Gene design, fusion technology and TEV cleavage conditions influence the purification of oxidized disulphide-rich venom peptides in Escherichia coli
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- 作者：Ana Filipa Sequeira ; Jeremy Turchetto ; Natalie J. Saez…
- 关键词：Venom peptides ; Gene design ; Recombinant expression ; Periplasm ; Disulphide ; rich peptides ; Fusion protein ; Escherichia coli (E. coli) ; High ; throughput expression
- 刊名：Microbial Cell Factories
- 出版年：2017
- 出版时间：December 2017
- 年：2017
- 卷：16
- 期：1
- 全文大小：3716KB
- 刊物类别：Chemistry and Materials Science
- 刊物主题：Applied Microbiology; Biotechnology; Microbiology; Microbial Genetics and Genomics; Enzymology; Genetic Engineering;
- 出版者：BioMed Central
- ISSN：1475-2859
- 卷排序：16
文摘
BackgroundAnimal venoms are large, complex libraries of bioactive, disulphide-rich peptides. These peptides, and their novel biological activities, are of increasing pharmacological and therapeutic importance. However, recombinant expression of venom peptides in Escherichia coli remains difficult due to the significant number of cysteine residues requiring effective post-translational processing. There is also an urgent need to develop high-throughput recombinant protocols applicable to the production of reticulated peptides to enable efficient screening of their drug potential. Here, a comprehensive study was developed to investigate how synthetic gene design, choice of fusion tag, compartment of expression, tag removal conditions and protease recognition site affect levels of solubility of oxidized venom peptides produced in E. coli.