Recent advances in engineering proteins for biocatalysis

Abstract

Protein engineers are increasingly able to rely on structure-function insights, computational methods, and deeper understanding of natural biosynthesis processes, to streamline the design and applications of enzymes. This review highlights recent successes in applying new or improved protein engineering strategies toward the design of improved enzymes and enzymes with new activities. We focus on three approaches: structure-guided protein design, computational design, and the use of novel scaffolding and compartmentalization techniques to improve performance of multienzyme systems. Examples described address problems relating to enzyme specificity, stability, and/or activity, or aim to balance sequential reactions and route intermediates by co-localizing multiple enzymes. Specific applications include improving production of biofuels using enzymes with altered cofactor specificity, production of high-value chiral compounds by enzymes with tailored substrate specificities, and accelerated cellulose degradation via multi-enzyme scaffold assemblies. Collectively, these studies demonstrate a growing variety of computational and molecular biology tools. Continued advances on these fronts coupled with better mindfulness of how to apply proteins in unique ways offer exciting prospects for future protein engineering and biocatalysis research.

Probing the Unfolded Configurations of a β-Hairpin Using Sketch-Map

“…The minima corresponding to the beta-hairpin configuration becomes deeper as do the satellite basins around it that correspond to the various partially unfolded configurations of this structure. 6 shows that the free energy difference between the perfect-alpha helix structure and the folded beta hairpin is larger for this mutated protein than it is in the wild type. Even so the alpha-helix still has a reasonably low free energy so the system would still expect to spend a considerable fraction of its time in alpha helical configurations.…”

Biocatalytic Epoxidation for Green Synthesis

“…Protein engineering, a design or optimization process of proteins, has become a powerful tool in biotechnology to generate engineered biocatalysts to meet the production requirements of pharmaceuticals, biofuels, and other chemicals [124][125][126][127][128]. SMO-catalyzed enantioselective epoxidation of (a) primary ally lic alcohols, (b) nonconjugated, and (c) conjugated secondary allylic alcohols.…”

Tuning the catalytic properties of lipases immobilized on divinylsulfone activated agarose by altering its nanoenvironment

“…Lipases, due to the flexibility of their active center may be very easily tuned via genetic modification [44][45][46][47], physical or chemical modification [48][49][50][51] or immobilization [34,43]. Immobilization has revealed itself as a potent tool to tuning lipase properties, even although the results are difficult to predict and are based on the building of a large library of biocatalysts immobilized following different protocols (trial and error) [34,43].…”