Immobilized Biocatalyst Engineering: High throughput enzyme immobilization for the integration of biocatalyst improvement strategies

Rodriguez-Nunez, Karen; Bernal, Claudia; Martinez, Ronny

Abstract

The increasing use of sustainable manufacturing technologies in the industry presents a constant challenge for the development of suitable biocatalysts. Traditionally, improved biocatalysts are developed either using protein engineering (PE) or enzyme immobilization (EI). However, these approaches are usually not simultaneously applied. In this work, we designed and validated an enzyme improvement platform. Immobilized Biocatalyst Engineering (IBE), which simultaneously integrates PE and EL with a unique combination of improvement through amino acid substitutions and attachment to a support material, allowing to select variants that would not be found through single or subsequent PE and EI improvement strategies. Our results show that there is a significant difference on the best performing variants identified through IBE, when compared to those that could be identified as soluble enzymes and then immobilized, especially when evaluating variants with low enzyme as soluble enzymes and high activity when immobilized. IBE allows evaluating thousands of variants in a short time through an integrated screening, and selection can be made with more information, resulting in the detection of highly stable and active heterogeneous biocatalysts. This novel approach can translate into a higher probability of finding suitable biocatalysts for highly demanding processes. (C) 2020 Elsevier B.V. All rights reserved.

Más información

Título según WOS: Immobilized Biocatalyst Engineering: High throughput enzyme immobilization for the integration of biocatalyst improvement strategies
Título de la Revista: INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Volumen: 170
Editorial: Elsevier
Fecha de publicación: 2021
Página de inicio: 61
Página final: 70
DOI:

10.1016/j.ijbiomac.2020.12.097

Notas: ISI