;结果证明脉冲强光用于枯草芽孢杆菌的诱变具有可行性。
Pulsed light was applied to mutagenize Bacillus subtilis.The resulting mutants were screened for improved tolerance.The extracellular enzymatic activity and genetic stability of the selected mutants were analyzed and compared with those of the original strain and the differential protein expression of the original strain and the mutant strains was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis（SDS-PAGE）.The results showed that 8 resistant mutant strains were generated under the conditions of pulse voltage of 2 450 V,distance of 5 cm and pulse number of 40.After primary and secondary screening,the mutant strains B3 and B7 with good tolerance to high temperature tolerance,acid and high concentration of bile salt were obtained.Their α-amylase activity,protease activity,lipase activity was increased by 67% and 77%（P 0.01）,56% and 71%（P 0.01）,and 34% and 42%（P 0.05）,compared with those of the original strain,respectively.The enzymatic activity of the obtained mutants was stable（P 0.05） and showed genetic variation.The protein expression level was different between before and after mutagenesis.These findings proved the feasibility of application of pulsed light to mutagenize B.subtilis.
作者
赵天惠
张佰清
ZHAO Tianhui;ZHANG Baiqing(College of Food Science, Shenyang Agricultural University, Shenyang 110866, China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2018年第2期192-197,共6页
Food Science
关键词
耐受性
枯草芽孢杆菌
脉冲强光
诱变筛选
产酶活力
tolerance
Bacillus subtilis
pulsed light
mutagenesis and screening
enzymatic activity
