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Organosoluble enzyme conjugates with poly(2-oxazoline)s via pyromellitic acid dianhydride

Paper ID Volume ID Publish Year Pages File Format Full-Text
23781 43472 2012 9 PDF Available
Title
Organosoluble enzyme conjugates with poly(2-oxazoline)s via pyromellitic acid dianhydride
Abstract

The use of enzymes in organic solvents offers a great opportunity for the synthesis of complex organic compounds and is therefore in focus of current research. In this work we describe the synthesis of poly(2-methyl-1,3-oxazoline) (PMOx) and poly(2-ethyl-1,3-oxazoline) (PEtOx) enzyme conjugates with hen-egg white lysozyme, RNase A and α-chymotrypsin using a new coupling technique. The POXylation was carried out reacting pyromellitic acid dianhydride subsequently with ethylenediamine terminated POx and then with the NH2-groups of the respective enzymes. Upon conjugation with the polymers, RNase A and lysozyme became fully soluble in DMF (1.4 mg/ml). These are the first examples of fully POXylated proteins, which become organosoluble. The synthesized enzyme conjugates were characterized by SDS-PAGE, isoelectric focusing, dynamic light scattering and size exclusion chromatography, which all indicated the full POXylation of the enzymes. The modified enzymes even partly retained their activity in water. With α-chymotrypsin as example we could demonstrate that the molecular weight of the attached polymer significantly influences the activity.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► POXylation is successful via pyromellitic acid dianhydride. ► Hen-egg white lysozyme and RNase A became fully DMF soluble by POXylation. ► POXylated enzymes partly retain their activity in water.

Keywords
CT, α-chymotrypsin; DM, degree of modification; DMF, N,N-dimethylformamide; DMSO, dimethylsulfoxide; DP, degree of polymerization; DTT, dithiothreitol; EDA, ethylenediamine; EtOx, 2-ethyl-1,3-oxazoline; GPC, gel permeation chromatography; IEF, isoelectric
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Organosoluble enzyme conjugates with poly(2-oxazoline)s via pyromellitic acid dianhydride
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 159, Issue 3, 15 June 2012, Pages 195–203
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