Part:BBa_K5384002
DDDDK
Usage and Biology
DDDDK enterokinase is a tool enzyme. Enterokinase specifically cleaves peptides or proteins containing recognition sequences and is commonly used in biotechnology to perform specific cleavages of fusion proteins to release the protein of interest. DDDDK enterokinase is highly effective and specific. In protein structure and function studies, it is used to accurately cleave the target protein from the fusion protein for subsequent analysis and experimentation. In the biopharmaceutical process, it can be used to prepare protein drugs with specific activities. The enterokinase recognition site on the plasmid serves as a recognition site, allowing the enterokinase to act on the induced proteins.[1,2]
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Application
The low-cost and high-efficiency one-step protein separation method of histidine (His) labeling at the end of the protein and affinity purification of Nickel metal chelating column is also widely used in the separation and purification of rEK, with a yield of more than 50%. For the His-tag site, Choi et al. found that the enzyme activity of rECL was not changed after the C-terminus was His-labeled, but the enzyme activity was lost after the N-terminus was labeled by the Nickel metal chelating column.[3] In this project, we designed a DDDDK site between His-tag and Vg segment, which can be recognized and cleaved by EK.
References
[1]ZHANG Xianghui,TAN Shuhua,LI Taiming. Research progress on the characteristics of enterokinase and its genetic engineering. CNKI, 2005
[2]Xu Liang, Wang Wenjing, Zu Xiangyang, Wang Xiaochun. Characteristics of enterokinase digestion of fusion protein Trx-Exendin-4. 《CNKI; WanFang", 2012
[3]Cao Zhifei, Li Xinping, Fan Kai, Li Jing, et al. Expression and purification of recombinant bovine enterokinase light chain gene in Pichia pastoris. 《CNKI; WanFang》,2006
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