The rise in multidrug-resistant bacteria defines the need for identification of new antibacterial agents that are less prone to resistance acquisition. Compounds that simultaneously inhibit multiple bacterial targets are more likely to suppress the evolution of target-based resistance than monotargeting compounds. The structurally similar ATP binding sites of DNA gyrase and topoisomerase Ⅳ offer an opportunity to accomplish this goal. Here we present the design and structure-activity relationship analysis of balanced, low nanomolar inhibitors of bacterial DNA gyrase and topoisomerase IV that show potent antibacterial activities against the ESKAPE pathogens. For inhibitor 31c, a crystal structure in complex with Staphylococcus aureus DNA gyrase B was obtained that confirms the mode of action of these compounds. The best inhibitor, 31h, does not show any in vitro cytotoxicity and has excellent potency against Gram-positive (MICs: range, 0.0078-0.0625 μg/mL) and Gram-negative pathogens (MICs: range, 1-2 μg/mL). Furthermore, 31h inhibits GyrB mutants that can develop resistance to other drugs. Based on these data, we expect that structural derivatives of 31h will represent a step toward clinically efficacious multitargeting antimicrobials that are not impacted by existing antimicrobial resistance.
- MeSH
- adenosintrifosfát chemická syntéza chemie farmakologie MeSH
- antibakteriální látky chemická syntéza chemie farmakologie MeSH
- DNA gyráza metabolismus MeSH
- DNA-topoisomerasa IV antagonisté a inhibitory metabolismus MeSH
- Escherichia coli účinky léků enzymologie patogenita MeSH
- krystalografie rentgenová MeSH
- mikrobiální testy citlivosti MeSH
- molekulární struktura MeSH
- simulace molekulového dockingu MeSH
- Staphylococcus aureus účinky léků enzymologie patogenita MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Publikační typ
- časopisecké články MeSH
Remote-controlled robotic systems are being used for analysis of various types of analytes in hostile environment including those called extraterrestrial. The aim of our study was to develop a remote-controlled robotic platform (ORPHEUS-HOPE) for bacterial detection. For the platform ORPHEUS-HOPE a 3D printed flow chip was designed and created with a culture chamber with volume 600 μL. The flow rate was optimized to 500 μL/min. The chip was tested primarily for detection of 1-naphthol by differential pulse voltammetry with detection limit (S/N = 3) as 20 nM. Further, the way how to capture bacteria was optimized. To capture bacterial cells (Staphylococcus aureus), maghemite nanoparticles (1 mg/mL) were prepared and modified with collagen, glucose, graphene, gold, hyaluronic acid, and graphene with gold or graphene with glucose (20 mg/mL). The most up to 50% of the bacteria were captured by graphene nanoparticles modified with glucose. The detection limit of the whole assay, which included capturing of bacteria and their detection under remote control operation, was estimated as 30 bacteria per μL.
- MeSH
- alkalická fosfatasa metabolismus MeSH
- design vybavení MeSH
- elektrochemické techniky přístrojové vybavení MeSH
- Escherichia coli enzymologie izolace a purifikace MeSH
- limita detekce MeSH
- magnetické nanočástice chemie MeSH
- mikrobiologie životního prostředí * MeSH
- mikrofluidní analytické techniky přístrojové vybavení MeSH
- naftoly izolace a purifikace MeSH
- robotika přístrojové vybavení MeSH
- Staphylococcus aureus enzymologie izolace a purifikace MeSH
- technologie dálkového snímání přístrojové vybavení MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Magnetic bead cellulose activated with divinyl sulfone was used for the immobilization of Staphylococcus aureus endoproteinase Glu-C (EC 3.4.21.19). The immobilized proteinase was characterized by increased thermostability, by decreased self-cleavage activity, and a possibility of repeated use. The prepared immobilized enzyme was applied for the proteolytic cleavage of α-casein and BSA under different conditions (different composition of buffers, different pH, and different time of digestion). The possibilities of the direct use of enzyme reaction products for MALDI TOF MS analysis were shown.
- MeSH
- bakteriální proteiny chemie MeSH
- celulosa chemie MeSH
- enzymy imobilizované chemie MeSH
- kaseiny chemie MeSH
- koncentrace vodíkových iontů MeSH
- magnetismus MeSH
- proteomika přístrojové vybavení metody MeSH
- serinové endopeptidasy chemie MeSH
- sérový albumin hovězí chemie MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- stabilita enzymů MeSH
- Staphylococcus aureus enzymologie MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Staphylococcus aureus ssp anaerobius strain S10 was isolated from an outbreak of sheep abscess disease. Sequence of the catalase gene of this strain showed 99% identity to the catalase gene (katB) sequence of the reference strain (S. aureus ssp. anaerobius strain MVF213) with mismatching of three base pairs. An important substitution located 1036 nucleotides upstream of the initiation codon from "C" in katB to "T" in the catalase gene of strain S10 originated a stop codon. The deduced protein (345 amino acids) is 105 amino acids shorter than that of katB. Partial sequence of the catalase gene of other 8 local isolates in addition to another reference strain (DSM 20714/ATCC 35844) revealed the same mutations in all local (African) strains, whereas the sequence of the reference (European) strain was typical to that of katB. Sequence of the catalase gene of S. aureus ssp. anaerobius strain S10 was deposited in GenBank under accession no. EU281993.
- MeSH
- absces epidemiologie mikrobiologie veterinární MeSH
- bakteriální proteiny genetika MeSH
- DNA bakterií genetika chemie MeSH
- epidemický výskyt choroby MeSH
- financování organizované MeSH
- katalasa genetika MeSH
- molekulární sekvence - údaje MeSH
- nemoci ovcí epidemiologie mikrobiologie MeSH
- nesmyslný kodon MeSH
- ovce MeSH
- polymorfismus genetický MeSH
- sekvenční analýza DNA MeSH
- sekvenční homologie nukleových kyselin MeSH
- stafylokokové infekce epidemiologie mikrobiologie veterinární MeSH
- Staphylococcus aureus enzymologie izolace a purifikace klasifikace MeSH
- substituce aminokyselin genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- MeSH
- antibakteriální látky farmakologie MeSH
- anticholesteremika farmakologie MeSH
- bakteriální proteiny antagonisté a inhibitory chemie metabolismus MeSH
- farnesyl-difosfofarnesyltransferasa antagonisté a inhibitory chemie metabolismus MeSH
- inhibitory enzymů farmakologie MeSH
- krystalografie rentgenová MeSH
- lidé MeSH
- Staphylococcus aureus enzymologie genetika účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- komentáře MeSH
- MeSH
- antibakteriální látky imunologie MeSH
- bakteriální léková rezistence genetika imunologie účinky záření MeSH
- bakteriocinové plazmidy genetika imunologie škodlivé účinky MeSH
- enterotoxiny izolace a purifikace MeSH
- finanční podpora výzkumu jako téma MeSH
- králíci imunologie mikrobiologie MeSH
- rezistence na ampicilin genetika imunologie účinky léků MeSH
- rezistence na methicilin genetika imunologie účinky léků MeSH
- Staphylococcus aureus enzymologie izolace a purifikace účinky léků MeSH
- zvířata MeSH
- Check Tag
- králíci imunologie mikrobiologie MeSH
- zvířata MeSH
The new method exploits the characteristic of staphylococcal protease V8, which specifically splits peptide bonds where l-glutamic or l-aspartic acid participate. Those peptide bonds where d-aspartic acid is present remain unsplit because of the stereospecifity of enzymes. In accordance with expectations, fewer peptide bonds are split by this protease at more advanced ages and larger peptide fragments are thus formed due to the higher content of d-amino acid residues in the proteins of older people. The samples of acid-extracted non-collagenous proteins from dentin were separated using high performance liquid chromatography after enzymatic hydrolysis. A peak with a retention time of 45.3 min was chosen and his enlarging area showed a linear correlation with increasing age. Although the linear correlation with age was proved, the scattering of values decreases the usefulness of the proposed method for age estimation.
- MeSH
- dentin chemie MeSH
- dospělí MeSH
- financování organizované MeSH
- kyselina aspartová analýza MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- peptidové mapování MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- soudní stomatologie metody MeSH
- Staphylococcus aureus enzymologie MeSH
- určení zubního věku metody MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- MeSH
- bakteriální toxiny izolace a purifikace MeSH
- dítě MeSH
- epidermis mikrobiologie patologie MeSH
- exfoliativní dermatitida mikrobiologie MeSH
- finanční podpora výzkumu jako téma MeSH
- lidé MeSH
- nemoci novorozenců mikrobiologie MeSH
- polymerázová řetězová reakce metody využití MeSH
- Staphylococcus aureus enzymologie izolace a purifikace patogenita MeSH
- Check Tag
- dítě MeSH
- lidé MeSH
