海洋微生物酯酶高产菌株的诱变选育研究  

Breeding of high yield esterase strains from protoplast of marine microorganism

作　　者：郑鸿飞[1] 孙谧[1] 王跃军[1] 郝建华[1] 宋永相[1] 

机构地区：[1]中国水产科学院黄海水产研究所海洋产物资源与酶工程实验室

出　　处：《食品工业科技》2008年第8期64-66,70,共4页Science and Technology of Food Industry

基　　金：国家高技术研究发展(863)计划项目(2007AA091602);国家自然科学基金项目(30571429)

摘　　要：以筛选自海洋的产酯酶Bacillus subtilis var.niger为出发菌株,在原生质体制备与再生的最佳条件下制得原生质体,并通过紫外线对其进行诱变处理,采用透明圈初筛和摇瓶复筛,获得高产稳定菌株EB-2,产酶活力由原来的22.8U/mL提高到了77.6U/mL;同时,实验确定了该菌株原生质体制备与再生的最佳条件为:以pH7.2的0.3mol/L KCl和0.3mol/L蔗糖为稳定剂,菌龄在16h时,用10mg/mL的溶菌酶酶液40℃下酶解30min后,原生质体制备率达到91.7%,再生率达到21.3%。Under the optimal conditions of formation and regeneration of protoplast, protoplasts of Bacillus subtilis var. niger were prepared. The highly-yield stable strain EB-2, of which enzymatic activity was elevated from 22.8U/mL to 77.6U/mL, was obtained by UV irradiation protoplasts and screened from many regenerative mutants with the methods of transparent zones and flask culture. Meanwhile, it turned out that the optimal conditions of the formation and regeneration of protoplast were ascertained as well： select 0.3mol/L KCI plus 0.3mol/L sucrose as osmotic stabilizer, incubation time of the strain EB-1 was 16h, concentration of lysozyme was 10mg/mL, and time of enzymolysis was 30min at 40℃. The rate of formation and regeneration of protoplast under the optimal conditions were 91.7/% and 21.3%, respectively.

关 键 词：海洋微生物 酯酶 原生质体 紫外诱变 

分 类 号：TS201.3[轻工技术与工程—食品科学]