作　　者： ; ; ; ;

机构地区： 华南理工大学生物科学与工程学院

出　　处： 《生物技术通报》 2012年第3期135-140,共6页

摘　　要： 将来自枯草芽孢杆菌的碱性脂肪酶基因经密码子优化,全基因合成后克隆到pPICZαA载体,构建了pPICZαA-bsl分泌型重组质粒,该重组质粒经限制性内切酶PmeI线性化后使用LiCl法转化到毕赤酵母X-33,经过筛选获得分泌表达碱性脂肪酶的重组毕赤酵母X-33/pPICZαA-bsl。摇瓶发酵液上清酶活最高可达4.78 U/mL,初步研究了该脂肪酶的酶学性质,其最适作用温度为40-60℃,最适pH9.0,且具有高度耐碱的特性。该重组脂肪酶对旧新闻纸具备较明显的脱墨能力。 The aim of the present study was to express and evaluate a previously cloned lipase gene.The alkaline lipase gene was optimized and subcloned into the secreted expression vector pPICZαA to construction a recombinant plasmid pPICZαA-bsl,then the recombinant plasmid was transformated into the Pichia pastoris X33.The recombinant yeast x33/pPICZαA-bsl could secrete alkaline lipase,and the maximum activity of the culture supernatant was 4.78 U/mL when the recombinant strain was cultured on a rotary shaker.The properties of the recombinant lipase was be tentatively researched,the results showed that the optimal temperature and pH of this lipase were 40-60℃ and 9.0,and the lipase was extreme alkaline-resistant.The recombinant lipase showed obvious deinking ability in old news paper deinking test.

关 键 词： 枯草芽孢杆菌 碱性脂肪酶 密码子优化 毕赤酵母 分泌表达 脱墨

领　　域： [生物学]