Enzymes are essential and ubiquitous biocatalysts involved in various metabolic pathways and used in many industrial processes. Here, we reframe enzymes not just as biocatalysts transforming bioproducts but also as sensitive probes for exploring the structure and composition of complex bioproducts, like meat tissue, dairy products and plant materials, in both food and non-food bioprocesses. This review details the global strategy and presents the most recent investigations to prepare and use enzymes as relevant probes, with a focus on glycoside-hydrolases involved in plant deconstruction and proteases and lipases involved in food digestion. First, to expand the enzyme repertoire to fit bioproduct complexity, novel enzymes are mined from biodiversity and can be artificially engineered. Enzymes are further characterized by exploring sequence/structure/dynamics/function relationships together with the environmental factors influencing enzyme interactions with their substrates. Then, the most advanced experimental and theoretical approaches developed for exploring bioproducts at various 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 or transformed bioproduct architecture and composition, and paves the way to mainstream the use of enzymes as probes.

酶是参与各种代谢途径并在许多工业过程中使用的必不可少且普遍存在的生物催化剂。在这里，我们不仅将酶重组为转化生物产物的生物催化剂，而且还将其作为敏感探针，用于探索食品和非食品生物过程中复杂生物产物的结构和组成，例如肉组织，乳制品和植物材料。这篇综述详细介绍了全球战略，并提出了有关制备和使用酶作为相关探针的最新研究，重点是与植物解构有关的糖苷水解酶和与食品消化有关的蛋白酶和脂肪酶。首先，为了扩展酶库以适应生物制品的复杂性，从生物多样性中提取了新的酶，可以对其进行人工设计。通过探索序列/结构/动力学/功能关系以及影响酶与其底物相互作用的环境因素来进一步表征酶。然后，举例说明了使用活性和非活性酶作为探针探索各种规模（从纳米到毫米）的生物产物的最先进的实验和理论方法。总体而言，将多模式和多尺度方法相结合，可以更好地理解天然形式或转化的生物产品的结构和组成，并为将酶用作探针的主流铺平道路。举例说明了使用活性和非活性酶作为探针探索各种规模（从纳米到毫米）的生物产物而开发的最先进的实验和理论方法。总体而言，将多模式和多尺度方法相结合，可以更好地理解天然形式或转化的生物产品的结构和组成，并为将酶用作探针的主流铺平道路。举例说明了使用活性和非活性酶作为探针探索各种规模（从纳米到毫米）的生物产物而开发的最先进的实验和理论方法。总体而言，将多模式和多尺度方法相结合，可以更好地理解天然形式或转化的生物产品的结构和组成，并为将酶用作探针的主流铺平道路。