Recombinant expression, purification and PEGylation of Paneth cell peptide (cryptdin-2) with value added attributes against Staphylococcus aureus
IR@IMTECH: CSIR-Institute of Microbial Technology, Chandigarh
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Recombinant expression, purification and PEGylation of Paneth cell peptide (cryptdin-2) with value added attributes against Staphylococcus aureus
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| Creator |
Kaur, Navneet
Dilwari, Rahul Kaur, Amrita Sahni, Girish Rishi, Praveen |
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QR Microbiology
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| Description |
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
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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.
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Nature publishing group
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2020-07-22
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Article
PeerReviewed |
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| Relation |
https://www.nature.com/articles/s41598-020-69039-2
http://crdd.osdd.net/open/2595/ |
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| Identifier |
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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