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Cloning and characterization of short‐chain N ‐acyl homoserine lactone‐producing Enterobacter asburiae strain L1 from lettuce leaves

In gram-negative bacteria, bacterial communication or quorum sensing (QS) is achieved using common signaling molecules known as N-acyl homoserine lactones (AHL). We have previously reported the genome of AHL-producing bacterium, Enterobacter asburiae strain L1. In silico analysis of the strain L1 ge...

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Main Authors: Lau, Yin Yin, How, Kah Yan, Yin, Wai Fong, Chan, Kok Gan
Format: Article
Published: Wiley Open Access 2018
Subjects:
Q Science (General)
QH Natural history
Online Access:http://eprints.um.edu.my/21544/
https://doi.org/10.1002/mbo3.610
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spelling my.um.eprints.215442019-06-26T04:39:18Z http://eprints.um.edu.my/21544/ Cloning and characterization of short‐chain N ‐acyl homoserine lactone‐producing Enterobacter asburiae strain L1 from lettuce leaves Lau, Yin Yin How, Kah Yan Yin, Wai Fong Chan, Kok Gan Q Science (General) QH Natural history In gram-negative bacteria, bacterial communication or quorum sensing (QS) is achieved using common signaling molecules known as N-acyl homoserine lactones (AHL). We have previously reported the genome of AHL-producing bacterium, Enterobacter asburiae strain L1. In silico analysis of the strain L1 genome revealed the presence of a pair of luxI/R genes responsible for AHL-type QS, designated as easIR. In this work, the 639 bp luxI homolog, encoding 212 amino acids, have been cloned and overexpressed in Escherichia coli BL21 (DE3)pLysS. The purified protein (~25 kDa) shares high similarity to several members of the LuxI family among different E asburiae strains. Our findings showed that the heterologously expressed EasI protein has activated violacein production by AHL biosensor Chromobacterium violaceum CV026 as the wild-type E. asburiae. The mass spectrometry analysis showed the production of N-butanoyl homoserine lactone and N–hexanoyl homoserine lactone from induced E. coli harboring the recombinant EasI, suggesting that EasI is a functional AHL synthase. E. asburiae strain L1 was also shown to possess biofilm-forming characteristic activity using crystal violet binding assay. This is the first report on cloning and characterization of the luxI homolog from E. asburiae. Wiley Open Access 2018 Article PeerReviewed Lau, Yin Yin and How, Kah Yan and Yin, Wai Fong and Chan, Kok Gan (2018) Cloning and characterization of short‐chain N ‐acyl homoserine lactone‐producing Enterobacter asburiae strain L1 from lettuce leaves. MicrobiologyOpen, 7 (6). e00610. ISSN 2045-8827 https://doi.org/10.1002/mbo3.610 doi:10.1002/mbo3.610
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
Lau, Yin Yin
How, Kah Yan
Yin, Wai Fong
Chan, Kok Gan
Cloning and characterization of short‐chain N ‐acyl homoserine lactone‐producing Enterobacter asburiae strain L1 from lettuce leaves
description In gram-negative bacteria, bacterial communication or quorum sensing (QS) is achieved using common signaling molecules known as N-acyl homoserine lactones (AHL). We have previously reported the genome of AHL-producing bacterium, Enterobacter asburiae strain L1. In silico analysis of the strain L1 genome revealed the presence of a pair of luxI/R genes responsible for AHL-type QS, designated as easIR. In this work, the 639 bp luxI homolog, encoding 212 amino acids, have been cloned and overexpressed in Escherichia coli BL21 (DE3)pLysS. The purified protein (~25 kDa) shares high similarity to several members of the LuxI family among different E asburiae strains. Our findings showed that the heterologously expressed EasI protein has activated violacein production by AHL biosensor Chromobacterium violaceum CV026 as the wild-type E. asburiae. The mass spectrometry analysis showed the production of N-butanoyl homoserine lactone and N–hexanoyl homoserine lactone from induced E. coli harboring the recombinant EasI, suggesting that EasI is a functional AHL synthase. E. asburiae strain L1 was also shown to possess biofilm-forming characteristic activity using crystal violet binding assay. This is the first report on cloning and characterization of the luxI homolog from E. asburiae.
format Article
author Lau, Yin Yin
How, Kah Yan
Yin, Wai Fong
Chan, Kok Gan
author_facet Lau, Yin Yin
How, Kah Yan
Yin, Wai Fong
Chan, Kok Gan
author_sort Lau, Yin Yin
title Cloning and characterization of short‐chain N ‐acyl homoserine lactone‐producing Enterobacter asburiae strain L1 from lettuce leaves
title_short Cloning and characterization of short‐chain N ‐acyl homoserine lactone‐producing Enterobacter asburiae strain L1 from lettuce leaves
title_full Cloning and characterization of short‐chain N ‐acyl homoserine lactone‐producing Enterobacter asburiae strain L1 from lettuce leaves
title_fullStr Cloning and characterization of short‐chain N ‐acyl homoserine lactone‐producing Enterobacter asburiae strain L1 from lettuce leaves
title_full_unstemmed Cloning and characterization of short‐chain N ‐acyl homoserine lactone‐producing Enterobacter asburiae strain L1 from lettuce leaves
title_sort cloning and characterization of short‐chain n ‐acyl homoserine lactone‐producing enterobacter asburiae strain l1 from lettuce leaves
publisher Wiley Open Access
publishDate 2018
url http://eprints.um.edu.my/21544/
https://doi.org/10.1002/mbo3.610
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score 13.211869

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