Esterases receive special attention because their wide distribution in biological systems and environments and their importance for physiology and chemical synthesis. The prediction of esterases substrate promiscuity level from sequence data and the molecular reasons why certain such enzymes are more promiscuous than others, remain to be elucidated. This limits the surveillance of the sequence space for esterases potentially leading to new versatile biocatalysts and new insights into their role in cellular function. Here we performed an extensive analysis of the substrate spectra of 145 phylogenetically and environmentally diverse microbial esterases, when tested with 96 diverse esters. We determined the primary factors shaping their substrate range by analyzing substrate range patterns in combination with structural analysis and protein-ligand simulations. We found a structural parameter that helps ranking (classifying) promiscuity level of esterases from sequence data at 94% accuracy. This parameter, the active site effective volume, exemplifies the topology of the catalytic environment by measuring the active site cavity volume corrected by the relative solvent accessible surface area (SASA) of the catalytic triad. Sequences encoding esterases with active site effective volumes (cavity volume/SASA) above a threshold show greater substrate spectra, which can be further extended in combination with phylogenetic data. This measure provides also a valuable tool for interrogating substrates capable of being converted. This measure, found to be transferred to phosphatases of the haloalkanoic acid dehalogenase superfamily and possibly other enzymatic systems, represents a powerful tool for low-cost bioprospecting for esterases with broad substrate ranges, in large scale sequence datasets.
Title: Female-biased attraction of Oriental fruit fly, bactrocera dorsalis (Hendel), to a blend of host fruit volatiles from Terminalia catappa L Siderhurst MS, Jang EB Ref: J Chem Ecol, 32:2513, 2006 : PubMed
Coupled gas chromatography-electroantennogram detection (GC-EAD) analysis of volatiles from tropical almond fruit, Terminalia catappa L., revealed 22 compounds that were detected by antennae of oriental fruit fly females, Bactrocera dorsalis (Hendel). Both solid-phase microextraction (SPME) and Porapak Q were used for sampling odors in fruit headspace, with SPME collections producing larger EAD responses from a greater number of compounds. Geranyl acetate and methyl eugenol elicited the largest EAD responses. A synthetic blend containing SPME collected, EAD stimulatory compounds showed female-biased attraction in laboratory wind tunnel bioassays, but heavily male-biased trap captures in a larger olfactometer arena. A nine-component subset of compounds eliciting relatively small EAD responses (EAD minor) and consisting of equal parts ethanol, ethyl acetate, ethyl hexanoate, hexyl acetate, linalyl acetate, ethyl nonanate, nonyl acetate, ethyl cinnamate, and (E)-beta-farnesene, attracted mainly females. This EAD minor blend was as attractive to females and much less attractive to males when compared to torula yeast in field cage experiments using glass McPhail traps. Similar results were obtained with outdoor rotating olfactometer tests in which the EAD minor blend was almost completely inactive for males.
The major component (>90% of volatiles) of the male rectal glandular extract of the nonpest speciesBactrocera visenda (Hardy) is 3-methyl2-butenyl acetate, with minor components being the isomeric 3-methyl-3-butenyl acetate, the homologous esters, 3-methyl-2-butenyl propanoate and 3-methyl-2-butenyl formate, along with 3-methyl-2-buten-1-ol, 3-methyl-2-butenal, and 3-methylbutyl acetate. None of these compounds has been identified previously from aBactrocera species, supporting the view thatBactrocera visenda is taxonomically distant from otherBactrocera species identified from the Australian mainland. This collection of compounds adds to the known types utilized by dipteran species and emphasizes their extensive biosynthetic capability.
