fulltext.study @t Gmail

Peptide disulfides CGC and RKCGC facilitate oxidative protein refolding

Paper ID Volume ID Publish Year Pages File Format Full-Text
3728 184 2011 7 PDF Available
Title
Peptide disulfides CGC and RKCGC facilitate oxidative protein refolding
Abstract

Oxidative protein refolding is a big challenge for the recovery of disulfide bonds containing recombinant proteins expressed as inclusion bodies. In this study, the disulfide form of a small peptide mimic of protein disulfide isomerase, CGC, was explored to facilitate oxidative protein refolding with residual dithiothreitol (DTT) as the reductant. Working with DTT, CGC gave higher yield and folding rate than oxidized glutathione. Then, a new pentapeptide, RKCGC disulfide, was designed. It was found that RKCGC was more effective than CGC in increasing the folding rate and final yield, as demonstrated in the oxidative refolding of lysozyme and ribonuclease A. It was because the pentapeptide has lower pKa and higher reduction potential than CGC. Moreover, it efficiently facilitated protein refolding not only at neutral pH but also in a weak alkaline buffer. With CGC or RKCGC as the oxidant, the DTT removal step and the addition of other thiol reductants were not needed. This would benefit in decreasing the processing time and cost in protein refolding processes. This design has thus provided more efficient oxidant, and it may pave the way for the design of more effective thiol-disulfide redox agents.

► A new oxidant, RKCGC disulfide, was designed to catalyze oxidative protein refolding. ► RKCGC facilitated oxidative protein refolding more effectively than CGC disulfide. ► RKCGC was effective not only in neutral pH but also in weak alkaline solutions. ► The design reveals that adding nearby positive charges to peptide disulfide is effective to improve its catalytic activity by lowering the pKa of the corresponding thiol.

Keywords
Bioseparation; Protein; Refolding; Kinetic parameters; Peptide disulfide; Redox system
First Page Preview
Peptide disulfides CGC and RKCGC facilitate oxidative protein refolding
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 55, Issue 3, 15 August 2011, Pages 169–175
Authors
, , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us