Inhibitory and bactericidal effect of Artilysin® Art-175 against colistin-resistant mcr-1-positive Escherichia coli isolates

Schirmeier, Eva; Zimmermann, Pia; Hofmann, Veronika; Biebl, Manfred; Gerstmans, Hans; Maervoet, Veerle E. T.; Briers, Yves

doi:10.1016/j.ijantimicag.2017.08.027

Cited by 48 publications

(38 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Quite recently, Art-175 was also shown to be bactericidal against colistin-resistant, mcr - 1 -positive E. coli isolates. This shows that the modifications of the lipid A moiety of LPS responsible for colistin resistance have no impact on the antibacterial activity of Art-175 [ 136 ].…”

Section: Improving the Potential Of Ples As Antibacterials Throughmentioning

confidence: 99%

Engineering of Phage-Derived Lytic Enzymes: Improving Their Potential as Antimicrobials

São-José

2018

Antibiotics

113

View full text Add to dashboard Cite

Lytic enzymes encoded by bacteriophages have been intensively explored as alternative agents for combating bacterial pathogens in different contexts. The antibacterial character of these enzymes (enzybiotics) results from their degrading activity towards peptidoglycan, an essential component of the bacterial cell wall. In fact, phage lytic products have the capacity to kill target bacteria when added exogenously in the form of recombinant proteins. However, there is also growing recognition that the natural bactericidal activity of these agents can, and sometimes needs to be, substantially improved through manipulation of their functional domains or by equipping them with new functions. In addition, often, native lytic proteins exhibit features that restrict their applicability as effective antibacterials, such as poor solubility or reduced stability. Here, I present an overview of the engineering approaches that can be followed not only to overcome these and other restrictions, but also to generate completely new antibacterial agents with significantly enhanced characteristics. As conventional antibiotics are running short, the remarkable progress in this field opens up the possibility of tailoring efficient enzybiotics to tackle the most menacing bacterial infections.

show abstract

Section: Improving the Potential Of Ples As Antibacterials Throughmentioning

confidence: 99%

Engineering of Phage-Derived Lytic Enzymes: Improving Their Potential as Antimicrobials

São-José

2018

Antibiotics

113

View full text Add to dashboard Cite

show abstract

“…Growth inhibition was determined as the lack of growth of a start inoculum treated with an Innolysin compared to the negative control (growth of ATCC11303 cells treated with supernatant lysates of cells carrying the empty vector, pVTSD2). A significant inhibitory activity was noticed when E. coli ATCC11303 was treated with Innolysin#6 similar to the positive control, Art-175, an engineered endolysin that was previously shown to have both an inhibitory and bactericidal effect against E. coli (20, 21). The remaining eight Innolysins, Pb5 or endolysin alone did not significantly affect the E. coli growth compared to the negative control (Fig.…”

Section: Resultsmentioning

confidence: 65%

Innolysins: A novel approach to engineer endolysins to kill Gram-negative bacteria

Zampara¹,

Sørensen²,

Grimon

et al. 2018

Preprint

Self Cite

View full text Add to dashboard Cite

13Bacteriophage-encoded endolysins degrading the essential peptidoglycan of bacteria are promising 14 alternative antimicrobials to handle the global threat of antibiotic resistant bacteria. However, 15 endolysins have limited use against Gram-negative bacteria, since their outer membrane prevents 16 access to the peptidoglycan. Here we present Innolysins, a novel concept for engineering endolysins 17 that allows the enzymes to pass through the outer membrane, hydrolyse the peptidoglycan and kill 18 the target bacterium. Innolysins combine the enzymatic activity of endolysins with the binding 19 capacity of phage receptor binding proteins (RBPs). As our proof of concept, we used phage T5 20 endolysin and receptor binding protein Pb5, which binds irreversibly to the phage receptor FhuA 21 involved in ferrichrome transport in Escherichia coli. In total, we constructed twelve Innolysins 22 fusing endolysin with Pb5 or the binding domain of Pb5 with or without flexible linkers in between. 23While the majority of the Innolysins maintained their muralytic activity, Innolysin#6 also showed 24 bactericidal activity against E. coli reducing the number of bacteria by 1 log, thus overcoming the 25 outer membrane barrier. Using an E. coli fhuA deletion mutant, we demonstrated that FhuA is 26 required for bactericidal activity, supporting that the specific binding of Pb5 to its receptor on E. 27 coli is needed for the endolysin to access the peptidoglycan. Accordingly, Innolysin#6 was able to 28 kill other bacterial species that carry conserved FhuA homologs such as Shigella sonnei and 29 Pseudomonas aeruginosa. In summary, the Innolysin approach expands recent protein engineering 30 strategies allowing customization of endolysins by exploiting phage RBPs to specifically target 31Gram-negative bacteria. 32 IMPORTANCE 33The extensive use of antibiotics has led to the emergence of antimicrobial resistant bacteria 34 responsible for infections causing more than 50,000 deaths per year across Europe and the US. In 35 response, the World Health Organization has stressed an urgent need to discover new antimicrobials 36 to control in particular Gram-negative bacterial pathogens, due to their extensive multi-drug 37 resistance. However, the outer membrane of Gram-negative bacteria limits the access of many 38 antibacterial agents to their targets. Here, we developed a new approach, Innolysins that enable 39 endolysins to overcome the outer membrane by exploiting the binding specificity of phage receptor 40 binding proteins. As proof of concept, we constructed Innolysins against E. coli using the endolysin 41 and the receptor binding protein of phage T5. Given the rich diversity of phage receptor binding 42 proteins and their different binding specificities, our proof of concept paves the route for creating an 43 arsenal of pathogen specific alternative antimicrobials. 44 intracellular targets (1). Bacteriophages (phages), viruses that infect bacteria, have naturally 48 evolved mechanisms to overcome the outer membrane to infect their ...

show abstract

“…The potential of purified recombinant bacteriophage lytic enzymes (lysins) has been regarded as an alternative method to control bacterial pathogens, including S. aureus [23,24,39,46,47], Streptococcus pneumoniae [19,20,22,25,26], group B streptococci [24], Bacillus anthracis [48]. Recently, a new engineered endolysin (Artilysin Art-175) has shown activity against gram-negative pathogens, such as Pseudomonas aeruginosa [49]. In addition, endolysins have advantageous characteristics that avoid most of the common resistance mechanisms against antibiotics [48].…”

Section: Discussionmentioning

confidence: 99%

Antimicrobial activity of Mycobacteriophage D29 Lysin B during Mycobacterium ulcerans infection

Fraga

Trigo

Dominguez

et al. 2018

Preprint

View full text Add to dashboard Cite

16Buruli Ulcer (BU) is a cutaneous disease caused by Mycobacterium ulcerans. The 17 pathogenesis of this disease is closely related to the secretion of the toxin mycolactone that 18 induces extensive destruction of the skin and soft tissues. Although the World Health Organization 19 recommends a combination of rifampicin and streptomycin for the treatment of BU, clinical 20 management of advanced stages often requires extensive surgical resection of the infected 21 tissue. Therefore, it is important to develop alternative strategies for the treatment of BU. 22 Endolysins (lysins) are phage encoded enzymes that degrade peptidoglycan of bacterial 23 cell walls. Over the past years, lysins have been emerging as alternative antimicrobial agents 24 against Gram-positive bacteria. Amongst Gram-positive bacteria, mycobacteria have an unusual 25 outer membrane that is covalently attached to the mycolylarabinogalactan-peptidoglycan 26 complex. To overcome this additional barrier to phage-mediated lysis, some mycobacteriophages 27 encode a lipolytic enzyme, Lysin B (Lys B). 28In this study, we demonstrated for the first time that recombinant Lys B displays lytic 29 activity against M. ulcerans isolates. Moreover, using a mouse model of M. ulcerans footpad 30 infection, we show that subcutaneous treatment with Lys B leads to a reduction in bacterial 31 burdens, associated with IFN-γ and TNF production in the draining lymph node. These findings 32 highlight the potential use of lysins as a novel therapeutic approach against this neglected tropical 33 disease. AUTHOR SUMMARY 35Buruli Ulcer (BU) is a necrotizing skin disease caused by Mycobacterium ulcerans. Standard 36 treatment for BU lesions consists of a combination of rifampicin and streptomycin for 8 weeks. 37However, clinical management of advanced stages of the disease often requires extensive 38 surgical resection of the infected tissue. Therefore, it is important to develop alternative strategies 39 for the treatment of BU. In that sense, we tested the efficacy of Lysin B (Lys B), a phage encoded 40 lipolytic enzyme that degrades the mycolylarabinogalactan-peptidoglycan complex present in the 41 mycobacterial cell wall. In this study, we show that Lys B not only displays lytic activity against M. 42 ulcerans isolates in vitro, but also leads to a reduction in bacterial burdens in M. ulcerans-infected 43 mouse footpads. These findings highlight the potential use of lysins as a novel therapeutic 44 approach against this neglected tropical disease. mycolactone that presents cytotoxic and immunosuppressive properties [4][5][6][7][8]. Early presentations 51 of active BU include a painless pre-ulcerative nodule, papule, plaque or edematous lesion, which 52 can evolve into typical ulcers or, in the most extreme cases, may result in extensive skin 53 destruction, multifocal lesions or bone involvement [1,3,7,9,10]. 54The antibiotic regimen recommended by the World Health Organization (WHO) (daily 55 administration of rifampicin (RIF) and streptomycin (STR) for 8 weeks) was p...

show abstract

Inhibitory and bactericidal effect of Artilysin® Art-175 against colistin-resistant mcr-1-positive Escherichia coli isolates

Cited by 48 publications

References 5 publications

Engineering of Phage-Derived Lytic Enzymes: Improving Their Potential as Antimicrobials

Engineering of Phage-Derived Lytic Enzymes: Improving Their Potential as Antimicrobials

Innolysins: A novel approach to engineer endolysins to kill Gram-negative bacteria

Antimicrobial activity of Mycobacteriophage D29 Lysin B during Mycobacterium ulcerans infection

Contact Info

Product

Resources

About