(316b) Combinatorial Experimental and Computational Approach for the Effective Entrapment of Glucose Oxidase in Hyaluronic Acid Nanogels

Authors: 
Chapman, J., West Virginia University
Ismail, A. E., West Virginia University
Dinu, C. Z., West Virginia University
Biocatalysis offers important advantages over traditional chemical catalysts when sustainable technology pathways are being considered. The advantages of high specificity, activity, mild reaction conditions, and high product selectivity of the biocatalysts are however outflanked by the limitations that an enzymatic approach will have in commercial-scale processes, namely loss of enzyme stability over time, narrow process operating conditions, and requirement of intensive and costly separation processes from products. Much work is currently done in the field of enzyme immobilization for enzymatic activity retention and reusability, as well as for improvement of product yield, all at low implementation costs. In particular, the immobilization of enzymes onto renewable biopolymers has recently gained attention for both its sustainable approach and its abundant, inexpensive starting materials. Herein we evaluate the suitability of a naturally occurring polysaccharide and main component of the extracellular matrix as both a support as well as an active and protective encapsulator during enzyme immobilization. For this, we combined hyaluronic acid (HA) with model enzyme glucose oxidase (GOx) and evaluated both the enzyme’s functionality at the interface with the polysaccharide as well as its ability to catalyze direct electron transfer controlled reactions. The reaction kinetics at the interface have been elucidated both experimentally using spectroscopic and atomic force microscopy assays and computationally using molecular dynamics simulations to offer both a viable mean for understanding transfer reactions when product formation is being considered as well as for evaluating the effectiveness of immobilization. Our combinatorial approaches provide a feasible strategy to control enzyme-based interface reactions that allow for fast product formation while preserving and extending enzyme functionality and stability. Further, our results provide a viable approach to implement enzyme-based immobilization for different applications ranging from glucose biosensors for the control of diabetes to stabilizers for food preservatives and colors.