Crystal structure of NucB, a biofilm-degrading endonuclease

Baslé, Arnaud; Hewitt, Lorraine; Koh, Alan; Lamb, Heather K.; Thompson, Paul R.; Burgess, J. Grant; Hall, Michael J.; Hawkins, Alastair R.; Murray, Heath; Lewis, Richard J.

doi:10.1093/nar/gkx1170

Cited by 21 publications

(7 citation statements)

References 65 publications

(93 reference statements)

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“…Furthermore, strategies to eliminate the biofilm during the infectious process will decrease the bacterial tolerance to antimicrobial agents, and therefore avoid further appearance of AMR. Enzymes can be used to prevent the biofilm formation by disrupting the inter-cell communications necessary for its formation or to directly target the biofilm structural compounds with different hydrolytic enzymes such as proteases, nucleases, and glycosyl hydrolases . On the other hand, different NP-based strategies have also shown promising results in the biofilm elimination. , …”

Section: Introductionmentioning

confidence: 99%

Metal–Enzyme Nanoaggregates Eradicate Both Gram-Positive and Gram-Negative Bacteria and Their Biofilms

Ferreres

Bassegoda

Hoyo

et al. 2018

ACS Appl. Mater. Interfaces

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To palliate the appearance of antimicrobial resistance (AMR), the use of bactericidal agents acting differently than conventional antibiotics and the elimination of bacterial biofilm, are the two most promising strategies. Here, we integrated these two complementary strategies into new antimicrobial metal–enzyme nanoaggregates (NAs) of α-amylase and silver (αAgNAs) that are able to eliminate bacteria and their biofilm. The nanoparticle (NP) synthesis approach applied protein desolvation and laccase-mediated NP stabilization to innovatively produce catalytically active α-amylase nanoparticles (αNPs) for the elimination of the bacterial biofilm. At the same time, αNPs efficiently reduced silver for the incorporation of bactericidal Ag0 and formation of the αAgNAs. The bactericidal and antibiofilm efficacies of αAgNAs were demonstrated by 5.4 and 6.1 log reduction of Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli, respectively, and more than 80% removal of their biofilms, coupled with high biocompatibility. The biofilm−αAgNA interaction was assessed by quartz crystal microbalance and atomic force microscopy revealing how the degradation of a settled biofilm by αAgNAs caused an increase of the biofilm water content, thus weakening the biofilm surface attachment and facilitating its removal. With the present work, we not only provide a new efficient antimicrobial material to face the AMR threat, but we also envisage that the newly established method for the synthesis of metal–enzyme NAs is potentially transferable to other biocatalysts to expand the enzyme NP toolbox.

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Section: Introductionmentioning

confidence: 99%

Metal–Enzyme Nanoaggregates Eradicate Both Gram-Positive and Gram-Negative Bacteria and Their Biofilms

Ferreres

Bassegoda

Hoyo

et al. 2018

ACS Appl. Mater. Interfaces

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“…To determine if the uptake deficiency of the ΔnucA strain was due to the nuclease activity of NucA, we inserted a D98A active site mutation at the native nucA locus. The design of this mutation was based on analysis of NucB (45), which exhibits 58% identity with NucA. D98A corresponds in position to residue D87A in NucB.…”

Section: Nucamentioning

confidence: 99%

Mechanisms of transforming DNA uptake to the periplasm ofBacillus subtilis

Hahn

DeSantis

Dubnau

2021

Preprint

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We demonstrate here that the acquisition of DNAase resistance by transforming DNA, often assumed to indicate transport to the cytoplasm, actually reflects uptake to the periplasm, requiring a re-evaluation of conclusions about the roles of several proteins in transformation. The new evidence suggests that the transformation pilus is needed for DNA binding to the cell surface near the cell poles and for the initiation of uptake. The cellular distribution of the membrane-anchored ComEA of B. subtilis does not noticeably change during DNA uptake as does the unanchored ComEA of Vibrio and Neisseria. Instead, our evidence suggests that ComEA stabilizes the attachment of transforming DNA at localized regions in the periplasm and then mediates uptake, probably by a Brownian ratchet mechanism. Following that, the DNA is transferred to periplasmic portions of the channel protein ComEC, which plays a previously unsuspected role in uptake to the periplasm. We show that the transformation endonuclease NucA also facilitates uptake to the periplasm and that the previously demonstrated role of ComFA in the acquisition of DNAase resistance actually derives from the instability of ComGA when ComFA is deleted. These results prompt a new understanding of the early stages of DNA uptake for transformation.

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“…The DNase can also disrupt biofilms by degrading the eDNA integrated with other EPS components for structural support . Recombinant human DNase I (dornase alfa) was shown in clinical trials to degrade microbial eDNA in the sputum of patients with cystic fibrosis and ventilator‐associated infection in preterm infants …”

Section: Development Of Anti‐biofilm Chemotherapymentioning

confidence: 99%