Recent progress in engineering α/β hydrolase‐fold family members

Abstract

The members of the alpha/beta hydrolase-fold family represent a functionally versatile group of enzymes with many important applications in biocatalysis. Given the technical significance of alpha/beta hydrolases in processes ranging from the kinetic resolution of enantiomeric precursors for pharmaceutical compounds to bulk products such as laundry detergent, optimizing and tailoring enzymes for these applications presents an ongoing challenge to chemists, biochemists, and engineers alike. A review of the recent literature on alpha/beta hydrolase engineering suggests that the early successes of "random processes" such as directed evolution are now being slowly replaced by more hypothesis-driven, focused library approaches. These developments reflect a better understanding of the enzymes' structure-function relationship and improved computational resources, which allow for more sophisticated search and prediction algorithms, as well as, in a very practical sense, the realization that bigger is not always better.

Widespread Head-to-Head Hydrocarbon Biosynthesis in Bacteria and Role of OleA

“…Thousands of homologous sequences were found. The sequence and catalytic diversity within each superfamily were sufficiently broad that standard sequence alignment tools did not align amino acid residues that are known to comprise the active sites in proteins for which X-ray structures are available (15,25,27,28,32,34,44,49,56,61,71,74). Thus, to properly align Ole protein sequences with other proteins in their respective superfamilies, it was necessary to generate structure-based alignments.…”

“…For each OleABCD alignment, 6 to 10 homologous proteins from previously described high-resolution X-ray structures were structurally superimposed, using the Match command in Chimera (42). Conserved residues within each superfamily of homologs were derived from the literature (15,25,27,28,32,34,44,49,56,61,71,74), and their locations were plotted onto the protein backbone to confirm alignments. Sequence alignments based on the structure alignments were exported.…”

Crystal Structure of the Cystic Fibrosis Transmembrane Conductance Regulator Inhibitory Factor Cif Reveals Novel Active-Site Features of an Epoxide Hydrolase Virulence Factor

“…In mammals, they metabolize potent chemical signal mediators (9). As a family, they also are of potential biocatalytic interest (46). EHs have not previously been reported as bacterial virulence factors, but Pseudomonas is adept at exploiting a wide variety of biochemical strategies to subvert host cell functions.…”

Improved triglyceride transesterification by circular permuted Candida antarctica lipase B

“…Higher productivity represents one approach for reducing costs and can be accomplished through the optimization of reaction conditions and the identification of better catalyst. The search for new enzymes in nature has been a promising strategy while engineering biocatalysts by rational design and directed evolution has produced enzymes with improved properties (Qian et al, 2007b).…”