Abstract
The ability to express heterologous proteins in microbial hosts is crucial for many areas of research and technology. In most cases, however, successful expression and purification of the desired protein require fusion to another protein. To date, all fusion partners have been chosen from natural sequences, which evolved for other purposes, and may not be optimal fusion partners. However, the rise of synthetic biology and protein design make it possible to design and optimize fusion proteins using novel sequences that did not arise in nature. Here, we describe a series of De novo Expression Enhancer Proteins (DEEPs) that facilitate high-level expression and facile purification of heterologous proteins and peptides. To test the DEEP system, a de novo protein was fused to several target proteins covering a range of sizes and solubilities. In all cases, fusions to DEEP outperformed fusions to SUMO, a commonly used natural fusion partner. The availability of novel proteins that can be engineered for specific fusion applications could be beneficial to enhance the expression of a wide range of heterologous proteins.
Original language | English (US) |
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Pages (from-to) | 1698-1706 |
Number of pages | 9 |
Journal | Protein Science |
Volume | 29 |
Issue number | 8 |
DOIs | |
State | Published - Aug 1 2020 |
All Science Journal Classification (ASJC) codes
- Molecular Biology
- Biochemistry
Keywords
- de novo protein design
- fusion tag
- protein expression
- purification tag