Disruption of bacterial biofilms using recombinant dispersin B

Dobrynina, O. Yu.; Bolshakova, Tatyana N.; Umyarov, A. M.; Boksha, Irina S.; Лаврова, Н. В.; Grishin, A. V.; Lyashchuk, A. M.; Галушкина, З. М.; Аветисян, Л Р; Chernukha, M.; Шагинян, И А; Лунин, В. Г.; Karyagina, A. S.

doi:10.1134/s0026261715040062

Cited by 19 publications

(9 citation statements)

References 10 publications

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“…For example, recombinant dispersin B has been shown to have antibiofilm activity against Staph. epidermidis (Dobrynina et al ., ) and Staph. aureus (Chaignon et al ., ).…”

Section: Biological Methodsmentioning

confidence: 96%

Bacterial biofilms in food processing environments: a review of recent developments in chemical and biological control

Phillips

2016

Int J of Food Sci Tech

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Biofilms are immobile communities of micro-organisms attached to any surface, such as stainless steel or a food matrix surface or on packaging material. They may be composed of a single species, but more generally, in the natural environment, they consist of mixed species together with an extracellular matrix. Biofilms provide a common mechanism of persistence for a number of bacterial species especially in food processing environments, and therefore, prevention of biofilm formation and the removal of preformed biofilms are an important issue for the food industry. This article reviews the current understanding on the formation of biofilms and recent developments in biological and chemical methods for prevention and removal.

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“…For example, recombinant dispersin B has been shown to have antibiofilm activity against Staph. epidermidis (Dobrynina et al ., ) and Staph. aureus (Chaignon et al ., ).…”

Section: Biological Methodsmentioning

confidence: 96%

Bacterial biofilms in food processing environments: a review of recent developments in chemical and biological control

Phillips

2016

Int J of Food Sci Tech

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“…Despite being considered a biofilm virulence factor, these enzymes may be engineered to be used in strategies for biofilm disassembly. For instance, recombinant dispersin B was produced by cloning in E. coli a synthetic gene encoding the protein from Aggregatibacter actinomycetemcomitans (Dobrynina et al, 2015). After purification, the enzyme was tested against biofilms of S. epidermidis, Burkholderia cenocepacia, and Achromobacter xylosoxidans.…”

Section: Enzymesmentioning

confidence: 99%

“…Dispersin B actively disrupted mature S. epidermidis biofilms at low concentrations (lower than 0.3 µg per sample). However, for the other two strains, a dispersin B concentration above 5 µg per sample was required to reduce the biofilm biomass (Dobrynina et al, 2015). Dispersin B is a glycosyl hydrolase able to specifically disrupt poly-N-acetylglucosamine (PNAG), which is the main exopolysaccharide of S. epidermidis biofilms (Chaignon et al, 2007).…”

Section: Enzymesmentioning

confidence: 99%

“…Dispersin B is a glycosyl hydrolase able to specifically disrupt poly-N-acetylglucosamine (PNAG), which is the main exopolysaccharide of S. epidermidis biofilms (Chaignon et al, 2007). Contrarily, PNAG is not a predominant component of Burkholderia cenocepacia and Achromobacter xylosoxidans biofilms, which may explain their higher resistance to the recombinant enzyme (Yakandawala et al, 2011;Dobrynina et al, 2015). In a more complex approach, Chen et al conjugated recombinant dispersin B with a silver-binding peptide, which promotes in situ formation of silver nanoparticles (AgNPs) in the presence of silver ions (Lee et al, 2008;Chen and Lee, 2018).…”

Section: Enzymesmentioning

confidence: 99%

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Innovative Strategies Toward the Disassembly of the EPS Matrix in Bacterial Biofilms

et al. 2020

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Bacterial biofilms represent a major concern at a worldwide level due to the high demand for implantable medical devices and the rising numbers of bacterial resistance. The complex structure of the extracellular polymeric substances (EPS) matrix plays a major role in this phenomenon, since it protects bacteria from antibiotics, avoiding drug penetration at bactericidal concentrations. Besides, this structure promotes bacterial cells to adopt a dormant lifestyle, becoming less susceptible to antibacterial agents. Currently, the available treatment for biofilm-related infections consists in the administration of conventional antibiotics at high doses for a long-term period. However, this treatment lacks efficiency against mature biofilms and for implant-associated biofilms it may be necessary to remove the medical device. Thus, biofilm-related infections represent an economical burden for the healthcare systems. New strategies focusing on the matrix are being highlighted as alternative therapies to eradicate biofilms. Here, we outline reported matrix disruptive agents, nanocarriers, and technologies, such as application of magnetic fields, photodynamic therapy, and ultrasounds, that have been under investigation to disrupt the EPS matrix of clinically relevant bacterial biofilms. In an ideal therapy, a synergistic effect between antibiotics and the explored innovated strategies is aimed to completely eradicate biofilms and avoid antimicrobial resistance phenomena.

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“…The production of recombinant biofilm-degrading enzymes has been attempted in microbes such as Escherichia coli ( Dobrynina et al, 2015 ; Cherdvorapong et al, 2020 ), but this is challenging because the inherent antibacterial properties of these products can interfere with host cell growth ( Oey et al, 2009 ). Plants such as Nicotiana benthamiana and tobacco ( Nicotiana tabacum ) are promising alternative hosts that benefit from inexpensive and rapidly scalable upstream production as well as product yields of more than 4 g kg −1 fresh leaf biomass ( Yamamoto et al, 2018 ) combined with high biomass yields of ∼100,000 kg ha −1 y −1 ( N. tabacum ) ( Stoger et al, 2002 ; Buyel et al, 2017 )].…”

Section: Introductionmentioning

confidence: 99%

Expression of Biofilm-Degrading Enzymes in Plants and Automated High-Throughput Activity Screening Using Experimental Bacillus subtilis Biofilms

Opdensteinen

Dietz

Gengenbach

et al. 2021

Front. Bioeng. Biotechnol.

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Biofilm-forming bacteria are sources of infections because they are often resistant to antibiotics and chemical removal. Recombinant biofilm-degrading enzymes have the potential to remove biofilms gently, but they can be toxic toward microbial hosts and are therefore difficult to produce in bacteria. Here, we investigated Nicotiana species for the production of such enzymes using the dispersin B-like enzyme Lysobacter gummosus glyco 2 (Lg2) as a model. We first optimized transient Lg2 expression in plant cell packs using different subcellular targeting methods. We found that expression levels were transferable to differentiated plants, facilitating the scale-up of production. Our process yielded 20 mg kg−1 Lg2 in extracts but 0.3 mg kg−1 after purification, limited by losses during depth filtration. Next, we established an experimental biofilm assay to screen enzymes for degrading activity using different Bacillus subtilis strains. We then tested complex and chemically defined growth media for reproducible biofilm formation before converting the assay to an automated high-throughput screening format. Finally, we quantified the biofilm-degrading activity of Lg2 in comparison with commercial enzymes against our experimental biofilms, indicating that crude extracts can be screened directly. This ability will allow us to combine high-throughput expression in plant cell packs with automated activity screening.

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Disruption of bacterial biofilms using recombinant dispersin B

Cited by 19 publications

References 10 publications

Bacterial biofilms in food processing environments: a review of recent developments in chemical and biological control

Bacterial biofilms in food processing environments: a review of recent developments in chemical and biological control

Innovative Strategies Toward the Disassembly of the EPS Matrix in Bacterial Biofilms

Expression of Biofilm-Degrading Enzymes in Plants and Automated High-Throughput Activity Screening Using Experimental Bacillus subtilis Biofilms

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