Strategies for GH functional annotation: toward harmonized and predictive CAZymology

Area of Science:

• Biochemistry
• Genomics
• Enzymology

Background:

• Functional annotation and biochemical characterization of glycoside hydrolases (GHs) for polysaccharide deconstruction present significant challenges.
• The field is transitioning from traditional "activity first" biochemical assays to modern "sequence first" discovery methods leveraging genomics.

Purpose of the Study:

• To address challenges in GH characterization, including substrate specificity, assay variability, and reporting inconsistencies.
• To provide a practical framework for characterizing newly identified GHs.

Main Methods:

• Compilation of a comprehensive list of substrates for each GH family's glycosidase activities.
• Review of techniques used for GH characterization.

Main Results:

• Reported 169 glycosidase activities, their substrates, and characterization techniques.
• Highlighted challenges arising from GH family diversity, overlapping specificities, and inconsistent methodologies.

Conclusions:

• Standardized substrates, protocols, and reporting practices are essential for harmonized GH functional annotation.
• Community-driven data sharing and adoption of best practices offer a roadmap for predictive enzymology.
• Acknowledged current limitations in data standardization and reporting heterogeneity.