摘要
从土壤宏基因组中筛选获得在芽孢杆菌中高效分泌重组蛋白的信号肽。通过数据分析,将预测到的信号肽DNA片段与蛋白酶基因融合,在枯草芽孢杆菌(Bacillus subtilis)WB800和地衣芽孢杆菌(Bacillus licheniformis)DL6中表达,进行胞外蛋白酶酶活测定。筛选得到25个可能来源于G^+菌的信号肽序列,胞外蛋白酶活性分析的结果显示,有14个信号肽具有较好的分泌蛋白酶的能力,其中引导效果最好的信号肽(sig20)是蛋白酶自身信号肽的1.64倍。将高分泌信号肽转化到地衣芽孢杆菌中,证实其在地衣芽孢杆菌同样具有较好的引导效果。应用生物信息学与宏基因组学相结合的方法,可有效获得高效的芽孢杆菌信号肽序列,为蛋白质高效分泌表达系统的构建提供丰富的表达元件。
Signal peptide, efficiently secreting recombinant proteins in Bacillus sp. was obtained from soil metagenome. Atter database analysis, the predicted signal peptide DNA fragment was fused with a protease gene, and then expressed in Bacillus subtilis WB800 and Bacillus licheniformis DL6 to measure the extracellular protease activity. Totally 25 signal peptide sequences which may derived from G＋ bacteria were screened, extracel- lular protease activity results showed that there were 14 signal peptides had excellent ability of secreting protease. One of the signal peptides, named sig20, was 1.64 times of its own protease signal peptide, which performed the optimal secretion ability. The high secretion signal peptides were trans- formed into B. licheniformis and the results confirmed the similar secretion ability in B. subtilis. By combining bioinformatics and metagenomics methods, effectively Bacillus signal peptipe sequence could be obtained, which provided a rich expression elements for expression and secretion of recombinant proteins.
出处
《中国酿造》
CAS
北大核心
2016年第6期138-142,共5页
China Brewing
基金
国家高技术研究发展计划‘863计划’项目(2014AA021303
2014AA021302)
天津市应用基础与前沿科学研究计划(13JCYBJC39500)
