Characterization of a novel N-acylhomoserine lactonase, AidP, from Antarctic Planococcus sp.

BACKGROUND: N-acylhomoserine lactones (AHLs) are well-studied signalling molecules produced by some Gram-negative Proteobacteria for bacterial cell-to-cell communication or quorum sensing. We have previously demonstrated the degradation of AHLs by an Antarctic bacterium, Planococcus versutus L10.15T...

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Main Authors: See-Too, Wah Seng, Convey, Peter, Pearce, David Anthony, Chan, Kok Gan
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Published: BMC 2018
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Online Access:http://eprints.um.edu.my/20752/
https://doi.org/10.1186/s12934-018-1024-6
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spelling my.um.eprints.207522019-03-19T04:36:55Z http://eprints.um.edu.my/20752/ Characterization of a novel N-acylhomoserine lactonase, AidP, from Antarctic Planococcus sp. See-Too, Wah Seng Convey, Peter Pearce, David Anthony Chan, Kok Gan Q Science (General) QH Natural history BACKGROUND: N-acylhomoserine lactones (AHLs) are well-studied signalling molecules produced by some Gram-negative Proteobacteria for bacterial cell-to-cell communication or quorum sensing. We have previously demonstrated the degradation of AHLs by an Antarctic bacterium, Planococcus versutus L10.15T, at low temperature through the production of an AHL lactonase. In this study, we cloned the AHL lactonase gene and characterized the purified novel enzyme. RESULTS: Rapid resolution liquid chromatography analysis indicated that purified AidP possesses high AHL-degrading activity on unsubstituted, and 3-oxo substituted homoserine lactones. Liquid chromatography-mass spectrometry analysis confirmed that AidP functions as an AHL lactonase that hydrolyzes the ester bond of the homoserine lactone ring of AHLs. Multiple sequence alignment analysis and phylogenetic analysis suggested that the aidP gene encodes a novel AHL lactonase enzyme. The amino acid composition analysis of aidP and the homologous genes suggested that it might be a cold-adapted enzyme, however, the optimum temperature is 28 °C, even though the thermal stability is low (reduced drastically above 32 °C). Branch-site analysis of several aidP genes of Planococcus sp. branch on the phylogenetic trees also showed evidence of episodic positive selection of the gene in cold environments. Furthermore, we demonstrated the effects of covalent and ionic bonding, showing that Zn2+ is important for activity of AidP in vivo. The pectinolytic inhibition assay confirmed that this enzyme attenuated the pathogenicity of the plant pathogen Pectobacterium carotovorum in Chinese cabbage. CONCLUSION: We demonstrated that AidP is effective in attenuating the pathogenicity of P. carotovorum, a plant pathogen that causes soft-rot disease. This anti-quorum sensing agent is an enzyme with low thermal stability that degrades the bacterial signalling molecules (AHLs) that are produced by many pathogens. Since the enzyme is most active below human body temperature (below 28 °C), and lose its activity drastically above 32 °C, the results of a pectinolytic inhibition assay using Chinese cabbage indicated the potential of this anti-quorum sensing agent to be safely applied in the field trials. BMC 2018 Article PeerReviewed See-Too, Wah Seng and Convey, Peter and Pearce, David Anthony and Chan, Kok Gan (2018) Characterization of a novel N-acylhomoserine lactonase, AidP, from Antarctic Planococcus sp. Microbial Cell Factories, 17 (1). p. 179. ISSN 1475-2859 https://doi.org/10.1186/s12934-018-1024-6 doi:10.1186/s12934-018-1024-6
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
topic Q Science (General)
QH Natural history
spellingShingle Q Science (General)
QH Natural history
See-Too, Wah Seng
Convey, Peter
Pearce, David Anthony
Chan, Kok Gan
Characterization of a novel N-acylhomoserine lactonase, AidP, from Antarctic Planococcus sp.
description BACKGROUND: N-acylhomoserine lactones (AHLs) are well-studied signalling molecules produced by some Gram-negative Proteobacteria for bacterial cell-to-cell communication or quorum sensing. We have previously demonstrated the degradation of AHLs by an Antarctic bacterium, Planococcus versutus L10.15T, at low temperature through the production of an AHL lactonase. In this study, we cloned the AHL lactonase gene and characterized the purified novel enzyme. RESULTS: Rapid resolution liquid chromatography analysis indicated that purified AidP possesses high AHL-degrading activity on unsubstituted, and 3-oxo substituted homoserine lactones. Liquid chromatography-mass spectrometry analysis confirmed that AidP functions as an AHL lactonase that hydrolyzes the ester bond of the homoserine lactone ring of AHLs. Multiple sequence alignment analysis and phylogenetic analysis suggested that the aidP gene encodes a novel AHL lactonase enzyme. The amino acid composition analysis of aidP and the homologous genes suggested that it might be a cold-adapted enzyme, however, the optimum temperature is 28 °C, even though the thermal stability is low (reduced drastically above 32 °C). Branch-site analysis of several aidP genes of Planococcus sp. branch on the phylogenetic trees also showed evidence of episodic positive selection of the gene in cold environments. Furthermore, we demonstrated the effects of covalent and ionic bonding, showing that Zn2+ is important for activity of AidP in vivo. The pectinolytic inhibition assay confirmed that this enzyme attenuated the pathogenicity of the plant pathogen Pectobacterium carotovorum in Chinese cabbage. CONCLUSION: We demonstrated that AidP is effective in attenuating the pathogenicity of P. carotovorum, a plant pathogen that causes soft-rot disease. This anti-quorum sensing agent is an enzyme with low thermal stability that degrades the bacterial signalling molecules (AHLs) that are produced by many pathogens. Since the enzyme is most active below human body temperature (below 28 °C), and lose its activity drastically above 32 °C, the results of a pectinolytic inhibition assay using Chinese cabbage indicated the potential of this anti-quorum sensing agent to be safely applied in the field trials.
format Article
author See-Too, Wah Seng
Convey, Peter
Pearce, David Anthony
Chan, Kok Gan
author_facet See-Too, Wah Seng
Convey, Peter
Pearce, David Anthony
Chan, Kok Gan
author_sort See-Too, Wah Seng
title Characterization of a novel N-acylhomoserine lactonase, AidP, from Antarctic Planococcus sp.
title_short Characterization of a novel N-acylhomoserine lactonase, AidP, from Antarctic Planococcus sp.
title_full Characterization of a novel N-acylhomoserine lactonase, AidP, from Antarctic Planococcus sp.
title_fullStr Characterization of a novel N-acylhomoserine lactonase, AidP, from Antarctic Planococcus sp.
title_full_unstemmed Characterization of a novel N-acylhomoserine lactonase, AidP, from Antarctic Planococcus sp.
title_sort characterization of a novel n-acylhomoserine lactonase, aidp, from antarctic planococcus sp.
publisher BMC
publishDate 2018
url http://eprints.um.edu.my/20752/
https://doi.org/10.1186/s12934-018-1024-6
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score 13.222552