The Importance of Porins and β-Lactamase in Outer Membrane Vesicles on the Hydrolysis of β-Lactam Antibiotics

Kim, Si Won; Lee, Jung Seok; Park, Seong Bin; Lee, Ae Rin; Jung, Jin Tae; Chun, Jin Hong; Lazarte, Jassy Mary S.; Kim, Jae-Sung; Seo, Jong‐Su; Kim, Jong-Hwan; Song, Jong-Wook; Ha, Min Woo; Thompson, Kim D.; Lee, Chang-Ro; Jung, Myunghwan; Jung, Tae Sung

doi:10.3390/ijms21082822

Cited by 36 publications

(24 citation statements)

References 60 publications

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“…In order to perform docking, the three-dimensional X-ray crystal structure of E. coli β-lactamase (PDB ID: 4E3K, resolution = 1.43Å) in complex with known inhibitor (4-tetrazolyl pyridine sulfonamide boronic acid) was retrieved from RCSB Protein Data Bank [ 67 ] and docking was conducted on Molecular Operating Environment (MOE version 2014.09). β-Lactamases are enzymes found in outer membrane vesicles of β-lactam-resistant Escherichia coli that are able to inactivate the antibacterial properties of β-lactam antibiotics [ 68 ].…”

Section: Methodsmentioning

confidence: 99%

Photocatalytic Decolorization and Biocidal Applications of Nonmetal Doped TiO2: Isotherm, Kinetic Modeling and In Silico Molecular Docking Studies

et al. 2020

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Textile dyes and microbial contamination of surface water bodies have been recognized as emerging quality concerns around the globe. The simultaneous resolve of such impurities can pave the route for an amicable technological solution. This study reports the photocatalytic performance and the biocidal potential of nitrogen-doped TiO2 against reactive black 5 (RB5), a double azo dye and E. coli. Molecular docking was performed to identify and quantify the interactions of the TiO2 with β-lactamase enzyme and to predict the biocidal mechanism. The sol-gel technique was employed for the synthesis of different mol% nitrogen-doped TiO2. The synthesized photocatalysts were characterized using thermal gravimetric analysis (TGA), scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), Brunauer–Emmett–Teller (BET) and diffuse reflectance spectroscopy (DRS). The effects of different synthesis and reaction parameters were studied. RB5 dye degradation was monitored by tracking shifts in the absorption spectrum and percent chemical oxygen demand (COD) removal. The best nanomaterial depicted 5.57 nm crystallite size, 49.54 m2 g−1 specific surface area, 11–40 nm particle size with spherical morphologies, and uniform distribution. The RB5 decolorization data fits well with the pseudo-first-order kinetic model, and the maximum monolayer coverage capacity for the Langmuir adsorption model was found to be 40 mg g−1 with Kads of 0.113 mg−1. The LH model yielded a higher coefficient KC (1.15 mg L−1 h−1) compared to the adsorption constant KLH (0.3084 L mg−1). 90% COD removal was achieved in 60 min of irradiation, confirmed by the disappearance of spectral peaks. The best-optimized photocatalysts showed a noticeable biocidal potential against human pathogenic strain E. coli in 150 min. The biocidal mechanism of best-optimized photocatalyst was predicted by molecular docking simulation against E. coli β-lactamase enzyme. The docking score (−7.6 kcal mol−1) and the binding interaction with the active site residues (Lys315, Thr316, and Glu272) of β-lactamase further confirmed that inhibition of β-lactamase could be a most probable mechanism of biocidal activity.

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Section: Methodsmentioning

confidence: 99%

Photocatalytic Decolorization and Biocidal Applications of Nonmetal Doped TiO2: Isotherm, Kinetic Modeling and In Silico Molecular Docking Studies

et al. 2020

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“…These enzymes are present in the periplasmic space. Several studies indicated that they might be sequestered into MVs, and exert their protective activity on susceptible bacteria [38][39][40]. We performed an experiment that proved the presence of β-lactam degrading enzymes in the MVs of the ampicillin resistant strains of Pectobacterium.…”

Section: Discussionmentioning

confidence: 93%

Membrane Vesicles of Pectobacterium as an Effective Protein Secretion System

Jońca

Waleron

Czaplewska

et al. 2021

IJMS

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Bacteria of genus Pectobacterium are Gram-negative rods of the family Pectobacteriaceae. They are the causative agent of soft rot diseases of crops and ornamental plants. However, their virulence mechanisms are not yet fully elucidated. Membrane vesicles (MVs) are universally released by bacteria and are believed to play an important role in the pathogenicity and survival of bacteria in the environment. Our study investigates the role of MVs in the virulence of Pectobacterium. The results indicate that the morphology and MVs production depend on growth medium composition. In polygalacturonic acid (PGA) supplemented media, Pectobacterium produces large MVs (100–300 nm) and small vesicles below 100 nm. Proteomic analyses revealed the presence of pectate degrading enzymes in the MVs. The pectate plate test and enzymatic assay proved that those enzymes are active and able to degrade pectates. What is more, the pathogenicity test indicated that the MVs derived from Pectobacterium were able to induce maceration of Zantedeschia sp. leaves. We also show that the MVs of β-lactamase producing strains were able to suppress ampicillin activity and permit the growth of susceptible bacteria. Those findings indicate that the MVs of Pectobacterium play an important role in host-pathogen interactions and niche competition with other bacteria. Our research also sheds some light on the mechanism of MVs production. We demonstrate that the MVs production in Pectobacterium strains, which overexpress a green fluorescence protein (GFP), is higher than in wild-type strains. Moreover, proteomic analysis revealed that the GFP was present in the MVs. Therefore, it is possible that protein sequestration into MVs might not be strictly limited to periplasmic proteins. Our research highlights the importance of MVs production as a mechanism of cargo delivery in Pectobacterium and an effective secretion system.

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“…For patients with GAS pharyngotonsillitis, this mechanism may be of a particular importance, since more than one fourth of children with GAS pharyngotonsillitis have their tonsils colonized with H. influenzae or M. catarrhalis [37], both of which can produce β-lactamase-carrying OMVs ( [22,23], this study). Mechanistically, the structural identity between the bacterial outer membrane and OMV membrane allows β-lactams to enter, through the porin channels, the OMV lumen, where β-lactamase, originating from the bacterial periplasm, is located and hydrolyzes the antibiotics [22][23][24]38]. Through this process, OMVs secreted outside bacterial cells serve as a first line of protection against β-lactams before the antibiotics reach the target bacterial population.…”

Section: Discussionmentioning

confidence: 99%

In Vivo Secretion of β-Lactamase-Carrying Outer Membrane Vesicles as a Mechanism of β-Lactam Therapy Failure

et al. 2021

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Outer membrane vesicles carrying β-lactamase (βLOMVs) protect bacteria against β-lactam antibiotics under experimental conditions, but their protective role during a patient’s treatment leading to the therapy failure is unknown. We investigated the role of βLOMVs in amoxicillin therapy failure in a patient with group A Streptococcus pyogenes (GAS) pharyngotonsillitis. The patient’s throat culture was examined by standard microbiological procedures. Bacterial vesicles were analyzed for β-lactamase by immunoblot and the nitrocefin assay, and in vivo secretion of βLOMVs was detected by electron microscopy. These analyses demonstrated that the patient’s throat culture grew, besides amoxicillin-susceptible GAS, an amoxicillin-resistant nontypeable Haemophilus influenzae (NTHi), which secreted βLOMVs. Secretion and β-lactamase activity of NTHi βLOMVs were induced by amoxicillin concentrations reached in the tonsils during therapy. The presence of NTHi βLOMVs significantly increased the minimal inhibitory concentration of amoxicillin for GAS and thereby protected GAS against bactericidal concentrations of amoxicillin. NTHi βLOMVs were identified in the patient’s pharyngotonsillar swabs and saliva, demonstrating their secretion in vivo at the site of infection. We conclude that the pathogen protection via βLOMVs secreted by the flora colonizing the infection site represents a yet underestimated mechanism of β-lactam therapy failure that warrants attention in clinical studies.

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The Importance of Porins and β-Lactamase in Outer Membrane Vesicles on the Hydrolysis of β-Lactam Antibiotics

Cited by 36 publications

References 60 publications

Photocatalytic Decolorization and Biocidal Applications of Nonmetal Doped TiO2: Isotherm, Kinetic Modeling and In Silico Molecular Docking Studies

Photocatalytic Decolorization and Biocidal Applications of Nonmetal Doped TiO2: Isotherm, Kinetic Modeling and In Silico Molecular Docking Studies

Membrane Vesicles of Pectobacterium as an Effective Protein Secretion System

In Vivo Secretion of β-Lactamase-Carrying Outer Membrane Vesicles as a Mechanism of β-Lactam Therapy Failure

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