| Issue |
E3S Web Conf.
Volume 185, 2020
2020 International Conference on Energy, Environment and Bioengineering (ICEEB 2020)
|
|
|---|---|---|
| Article Number | 04050 | |
| Number of page(s) | 4 | |
| Section | Chemical Engineering and Food Biotechnology | |
| DOI | https://doi.org/10.1051/e3sconf/202018504050 | |
| Published online | 01 September 2020 | |
Expression of recombinant mouse cytosolic carboxypeptidase 6 in Escherichia coli
School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072, China
* Corresponding author: 1336564890@qq.com
* Corresponding author: huiyuan.wu@tju.edu.cn
Cytosolic carboxypeptidase 6 (CCP6) is a member of cytosolic carboxypeptidase (CCP) family that catalyze the removal of polyglutamate side chains from protein substrates. Biochemical and biophysical characterization of CCPs requires large quantities of purified proteins. However, no method describing the expression of any mammalian CCP family member from bacteria has been published to our best knowledge. After considerable efforts to improve the solubility of mammalian CCPs expressed in bacteria, including the optimization of induction temperature and by using different receptive cells, we were able to get less expression of mouse CCP6 in soluble fraction of bacterial lysates. We report in this article, the bacterial expression of CCP6 using Arctic Express (DE3) competent cells that co-express the chaperonin system GroEL and GroES from Oleispira antarctica. However, to achieve a large number of soluble target proteins, the expression conditions still need to be further optimized.
© The Authors, published by EDP Sciences, 2020
This is an Open Access article distributed under the terms of the Creative Commons Attribution License 4.0, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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