scholarly journals Fusion with a cell wall binding domain renders autolysin LytM a potent anti-Staphylococcus aureus agent

2014 ◽  
Vol 362 (2) ◽  
pp. 1-7 ◽  
Author(s):  
Daniel C. Osipovitch ◽  
Karl E. Griswold
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Binding Domain ◽  
Cell Wall Binding ◽  
A Cell ◽  
Cell Wall Binding Domain

scholarly journals LysPBC2, a Novel Endolysin Harboring aBacillus cereusSpore Binding Domain

2018 ◽  
Vol 85 (5) ◽  
Author(s):  
Minsuk Kong ◽  
Hongjun Na ◽  
Nam-Chul Ha ◽  
Sangryeol Ryu
Keyword(s):  
Cell Wall ◽  
Catalytic Domain ◽  
Binding Activity ◽  
Binding Domain ◽  
Lytic Activity ◽  
Content Type ◽  
Cell Wall Binding ◽  
A Cell ◽  
Cell Wall Binding Domain

ABSTRACTTo control the spore-forming human pathogenBacillus cereus, we isolated and characterized a novel endolysin, LysPBC2, from a newly isolatedB. cereusphage, PBC2. Compared to the narrow host range of phage PBC2, LysPBC2 showed very broad lytic activity against allBacillus,Listeria, andClostridiumspecies tested. In addition to a catalytic domain and a cell wall binding domain, LysPBC2 has a spore binding domain (SBD) partially overlapping its catalytic domain, which specifically binds toB. cereusspores but not to vegetative cells ofB. cereus. Both immunogold electron microscopy and a binding assay indicated that the SBD binds the external region of the spore cortex layer. Several amino acid residues required for catalytic or spore binding activity of LysPBC2 were determined by mutagenesis studies. Interestingly, LysPBC2 derivatives with impaired spore binding activity showed an increased lytic activity against vegetative cells ofB. cereuscompared with that of wild-type LysPBC2. Further biochemical studies revealed that these LysPBC2 derivatives have lower thermal stability, suggesting a stabilizing role of SBD in LysPBC2 structure.IMPORTANCEBacteriophages produce highly evolved lytic enzymes, called endolysins, to lyse peptidoglycan and release their progeny from bacterial cells. Due to their potent lytic activity and specificity, the use of endolysins has gained increasing attention as a natural alternative to antibiotics. Since most endolysins from Gram-positive-bacterium-infecting phages have a modular structure, understanding the function of each domain is crucial to make effective endolysin-based therapeutics. Here, we report the functional and biochemical characterization of aBacillus cereusphage endolysin, LysPBC2, which has an unusual spore binding domain and a cell wall binding domain. A single point mutation in the spore binding domain greatly enhanced the lytic activity of endolysin at the cost of reduced thermostability. This work contributes to the understanding of the role of each domain in LysPBC2 and will provide insight for the rational design of efficient antimicrobials or diagnostic tools for controllingB. cereus.

scholarly journals Endolysin of bacteriophage BFK20: evidence of a catalytic and a cell wall binding domain

2011 ◽  
Vol 321 (2) ◽  
pp. 83-91 ◽  
Author(s):  
Martina Gerova ◽  
Nora Halgasova ◽  
Jana Ugorcakova ◽  
Gabriela Bukovska
Keyword(s):  
Cell Wall ◽  
Binding Domain ◽  
Cell Wall Binding ◽  
A Cell ◽  
Cell Wall Binding Domain

scholarly journals An Engineered Multimodular Enzybiotic against Methicillin-Resistant Staphylococcus aureus

Life ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 1384
Author(s):  
Salim Manoharadas ◽  
Mohammad Altaf ◽  
Abdulwahed Fahad Alrefaei ◽  
Naushad Ahmad ◽  
Shaik Althaf Hussain ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Pathogenic Bacteria ◽  
Antimicrobial Agents ◽  
Methicillin Resistant Staphylococcus Aureus ◽  
Binding Domain ◽  
Strong Activity ◽  
Methicillin Resistant ◽  
Cell Wall Binding ◽  
Cell Wall Binding Domain

Development of multidrug antibiotic resistance in bacteria is a predicament encountered worldwide. Researchers are in a constant hunt to develop effective antimicrobial agents to counter these dreadful pathogenic bacteria. Here we describe a chimerically engineered multimodular enzybiotic to treat a clinical isolate of methicillin-resistant Staphylococcus aureus (S. aureus). The cell wall binding domain of phage ϕ11 endolysin was replaced with a truncated and more potent cell wall binding domain from a completely unrelated protein from a different phage. The engineered enzybiotic showed strong activity against clinically relevant methicillin-resistant Staphylococcus aureus. In spite of a multimodular peptidoglycan cleaving catalytic domain, the engineered enzybiotic could not exhibit its activity against a veterinary isolate of S. aureus. Our studies point out that novel antimicrobial proteins can be genetically engineered. Moreover, the cell wall binding domain of the engineered protein is indispensable for a strong binding and stability of the proteins.

scholarly journals Characterization of an Endolysin Targeting Clostridioides difficile That Affects Spore Outgrowth

2021 ◽  
Vol 22 (11) ◽  
pp. 5690
Author(s):  
Shakhinur Islam Mondal ◽  
Arzuba Akter ◽  
Lorraine A. Draper ◽  
R. Paul Ross ◽  
Colin Hill
Keyword(s):  
Cell Wall ◽  
Therapeutic Potential ◽  
Bacterial Species ◽  
Binding Domain ◽  
Cell Wall Binding ◽  
Clostridioides Difficile ◽  
Life Threatening ◽  
A Cell ◽  
Cell Wall Binding Domain ◽  
Spore Outgrowth

Clostridioides difficile is a spore-forming enteric pathogen causing life-threatening diarrhoea and colitis. Microbial disruption caused by antibiotics has been linked with susceptibility to, and transmission and relapse of, C. difficile infection. Therefore, there is an urgent need for novel therapeutics that are effective in preventing C. difficile growth, spore germination, and outgrowth. In recent years bacteriophage-derived endolysins and their derivatives show promise as a novel class of antibacterial agents. In this study, we recombinantly expressed and characterized a cell wall hydrolase (CWH) lysin from C. difficile phage, phiMMP01. The full-length CWH displayed lytic activity against selected C. difficile strains. However, removing the N-terminal cell wall binding domain, creating CWH351—656, resulted in increased and/or an expanded lytic spectrum of activity. C. difficile specificity was retained versus commensal clostridia and other bacterial species. As expected, the putative cell wall binding domain, CWH1—350, was completely inactive. We also observe the effect of CWH351—656 on preventing C. difficile spore outgrowth. Our results suggest that CWH351—656 has therapeutic potential as an antimicrobial agent against C. difficile infection.

scholarly journals The structure of Rv3717 reveals a novel amidase fromMycobacterium tuberculosis

2013 ◽  
Vol 69 (12) ◽  
pp. 2543-2554 ◽  
Author(s):  
Atul Kumar ◽  
Sanjiv Kumar ◽  
Dilip Kumar ◽  
Arpit Mishra ◽  
Rikeshwer P. Dewangan ◽  
...  
Keyword(s):  
Cell Wall ◽  
Bartonella Henselae ◽  
General Mechanism ◽  
Isolation And Identification ◽  
Cell Wall Binding ◽  
New Family ◽  
A Cell ◽  
Sad Phasing ◽  
Cell Wall Binding Domain ◽  
Net Positive Charge

BacterialN-acetylmuramoyl-L-alanine amidases are cell-wall hydrolases that hydrolyze the bond betweenN-acetylmuramic acid and L-alanine in cell-wall glycopeptides. Rv3717 ofMycobacterium tuberculosishas been identified as a unique autolysin that lacks a cell-wall-binding domain (CBD) and its structure has been determined to 1.7 Å resolution by the Pt-SAD phasing method. Rv3717 possesses an α/β-fold and is a zinc-dependent hydrolase. The structure reveals a short flexible hairpin turn that partially occludes the active site and may be involved in autoregulation. This type of autoregulation of activity of PG hydrolases has been observed inBartonella henselaeamidase (AmiB) and may be a general mechanism used by some of the redundant amidases to regulate cell-wall hydrolase activity in bacteria. Rv3717 utilizes its net positive charge for substrate binding and exhibits activity towards a broad spectrum of substrate cell walls. The enzymatic activity of Rv3717 was confirmed by isolation and identification of its enzymatic products by LC/MS. These studies indicate that Rv3717, anN-acetylmuramoyl-L-alanine amidase fromM. tuberculosis, represents a new family of lytic amidases that do not have a separate CBD and are regulated conformationally.

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