Premium
Charge‐directed targeting of antimicrobial protein‐nanoparticle conjugates
Author(s) -
Satishkumar Rohan,
Vertegel Alexey
Publication year - 2008
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21782
Subject(s) - lysozyme , chemistry , nanoparticle , conjugated system , zeta potential , lysis , surface charge , bacterial cell structure , substrate (aquarium) , bacteria , enzyme , covalent bond , biophysics , biochemistry , organic chemistry , nanotechnology , materials science , biology , polymer , ecology , genetics
Use of antimicrobial enzymes covalently attached to nanoparticles is of great interest as an antibiotic‐free approach to treat microbial infections. Intrinsic properties of nanoparticles can also be used to add functionality to their conjugates with biomolecules. Here, we show in a model system that nanoparticle charge can be used to enhance delivery and increase bactericidal activity of an antimicrobial enzyme, lysozyme. Hen egg lysozyme was covalently attached to two types of polystyrene latex nanoparticles: positively charged, containing aliphatic amine surface groups, and negatively charged, containing sulfate and chloromethyl surface groups. In the case of bacterial lysis assay with a Gram‐positive bacteria Micrococcus lysodeikticus , activity of lysozyme conjugated to positively charged nanoparticles was approximately twice as large as that of free lysozyme, while lysozyme conjugated to negatively charged nanoparticles showed little detectable activity. At the same time, when assayed using a low‐molecular weight oligosaccharide substrate, lysozyme attached to both positively and negatively charged nanoparticles showed slightly lower activity than free enzyme. A possible explanation of these results is that lysozyme attached to negatively charged nanoparticles cannot be effectively targeted to the bacteria because of the electrostatic Coulombic repulsion from the negatively charged bacterial cell walls, whereas lysozyme conjugated to positively charged nanoparticles was targeted better than free enzyme due to stronger electrostatic attraction to bacteria. Zeta potential measurements confirmed the validity of this hypothesis. Thus, nanoparticle charge is an important factor that can be used to control targeting and activity of protein‐nanoparticle conjugates. Biotechnol. Bioeng. 2008;100: 403–412. © 2008 Wiley Periodicals, Inc.