Molecularly imprinted polymer-assisted refolding of lysozyme.
For production of active proteins using heterologous expression systems, refolding of proteins from inclusion bodies often creates a bottleneck due to its poor yield. In this study, we show that molecularly imprinted polymer (MIP) toward native lysozyme promotes the folding of chemically denatured lysozyme. The MIP, which was prepared with 1 M acrylamide, 1 M methacrylic acid, 1 M 2-(dimethylamino)ethyl methacrylate, and 5 mg/mL lysozyme, successfully promoted the refolding of lysozyme, whereas the non-imprinted polymer did not. The refolding yield of 90% was achieved when 15 mg of the MIP was added to 0.3 mg of the unfolded lysozyme. The parallel relationship between the refolding yield and the binding capacity of the MIP suggests that MIP promotes refolding through shifting the folding equilibrium toward the native form by binding the refolded protein.
Department of Materials Chemistry and Engineering, College of Engineering, Nihon University, Koriyama, Fukushima 963-8642, Japan. haruki@chem.ce.nihon-u.ac.jp
Mitsuru
M
Haruki
Yoshihiro
Y
Konnai
Ayumi
A
Shimada
Hirofumi
H
Takeuchi
D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D004789:Enzyme Activation; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry; D008689:Methacrylates / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry; D017510:Protein Folding; D013499:Surface Properties
D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D008689:Methacrylates; D011108:Polymers; D009113:Muramidase