In the last decades, cascade biocatalysis has attracted significant interest from both academia and industry as a promising technology for green and sustainable chemical production. Unlike the step-by-step synthesis, cascade biocatalysts accomplish two or more consecutive reactions in one-pot, thus avoiding isolation and purification of intermediates. Nowadays, numerous drugs and fine chemicals, as well as some polymers are synthesized through cascade bio-catalysis. However, the industrial use of biocatalysts based on free enzymes is problematic, due to their poor stability and difficult reuse. A possible way to overcome these limitations is the application of immobilization technologies, in which two or more enzymes are co-localized close to one another on a suitable support. Major advantage of these carrier-multienzyme complexes is that the enzymes´ active sites are in close proximity, minimizing the diffusion of the intermediates thereby enhancing the overall reaction efficiency. In this work various highly porous polyamide microparticles (PAM) based on Polyamide 4 (PA4), Polyamide 6 (PA6), Polyamide 12 (PA12) as well as their co-polymers with and without magnetic properties were synthesized by proper method. PAM were used as scaffolds for ordered sequential co-immobilization of Glucose oxidase (GOx) and Horse radish peroxifdase (HRP). The catalytic efficiency of these hybrid bi-enzyme biocatalysts was assessed in the cascade reaction for glucose detection. The kinetic parameters, as well as the reusability and storage stability were also studied.  

Nadya Vasileva Dencheva has completed his PhD at the age of 45 years and postdoctoral studies from the University of Minho, Department of Polymer Engineering, Portugal. She is Junior Researcher at the Institute for Polymers and Composites research center at the University of Minho. She has published more than 55 papers in international peer reviewed journals, 3 book chapters and 6 patents (Index h 18; Index i10 – 32, Google scholar) http://orcid.org/0000-0003-1698-553X