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Nina Schleimer, Ursula Kaspar, Dennis Knaack, Christof von Eiff, Sonja Molinaro, Holger Grallert, Evgeny A Idelevich, Karsten Becker
Nasal carriage of methicillin-susceptible (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) represents both a source and a risk factor for subsequent infections. However, existing MRSA decolonization strategies and antibiotic treatment options are hampered by the duration of administration and particularly by the emergence of resistance. Moreover, beyond classical resistance mechanisms, functional resistance as the formation of the small-colony variant (SCV) phenotype may also impair the course and treatment of S ...
February 7, 2019: International Journal of Molecular Sciences
Vera Bischoff, Boyke Bunk, Jan P Meier-Kolthoff, Cathrin Spröer, Anja Poehlein, Marco Dogs, Mary Nguyen, Jörn Petersen, Rolf Daniel, Jörg Overmann, Markus Göker, Meinhard Simon, Thorsten Brinkhoff, Cristina Moraru
Bacteriophages are widely considered to influence bacterial communities, however most phages are still unknown or not studied well enough to understand their ecological roles. We have isolated two phages infecting Lentibacter sp. SH36, affiliated with the marine Roseobacter group, and retrieved similar phage genomes from publicly available metagenomics databases. Phylogenetic analysis placed the new phages within the Cobavirus group, in the here newly proposed genus Siovirus and subfamily Riovirinae of the Podoviridae...
February 4, 2019: ISME Journal
Magdalena Plotka, Enea Sancho-Vaello, Sebastian Dorawa, Anna-Karina Kaczorowska, Lukasz P Kozlowski, Tadeusz Kaczorowski, Kornelius Zeth
To escape from hosts after completing their life cycle, bacteriophages often use endolysins, which degrade bacterial peptidoglycan. While mesophilic phages have been extensively studied, their thermophilic counterparts are not well characterized. Here, we present a detailed analysis of the structure and function of Ts2631 endolysin from thermophilic phage vB_Tsc2631, which is a zinc-dependent amidase. The active site of Ts2631 consists of His30, Tyr58, His131 and Cys139, which are involved in Zn2+ coordination and catalysis...
February 4, 2019: Scientific Reports
Meenakshi Sharama, Nancy Jaswal, Dinesh Kumar, Krishna Mohan Poluri
Proteins are dynamic in nature and exists in a set of equilibrium conformations on various timescale motions. The flexibility of proteins governs various biological functions, and therefore elucidation of such functional dynamics is essential. In this context, we have studied the structure-dynamics-stability-activity relationship of bacteriophage T7 lysozyme/endolysin (T7L) native state ensemble in the pH range 6-8. Our studies established that T7L native state is conformationally heterogeneous, as several residues of its C-terminal half are present in two conformations (major and minor) in the slow exchange time scale of NMR...
January 25, 2019: Biochemical Journal
Ji Yun Bae, Kang Il Jun, Chang Kyung Kang, Kyoung-Ho Song, Pyoeng Gyun Choe, Ji-Hwan Bang, Eu Suk Kim, Sang Won Park, Hong Bin Kim, Nam-Joong Kim, Wan Beom Park, Myoung-Don Oh
SAL200 is derived from a phage endolysin and is a novel candidate drug for the treatment of Staphylococcus aureus infection. We investigated the efficacy of the recombinant endolysin SAL200 in a lethal murine pneumonia model. Lethal pneumonia was established by intranasally administering a methicillin-susceptible (Newman) or methicillin-resistant (LAC) S. aureus strain into BALB/c mice. The mice were treated with single intranasal administration of SAL200 or phosphate-buffered saline at 2 h after S. aureus infection...
January 22, 2019: Antimicrobial Agents and Chemotherapy
Jesse Cahill, Ry Young
The first steps in phage lysis involve a temporally controlled permeabilization of the cytoplasmic membrane followed by enzymatic degradation of the peptidoglycan. For Caudovirales of Gram-negative hosts, there are two different systems: the holin-endolysin and pinholin-SAR endolysin pathways. In the former, lysis is initiated when the holin forms micron-scale holes in the inner membrane, releasing active endolysin into the periplasm to degrade the peptidoglycan. In the latter, lysis begins when the pinholin causes depolarization of the membrane, which activates the secreted SAR endolysin...
2019: Advances in Virus Research
Lorena Rodríguez-Rubio, David M Donovan, Beatriz Martínez, Ana Rodríguez, Pilar García
Zymogram or zymography is an electrophoretic technique based on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), which enables visualization of enzymatically active protein species separated by molecular mass. The strategy is to perform SDS-PAGE on the proteins in question while including an opaque substrate of the enzyme embedded within the polyacrylamide gel. Here, we describe a zymogram protocol for phage lytic proteins (peptidoglycan hydrolases) using peptidoglycan (or whole cells) from a susceptible gram-positive bacterial species as substrate...
2019: Methods in Molecular Biology
Dennis Knaack, Evgeny A Idelevich, Nina Schleimer, Sonja Molinaro, André Kriegeskorte, Georg Peters, Karsten Becker
Methicillin-resistant Staphylococcus aureus (MRSA) decolonization is expensive and time consuming, and new agents are necessary due to increasing resistance rates. The administration of bacteriophages or particularly their endolysins may offer an alternative treatment strategy and could provide a solution to overcome the selection pressure due to classical antibiotics. Here, the bactericidal activity was characterized for the recombinant chimeric bacteriophage endolysin HY-133 in comparison to other antimicrobials...
November 20, 2018: Diagnostic Microbiology and Infectious Disease
Minsuk Kong, Hongjun Na, Nam-Chul Ha, Sangryeol Ryu
To control a spore-forming human pathogen Bacillus cereus , we isolated and characterized a novel endolysin, LysPBC2, from a newly isolated B. cereus phage PBC2. Compared to the narrow host range of phage PBC2, LysPBC2 showed very broad lytic activity against all Bacillus , Listeria , and Clostridium species tested. In addition to a catalytic domain and a cell wall binding domain, LysPBC2 has a spore binding domain (SBD) partially overlapped with its catalytic domain, which specifically binds to B. cereus spores but not to vegetative cells of B...
December 14, 2018: Applied and Environmental Microbiology
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
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