Enzymes are typically manipulated as dilute aqueous solutions. This is however remote from their natural environment. For example, it is well-recognized that the cell is a highly crowded milieu, with a total molecule concentration estimated to ~400 mg/mL. Furthermore, many enzymes are secreted by microorganisms in the environment (<i>e.g.</i> cellulases, chitinases, and cutinases), and work directly on moist, solid surfaces, in the absence of bulk water. From this perspective, we envisaged that some enzymes might have evolved to work optimally in moist solid mixtures. This led to the development of an unconventional and highly efficient method for biocatalysis without bulk water. Our process, coined reactive aging or RAging, is based on alternating periods of brief, gentle ball milling, followed by aging (<i>i.e.</i> static incubation). RAging is especially promising for poorly soluble and chemically recalcitrant substrates such as biomass and plastic waste.

Enzymatic reactions in moist solids for the depolymerization of biowastes and plastics

ACS Spring 2022

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