Engineering the Performance of Artificial Inclusion Bodies Built of Catalytic β‑Galactosidase

JULIETA M. SANCHEZ; HECTOR LOPEZ LAGUNA; NAROA SERNA; UGUTZ UNZUETA ,; PEDRO D CLOP; ANTONIO VILLAVERDE; ESTHER VÁZQUEZ

ACS Sustainable Chemistry and Engineering

ACS publications

Año: 2021

2168-0485

One of the most critical bottlenecks in the application of industrial enzymesis the preservation of protein stability throughout the catalytic reaction, which oftenrequires protein engineering and/or process optimization. In this context, we havedesigned and deeply characterized an efficient, stable, and reusable enzymatic platformbased on the Escherichia coli β-galactosidase. The enzyme was assembled in vitro, by usingdivalent cations as molecular linkers, as stable protein microparticles showing catalyticactivity. In this assembled microstructure, β-galactosidase exhibits a particularconformation within the microparticles, sharing structural traits (a high cross-parallelbeta-sheet content) with the bacterial inclusion bodies and secretory amyloids from themammalian endocrine system. This fact confers enhanced thermal stability compared to the soluble protein version and ensures highreusability in industry-oriented processes. On the other hand, among the catalog of cations tested as molecular linkers, a mixture ofCa2+ and Mg2+ offers the best performance to the catalytic particle. Altogether, these data offer clues for the application of a selfimmobilizedenzymatic platform with transversal applicability and enormous potential in biotechnology and biomedicine.