Exploring Metagenomic Enzymes: A Novel Esterase Useful for Short-Chain Ester Synthesis

Abstract

Enzyme-mediated esterification reactions can be a promising alternative to produce esters of commercial interest, replacing conventional chemical processes. The aim of this work was to verify the potential of an esterase for ester synthesis. For that, recombinant lipolytic enzyme EST5 was purified and presented higher activity at pH 7.5, 45 °C, with a Tm of 47 °C. Also, the enzyme remained at least 50% active at low temperatures and exhibited broad substrate specificity toward p-nitrophenol esters with highest activity for p-nitrophenyl valerate with a Kcat/Km of 1533 s−1 mM−1. This esterase exerted great properties that make it useful for industrial applications, since EST5 remained stable in the presence of up to 10% methanol and 20% dimethyl sulfoxide. Also, preliminary studies in esterification reactions for the synthesis of methyl butyrate led to a specific activity of 127.04 U·mg−1. The enzyme showed higher esterification activity compared to other literature results, including commercial enzymes such as LIP4 and CL of Candida rugosa assayed with butyric acid and propanol which showed esterification activity of 86.5 and 15.83 U·mg−1, respectively. In conclusion, EST5 has potential for synthesis of flavor esters, providing a concept for its application in biotechnological processes.

Structural Insights into Carboxylic Polyester-Degrading Enzymes and Their Functional Depolymerizing Neighbors

“…The most well studied system is the PETase isolated from the marine bacterium I. sakaiensis , which showed an increased degradation yield when compared with other similar enzymes [ 20 ]. However, the number and diversity of polyester-degrading enzymes is expected to be expanded in the following years with the onset of the high-throughput metagenomic analysis of natural samples [ 21 , 52 ].…”

Unravelling the hidden power of esterases for biomanufacturing of short-chain esters

“…A positive Gibbs free energy for biological esterification has been attributed to this limited interest 1 , 2 . However, there are pieces of evidence for biological esterification by esterases, which contradict these estimations 19 – 21 . The Gibbs free energy of reaction estimated in these earlier reports were based on an online interface, eQuilibrator 1 , 2 , 13 .…”

Enhanced Activity and Stability of an Acetyl Xylan Esterase in Hydrophilic Alcohols through Site-Directed Mutagenesis

“…Esterases with novel properties are widely needed due to their potential in catalysing diverse reactions such as deacetylation of the xylan backbone, degradation of poly(ethylene terephthalate), and the synthesis of short-chain esters [40,41]. Biocatalysis in organic solvents is desired due to the possibility of performing reactions that are restricted kinetically or thermodynamically in an aqueous environment, improved solubility of substrates, easier product recovery, and because the possibility of side-reactions and inhibitory compounds occurrence is reduced [42].…”