Mechanoenzymatic Breakdown of Chitinous Material to N-Acetylglucosamine: The Benefits of a Solventless Environment.

Chitin is not only the most abundant nitrogen-containing biopolymer on the planet, but also a renewable feedstock that is often treated as a waste. Current chemical methods to break down chitin typically employ harsh conditions, large volumes of solvent, and generate a mixture of products. Although enzymatic methods have been reported, they require a harsh chemical pretreatment of the chitinous substrate and rely on dilute solution conditions that are remote from the natural environment of microbial chitinase enzymes, which typically consists of surfaces exposed to air and moisture. We report an innovative and efficient mechanoenzymatic method to hydrolyze chitin to the N-acetylglucosamine monomer by using chitinases under the recently developed reactive aging (RAging) methodology, based on repeating cycles of brief ball-milling followed by aging, in the absence of bulk solvent. Our results demonstrate that the activity of chitinases increases several times by switching from traditional solution-based conditions of enzymatic catalysis to solventless RAging, which operates on moist solid substrates. Importantly, RAging is also highly efficient for the production of N-acetylglucosamine directly from shrimp and crab shell biomass without any other processing except for a gentle wash with aqueous acetic acid.