Engineering Commons^™

Influence Of Macromolecular Crowding And Confinement On Enzyme Activity And Structure Under Native And Denaturing Conditions, Steven Long, Jeffrey Kunkel, Prashanth Asuri

Bioengineering

It is now well appreciated that both crowding and confinement influence enzyme structure and function due to excluded volume effects; however, the relative efficacies of these environments on protein fates remain unclear due to a lack of direct comparison studies. In this study, we explore the use of the biopolymer alginate to develop an in vitro platform to investigate the effects of both crowding and confinement on the behavior of two model enzymes - horseradish peroxidase and β-galactosidase. Alginate, in its solution phase, can be used as a crowding agent and, in its gel phase by crosslinking using divalent cations, …