高温α-淀粉酶高表达的研究  

Integrated Expression of Bacillus licheniformis α-amylase

作　　者：黄春敏[1] 沈微[1] 王正祥[1] 

机构地区：[1]江南大学生物工程学院生物资源与生物能源研究中心和教育部工业生物技术教育部重点实验室,江苏无锡214122

出　　处：《微生物学杂志》2010年第5期36-40,共5页Journal of Microbiology

基　　金：国家高技术研究发展计划(2006AA020204)

摘　　要：为了进一步提高工业上重要的地衣芽胞杆菌高温α-淀粉酶（BLA）的发酵生产性能,以高温α-淀粉酶基因（amyL）为目的基因,构建地衣芽胞杆菌高温α-淀粉酶基因产生菌的整合表达通用性质粒pB li16 s-amyL-EryR,采用原生质体转化法将此整合型表达质粒导入B.licheniformis CBBD302,在整合表达质粒中的16SrDNA序列的介导下实现了同源整合。挑选20株阳性转化子,分别通过提高培养基中红霉素的使用浓度,增加染色体上目的基因amyL的拷贝数,由此获得的重组菌的BLA生产水平提高了56.37%-80.29%。In order to further improve fermentation productivity of the important industry Bacillus licheniformis thermophilic α-amylase（BLA）,the common integrated expression plasmid pBli16s-amyL-EryR of BLA producing strain was constructed using thermophilic α-amylase gene（amyL） as the purpose gene,This integrated expression plasmid was transformed into B.licheniformis CBBD302 by protoplast transformation,realizing homologous integration under 16S rDNA sequence of the integrated expression plasmid.Then 20 positive transformants were chosen to increase the copies of the purpose gene amyL on chromosome through the use of concentration of erythromycin in the medium,thus,the obtained recombinant BLA production level increased by 56.37%～80.29%.

关 键 词：高温Α-淀粉酶 地衣芽胞杆菌 基因倍增 整合表达 

分 类 号：Q786[生物学—分子生物学]