Kaur, Navneet and Dilwari, Rahul and Kaur, Amrita and Sahni, Girish and Rishi, Praveen (2020) Recombinant expression, purification and PEGylation of Paneth cell peptide (cryptdin-2) with value added attributes against Staphylococcus aureus. Scientific reports, 10 (12164). pp. 1-14. ISSN 2045-2322
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Article Open Access Published: 22 July 2020 Recombinant expression, purification and PEGylation of Paneth cell peptide (cryptdin-2) with value added attributes against Staphylococcus aureus Navneet Kaur, Rahul Dilawari, Amrita Kaur, Girish Sahni & Praveen Rishi Scientific Reports volume 10, Article number: 12164 (2020) Cite this article 140 Accesses 1 Altmetric Metricsdetails Abstract Cryptdins are disulfide-rich cationic antimicrobial peptides secreted by mouse Paneth cells and are known to exhibit potent antimicrobial activity against various deadly pathogens. Keeping in view the extremely low yield obtained from mouse Paneth cells and high cost of synthetic peptide(s), herein, we have attempted to produce cryptdin-2 in Escherichia coli using recombinant technology. To avoid lethal effects of peptide on the host cells, cryptdin-2 was expressed as a fusion protein with thioredoxin as fusion partner which yielded 40 mg/L protein in the soluble fraction. Subsequently, mature cryptdin-2 was cleaved from the fusion partner and purified by cation exchange chromatography. Since conjugation of poly(ethylene) glycol (PEG) has been known to improve the biological properties of biomolecules, therefore, we further attempted to prepare PEG-conjugated variant of cryptdin-2 using thiol specific PEGylation. Though the antimicrobial activity of PEGylated cryptdin-2 was compromised to some extent, but it was found to have enhanced serum stability for longer duration as compared to its un-modified forms. Also, it was found to exhibit reduced toxicity to the host cells. Further, its synergism with gentamicin suggests that PEGylated cryptdin-2 can be used with conventional antibiotics, thereby indicating its possibility to be used as an adjunct therapy.
| Item Type: | Article |
|---|---|
| Additional Information: | Open Access |
| Subjects: | Q Science > QR Microbiology |
| Depositing User: | Dr. K.P.S.Sengar |
| Date Deposited: | 30 Jul 2020 16:05 |
| Last Modified: | 30 Jul 2020 16:05 |
| URI: | http://crdd.osdd.net/open/id/eprint/2595 |
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