Improving Protein Yield and Solubility with Wheat Germ Cell-Free Expression System
Author Information
Author(s): Guild Katherine, Zhang Yang, Stacy Robin, Mundt Elizabeth, Benbow Sarah, Green Amanda, Myler Peter J.
Primary Institution: Seattle Biomedical Research Institute
Hypothesis
Can the wheat germ cell-free expression system improve the yield and solubility of proteins that are difficult to express in E. coli?
Conclusion
The wheat germ cell-free expression system significantly increases the yield of soluble proteins that are insoluble when expressed in E. coli.
Supporting Evidence
- 87% of protein targets expressed detectable protein in the wheat germ cell-free system.
- 83% of targets that were insoluble in E. coli became soluble in the wheat germ system.
- NVoy polymer increased the percentage of soluble protein yield in most tests.
Takeaway
Scientists found that using a special system from wheat can help make more proteins that are usually hard to get, and adding a special ingredient can make them even better.
Methodology
The study tested 44 protein targets using a wheat germ cell-free expression system and assessed their solubility and expression levels.
Limitations
The quantities of soluble protein produced were insufficient for standard crystallization studies.
Digital Object Identifier (DOI)
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