Novel Bifunctional Acylase from Actinoplanes utahensis: A Versatile Enzyme to Synthesize Antimicrobial Compounds and Use in Quorum Quenching Processes.

Lara Serrano-Aguirre,Rodrigo Velasco-Bucheli,Ana Saborido,Begoña García-Álvarez,Miguel Arroyo,Isabel De La Mata

doi:10.3390/antibiotics10080922

Lara Serrano-Aguirre, Rodrigo Velasco-Bucheli + Show 4 more

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https://doi.org/10.3390/antibiotics10080922

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Journal: Antibiotics	Publication Date: Jul 29, 2021
Citations: 6	License type: CC BY 4.0

Affiliation: Universidad Complutense de Madrid

Abstract

Many intercellular communication processes, known as quorum sensing (QS), are regulated by the autoinducers N-acyl-l-homoserine lactones (AHLs) in Gram-negative bacteria. The inactivation of these QS processes using different quorum quenching (QQ) strategies, such as enzymatic degradation of the autoinducers or the receptor blocking with non-active analogs, could be the basis for the development of new antimicrobials. This study details the heterologous expression, purification, and characterization of a novel N-acylhomoserine lactone acylase from Actinoplanes utahensis NRRL 12052 (AuAHLA), which can hydrolyze different natural penicillins and N-acyl-homoserine lactones (with or without 3-oxo substitution), as well as synthesize them. Kinetic parameters for the hydrolysis of a broad range of substrates have shown that AuAHLA prefers penicillin V, followed by C12-HSL. In addition, AuAHLA inhibits the production of violacein by Chromobacterium violaceum CV026, confirming its potential use as a QQ agent. Noteworthy, AuAHLA is also able to efficiently synthesize penicillin V, besides natural AHLs and phenoxyacetyl-homoserine lactone (POHL), a non-natural analog of AHLs that could be used to block QS receptors and inhibit signal of autoinducers, being the first reported AHL acylase capable of synthesizing AHLs.

Highlights

Published: 29 July 2021N-acyl-L-homoserine lactones (AHLs) are small signal molecules involved in intercellular communication processes, known as quorum sensing (QS), in Gram-negative bacteria.QS allows a bacterial community to synchronize the expression of a set of target genes in order to respond to environmental changes in a density-dependent manner [1]
Among Quorum quenching (QQ) enzymes, the main groups of AHL-degrading or modifying enzymes are lactonases, which catalyze the hydrolysis of the lactone ring; acylases, which catalyze the hydrolysis of the amide bond to form the homoserine lactone ring (HSL) and the corresponding fatty acid; and oxidoreductases, which reduce the carbonyl to hydroxyl [7,8,9]
We demonstrate its capacity to carry out the enzymatic synthesis of penicillin V, as well as natural AHLs and non-natural analogs of AHLs that could be used in QQ processes, being the first example of AHLs enzymatic synthesis catalyzed by an acylase

Summary

Introduction

N-acyl-L-homoserine lactones (AHLs) are small signal molecules involved in intercellular communication processes, known as quorum sensing (QS), in Gram-negative bacteria. QS allows a bacterial community to synchronize the expression of a set of target genes in order to respond to environmental changes in a density-dependent manner [1]. Several biological functions, such as secretion of virulence factors and biofilm formation, are regulated by this mechanism in many pathogenic microorganisms [2,3,4]. Quorum quenching (QQ) processes consist of the interference of QS by inhibition of the biosynthesis or detection of signal molecules or by enzymatic modification or degradation of the autoinducers [6]. QQ enzymes could be used for biotechnological applications such as antivirulence tool in the control and treatment of bacterial infections [10] and decreasing of biofouling in membrane bioreactors [11,12,13]

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