Improved Complementary Polymer Pair System: Switching for Enzyme Activity by PEGylated Polymers

Abstract

The development of technology for on/off switching of enzyme activity is expected to expand the applications of enzyme in a wide range of research fields. We have previously developed a complementary polymer pair system (CPPS) that enables the activity of several enzymes to be controlled by a pair of oppositely charged polymers. However, it failed to control the activity of large and unstable α-amylase because the aggregation of the complex between anionic α-amylase and cationic poly(allylamine) (PAA) induced irreversible denaturation of the enzyme. To address this issue, we herein designed and synthesized a cationic copolymer with a poly(ethylene glycol) backbone, poly(N,N-diethylaminoethyl methacrylate)-block-poly(ethylene glycol) (PEAMA-b-PEG). In contrast to PAA, α-amylase and β-galactosidase were inactivated by PEAMA-b-PEG with the formation of soluble complexes. The enzyme/PEAMA-b-PEG complexes were then successfully recovered from the complex by the addition of anionic poly(acrylic acid) (PAAc). Thus, dispersion of the complex by PEG segment in PEAMA-b-PEG clearly plays a crucial role for regulating the activities of these enzymes, suggesting that PEGylated charged polymer is a new candidate for CPPS for large and unstable enzymes.

Polymer Directed Protein Assemblies

“…This was demonstrated by Kurinomaru et al who encapsulated/complexed α-amylase and β-galactosidase with poly-allylamine [24]. In order to recover the protein as a "free" structure, poly-acrylic acid was added to interact with the cationic poly-allylamine and thereby setting free the protein.…”

Noncovalent PEGylation via Lectin–Glycopolymer Interactions

“…[16][17][18][19][20][21][22][23][24][25][26] Examples include polymer conjugation via: co-factor reconstitution, 16 the biotin-streptavidin complex, 18 supramolecular complexation with cucurbituril, 19 charge-charge 20 and metal affinity 21,24 interactions, as well as lectin-mediated complexation. 25,27,28 The advantage of noncovalent systems is that temporal control of conjugate assembly and disassembly can be asserted by the presence of competitor molecules.…”

The Use of an Enzyme-based Sensor Array to Fingerprint Proteomic Signatures of Sera from Different Mammalian Species

“…8,9,17 The catalytic activity of enzymes was reversibly inhibited without denaturing by non-covalent PIC formation with PEGylated polyamines. 18 Since each PIC was expected to possess different affinities for serum proteins, the addition of mammalian sera to PICs would render unique patterns of changes in enzyme activity through competitive interactions between PICs and serum proteins. Generated patterns possibly reflect proteomic signatures of individual sera, and are therefore available for discrimination using a chemometric method.…”