Many Gram-negative bacteria employ N-acylhomoserine lactones (AHLs) as quorum-sensing (QS) signal molecules to regulate virulence expression in a density-dependent manner. Quorum quenching (QQ) via enzymatic inactivation of AHLs is a promising strategy to reduce bacterial infections and drug resistance. Herein, a thermostable AHL lactonase (AidB), which could degrade different AHLs, with or without a substitution of carbonyl or hydroxyl at the C-3 position, was identified from the soil bacterium Bosea sp. strain F3-2. Ultrahigh-performance liquid chromatography analysis demonstrated that AidB is an AHL lactonase that hydrolyzes the ester bond of the homoserine lactone (HSL) ring. AidB was thermostable in the range 30 to 80°C and showed maximum activity after preincubation at 60°C for 30 min. The optimum temperature of AidB was 60°C, and the enzyme could be stably stored in double-distilled water (ddH2O) at 4°C or room temperature. AidB homologs were found only in Rhizobiales and Rhodospirillales of the Alphaproteobacteria. AidB from Agrobacterium tumefaciens and AidB from Rhizobium multihospitium (with amino acid identities of 50.6% and 52.8% to AidB, respectively) also showed thermostable AHL degradation activity. When introduced into bacteria, plasmid-expressed AidB attenuated pyocyanin production by Pseudomonas aeruginosa PAO1 and the pathogenicity of Pectobacterium carotovorum subsp. carotovorum Z3-3, suggesting that AidB is a potential therapeutic agent by degrading AHLs.
IMPORTANCE A quorum-sensing system using AHLs as the signal in many bacterial pathogens is a critical virulence regulator and an attractive target for anti-infective drugs. In this work, we identified a novel AHL lactonase, AidB, from a soil bacterial strain, Bosea sp. F3-2. The expression of aidB reduced the production of AHL signals and QS-dependent virulence factors in Pseudomonas aeruginosa and Pectobacterium carotovorum. The homologs of AidB with AHL-degrading activities were found only in several genera belonging to the Alphaproteobacteria. Remarkably, AidB is a thermostable enzyme that retained its catalytic activity after treatment at 80°C for 30 min and exhibits reliable storage stability at both 4°C and room temperature. These properties might make it more suitable for practical application.
Nanoplastic (NP) contamination is becoming a pervasive issue as NPs, originating from microplastic particles, pose potentially harmful environmental impacts on aquatic ecosystems. The environmental hazards of NPs on microorganisms have been well documented in recent studies; however, little is known about their ecotoxicity effects on freshwater biofilms, which serve as important primary producers and decomposers and are highly connected with other ecosystem components. We investigated the effects of NPs on the microbial metabolic functions of freshwater biofilms in terms of carbon source utilization ability. Biofilm samples were collected, cultivated in a hydrodynamic flume for six weeks, and then exposed in polystyrene (PS) beads (100 nm in size) with different NP concentrations (1, 5, and 10 mg/L). BIOLOG ECO microplates were used to quantify carbon source utilization characteristics. The data were analyzed using average well-color development (AWCD), functional diversity indices, and principle component analysis (PCA). Results showed that the total carbon metabolic functions (represented by AWCD) remained constant (p > 0.05) with elevated NP concentrations, but some specific carbon sources (e.g., esters) changed in their utilization ability (p < 0.05). The microbial functional diversity (Shannon–Wiener diversity index, Simpson diversity index, and Shannon evenness index) was significantly reduced under 10 mg/L NPs (p < 0.05), indicating an inhibiting effect of NPs on biofilm metabolic diversity. This study examined NP ecotoxicity effects on microbial metabolic activities at the community level, but further studies are required to fully understand the mechanisms driving this change.
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