Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation
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...
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Frontiers Media S.A.
2017-10-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fmicb.2017.01950/full |
| _version_ | 1818600859642101760 |
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| author | Xinjiong Fan Mingjun Liang Lei Wang Ruo Chen He Li Xiaolong Liu |
| author_facet | Xinjiong Fan Mingjun Liang Lei Wang Ruo Chen He Li Xiaolong Liu |
| author_sort | Xinjiong Fan |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-16T12:42:11Z |
| format | Article |
| id | doaj.art-4148e5b189314eeaa9b6371fd74d45f7 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-16T12:42:11Z |
| publishDate | 2017-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-4148e5b189314eeaa9b6371fd74d45f72022-12-21T22:31:23ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2017-10-01810.3389/fmicb.2017.01950284902Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm FormationXinjiong Fan0Mingjun Liang1Lei Wang2Ruo Chen3He Li4Xiaolong Liu5School of Basic Medical Sciences, Anhui Medical University, Hefei, ChinaSchool of Basic Medical Sciences, Anhui Medical University, Hefei, ChinaSchool of Basic Medical Sciences, Anhui Medical University, Hefei, ChinaSchool of Basic Medical Sciences, Anhui Medical University, Hefei, ChinaSchool of Basic Courses, Guangdong Pharmaceutical University, Guangzhou, ChinaSchool of Basic Medical Sciences, Anhui Medical University, Hefei, ChinaThe 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.http://journal.frontiersin.org/article/10.3389/fmicb.2017.01950/fullbiofilmN-acylhomoserine lactonasequorum sensingPseudomonas aeruginosacold adaptation |
| spellingShingle | Xinjiong Fan Mingjun Liang Lei Wang Ruo Chen He Li Xiaolong Liu Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation Frontiers in Microbiology biofilm N-acylhomoserine lactonase quorum sensing Pseudomonas aeruginosa cold adaptation |
| title | Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation |
| title_full | Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation |
| title_fullStr | Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation |
| title_full_unstemmed | Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation |
| title_short | Aii810, a Novel Cold-Adapted N-Acylhomoserine Lactonase Discovered in a Metagenome, Can Strongly Attenuate Pseudomonas aeruginosa Virulence Factors and Biofilm Formation |
| title_sort | aii810 a novel cold adapted n acylhomoserine lactonase discovered in a metagenome can strongly attenuate pseudomonas aeruginosa virulence factors and biofilm formation |
| topic | biofilm N-acylhomoserine lactonase quorum sensing Pseudomonas aeruginosa cold adaptation |
| url | http://journal.frontiersin.org/article/10.3389/fmicb.2017.01950/full |
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