Abstract
Protein refolding is a crucial procedure in bacterial recombinant expression. Aggregation and misfolding are the two challenges that can affect the overall yield and specific activity of the folded proteins. We demonstrated the in vitro use of nanoscale “thermostable exoshells” (tES) to encapsulate, fold and release diverse protein substrates. With tES, the soluble yield, functional yield, and specific activity increased from 2-fold to >100-fold when compared to folding in its absence. On average, the soluble yield was determined to be 6.5 mg/100 mg of tES for a set of 12 diverse substrates evaluated. The electrostatic charge complementation between the tES interior and the protein substrate was considered as the primary determinant for functional folding. We thus describe a useful and simple method for in vitro folding that has been evaluated and implemented in our laboratory.
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Acknowledgments
This research was funded by NMRC-Clinician Scientist Award, CSAINV17nov012, and A*STAR-AME, A2083c0055. We thank Kong Shik Nie and Siddhesh Sujit Vaidya for their recent contributions to the study.
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Vallerinteavide Mavelli, G., Sadeghi, S., Drum, C.L. (2023). Laboratory Scale Production of Complex Proteins Using Charge Complimentary Nanoenvironments. In: Ueno, T., Lim, S., Xia, K. (eds) Protein Cages. Methods in Molecular Biology, vol 2671. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3222-2_23
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DOI: https://doi.org/10.1007/978-1-0716-3222-2_23
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