Characterization of a novel thermostable N-acylhomoserine lactonase from the thermophilic bacterium Thermaerobacter marianensis

Morohoshi, Toshio; Tominaga, Yoshiaki; Someya, Nobutaka; Ikeda, Tsukasa

doi:10.1016/j.jbiosc.2014.11.014

Cited by 19 publications

(17 citation statements)

References 27 publications

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“…GcL shares 28.5% sequence identity with AiiA and 44.3% sequence identity with the closest known crystallized lactonase (AiiB). Previous work, possibly on the exact same protein (only identified by N-terminal sequencing), identified a thermophilic enzyme from G. caldoxylosilyticus and demonstrated the catalytic activity of this enzyme against a few lactone substrates (Morohoshi et al, 2015;Seo et al, 2011). Here, we report the protein production, purification, crystallization and preliminary X-ray diffraction data of this new thermophilic representative of the MLL family.…”

Section: Introductionmentioning

confidence: 89%

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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Section: Introductionmentioning

confidence: 89%

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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“…Thermostable AHL lactonase enzymes with maximum activity at high temperature have also been isolated from Tenacibaculum sp. 20J [108] and Thermaerobacter marianensis [109], both of which are marine bacteria. These can be potential candidates for the isolation of related genes that express useful thermostable QQ enzymes.…”

Section: Introductionmentioning

confidence: 99%

Quorum Sensing and the Use of Quorum Quenchers as Natural Biocides to Inhibit Sulfate-Reducing Bacteria

2016

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Sulfate-reducing bacteria (SRB) are one of the main protagonist groups of biocorrosion in the seawater environment. Given their principal role in biocorrosion, it remains a crucial task to develop strategies to reduce the abundance of SRBs. Conventional approaches include the use of biocides and antibiotics, which can impose health, safety, and environmental concerns. This review examines an alternative approach to this problem. This is achieved by reviewing the role of quorum sensing (QS) in SRB populations and its impact on the biofilm formation process. Genome databases of SRBs are mined to look for putative QS systems and homologous protein sequences representative of autoinducer receptors or synthases. Subsequently, this review puts forward the potential use of quorum quenchers as natural biocides against SRBs and outlines the potential strategies for the implementation of this approach.

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“…Aii810 showed stronger cold adaptability than any other AHL-degrading enzymes reported previously, such as Est816 ( Fan et al, 2012 ), AiiAAI96 ( Cao et al, 2012 ), and AiiAB546 ( Chen et al, 2010 ), which were most active at 30–60°C. A thermostable N -acylhomoserine lactonase AiiT showed maximal activity at higher temperatures ranging from 60 to 80°C ( Morohoshi et al, 2015 ). However, temperature properties of other N -acylhomoserine lactonases were not quantified ( Dong et al, 2000 ; Cao et al, 2012 ; Mayer et al, 2015 ; Tang et al, 2015 ; Valera et al, 2016 ).…”

Section: Resultsmentioning

confidence: 99%

“…Until now, many AHL-degrading enzymes have been discovered from the metagenome and from several bacteria. On the basis of their catalytic mechanisms, AHL-degrading enzymes could be divided into three families: AHL-lactonases ( Dong et al, 2000 ; Bijtenhoorn et al, 2011b ; Fan et al, 2012 ; Mayer et al, 2015 ; Morohoshi et al, 2015 ), AHL-acylases ( Lin et al, 2003 ), and AHL-oxidases ( Bijtenhoorn et al, 2011a ). AHL-lactonases degrade AHLs by breaking the lactone bond ( Dong et al, 2000 ); AHL-acylases hydrolyze AHLs by degrading the amide linkage and separating the AHL into homoserine lactone and fatty acid ( Lin et al, 2003 ); AHL-oxidases degrade signal molecules by oxidizing the u-1, u-2, and u-3 carbons of the acyl chain of AHL ( Chowdhary et al, 2007 ).…”

Section: Introductionmentioning

confidence: 99%

Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation

et al. 2017

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The pathogen Pseudomonas aeruginosa uses quorum sensing (QS) to control virulence and biofilm formation. Enzymatic disruption of quorum sensing is a promising anti-infection therapeutic strategy that does not rely on antibiotics. Here, a novel gene (aii810) encoding an N-acylhomoserine lactonase was isolated from the Mao-tofu metagenome for the first time. Aii810 encoded a protein of 269 amino acids and was expressed in Escherichia coli BL21 (DE3) in soluble form. It showed the highest activity at 20°C, and it maintained 76.5% of activity at 0°C and more than 50% activity at 0–40°C. The optimal pH was 8.0. It was stable in both neutral and slightly alkaline conditions and at temperatures below 40°C. The enzyme hydrolyzed several ρ-nitrophenyl esters, but its best substrate was ρ-nitrophenyl acetate. Its kcat and Km values were 347.7 S-1 and 205.1 μM, respectively. It efficiently degraded N-butyryl-L-homoserine lactone and N-(3-oxododecanoyl)-L-homoserine lactone, exceeding hydrolysis rates of 72.3 and 100%, respectively. Moreover, Aii810 strongly attenuated P. aeruginosa virulence and biofilm formation. This enzyme with high anti-QS activity was the most cold-adapted N-acylhomoserine lactonase reported, which makes it an attractive enzyme for use as a therapeutic agent against P. aeruginosa infection.

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Characterization of a novel thermostable N-acylhomoserine lactonase from the thermophilic bacterium Thermaerobacter marianensis

Cited by 19 publications

References 27 publications

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

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

Quorum Sensing and the Use of Quorum Quenchers as Natural Biocides to Inhibit Sulfate-Reducing Bacteria

Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation

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