Sac Pox from the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius is a proficient lactonase

Bzdrenga, Janek; Hiblot, Julien; Gotthard, Guillaume; Champion, Charlotte; Elias, Mikael; Chabrière, Éric

doi:10.1186/1756-0500-7-333

Cited by 27 publications

(18 citation statements)

References 61 publications

(127 reference statements)

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“…Notably, the limited dynamic range of the pH indicator based assay used could limit the observation of lower K M values. The observed K M value for GcL is in contrast to values observed for other MLL lactonases, such as AiiA and AiiB ( K M ≈1600–5600 μ m ), and other classes of lactonases (e.g., PLLs and PONs; K M ≈50–500 μ m ). Yet, these low K M values are similar to those observed for AidC and AaL .…”

Section: Resultssupporting

confidence: 91%

“…The phosphotriesterase‐like lactonases (PLLs) exhibit an (α/β) 8 fold and are found in archaea and bacteria. PLL representatives were found to hydrolyze δ‐lactones, γ‐lactones, and AHLs and showed substrate preference for long acyl chain AHLs . A second family of lactonases are the paraoxonases (PONs), primarily isolated from mammals, exhibit a six‐bladed β‐propeller fold .…”

Section: Introductionmentioning

confidence: 99%

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The Structural Determinants Accounting for the Broad Substrate Specificity of the Quorum Quenching Lactonase GcL

et al. 2019

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Quorum quenching lactonases are enzymes capable of hydrolyzing lactones, including N‐acyl homoserine lactones (AHLs). AHLs are molecules known as signals in bacterial communication dubbed quorum sensing. Bacterial signal disruption by lactonases was previously reported to inhibit behavior regulated by quorum sensing, such as the expression of virulence factors and the formation of biofilms. Herein, we report the enzymatic and structural characterization of a novel lactonase representative from the metallo‐β‐lactamase superfamily, dubbed GcL. GcL is a broad spectrum and highly proficient lactonase, with kcat/KM values in the range of 104 to 106 m−1 s−1. Analysis of free GcL structures and in complex with AHL substrates of different acyl chain length, namely, C4‐AHL and 3‐oxo‐C12‐AHL, allowed their respective binding modes to be elucidated. Structures reveal three subsites in the binding crevice: 1) the small subsite where chemistry is performed on the lactone ring; 2) a hydrophobic ring that accommodates the amide group of AHLs and small acyl chains; and 3) the outer, hydrophilic subsite that extends to the protein surface. Unexpectedly, the absence of structural accommodation for long substrate acyl chains seems to relate to the broad substrate specificity of the enzyme.

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Section: Resultssupporting

confidence: 91%

Section: Introductionmentioning

confidence: 99%

The Structural Determinants Accounting for the Broad Substrate Specificity of the Quorum Quenching Lactonase GcL

et al. 2019

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“…Lactonases are naturally occurring enzymes and can be found in a variety of organisms, including bacteria, archaea, plants, fungi, and mammals (Elias and Tawfik, 2012;LaSarre and Federle, 2013). Lactonases can be found in various protein families, including the paraoxonases (PONs) (Khersonsky and Tawfik, 2005;Ben-David et al, 2012, the phosphotriesteraselike lactonases (PLLs) (Afriat et al, 2006;Elias and Tawfik, 2012;Hiblot et al, 2013Hiblot et al, , 2015Bzdrenga et al, 2014) and the metallo-β-lactamases lactonases (MLLs) (Liu et al, 2007(Liu et al, , 2008Momb et al, 2008;Mascarenhas et al, 2015;Tang et al, 2015;Bergonzi et al, 2016Bergonzi et al, , 2018. Remarkably, while AHLs vary considerably in their chemical structure, and in particular, the nature and length of their acyl chain, recent work on lactonase kinetic properties suggest, in contrast, a low variety in the lactonase's substrate specificities.…”

Section: Introductionmentioning

confidence: 99%

Effects of Signal Disruption Depends on the Substrate Preference of the Lactonase

et al. 2020

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Many bacteria produce and use extracellular signaling molecules such as acyl homoserine lactones (AHLs) to communicate and coordinate behavior in a cell-density dependent manner, via a communication system called quorum sensing (QS). This system regulates behaviors including but not limited to virulence and biofilm formation. We focused on Pseudomonas aeruginosa, a human opportunistic pathogen that is involved in acute and chronic lung infections and which disproportionately affects people with cystic fibrosis. P. aeruginosa infections are becoming increasingly challenging to treat with the spread of antibiotic resistance. Therefore, QS disruption approaches, known as quorum quenching, are appealing due to their potential to control the virulence of resistant strains. Interestingly, P. aeruginosa is known to simultaneously utilize two main QS circuits, one based on C4-AHL, the other with 3-oxo-C12-AHL. Here, we evaluated the effects of signal disruption on 39 cystic fibrosis clinical isolates of P. aeruginosa, including drug resistant strains. We used two enzymes capable of degrading AHLs, known as lactonases, with distinct substrate preference: one degrading 3-oxo-C12-AHL, the other degrading both C4-AHL and 3-oxo-C12-AHL. Two lactonases were used to determine the effects of signal disruption on the clinical isolates, and to evaluate the importance of the QS circuits by measuring effects on virulence factors (elastase, protease, and pyocyanin) and biofilm formation. Signal disruption results in at least one of these factors being inhibited for most isolates (92%). Virulence factor activity or production were inhibited by up to 100% and biofilm was inhibited by an average of 2.3 fold. Remarkably, the treatments led to distinct inhibition profiles of the isolates; the treatment with the lactonase degrading both signaling molecules resulted in a higher fraction of inhibited isolates (77% vs. 67%), and the simultaneous inhibition of more virulence factors per strain (2 vs. 1.5). This finding suggests that the lactonase AHL preference is key to its inhibitory spectrum and is an essential parameter to improve quorum quenching strategies.

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“…They belong to the amidohydrolase superfamily and adopt a (/) 8 fold (Afriat et al, 2006;Elias et al, 2008;Del Vecchio et al, 2009). PLLs have been isolated from numerous sources (Xiang et al, 2009;Hawwa et al, 2009), including extremophilic archaea (Hiblot et al, 2012(Hiblot et al, , 2015Bzdrenga et al, 2014;. PLLs have been subdivided into two classes: PLL-As are proficient against -lactones, -lactones and AHLs, whereas PLL-Bs prefer -lactones and -lactones (Hiblot et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

The quorum-quenching lactonase fromGeobacillus caldoxylosilyticus: purification, characterization, crystallization and crystallographic analysis

Bergonzi¹,

Schwab²,

Elias³

2016

Acta Crystallogr F Struct Biol Commun

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Lactonases are enzymes that are capable of hydrolyzing various lactones such as aliphatic lactones or acyl-homoserine lactones (AHLs), with the latter being used as chemical signaling molecules by numerous Gram-negative bacteria. Lactonases therefore have the ability to quench the chemical communication, also known as quorum sensing, of numerous bacteria, and in particular to inhibit behaviors that are regulated by this system, such as the expression of virulence factors or the production of biofilms. A novel representative from the metallo-β-lactamase superfamily, dubbed GcL, was isolated from the thermophilic bacterium Geobacillus caldoxylosilyticus. Because of its thermophilic origin, GcL may constitute an interesting candidate for the development of biocontrol agents. Here, we show that GcL is a thermostable enzyme with a half-life at 75°C of 152.5 ± 10 min. Remarkably, it is also shown that GcL is among the most active lactonases characterized to date, with catalytic efficiencies (kcat/Km) against AHLs of greater than 10(6) M(-1) s(-1). The structure of GcL is expected to shed light on the catalytic mechanism of the enzyme and the molecular determinants for the substrate specificity in this class of lactonases. Here, the expression, purification, characterization, crystallization and X-ray diffraction data collection to 1.6 Å resolution of GcL are reported.

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Sac Pox from the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius is a proficient lactonase

Cited by 27 publications

References 61 publications

The Structural Determinants Accounting for the Broad Substrate Specificity of the Quorum Quenching Lactonase GcL

The Structural Determinants Accounting for the Broad Substrate Specificity of the Quorum Quenching Lactonase GcL

Effects of Signal Disruption Depends on the Substrate Preference of the Lactonase

The quorum-quenching lactonase fromGeobacillus caldoxylosilyticus: purification, characterization, crystallization and crystallographic analysis

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