A Selective Phenyl Azo Acetate Substrate for Enhanced Carbonic Anhydrase Functional Analysis.

Authors

Justinas Babinskas, Inga Matijošytė

Published in

Analytical science advances. Volume 7. Issue 1. Pages e70091. Epub May 26, 2026.

Abstract

Carbon dioxide (CO₂) sequestration is a key strategy for mitigating climate change, and biocatalytic approaches offer sustainable, cost-effective alternatives to chemical and physical methods. Carbonic anhydrases (CAs) efficiently catalyse the reversible hydration of CO₂ to bicarbonate, yet their large-scale application is constrained by limited stability and the lack of robust analytical tools for activity and/or inhibition profiling. In this work, we synthesised and characterised a novel phenyl azo dye acetate to address these limitations. The substrate was optimised for spectrophotometric assays, enabling reliable kinetic and inhibition analyses of CAs. Compared with conventional substrates such as p-nitrophenyl acetate and indoxyl acetate, the new assay demonstrated more than 4.5-fold higher sensitivity, substantial thermal stability and wider applicability across tested hydrolases. These findings provide a more robust platform for evaluating CA activity, supporting the development of more efficient biocatalytic systems for CO₂ sequestration.

PMID:
42255016
Bibliographic data and abstract were imported from PubMed on 14 Sep 2026.

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