Improving aquaporin Z expression in Escherichia coli by fusion partners and subsequent condition optimization

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• 作者：Jiazhang Lian ; Shinghua Ding ; Jin Cai ; Danping Zhang ; Zhinan Xu and Xiaoning Wang
• 关键词：Aquaporin Z ; MBP ; GST ; DsbA ; Membrane protein
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2009
• 出版时间：March, 2009
• 年：2009
• 卷：82
• 期：3
• 页码：463-470
• 全文大小：264.1 KB

文摘

Aquaporin Z (AqpZ), a typical orthodox aquaporin with six transmembrane domains, was expressed as a fusion protein with TrxA in E. coli in our previous work. In the present study, three fusion partners (DsbA, GST and MBP) were employed to improve the expression level of this channel protein in E. coli. The result showed that, compared with the expression level of TrxA-AqpZ, five- to 40-fold increase in the productivity of AqpZ with fusion proteins was achieved by employing these different fusion partners, and MBP was the most efficient fusion partner to increase the expression level. By using E. coli C43 (DE3)/pMAL-AqpZ, the effects of different expression conditions were investigated systematically to improve the expression level of MBP-AqpZ in E. coli. The high productivity of MBP-AqpZ (200 mg/l) was achieved under optimized conditions. The present work provides a novel approach to improve the expression level of membrane proteins in E. coli.