不同信号肽及其组合对果聚糖蔗糖酶异源表达的影响  

Effects of Different Signal Peptides and Their Combinations on Heterologous Expression of Levansucrase

作　　者：王茂军 色依德·斯马依 蔡逸安 李庆刚 路福平[1] 李玉[1] WANG Mao-jun;SIMAYI Seyide;CAI Yi-an;LI Qing-gang;LU Fu-ping;LI Yu(Key Laboratory of Industrial Fermentation Microbiology,Ministry of Education,College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China)

机构地区：[1]工业发酵微生物教育部重点实验室,天津科技大学生物工程学院,天津300457

出　　处：《中国生物工程杂志》2023年第5期37-44,共8页China Biotechnology

基　　金：国家重点研发计划(2021YFC2100402)资助项目。

摘　　要：为了实现果聚糖蔗糖酶在解淀粉芽孢杆菌Bacillus amyloliquefaciens 018(简称G3)中的高效表达,选择来源于不同芽孢杆菌的4个果聚糖蔗糖酶基因lsLich、lsAmy、lsSub和lsMega进行异源表达,并将课题组前期确定的对碱性蛋白酶酶活提升较高的5种信号肽进行筛选和组合。其中来源于地衣芽孢杆菌Bacillus licheniformis RN-01的lsLich在重组菌株G3/pLY-2-lsLich中的酶活力最高,酶活力为62.73 U/mL。以LS-Lich作为目的蛋白筛选单信号肽和双信号肽,其中信号肽SP_(DacB)和SP_(AmyE)组合的重组菌株G3/pLY-2-SDA-ls酶活最高,胞外酶活达到125.76 U/mL,较重组菌G3/pLY-2-SD-ls和G3/pLY-2-SA-ls分别提高了31.3%和39.2%,较原始菌株提高了100.49%。该结果表明双信号肽较单信号肽有助于提高LS-Lich的分泌量,同时信号肽组合顺序不同也产生一定的差异。To express levansucrase in Bacillus amyloliquefaciens 018(G3)efficiently.Four levansucrase genes lsLich,lsAmy,lsSub and lsMega from different Bacillus species were heterologously expressed,and five signal peptides with relatively high levels of alkaline protease identified by the research group were screened and combined.lsLich derived from Bacillus licheniformis RN-01 had the highest enzyme activity in the recombinant strain G3/pLY-2-lsLich,with an enzyme activity of 62.73 U/mL.LS-Lich was used as the target protein to screen single signal peptide and double signal peptide.The recombinant strain G3/pLY-2-SDA-ls combined with SP_(DacB)and SP_(AmyE)had the highest enzyme activity,and the extracellular enzyme activity reached 125.76 U/mL.Compared with the recombinant strain G3/pLY-2-SD-ls and G3/pLY-2-SA-ls,they increased by 31.3%and 39.2%,respectively,and increased by 100.49%compared with the original strain.The results indicated that the double signal peptide was helpful in increasing the secretion of LS-Lich compared with the single signal peptide,and the combination order of signal peptides also produced some differences.

关 键 词：信号肽 果聚糖蔗糖酶 解淀粉芽孢杆菌 异源表达 

分 类 号：Q819[生物学—生物工程]