Publication

PA0305 of Pseudomonas aeruginosa is a quorum quenching acylhomoserine lactone acylase belonging to the Ntn hydrolase superfamily

Wahjudi, M., Papaioannou, E., Hendrawati, O., van Assen, A. H. G., van Merkerk, R., Cool, R. H., Poelarends, G. J. & Quax, W., Jul-2011, In : Microbiology-Sgm. 157, 7, p. 2042-2055 14 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Wahjudi, M., Papaioannou, E., Hendrawati, O., van Assen, A. H. G., van Merkerk, R., Cool, R. H., ... Quax, W. (2011). PA0305 of Pseudomonas aeruginosa is a quorum quenching acylhomoserine lactone acylase belonging to the Ntn hydrolase superfamily. Microbiology-Sgm, 157(7), 2042-2055. https://doi.org/10.1099/mic.0.043935-0

Author

Wahjudi, Mariana ; Papaioannou, Evelina ; Hendrawati, Oktavia ; van Assen, Aart H. G. ; van Merkerk, Ronald ; Cool, Robbert H. ; Poelarends, Gerrit J. ; Quax, Wim. / PA0305 of Pseudomonas aeruginosa is a quorum quenching acylhomoserine lactone acylase belonging to the Ntn hydrolase superfamily. In: Microbiology-Sgm. 2011 ; Vol. 157, No. 7. pp. 2042-2055.

Harvard

Wahjudi, M, Papaioannou, E, Hendrawati, O, van Assen, AHG, van Merkerk, R, Cool, RH, Poelarends, GJ & Quax, W 2011, 'PA0305 of Pseudomonas aeruginosa is a quorum quenching acylhomoserine lactone acylase belonging to the Ntn hydrolase superfamily', Microbiology-Sgm, vol. 157, no. 7, pp. 2042-2055. https://doi.org/10.1099/mic.0.043935-0

Standard

PA0305 of Pseudomonas aeruginosa is a quorum quenching acylhomoserine lactone acylase belonging to the Ntn hydrolase superfamily. / Wahjudi, Mariana; Papaioannou, Evelina; Hendrawati, Oktavia; van Assen, Aart H. G.; van Merkerk, Ronald; Cool, Robbert H.; Poelarends, Gerrit J.; Quax, Wim.

In: Microbiology-Sgm, Vol. 157, No. 7, 07.2011, p. 2042-2055.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Wahjudi M, Papaioannou E, Hendrawati O, van Assen AHG, van Merkerk R, Cool RH et al. PA0305 of Pseudomonas aeruginosa is a quorum quenching acylhomoserine lactone acylase belonging to the Ntn hydrolase superfamily. Microbiology-Sgm. 2011 Jul;157(7):2042-2055. https://doi.org/10.1099/mic.0.043935-0


BibTeX

@article{d0db1c3d85ef44fbbb38e190a553192b,
title = "PA0305 of Pseudomonas aeruginosa is a quorum quenching acylhomoserine lactone acylase belonging to the Ntn hydrolase superfamily",
abstract = "The Pseudomonas aeruginosa PAO1 genome has at least two genes, pvdQ and quiP, encoding acylhomoserine lactone (AHL) acylases. Two additional genes, pa 1893 and pa0305, have been predicted to encode penicillin acylase proteins, but have not been characterized. Initial studies on a pa0305 transposon insertion mutant suggested that the gene is not related to the AHL growth phenotype of P. aeruginosa. The close similarity (6 7 {\%}) of pa0305 to HacB, an AHL acylase of Pseudomonas syringae, prompted us to investigate whether the PA0305w protein might also function as an AHL acylase. The pa0305 gene has been cloned and the protein (PA0305) has been overproduced, purified and subjected to functional characterization. Analysis of the purified protein showed that, like beta-lactam acylases, PA0305 undergoes post-translational processing resulting in alpha- and beta-subunits, with the catalytic serine as the first amino acid of the beta-subunit, strongly suggesting that PA0305 is a member of the N-terminal nucleophile hydrolase superfamily. Using a biosensor assay, PA0305 his was shown to degrade AHLs with acyl side chains ranging in length from 6 to 14 carbons. Kinetics studies using N-octanoyl-L-homoserine lactone (C(8)-HSL) and N-(3-oxo-dodecanoyl)-L-homoserine lactone (3-oxo-C(12)-HSL) as substrates showed that the enzyme has a robust activity towards these two AHLs, with apparent K(cat)/K(m) values of 0.14x10(4) M(-1) s(-1) towards C(8)-HSL and 7.8x10(4) M(-1) s(-1) towards 3-oxo-C(12)-HSL. Overexpression of the pa0305 gene in P. aeruginosa showed significant reductions in both accumulation of 3-oxo-C(12)-HSL and expression of virulence factors. A mutant P. aeruginosa strain with a deleted pa 0305 gene showed a slightly increased capacity to kill Caenorhabditis elegans compared with the P. aeruginosa PA01 wild-type strain and the PA01 strain carrying a plasmid overexpressing pa0305. The harmful effects of the Delta pa0305 strain on the animals were most visible at 5 days post-exposure and the mortality rate of the animals fed on the Delta pa0305 strain was faster than for the animals fed on either the wild-type strain or the strain overexpressing pa0305. In conclusion, the pa0305 gene encodes an efficient acylase with activity towards long-chain homoserine lactones, including 3-oxo-C(12)-HSL, the natural quorum sensing signal molecule in P. aeruginosa, and we propose to name this acylase HacB.",
keywords = "CEPHALOSPORIN ACYLASE, PENICILLIN ACYLASE, MOLECULAR-CLONING, IDENTIFICATION, GENES, PAO1, GROWTH, HOST, CONSTRUCTION, SPECIFICITY",
author = "Mariana Wahjudi and Evelina Papaioannou and Oktavia Hendrawati and {van Assen}, {Aart H. G.} and {van Merkerk}, Ronald and Cool, {Robbert H.} and Poelarends, {Gerrit J.} and Wim Quax",
year = "2011",
month = "7",
doi = "10.1099/mic.0.043935-0",
language = "English",
volume = "157",
pages = "2042--2055",
journal = "Microbiology",
issn = "1465-2080",
publisher = "MAIK NAUKA/INTERPERIODICA/SPRINGER",
number = "7",

}

RIS

TY - JOUR

T1 - PA0305 of Pseudomonas aeruginosa is a quorum quenching acylhomoserine lactone acylase belonging to the Ntn hydrolase superfamily

AU - Wahjudi, Mariana

AU - Papaioannou, Evelina

AU - Hendrawati, Oktavia

AU - van Assen, Aart H. G.

AU - van Merkerk, Ronald

AU - Cool, Robbert H.

AU - Poelarends, Gerrit J.

AU - Quax, Wim

PY - 2011/7

Y1 - 2011/7

N2 - The Pseudomonas aeruginosa PAO1 genome has at least two genes, pvdQ and quiP, encoding acylhomoserine lactone (AHL) acylases. Two additional genes, pa 1893 and pa0305, have been predicted to encode penicillin acylase proteins, but have not been characterized. Initial studies on a pa0305 transposon insertion mutant suggested that the gene is not related to the AHL growth phenotype of P. aeruginosa. The close similarity (6 7 %) of pa0305 to HacB, an AHL acylase of Pseudomonas syringae, prompted us to investigate whether the PA0305w protein might also function as an AHL acylase. The pa0305 gene has been cloned and the protein (PA0305) has been overproduced, purified and subjected to functional characterization. Analysis of the purified protein showed that, like beta-lactam acylases, PA0305 undergoes post-translational processing resulting in alpha- and beta-subunits, with the catalytic serine as the first amino acid of the beta-subunit, strongly suggesting that PA0305 is a member of the N-terminal nucleophile hydrolase superfamily. Using a biosensor assay, PA0305 his was shown to degrade AHLs with acyl side chains ranging in length from 6 to 14 carbons. Kinetics studies using N-octanoyl-L-homoserine lactone (C(8)-HSL) and N-(3-oxo-dodecanoyl)-L-homoserine lactone (3-oxo-C(12)-HSL) as substrates showed that the enzyme has a robust activity towards these two AHLs, with apparent K(cat)/K(m) values of 0.14x10(4) M(-1) s(-1) towards C(8)-HSL and 7.8x10(4) M(-1) s(-1) towards 3-oxo-C(12)-HSL. Overexpression of the pa0305 gene in P. aeruginosa showed significant reductions in both accumulation of 3-oxo-C(12)-HSL and expression of virulence factors. A mutant P. aeruginosa strain with a deleted pa 0305 gene showed a slightly increased capacity to kill Caenorhabditis elegans compared with the P. aeruginosa PA01 wild-type strain and the PA01 strain carrying a plasmid overexpressing pa0305. The harmful effects of the Delta pa0305 strain on the animals were most visible at 5 days post-exposure and the mortality rate of the animals fed on the Delta pa0305 strain was faster than for the animals fed on either the wild-type strain or the strain overexpressing pa0305. In conclusion, the pa0305 gene encodes an efficient acylase with activity towards long-chain homoserine lactones, including 3-oxo-C(12)-HSL, the natural quorum sensing signal molecule in P. aeruginosa, and we propose to name this acylase HacB.

AB - The Pseudomonas aeruginosa PAO1 genome has at least two genes, pvdQ and quiP, encoding acylhomoserine lactone (AHL) acylases. Two additional genes, pa 1893 and pa0305, have been predicted to encode penicillin acylase proteins, but have not been characterized. Initial studies on a pa0305 transposon insertion mutant suggested that the gene is not related to the AHL growth phenotype of P. aeruginosa. The close similarity (6 7 %) of pa0305 to HacB, an AHL acylase of Pseudomonas syringae, prompted us to investigate whether the PA0305w protein might also function as an AHL acylase. The pa0305 gene has been cloned and the protein (PA0305) has been overproduced, purified and subjected to functional characterization. Analysis of the purified protein showed that, like beta-lactam acylases, PA0305 undergoes post-translational processing resulting in alpha- and beta-subunits, with the catalytic serine as the first amino acid of the beta-subunit, strongly suggesting that PA0305 is a member of the N-terminal nucleophile hydrolase superfamily. Using a biosensor assay, PA0305 his was shown to degrade AHLs with acyl side chains ranging in length from 6 to 14 carbons. Kinetics studies using N-octanoyl-L-homoserine lactone (C(8)-HSL) and N-(3-oxo-dodecanoyl)-L-homoserine lactone (3-oxo-C(12)-HSL) as substrates showed that the enzyme has a robust activity towards these two AHLs, with apparent K(cat)/K(m) values of 0.14x10(4) M(-1) s(-1) towards C(8)-HSL and 7.8x10(4) M(-1) s(-1) towards 3-oxo-C(12)-HSL. Overexpression of the pa0305 gene in P. aeruginosa showed significant reductions in both accumulation of 3-oxo-C(12)-HSL and expression of virulence factors. A mutant P. aeruginosa strain with a deleted pa 0305 gene showed a slightly increased capacity to kill Caenorhabditis elegans compared with the P. aeruginosa PA01 wild-type strain and the PA01 strain carrying a plasmid overexpressing pa0305. The harmful effects of the Delta pa0305 strain on the animals were most visible at 5 days post-exposure and the mortality rate of the animals fed on the Delta pa0305 strain was faster than for the animals fed on either the wild-type strain or the strain overexpressing pa0305. In conclusion, the pa0305 gene encodes an efficient acylase with activity towards long-chain homoserine lactones, including 3-oxo-C(12)-HSL, the natural quorum sensing signal molecule in P. aeruginosa, and we propose to name this acylase HacB.

KW - CEPHALOSPORIN ACYLASE

KW - PENICILLIN ACYLASE

KW - MOLECULAR-CLONING

KW - IDENTIFICATION

KW - GENES

KW - PAO1

KW - GROWTH

KW - HOST

KW - CONSTRUCTION

KW - SPECIFICITY

U2 - 10.1099/mic.0.043935-0

DO - 10.1099/mic.0.043935-0

M3 - Article

VL - 157

SP - 2042

EP - 2055

JO - Microbiology

JF - Microbiology

SN - 1465-2080

IS - 7

ER -

ID: 5373370