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

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.

Design and Preparation of Antimicrobial Agents

“…Serrano-Aguirre and co-workers [ 3 ] focused on the heterologous expression, purification, and characterization of a new N -acylhomoserine lactone (AHL) acylase from Actinoplanes utahensis NRRL 12,052 ( Au AHLA) that can hydrolyze a wide range of substrates, including different natural penicillins and several N -acyl-L-homoserine lactones, both aliphatic and 3-oxo substituted. Moreover, this enzyme exhibits quorum-quenching activity by inhibiting the production of violacein by C. violaceum CV026.…”

Bioassay-Guided Fractionation Leads to the Detection of Cholic Acid Generated by the Rare Thalassomonas sp.

“…The evolutionary relationship between the predicted penicillin acylases from Thalassomonas and other members of the Ntn-hydrolase family was further assessed through a comparison with 32 enzymes with reported AHL acylase activity, penicillin acylase activity and bile salt hydrolase activity [ 48 , 49 ] ( Table S8 and Figure 5 ).…”

A Novel Lipase from Streptomyces exfoliatus DSMZ 41693 for Biotechnological Applications

“…T104. The suitability of this system relies mainly on the similar codon usage of several actinomycetes and its ability to recognize and cleave native signal peptides, as reported for the expression of some polyhydroxyalkanoate depolymerizes from Streptomyces exfoliatus [35,36] and Streptomyces ascomycinicus [37], as well as an N-acylhomoserine lactone acylase from Actinoplanes utahensis [38]. In this work, successful extracellular production of two lipases (SeLipA and SeLipC) from S. exfoliatus has been achieved since enzymatic activity was detected in culture broths of recombinant Rhodococcus sp.…”