现代食品科技2017，Vol.33Issue(3)：105-109,87,6.DOI:10.13982/j.mfst.1673-9078.2017.3.017
人溶菌酶基因在乳酸克鲁维酵母中的表达
Expression of the Human Lysozyme Gene in Kluyveromyces lactis
摘要
Abstract
Human lysozyme,an enzyme with natural broad-spectrum antimicrobial activity,has potential applications in the food and pharmaceutical industries.To obtain human lysozyme preparations with high activity,a Kluyveromyces lactis yeast expression system was used for soluble expression of the human lysozyme gene (hLYZ) with optimized codons.An artificial hLYZ gene was cloned into the K.lactis expression vector pKLAC 1,resulting in the recombinant vector pKLAC 1-hLZY,which was then linearized with SacⅡ and transformed into K.lactis GG799 by electroporation.Following whole-cell PCR,a multicopy recombinant strain (named hLYZ1) was obtained.The engineered strain secreted a target protein of approximately 14 ku in molecular weight,which agreed with that of the expected product.The maximum activity of hLZY in the culture supematant reached 1430 U/mL after 128 h of culture.Antibacterial activity testing confirmed that the recombinant human lysozyme was effective against Micrococcus luteus,Escherichia coli,and Bacillus subtilis.In conclusion,a recombinant hLYZ with high enzyme activity was successfully expressed in K.lactis GG799,thereby providing a basis for studies on the large-scale production of recombinant human lysozyme using the K.lactis expression system.关键词
人溶菌酶/乳酸克鲁维酵母/分泌表达/抗菌活性Key words
human lysozyme/Kluyveromyces lactis/secretory expression/antibiotic activity引用本文复制引用
刘思彤,徐龙龙,衣泽浩,刘晓静,惠丰立..人溶菌酶基因在乳酸克鲁维酵母中的表达[J].现代食品科技,2017,33(3):105-109,87,6.基金项目
国家自然科学基金项目(31370073) （31370073）
河南省基础与前沿技术研究计划(102300410146) （102300410146）
