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Application of poly-arginine fused minichaperone to renaturation of cyclodextrin glycosyltransferase expressed in recombinant Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
18443 42723 2006 7 PDF Available
Title
Application of poly-arginine fused minichaperone to renaturation of cyclodextrin glycosyltransferase expressed in recombinant Escherichia coli
Abstract

Minichaperone, a monomeric polypeptide binding domain fragment of GroEL, has chaperone activities in vivo and in vitro. Fed-batch cultures to optimize production of poly-arginine fused minichaperone (MiniELR10) in recombinant Escherichia coli were done to produce dry cell weight of 75.2 g/L, MiniELR10 expression level of 30.7%, and MiniELR10 concentration of 11.1 g/L in yeast extract supplemented conditions. MiniELR10 was simply purified through a single step of cation exchange chromatography with high purity. In a prewashing step and an adsorption process by controlling NaCl concentration, removal of contaminant intracellular E. coli proteins and selective binding of MiniELR10 on a cation exchanger were achieved without loss of the target protein. The purified MiniELR10 was efficiently immobilized on the cation exchanger and applied to the refolding of Bacillus macerans cyclodextrin glycosyltransferase (CGTase), which was expressed as inclusion body in recombinant E. coli. The refolding yield of CGTase was enhanced by 71% by the immobilized MiniELR10 compared with a conventional refolding method. Thus, minichaperone immobilized by poly-arginine fusion could assist refolding of heterologous proteins expressed as inclusion body.

Keywords
Minichaperone; Poly-arginine fusion; Cyclodextrin glycosyltransferase; Refolding
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Application of poly-arginine fused minichaperone to renaturation of cyclodextrin glycosyltransferase expressed in recombinant Escherichia coli
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 39, Issue 3, 3 July 2006, Pages 459–465
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