Organosoluble enzyme conjugates with poly(2-oxazoline)s via pyromellitic acid dianhydride.
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.
Chair of Biomaterials and Polymer Science, Department of Biochemical and Chemical Engineering, TU Dortmund, Emil-Figge-Str. 66, 44227 Dortmund, Germany.
Stefan
S
Konieczny
Christoph P
CP
Fik
Nils J H
NJ
Averesch
Joerg C
JC
Tiller
D000818:Animals; D001565:Benzoates / Q000737:chemistry*; D001709:Biotechnology; D002417:Cattle; D002645:Chickens; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006867:Hydrolases / Q000737:chemistry* / Q000378:metabolism*; D007525:Isoelectric Focusing; D008970:Molecular Weight; D010080:Oxazoles / Q000737:chemistry*; D011073:Polyamines / Q000737:chemistry*; D012995:Solubility
D001565:Benzoates; D010080:Oxazoles; D011073:Polyamines; C012019:pyromellitic dianhydride; D006867:Hydrolases