Abstract
Enzymes are essential and ubiquitous biocatalysts involved in various metabolic pathways and used in manyindustrial processes. Here, we reframe enzymes not just as biocatalysts transforming bioproducts but also assensitive probes for exploring the structure and composition of complex bioproducts, like meat tissue, dairyproducts and plant materials, in both food and non-food bioprocesses. This review details the global strategy andpresents the most recent investigations to prepare and use enzymes as relevant probes, with a focus on glycosidehydrolases involved in plant deconstruction and proteases and lipases involved in food digestion. First, to expandthe enzyme repertoire to fit bioproduct complexity, novel enzymes are mined from biodiversity and can beartificially engineered. Enzymes are further characterized by exploring sequence/structure/dynamics/functionrelationships together with the environmental factors influencing enzyme interactions with their substrates.Then, the most advanced experimental and theoretical approaches developed for exploring bioproducts atvarious scales (from nanometer to millimeter) using active and inactive enzymes as probes are illustrated.Overall, combining multimodal and multiscale approaches brings a better understanding of native-form ortransformed bioproduct architecture and composition, and paves the way to mainstream the use of enzymes asprobes