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Statistical optimization and multiple objective programming of lysozyme refolding catalyzed by recombinant DsbA in vitro

Paper ID Volume ID Publish Year Pages File Format Full-Text
35083 45073 2012 9 PDF Available
Title
Statistical optimization and multiple objective programming of lysozyme refolding catalyzed by recombinant DsbA in vitro
Abstract

DsbA (disulfide bond formation protein A) is essential for disulfide bond formation directly affecting the nascent peptides folding to the correct conformation in vivo. In this paper, recombinant DsbA protein was employed to catalyze denatured lysozyme refolding and inhibit the aggregation of folding intermediates in vitro. Statistical methods, i.e., Plackett–Burman design and small central composite design, were adopted to screen out important factors affecting the refolding process and correlating these parameters with the refolding efficiency including both protein recovery and specific activity of refolded lysozyme. Four important parameters: initial lysozyme concentration, urea concentration, KCl concentration and GSSG (glutathione disulfide) concentration were picked out and operating conditions were optimized by introducing the effectiveness coefficient method and transforming the multiple objective programming into an ordinary constrained optimization issue. Finally, 99.7% protein recovery and 25,600 U/mg specific activity of lysozyme were achieved when 281.35 μg/mL denatured lysozyme refolding was catalyzed by an equivalent molar of DsbA at the optimal settings. The results indicated that recombinant DsbA protein could effectively catalyze the oxidized formation and reduced isomerization of intramolecular disulfide bonds in the refolding of lysozyme in vitro.

► Foldase DsbA was introduced to an in vitro refolding system of denatured lysozyme. ► Statistical design was successfully applied to the multiple object programming issue. ► DsbA accelerates the oxidized formation and reduced isomerization of disulfide bonds. ► Activity recovery of lysozyme was increased with the assistance of recombinant DsbA.

Keywords
DsbA; Lysozyme; Refolding; Plackett–Burman design; Small central composite design; Multiple objective programming
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Statistical optimization and multiple objective programming of lysozyme refolding catalyzed by recombinant DsbA in vitro
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 47, Issue 8, August 2012, Pages 1268–1276
Authors
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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