Complex coacervate core micelles with a lysozyme-modified corona.
This paper describes the preparation, characterization, and enzymatic activity of complex coacervate core micelles (C3Ms) composed of poly(acrylic acid) (PAA) and poly(N-methyl-2-vinyl pyridinium iodide)-b-poly(ethylene oxide) (PQ2VP-PEO) to which the antibacterial enzyme lysozyme is end-attached. C3Ms were prepared by polyelectrolyte complex formation between PAA and mixtures containing different ratios of aldehyde and hydroxyl end-functionalized PQ2VP-PEO. This resulted in the formation of C3Ms containing 0-40% (w/w) of the aldehyde end-functionalized PQ2VP-PEO block copolymer (PQ2VP-PEO-CHO). Chemical conjugation of lysozyme was achieved via reductive amination of the aldehyde groups, which are exposed at the surface of the C3M, with the amine groups present in the side chains of the lysine residues of the protein. Dynamic and static light scattering indicated that the conjugation of lysozyme to C3Ms prepared using 10 and 20% (w/w) PQ2VP-PEO-CHO resulted in the formation of unimicellar particles. Multimicellar aggregates, in contrast, were obtained when lysozyme was conjugated to C3Ms prepared using 30 or 40% (w/w) PQ2VP-PEO-CHO. The enzymatic activity of the unimicellar lysozyme-C3M conjugates toward the hydrolysis of the bacterial substrate Micrococcus lysodeikticus was comparable to that of free lysozyme. For the multimicellar particles, in contrast, significantly reduced enzymatic rates of hydrolysis, altered circular dichroism, and red-shifted tryptophan fluorescence spectra were measured. These results are attributed to the occlusion of lysozyme in the interior of the multimicellar conjugates.
Danial
Maarten
M
Laboratory for Physical Chemistry and Colloid Science, Wageningen University, Dreijenplein 6, 6703 HB Wageningen, The Netherlands.
Klok
Harm-Anton
HA
Norde
Willem
W
Stuart
Martien A Cohen
MA
D000180:Acrylic Resins / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000737:chemistry; D006868:Hydrolysis; D008823:Micelles; D008837:Micrococcus / Q000737:chemistry*; D009113:Muramidase / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D014753:Vinyl Compounds / Q000737:chemistry*
D000180:Acrylic Resins; D004800:Enzymes, Immobilized; D008823:Micelles; D014753:Vinyl Compounds; C523649:poly(N-methyl-2-vinyl pyridinium iodide)-b-poly(ethylene oxide); D011092:Polyethylene Glycols; C006903:carbopol 940; D009113:Muramidase