From Protein Engineering to Immobilization: Promising Strategies for the Upgrade of Industrial Enzymes

Abstract

Enzymes found in nature have been exploited in industry due to their inherent catalytic properties in complex chemical processes under mild experimental and environmental conditions. The desired industrial goal is often difficult to achieve using the native form of the enzyme. Recent developments in protein engineering have revolutionized the development of commercially available enzymes into better industrial catalysts. Protein engineering aims at modifying the sequence of a protein, and hence its structure, to create enzymes with improved functional properties such as stability, specific activity, inhibition by reaction products, and selectivity towards non-natural substrates. Soluble enzymes are often immobilized onto solid insoluble supports to be reused in continuous processes and to facilitate the economical recovery of the enzyme after the reaction without any significant loss to its biochemical properties. Immobilization confers considerable stability towards temperature variations and organic solvents. Multipoint and multisubunit covalent attachments of enzymes on appropriately functionalized supports via linkers provide rigidity to the immobilized enzyme structure, ultimately resulting in improved enzyme stability. Protein engineering and immobilization techniques are sequential and compatible approaches for the improvement of enzyme properties. The present review highlights and summarizes various studies that have aimed to improve the biochemical properties of industrially significant enzymes.

Effects of Temperature and pH on Immobilized Laccase Activity in Conjugated Methacrylate-Acrylate Microspheres

“…Immobilization is always a successful strategy for stabilizing enzymes or improving their performance [8]. Bezerra et al (2015) gave guidelines on the selection of suitable solid supports and immobilization conditions [9].…”

Role of Glutaraldehyde in Imparting Stability to Immobilized β-Galactosidase Systems

“…Therefore, numerous technologies and novel carrier molecules have been adopted to improve βG immobilization and enhance enzyme loading, activity and stability in order to reduce overall cost of biocatalytic activity at industrial scale 51 . These approaches encompass formation of crosslinked enzyme aggregates wherein active enzyme aggregates are immobilized using a multifunctional crosslinker that provides extra stability and enhanced their recyclability during their use as biocatalysts 52 .…”

Enantioselective Transamination in Continuous Flow Mode with Transaminase Immobilized in a Macrocellular Silica Monolith

“…Indeed, the use of free enzymes in homogeneous batch processes suffers from severe limitations related to the impossibility to reuse and recover the biocatalysts [1]. Therefore, one particular challenge is to make them recyclable while maintaining their activity [2]. This is typically achieved by immobilizing the enzymes on an appropriate support-inorganic, biological, polymer or hybrid materials [3,4].…”