Potential Role of the Host-Derived Cell-Wall Binding Domain of Endolysin CD16/50L as a Molecular Anchor in Preservation of Uninfected Clostridioides difficile for New Rounds of Phage Infection

Phothichaisri, Wichuda; Chankhamhaengdecha, Surang; Janvilisri, Tavan; Nuadthaisong, Jirayu; Phetruen, Tanaporn; Fagan, Robert P.; Chanarat, Sittinan

doi:10.1128/spectrum.02361-21

Cited by 14 publications

(13 citation statements)

References 100 publications

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“…The stationary phase cultures were then harvested by centrifugation at 6000× g for 5 min at 25 °C, washed once with sterile distilled water, and resuspended in 50 mM potassium phosphate (pH 7.5) and 10 mM 2-mercaptoethanol. Zymolyase-100T (0.4 units) (Zymo Research, Irvine, CA, USA) was added to cell suspensions and the absorbance at 600 nm was measured using a Multiskan SkyHigh (Thermo Fisher Scientific) every 5 min for 1 h [ 26 , 27 ].…”

Section: Methodsmentioning

confidence: 99%

Andrographolide Induces ROS-Mediated Cytotoxicity, Lipid Peroxidation, and Compromised Cell Integrity in Saccharomyces cerevisiae

Phetruen,

van Dam,

Chanarat

2023

Antioxidants

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Andrographolide, a bioactive compound found in Andrographis paniculata, has gained significant attention for its potential therapeutic properties. Despite its promising benefits, the understanding of its side effects and underlying mechanisms remains limited. Here, we investigated the impact of andrographolide in Saccharomyces cerevisiae and observed that andrographolide induced cytotoxicity, particularly when oxidative phosphorylation was active. Furthermore, andrographolide affected various cellular processes, including vacuole fragmentation, endoplasmic reticulum stress, lipid droplet accumulation, reactive oxygen species levels, and compromised cell integrity. Moreover, we unexpectedly observed that andrographolide induced the precipitation of biomolecules secreted from yeast cells, adding an additional source of stress. Overall, this study provides insights into the cellular effects and potential mechanisms of andrographolide in yeast, shedding light on its side effects and underlying cytotoxicity pathways.

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Section: Methodsmentioning

confidence: 99%

Andrographolide Induces ROS-Mediated Cytotoxicity, Lipid Peroxidation, and Compromised Cell Integrity in Saccharomyces cerevisiae

Phetruen,

van Dam,

Chanarat

2023

Antioxidants

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“…On the one hand, CBD may allow localization of the substrate (peptidoglycan) in the proximity of the catalytic site of the enzyme 132 ; on the other hand, it may anchor endolysins to peptidoglycan of post-lytic cell remnants, thus limiting their availability for other cells. 133 Accordingly, some recombinant endolysins have higher lytic activity when they lack CBD, whereas others have lower or no lytic activity. 129 , 134 Also, some studies indicate that EAD alone determines endolysin specificity, 135 which is somewhat broader compared with endolysins containing CBD.…”

Section: Phage-based Therapies Against C Difficile Infectionsmentioning

confidence: 99%

New treatment approaches for Clostridioides difficile infections: alternatives to antibiotics and fecal microbiota transplantation

Bratkovič,

Zahirović,

Bizjak

et al. 2024

Gut Microbes

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Clostridioides difficile causes a range of debilitating intestinal symptoms that may be fatal. It is particularly problematic as a hospital-acquired infection, causing significant costs to the health care system. Antibiotics, such as vancomycin and fidaxomicin, are still the drugs of choice for C. difficile infections, but their effectiveness is limited, and microbial interventions are emerging as a new treatment option. This paper focuses on alternative treatment approaches, which are currently in various stages of development and can be divided into four therapeutic strategies. Direct killing of C. difficile (i) includes beside established antibiotics, less studied bacteriophages, and their derivatives, such as endolysins and tailocins. Restoration of microbiota composition and function (ii) is achieved with fecal microbiota transplantation, which has recently been approved, with standardized defined microbial mixtures, and with probiotics, which have been administered with moderate success. Prevention of deleterious effects of antibiotics on microbiota is achieved with agents for the neutralization of antibiotics that act in the gut and are nearing regulatory approval. Neutralization of C. difficile toxins (iii) which are crucial virulence factors is achieved with antibodies/antibody fragments or alternative binding proteins. Of these, the monoclonal antibody bezlotoxumab is already in clinical use. Immunomodulation (iv) can help eliminate or prevent C. difficile infection by interfering with cytokine signaling. Small-molecule agents without bacteriolytic activity are usually selected by drug repurposing and can act via a variety of mechanisms. The multiple treatment options described in this article provide optimism for the future treatment of C. difficile infection.

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“…Of note, one common observation that has been reported with all C. difficile phage endolysins is that the catalytic domain of the enzyme alone (EAD) is sufficient for full activity. In fact, the removal of the CBD was reported to increase the lytic activity of C. difficile ( 30 – 32 , 36 , 38 ). It was also shown that the CBD from the CD16/50L endolysin was responsible for trapping the endolysin within cellular debris after lysis.…”

Section: Cell Wall-degrading Enzymes As Antimicrobialsmentioning

confidence: 99%

“…It was also shown that the CBD from the CD16/50L endolysin was responsible for trapping the endolysin within cellular debris after lysis. It was hypothesized that the CBD domain prevents the endolysin from being released freely into the environment upon cell lysis, therefore preserving uninfected bacteria that can serve as hosts for subsequent phage infection ( 36 ). Full-length endolysins were shown to be very specific toward C. difficile , as little or no activity was noted on other commensal bacteria, including related Clostridia.…”

Section: Cell Wall-degrading Enzymes As Antimicrobialsmentioning

confidence: 99%

The long and sinuous road to phage-based therapy of Clostridioides difficile infections

Umansky,

Fortier

2023

Front. Med.

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With the antibiotic crisis and the rise in antimicrobial resistance worldwide, new therapeutic alternatives are urgently needed. Phage therapy represents one of the most promising alternatives but for some pathogens, such as Clostridioides difficile, important challenges are being faced. The perspective of phage therapy to treat C. difficile infections is complicated by the fact that no strictly lytic phages have been identified so far, and current temperate phages generally have a narrow host range. C. difficile also harbors multiple antiphage mechanisms, and the bacterial genome is often a host of one or multiple prophages that can interfere with lytic phage infection. Nevertheless, due to recent advances in phage host receptor recognition and improvements in genetic tools to manipulate phage genomes, it is now conceivable to genetically engineer C. difficile phages to make them suitable for phage therapy. Other phage-based alternatives such as phage endolysins and phage tail-like bacteriocins (avidocins) are also being investigated but these approaches also have their own limitations and challenges. Last but not least, C. difficile produces spores that are resistant to phage attacks and all current antibiotics, and this complicates therapeutic interventions. This mini-review gives a brief historical overview of phage work that has been carried out in C. difficile, presents recent advances in the field, and addresses the most important challenges that are being faced, with potential solutions.

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Potential Role of the Host-Derived Cell-Wall Binding Domain of Endolysin CD16/50L as a Molecular Anchor in Preservation of Uninfected Clostridioides difficile for New Rounds of Phage Infection

Abstract: Endolysin is a peptidoglycan hydrolase encoded in a phage genome. The enzyme is attractive due to its potential use as antibacterial treatment.

Cited by 14 publications

References 100 publications

Andrographolide Induces ROS-Mediated Cytotoxicity, Lipid Peroxidation, and Compromised Cell Integrity in Saccharomyces cerevisiae

Andrographolide Induces ROS-Mediated Cytotoxicity, Lipid Peroxidation, and Compromised Cell Integrity in Saccharomyces cerevisiae

New treatment approaches for Clostridioides difficile infections: alternatives to antibiotics and fecal microbiota transplantation

The long and sinuous road to phage-based therapy of Clostridioides difficile infections

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