ARTP诱变联合抗生素抗性选育纳豆激酶高产菌株  

Breeding of Nattokinase High-producing Strains by ARTP Mutagenesis Combined with Antibiotic Resistance

作　　者：薛莹莹 林福兴 别小妹[1] 吕凤霞[1] 赵海珍[1] 陆兆新[1] XUE Ying-ying;LIN Fu-xing;BIE Xiao-mei;LV Feng-xia;ZHAO Hai-zhen;LU Zhao-xin(College of Food Science and Technology,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学食品科技学院

出　　处：《食品工业科技》2019年第23期93-97,共5页Science and Technology of Food Industry

基　　金：国家自然科学基金(31571887)

摘　　要：为获得产纳豆激酶(nattokinase,NK)活力增强的菌株,以纤维蛋白平板法测得酶活力值为判定指标,采用常压室温等离子体诱变系统(ARTP)对枯草芽孢杆菌XZI125进行诱变,通过酪蛋白平板法和利福平、卡那霉素、链霉素、氯霉素四种抗生素抗性筛选法获得高产NK突变菌株。结果表明,经6轮摇瓶发酵验证,获得了3株遗传稳定的高产突变菌株A27,Ar41和Ac35,摇瓶发酵5 d的产酶活力分别比原始菌株(2490 IU/mL)提高了23.0%、25.1%、26.5%。比较酪蛋白平板筛选方法和0.7μg/mL利福平、50μg/mL卡那霉素、60μg/mL链霉素、300μg/mL氯霉素四种抗性筛选方法,抗生素抗性筛选方法更有利于NK高产菌株的筛选。In order to obtain high-yield nattokinase( NK) strains,the enzyme activity was determined by fibrin plate method,Bacillus subtilis XZI125 was mutagenized by atmospheric pressure room temperature plasma mutagenesis system( ARTP).High-yield NK mutant strains were obtained by casein plate method and four antibiotic resistance screening methods: rifampicin,kanamycin,streptomycin and chloramphenicol.The results showed that three high-yield mutant strains A27,Ar41 and Ac35 were obtained by six rounds of shaking flask fermentation. The enzyme-producing activity of the three strains after 5 d of shaking flask fermentation was 23.0%,25.1% and 26.5% higher than that of the original strain( 2490 IU/mL),respectively.Compared with casein plate screening method and 0.7 μg/mL rifampicin,50 μg/mL kanamycin,60 μg/mL streptomycin and 300 μg/mL chloramphenicol,antibiotic resistance screening method was more conducive to the screening of NK high-yield strains.

关 键 词：纳豆激酶 枯草芽孢杆菌 ARTP 诱变 抗生素抗性 突变株筛选 

分 类 号：TS201.3[轻工技术与工程—食品科学]