A fusion protein designed for soluble expression, rapid purification, and enhanced stability of parasporin-2 with potential therapeutic applications
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Issued Date
2024-09-01
Resource Type
eISSN
2215017X
Scopus ID
2-s2.0-85200647295
Journal Title
Biotechnology Reports
Volume
43
Rights Holder(s)
SCOPUS
Bibliographic Citation
Biotechnology Reports Vol.43 (2024)
Suggested Citation
Srisaisap M., Suwankhajit T., Boonserm P. A fusion protein designed for soluble expression, rapid purification, and enhanced stability of parasporin-2 with potential therapeutic applications. Biotechnology Reports Vol.43 (2024). doi:10.1016/j.btre.2024.e00851 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/100480
Title
A fusion protein designed for soluble expression, rapid purification, and enhanced stability of parasporin-2 with potential therapeutic applications
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Abstract
Bacillus thuringiensis parasporin-2 (PS2Aa1 or Mpp46Aa1) selectively destroys human cancer cells, making it a promising anticancer agent. PS2Aa1 protoxin expression in Escherichia coli typically results in inclusion bodies that must be solubilized and digested by proteinase K to become active. Here, maltose-binding protein (MBP) was fused to the N-terminus of PS2Aa1, either full-length (MBP-fPS2) or truncated (MBP-tPS2), to increase soluble protein expression in E. coli and avoid solubilization and proteolytic activation. Soluble MBP-fPS2 and MBD-tPS2 proteins were produced in E. coli and purified with endotoxin levels below 1 EU/μg. MBP-fPS2 was cytotoxic against T cell leukemia MOLT-4 and Jurkat cell lines after proteinase-K digestion. However, MBP-tPS2 was cytotoxic immediately without MBP tag removal or activation. MBP-tPS2′s thermal stability also makes it appropriate for bioproduction and therapeutic applications.