Influence of bacterial culture medium on peptidoglycan binding of cell wall lytic enzymes

Bhagwat, Amala; Zhang, Fuming; Collins, Cynthia H.; Dordick, Jonathan S.

doi:10.1016/j.jbiotec.2021.02.010

Cited by 7 publications

(11 citation statements)

References 46 publications

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“…Two key questions that still require further elucidation are: (i) what are the determinants and mechanisms responsible for tolerance, and (ii) what are the exact mechanisms by which AMPs abolish tolerance and strongly stimulate endolysin lytic activity. In agreement with previous reports involving endolysins and autolysins 17 – 19 , 29 , we verified that WTA were important for S. aureus tolerance to Lys11. Based on the observation that in rich media WTA can hinder the binding of peptidoglycan-degrading enzymes to bacterial cells, namely to S. aureus , it was proposed that tolerance relied on a WTA-dependent shielding of the CW 17 – 19 .…”

Section: Discussionsupporting

confidence: 93%

“…In agreement with previous reports involving endolysins and autolysins 17 – 19 , 29 , we verified that WTA were important for S. aureus tolerance to Lys11. Based on the observation that in rich media WTA can hinder the binding of peptidoglycan-degrading enzymes to bacterial cells, namely to S. aureus , it was proposed that tolerance relied on a WTA-dependent shielding of the CW 17 – 19 . We have also observed such inhibitory role of WTA on Lys11 binding.…”

Section: Discussionsupporting

confidence: 93%

“…One aspect we started to explore in this work was the identification of cellular determinants of tolerance to endolysins. WTA were previously implicated in tolerance based on their capacity to hamper the binding of some lytic enzymes to target bacteria 17 – 19 , but for some endolysins they were also shown to work as the ligands required for specific recognition and binding to the CW 20 . We have therefore studied the impact of WTA on S. aureus susceptibility to Lys11 lytic action and on endolysin binding to cells.…”

Section: Resultsmentioning

confidence: 99%

“…Other studies have shown that WTA can be responsible for restricting access of autolysins and endolysins to certain regions of the CW, therefore coordinating their spatial distribution 15 , 16 . More recently, it was proposed that WTA can protect bacteria from the attack of lytic enzymes by hindering their binding to the CW, with this shielding effect being stronger when cells are in rich nutritional media 17 – 19 . For other enzymes however, binding to WTA appears to be a requirement for lytic activity 20 .…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Synthetic antimicrobial peptides as enhancers of the bacteriolytic action of staphylococcal phage endolysins

Gouveia

Pinto

Veiga

et al. 2022

Sci Rep

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Bacteriophage endolysins degrade the bacterial cell wall and are therefore considered promising antimicrobial alternatives to fight pathogens resistant to conventional antibiotics. Gram-positive bacteria are usually considered easy targets to exogenously added endolysins, since their cell walls are not shielded by an outer membrane. However, in nutrient rich environments these bacteria can also tolerate endolysin attack if they keep an energized cytoplasmic membrane. Hence, we have hypothesized that the membrane depolarizing action of antimicrobial peptides (AMPs), another attractive class of alternative antibacterials, could be explored to overcome bacterial tolerance to endolysins and consequently improve their antibacterial potential. Accordingly, we show that under conditions supporting bacterial growth, Staphylococcus aureus becomes much more susceptible to the bacteriolytic action of endolysins if an AMP is also present. The bactericidal gain resulting from the AMP/endolysin combined action ranged from 1 to 3 logs for different S. aureus strains, which included drug-resistant clinical isolates. In presence of an AMP, as with a reduced content of cell wall teichoic acids, higher endolysin binding to cells is observed. However, our results indicate that this higher endolysin binding alone does not fully explain the higher susceptibility of S. aureus to lysis in these conditions. Other factors possibly contributing to the increased endolysin susceptibility in presence of an AMP are discussed.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionsupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Synthetic antimicrobial peptides as enhancers of the bacteriolytic action of staphylococcal phage endolysins

Gouveia

Pinto

Veiga

et al. 2022

Sci Rep

View full text Add to dashboard Cite

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“…On the other hand, considering that the clinical application of endolysins is not only environmental disinfection, but also food disinfection and in vivo sterilization, it is important to have stable bactericidal activity under eutrophic conditions. However, reports showed that the killing activity of most endolysins is significantly reduced or lost under eutrophic conditions such as LB medium and serum [ 35 , 36 ]. To our knowledge, anti-staphylococcal Lysin CF301 is the only lysin that exhibits substantially greater potency in human blood [ 37 , 38 ].…”

Section: Introductionmentioning

confidence: 99%

Characterization of Salmonella endolysin XFII produced by recombinant Escherichia coli and its application combined with chitosan in lysing Gram-negative bacteria

et al. 2022

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Background Salmonella is a common foodborne pathogen, which can cause intestinal diseases. In the last decades, the overuse of antibiotics has led to a pandemic of drug-resistant bacterial infections. To tackle the burden of antimicrobial resistant pathogens, it is necessary to develop new antimicrobial drugs with novel modes of action. However, the research and development of antibiotics has encountered bottlenecks, scientific hurdles in the development process, as well as safety and cost challenges. Phages and phage endolysins are promising antibacterial agents that can be used as an alternative to antibiotics. In this context, the expression of endolysin derived from different phages through microbial cells as a chassis seems to be an attractive strategy. Results In this study, a new endolysin from the Salmonella phage XFII-1, named XFII, was screened and obtained. The endolysin yield exceeded 100 mg/mL by heterologous expression from E. coli BL21 and short induction. The endolysin XFII exhibited high bactericidal activity at a concentration of 0.5 μg/mL and reduced the OD600 nm of EDTA-pretreated E. coli JM109 from 0.8 to 0.2 within 5 min. XFII exhibited good thermo-resistance, as it was very stable at different temperatures from 20 to 80℃. Its bactericidal activity could keep constant at 4 °C for 175 days. In addition, the endolysin was able to exert lytic activity in eutrophic conditions, including LB medium and rabbit serum, and the lytic activity was even increased by 13.8% in 10% serum matrices. XFII also showed bactericidal activity against many Gram-negative bacteria, including Salmonella, E. coli, Acinetobacter baumannii, and Klebsiella pneumoniae. Surprisingly, the combination of endolysin XFII and chitosan showed a strong synergy in lysing E. coli and Salmonella without EDTA-pretreatment, and the OD600 nm of E. coli decreased from 0.88 to 0.58 within 10 min. Conclusions The novel globular endolysin XFII was screened and successfully expressed in E. coli BL21. Endolysin XFII exhibits a broad lysis spectrum, a rapid and strong bactericidal activity, good stability at high temperatures and under eutrophic conditions. Combined with chitosan, XFII could spontaneously lyse Gram-negative bacteria without pretreatment. This work presented the first characterization of combining endolysin and chitosan in spontaneously lysing Gram-negative bacteria in vitro.

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Influence of NaCl and pH on lysostaphin catalytic activity, cell binding, and bacteriolytic activity

Konstantinova

Grishin

Lyashchuk

et al. 2022

Appl Microbiol Biotechnol

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Influence of bacterial culture medium on peptidoglycan binding of cell wall lytic enzymes

Cited by 7 publications

References 46 publications

Synthetic antimicrobial peptides as enhancers of the bacteriolytic action of staphylococcal phage endolysins

Synthetic antimicrobial peptides as enhancers of the bacteriolytic action of staphylococcal phage endolysins

Characterization of Salmonella endolysin XFII produced by recombinant Escherichia coli and its application combined with chitosan in lysing Gram-negative bacteria

Influence of NaCl and pH on lysostaphin catalytic activity, cell binding, and bacteriolytic activity

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