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Johan Seijsing, Anna M Sobieraj, Nadia Keller, Yang Shen, Annelies S Zinkernagel, Martin J Loessner, Mathias Schmelcher
The increasing number of multidrug-resistant bacteria intensifies the need to develop new antimicrobial agents. Endolysins are bacteriophage-derived enzymes that degrade the bacterial cell wall and hold promise as a new class of highly specific and versatile antimicrobials. One major limitation to the therapeutic use of endolysins is their often short serum circulation half-life, mostly due to kidney excretion and lysosomal degradation. One strategy to increase the half-life of protein drugs is fusion to the albumin-binding domain (ABD)...
2018: Frontiers in Microbiology
Natalia Gómez-Torres, Marta Ávila, Arjan Narbad, Melinda J Mayer, Sonia Garde
Clostridium tyrobutyricum is a bacteria of concern in the cheese industry, capable of surviving the manufacturing process and causing butyric acid fermentation and late blowing defect of cheese. In this work, we implement a method based on the cell wall-binding domain (CBD) of endolysin CTP1L, which detects C. tyrobutyricum, to monitor its evolution in cheeses challenged with clostridial spores and in the presence or absence of reuterin, an anti-clostridial agent. For this purpose, total bacteria were extracted from cheese samples and C...
April 2019: Food Microbiology
Muhsin Jamal, Sayed M A U S Bukhari, Saadia Andleeb, Muhammad Ali, Sana Raza, Muhammad A Nawaz, Tahir Hussain, Sadeeq U Rahman, Syed S A Shah
Bacteriophages (phages/viruses) need host bacteria to replicate and propagate. Primarily, a bacteriophage contains a head/capsid to encapsidate the genetic material. Some phages contain tails. Phages encode endolysins to hydrolyze bacterial cell wall. The two main classes of phages are lytic or virulent and lysogenic or temperate. In comparison with antibiotics, to deal with bacterial infections, phage therapy is thought to be more effective. In 1921, the use of phages against bacterial infections was first demonstrated...
November 28, 2018: Journal of Basic Microbiology
Hayder Nsaif Jasim, Rand Riadh Hafidh, Ahmed Sahib Abdulamir
Objectives: Phage therapy is a potential alternative treatment for infections caused by Acinetobacter baumannii , a significant nosocomial pathogen, which has evolved resistance to almost all conventional antimicrobial drugs in poor hygiene and conflicts areas such as Iraq. Materials and Methods: Bacteriophages were isolated to highly resistant isolates of A. baumannii to form therapeutic phage cocktail, and to extract and evaluate native endolysin activity. Bacterial samples were collected in Al-Imamein Al-kadhimein Medical City Hospital...
November 2018: Iranian Journal of Basic Medical Sciences
Marek Harhala, Daniel C Nelson, Paulina Miernikiewicz, Ryan D Heselpoth, Beata Brzezicka, Joanna Majewska, Sara B Linden, Xiaoran Shang, Aleksander Szymczak, Dorota Lecion, Karolina Marek-Bukowiec, Marlena Kłak, Bartosz Wojciechowicz, Karolina Lahutta, Andrzej Konieczny, Krystyna Dąbrowska
Bacteriophage-derived endolysins have gained increasing attention as potent antimicrobial agents and numerous publications document the in vivo efficacy of these enzymes in various rodent models. However, little has been documented about their safety and toxicity profiles. Here, we present preclinical safety and toxicity data for two pneumococcal endolysins, Pal and Cpl-1. Microarray, and gene profiling was performed on human macrophages and pharyngeal cells exposed to 0.5 µM of each endolysin for six hours and no change in gene expression was noted...
November 15, 2018: Viruses
Jan W Kretzer, Mathias Schmelcher, Martin J Loessner
The genus Listeria includes foodborne pathogens that cause life-threatening infections in those at risk, and sensitive and specific methods for detection of these bacteria are needed. Based on their unrivaled host specificity and ability to discriminate viable cells, bacteriophages represent an ideal toolbox for the development of such methods. Here, the authors describe an ultrasensitive diagnostic protocol for Listeria by combining two phage-based strategies: (1) specific capture and concentration of target cells by magnetic separation, harnessing cell wall-binding domains from Listeria phage endolysins (CBD-MS); and (2) highly sensitive detection using an adaptation of the A511:: luxAB bioluminescent reporter phage assay in a microwell plate format...
November 13, 2018: Viruses
Denise Mehner-Breitfeld, Claudia Rathmann, Thomas Riedel, Ingo Just, Ralf Gerhard, Jörg Overmann, Thomas Brüser
The pathogenicity locus (PaLoc) of Clostridioides difficile usually comprises five genes ( tcdR, tcdB, tcdE, tcdA, tcdC ). While the proteins TcdA and TcdB represent the main toxins of this pathogen, TcdR and TcdC are involved in the regulation of their production. TcdE is a holin family protein, members of which are usually involved in the transport of cell wall-degrading enzymes (endolysins) for phage-induced lysis. In the past, TcdE has been shown to contribute to the release of TcdA and TcdB, but it is unclear whether it mediates a specific transport or rather a lysis of cells...
2018: Frontiers in Microbiology
Yihui Yuan, Qin Peng, Shuo Yang, Shaowen Zhang, Yajuan Fu, Yan Wu, Meiying Gao
Phages, the parasites of bacteria, are considered as a new kind of antimicrobial agent due to their ability to lyse pathogenic bacteria. Due to the increase of available phage isolates, the newly isolated phage showed increasing genomic similarities with previously isolated phages. In this study, the novel phage vB_BthS_BMBphi, infecting the Bacillus thuringiensis strain BMB171, is isolated and characterized together with its endolysin. This phage is the first tadpole-like phage infecting the Bacillus strains...
November 6, 2018: Viruses
Masanori Toyofuku, Nobuhiko Nomura, Leo Eberl
Most bacteria release membrane vesicles (MVs) that contain specific cargo molecules and have diverse functions, including the transport of virulence factors, DNA transfer, interception of bacteriophages, antibiotics and eukaryotic host defence factors, cell detoxification and bacterial communication. MVs not only are abundant in nature but also show great promise for applications in biomedicine and nanotechnology. MVs were first discovered to originate from controlled blebbing of the outer membrane of Gram-negative bacteria and are therefore often called outer-membrane vesicles (OMVs)...
January 2019: Nature Reviews. Microbiology
Peiling Geng, Xiaofu Wan, Jiao Cheng, Zhiming Yuan, Xiaomin Hu
Lysinibacillus sphaericus has great application potential not only in the biocontrol of mosquitoes but also in the bioremediation of toxic metals. Phages contribute to the genetic diversity and niche adaptation of bacteria, playing essential roles in their life cycle, but may also cause economic damage for industrially important bacteria through phage contamination during fermentation. In this study, the L. sphaericus phage vB_LspM-01, which belongs to the Myoviridae family, was isolated and characterized. Results showed that vB_LspM-01 could specifically infect most tested L...
October 25, 2018: Applied Microbiology and Biotechnology
Peng Luo, Long Yun, Yingying Li, Yushun Tian, Qiuting Liu, Wen Huang, Chaoqun Hu
BACKGROUND: Biocontrol of bacterial pathogens by bacteriophages (phages) represents a promising strategy. Vibrio alginolyticus, a gram-negative bacterium, is a notorious pathogen responsible for the loss of economically important farmed marine animals. To date, few V. alginolyticus phages have been successfully isolated, and only three complete genome sequences of them have been released. The limited available phage resources and poor genomic data hamper research on V. alginolyticus phages and their applications for the biocontrol of V...
October 11, 2018: BMC Genomics
Xia Wu, Keith Fraser, Jian Zha, Jonathan S Dordick
Chemical linkers are frequently used in enzyme immobilization to improve enzyme flexibility and activity, whereas peptide linkers, although ubiquitous in protein engineering, are much less explored in enzyme immobilization. Here, we report peptide-linker-assisted noncovalent immobilization of the bacteriolytic enzyme lysostaphin (Lst) to generate anti- Staphylococcus aureus surfaces. Lst was immobilized through affinity tags onto a silica surface (glass slides) and nickel nitrilotriacetic acid (NiNTA) agarose beads via silica-binding peptides (SiBPs) or a hexahistidine tag (His-tag) fused at the C-terminus of Lst, respectively...
October 31, 2018: ACS Applied Materials & Interfaces
Patricia Letrado, Bruno Corsini, Roberto Díez-Martínez, Noemí Bustamante, José E Yuste, Pedro García
AIM: To test the synergistic effect of Cpl-711 endolysin and antibiotics for antipneumococcal activity. MATERIALS & METHODS: A combination of Cpl-711 and different antibiotics (amoxicillin, cefotaxime, levofloxacin and vancomycin) was tested in a checkerboard assay against several multidrug-resistant Streptococcus pneumoniae strains. Mouse and zebrafish models of pneumococcal sepsis were used to confirm the in vitro data. RESULTS: The activity of Cpl-711 combined with amoxicillin or cefotaxime was synergistic in the bactericidal effect against a serotype 23F multiresistant clinical isolate of S...
September 2018: Future Microbiology
Sebastian S Broendum, Ashley M Buckle, Sheena McGowan
Bacteriophage-encoded endolysins can recognize and bind specific bacteria, and act to cleave the glycosidic and/or amide bonds in the peptidoglycan bacterial cell wall. Cleavage of the cell wall generally results in the death of the bacteria. Their utility as bacteriolytic agents could be exploited for human and veterinary medicines and as well as various biotechnological applications. As interest grows in the commercial uses of these proteins, there has been much effort to successfully employ rational design and engineering to produce endolysins with bespoke properties...
September 19, 2018: Molecular Microbiology
Rohit Kongari, Jeffrey Snowden, Joel D Berry, Ry Young
Spanins are bacteriophage lysis proteins responsible for disruption of the outer membrane, the final step of Gram-negative host lysis. The absence of spanins results in a terminal phenotype of fragile spherical cells. The phage T1 employs a unimolecular spanin gp 11 that has an N-terminal lipoylation signal and a C-terminal transmembrane domain. Upon maturation and localization, gp 11 ends up as an outer membrane lipoprotein with a C-terminal transmembrane domain embedded in the inner membrane, thus connecting both membranes as a covalent polypeptide chain...
November 15, 2018: Journal of Virology
Yuhui Yang, Shuai Le, Wei Shen, Qian Chen, Youying Huang, Shuguang Lu, Yinling Tan, Ming Li, Fuquan Hu, Yang Li
Multidrug-resistant Pseudomonas aeruginosa is one of the most life-threatening pathogens for global health. In this regard, phage encoded lytic proteins, including endolysins and virion-associated peptidoglycan hydrolases (VAPGH), have been proposed as promising antimicrobial agents to treat P. aeruginosa . Most dsDNA phages use VAPGH to degrade peptidoglycan (PG) during infection, and endolysin to lyse the host cells at the end of lytic cycle. By contrast, dsRNA phage encodes only one lytic protein, which is located in the viral membrane to digest the PG during penetration, and also serves as an endolysin to release the phage...
2018: Frontiers in Microbiology
Nanna M C Olsen, Elowine Thiran, Tobias Hasler, Thomas Vanzieleghem, Georgios N Belibasakis, Jacques Mahillon, Martin J Loessner, Mathias Schmelcher
Staphylococcus aureus is an important pathogen and biofilm former. Biofilms cause problems in clinics and food production and are highly recalcitrant to antibiotics and sanitizers. Bacteriophage endolysins kill bacteria by degrading their cell wall and are therefore deemed promising antimicrobials and anti-biofilm agents. Depolymerases targeting polysaccharides in the extracellular matrix have been suggested as parts of a multi-enzyme approach to eradicate biofilms. The efficacy of endolysins and depolymerases against S...
August 18, 2018: Viruses
Colin Buttimer, Alan Lucid, Horst Neve, Charles M A P Franz, Jim O'Mahony, Dann Turner, Rob Lavigne, Aidan Coffey
Pectobacterium atrosepticum is a phytopathogen of economic importance as it is the causative agent of potato blackleg and soft rot. Here we describe the Pectobacterium phage vB_PatP_CB5 (abbreviated as CB5), which specifically infects the bacterium. The bacteriophage is characterized in detail and TEM micrographs indicate that it belongs to the Podoviridae family . CB5 shares significant pairwise nucleotide identity (≥80%) with P. atrosepticum phages φM1, Peat1, and PP90 and also shares common genome organization...
July 26, 2018: Viruses
Nak-Hyun Kim, Wan Beom Park, Jeong Eun Cho, Yoon Jeong Choi, Su Jin Choi, Soo Youn Jun, Chang Kyung Kang, Kyoung-Ho Song, Pyoeng Gyun Choe, Ji-Hwan Bang, Eu Suk Kim, Sang Won Park, Nam-Joong Kim, Myoung-Don Oh, Hong Bin Kim
Phages and their derivatives are increasingly being reconsidered for use in the treatment of bacterial infections due to the rising rates of antibiotic resistance. We assessed the antistaphylococcal effect of the endolysin SAL200 in combination with standard-of-care (SOC) antibiotics. The activity of SAL200 when it was combined with SOC antibiotics was assessed in vitro by checkerboard and time-kill assays and in vivo with murine bacteremia and Galleria mellonella infection models. SAL200 reduced the SOC antibiotic MICs and showed a ≥3-log10 -CFU/ml reduction of Staphylococcus aureus counts within 30 min in time-kill assays...
October 2018: Antimicrobial Agents and Chemotherapy
Casey Stamereilers, Christopher P Fajardo, Jamison K Walker, Katterinne N Mendez, Eduardo Castro-Nallar, Julianne H Grose, Sandra Hope, Philippos K Tsourkas
The antibiotic-resistant bacterium Paenibacillus larvae is the causative agent of American foulbrood (AFB), currently the most destructive bacterial disease in honeybees. Phages that infect P. larvae were isolated as early as the 1950s, but it is only in recent years that P. larvae phage genomes have been sequenced and annotated. In this study we analyze the genomes of all 48 currently sequenced P. larvae phage genomes and classify them into four clusters and a singleton. The majority of P. larvae phage genomes are in the 38⁻45 kbp range and use the cohesive ends (cos) DNA-packaging strategy, while a minority have genomes in the 50⁻55 kbp range that use the direct terminal repeat (DTR) DNA-packaging strategy...
July 19, 2018: Viruses
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