Epoxide hydrolases are attractive and industrially important biocatalysts. They can catalyze the enantioselective hydrolysis of epoxides to the corresponding diols as chiral building blocks for bioactive compounds and drugs. In this review article, we discuss the state of the art and development potential of epoxide hydrolases as biocatalysts based on the most recent approaches and techniques. The review covers new approaches to discover epoxide hydrolases using genome mining and enzyme metagenomics, as well as improving enzyme activity, enantioselectivity, enantioconvergence, and thermostability by directed evolution and a rational design. Further improvements in operational and storage stabilization, reusability, pH stabilization, and thermal stabilization by immobilization techniques are discussed in this study. New possibilities for expanding the synthetic capabilities of epoxide hydrolases by their involvement in non-natural enzyme cascade reactions are described.

The inhibition effects (I₅₀) of a series of dyes (Cibacrom Blue 3G-A, Procion Blue MX-R, Remazo Brilliant Blue and Ostazin Brilliant Red S5-B) covalently linked to dextran T 70 were the criterion of the relationship between the structure of antraquinone-triazine compounds and their affinity to rabbit muscle lactate dehydrogenase (LDH). The I₅₀ values indicate a substantial importance of the terminal benzenesulfonane moiety in the structure of Cibacron Blue. Also the degree of substitution of polysaccharide plays an important role in the affinity; its influence upon I₅₀ can be expressed for Cibacron Blue-Dextrans T 70 by an equation of line. The influence of the structure of the polysaccharide constituent on the affinity of Cibacron Blue-(α-glucans) towards LDH is discussed, as well.

A novel immobilization matrix for the entrapment of viable whole-cell Baeyer–Villiger monooxygenase was developed. Viable recombinant Escherichia coli cells overexpressing cyclohexanone monooxygenase were entrapped in polyelectrolyte complex beads prepared by a two-step reaction of oppositely-charged polymers including highly defined cellulose sulphate. Immobilized cells exhibited higher operational stability than free cells during 10 repeated cycles of Baeyer–Villiger biooxidations of rac-bicyclo[3.2.0]hept-2-en-6-one to the corresponding lactones (1R,5S)-3-oxabicyclo-[3.3.0]oct-6-en-3-one and (1S,5R)-2-oxabicyclo-[3.3.0]oct-6-en-3-one. The morphology of polyelectrolyte complex beads was characterised by environmental scanning electron microscopy; the spatial distribution of polymers in the beads and cell viability were examined using confocal laser scanning microscopy, and the texture was characterised by the mechanical resistance measurements.