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This review highlights advances from 2020\u20132025 that demonstrate how expanding the amino acid repertoire unlocks new functionalities. We examine how genetically encoded ncAAs diversify and tune metal-binding properties, enabling programmable coordination and redox behavior in engineered enzymes. Further, we explore biocatalytic applications, including multi-fold activity enhancements in natural enzymes and the introduction of entirely novel reactivity in artificial systems. Finally, we discuss the growing use of ncAAs as intrinsic biophysical reporters, which support a wide range of spectroscopic methods for tracking structure, dynamics, and interactions at residue-level resolution. These capabilities establish ncAAs as essential tools that can be deployed at any stage of the protein design process, from constructing new catalytic centers to quantifying molecular behaviors in real time.<\/jats:p>","DOI":"10.1093\/protein\/gzag007","type":"journal-article","created":{"date-parts":[[2026,3,4]],"date-time":"2026-03-04T12:51:23Z","timestamp":1772628683000},"source":"Crossref","is-referenced-by-count":3,"title":["Bind, catalyze, and quantify: a modern protein and enzyme engineering toolbox of genetically encoded non-canonical amino acids"],"prefix":"10.1093","volume":"39","author":[{"given":"S\u00f8ren C","family":"Spina","sequence":"first","affiliation":[{"name":"Department of Chemical and Biomolecular Engineering, The Ohio State University , 151 W. 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