人溶菌酶基因在乳酸克鲁维酵母中的表达
发布时间：2019-04-17 18:23
【摘要】：人溶菌酶是一种天然广谱抑菌物质,在食品和医药工业有潜在应用前景。为获得高活性的人溶菌酶制剂,采用乳酸克鲁维酵母表达系统,对经密码子优化的人溶菌酶基因(h LYZ)进行分泌表达。将人工合成h LYZ插入到乳酸克鲁维酵母表达载体p KLAC1,构建重组载体p KLAC1-h LYZ,并用电脉冲法将SacⅡ线性化的重组质粒转化到乳酸克鲁维酵母GG799中。通过全细胞PCR鉴定,最后获得了一株多拷贝整合的基因工程菌h LYZ1。工程菌可以分泌表达分子量约14 ku的目的蛋白质,与预期大小相符。摇瓶发酵培养128 h,酶活最高达到1430 U/mL。抗菌活性检测结果显示,重组人溶菌酶对溶壁微球菌、大肠杆菌和枯草芽孢杆菌有较好的溶菌活性。本研究成功地在乳酸克鲁维酵母中表达了重组人溶菌酶,表达的蛋白具有较高的酶活性,试验结果为利用乳酸克鲁维酵母表达系统规模化生产重组人溶菌酶奠了基础。
[Abstract]:Human lysozyme is a kind of natural broad-spectrum bacteriostatic substance, which has potential application prospect in food and medicine industry. In order to obtain highly active human lysozyme preparation, the codon-optimized human lysozyme gene (h LYZ) was secreted and expressed by Kruyveromyces lactis expression system. The synthetic h-LYZ was inserted into the expression vector p-KLAC1, to construct the recombinant vector p-KLAC1-h LYZ, and the linearized recombinant plasmid of Sac 鈪，
本文编号：2459680
[Abstract]:Human lysozyme is a kind of natural broad-spectrum bacteriostatic substance, which has potential application prospect in food and medicine industry. In order to obtain highly active human lysozyme preparation, the codon-optimized human lysozyme gene (h LYZ) was secreted and expressed by Kruyveromyces lactis expression system. The synthetic h-LYZ was inserted into the expression vector p-KLAC1, to construct the recombinant vector p-KLAC1-h LYZ, and the linearized recombinant plasmid of Sac 鈪，
本文编号：2459680
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