Determinants and Prediction of Esterase Substrate Promiscuity Patterns.

Area of Science:

• Biochemistry
• Enzymology
• Structural Biology

Background:

• Esterases are crucial enzymes in biological systems and chemical synthesis.
• Predicting esterase substrate promiscuity and understanding its molecular basis are key challenges.
• Current limitations hinder the discovery of versatile biocatalysts and insights into esterase function.

Purpose of the Study:

• To analyze the substrate spectra of diverse microbial esterases.
• To identify factors influencing esterase substrate range.
• To develop a predictive method for esterase promiscuity from sequence data.

Main Methods:

• Extensive analysis of 145 microbial esterases tested with 96 diverse esters.
• Combination of substrate range pattern analysis, structural analysis, and protein-ligand simulations.
• Identification and validation of a structural parameter: active site effective volume.

Main Results:

• A structural parameter, active site effective volume (cavity volume/SASA), accurately predicts esterase promiscuity (94% accuracy).
• Esterases with higher active site effective volumes exhibit broader substrate spectra.
• This parameter is transferable to other enzyme families, like phosphatases.

Conclusions:

• Active site effective volume is a reliable predictor of esterase substrate range from sequence data.
• This finding facilitates low-cost bioprospecting for esterases with broad substrate specificity.
• The method offers new insights into enzyme structure-function relationships and biocatalyst design.