Effect of Low Molecular Weight Additives on Immobilization Strength, Activity, and Conformation of Protein Immobilized on PVC and UHMWPE

Kondyurin, Alexey; Nosworthy, Neil J.; Bilek, M.M.M.

doi:10.1021/la200376f

Cited by 30 publications

(21 citation statements)

References 67 publications

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“…Covalent attachment of the peptides on glass was confirmed with SDS treatment (Bilek and McKenzie, 2010). SDS can disrupt physical interactions such a hydrophobic binding between a protein and a surface (Bilek and McKenzie, 2010;Kondyurin et al, 2011). Both melimine and Mel4-coated surfaces lost only a small amount of amide nitrogen after SDS washing ( Table 1) confirming succesfull covalent attachment of the peptides.…”

Section: Discussionmentioning

confidence: 85%

Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa

et al. 2020

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Bacterial colonization and biofilm development on medical devices can lead to infection. Antimicrobial peptide-coated surfaces may prevent such infections. Melimine and Mel4 are chimeric cationic peptides showing broad-spectrum antimicrobial activity once attached to biomaterials and are highly biocompatible in animal models and have been tested in Phase I and II/III human clinical trials. These peptides were covalently attached to glass using an azidobenzoic acid linker. Peptide attachment was confirmed using X-ray photoelectron spectroscopy and amino acid analysis. Mel4 when bound to glass was able to adopt a more ordered structure in the presence of bacterial membrane mimetic lipids. The ability of surface bound peptides to neutralize endotoxin was measured along with their interactions with the bacterial cytoplasmic membrane which were analyzed using DiSC(3)-5 and Sytox green, Syto-9, and PI dyes with fluorescence microscopy. Leakage of ATP and nucleic acids from cells were determined by analyzing the surrounding fluid. Attachment of the peptides resulted in increases in the percentage of nitrogen by 3.0% and 2.4%, and amino acid concentrations to 0.237 nmole and 0.298 nmole per coverslip on melimine and Mel4 coated surfaces, respectively. The immobilized peptides bound lipopolysaccharide and disrupted the cytoplasmic membrane potential of Pseudomonas aeruginosa within 15 min. Membrane depolarization was associated with a reduction in bacterial viability by 82% and 63% for coatings melimine and Mel4, respectively (p < 0.001). Disruption of membrane potential was followed by leakage of ATP from melimine (1.5 ± 0.4 nM) or Mel4 (1.3 ± 0.2 nM) coated surfaces compared to uncoated glass after 2 h (p < 0.001). Sytox green influx started after 3 h incubation with either peptide. Melimine coatings yielded 59% and Mel4 gave 36% PI stained cells after 4 h. Release of the larger molecules (DNA/RNA) commenced after 4 h for melimine (1.8 ± 0.9 times more than control; p = 0.008) and after 6 h with Mel4 (2.1 ± 0.2 times more than control; p < 0.001). The mechanism of action of surface bound melimine and Mel4 was similar to that of the peptides in solution, however, their immobilization resulted in much slower (approximately 30 times) kinetics.

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Section: Discussionmentioning

confidence: 85%

Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa

et al. 2020

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“…These additives have been used to protect the enzyme from external denaturing agents by providing additional sites for hydrogen bonding with the enzyme surface that decreases dehydration and distortion of its three-dimensional structure [39]. However, these additives can interact with nucleophilic groups in the enzyme structure (e.g., –NH 2 and –OH) that influence the orientation of enzyme molecules on the support surface [40, 41]. Similar results were reported in previous studies carried out in our lab concerning the immobilization of Thermomyces lanuginosus lipase from different commercial formulations supplied by Novozymes (Lipolase® and Lipex® 100 L) by different protocols on activated organic supports (glyoxyl-agarose and epoxy-chitosan/alginate beads) [23, 24].…”

Section: Resultsmentioning

confidence: 99%

Selection of Lipases for the Synthesis of Biodiesel from Jatropha Oil and the Potential of Microwave Irradiation to Enhance the Reaction Rate

Souza

Mendes

Castro

2016

BioMed Research International

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The present study deals with the enzymatic synthesis of biodiesel by transesterification of Jatropha oil (Jatropha curcas L.) with ethanol in a solvent-free system. Seven commercial lipase preparations immobilized by covalent attachment on epoxy-polysiloxane-polyvinyl alcohol composite (epoxy-SiO2-PVA) were tested as biocatalysts. Among them, immobilized lipases from Pseudomonas fluorescens (lipase AK) and Burkholderia cepacia (lipase PS) were the most active biocatalysts in biodiesel synthesis, reaching ethyl ester yields (FAEE) of 91.1 and 98.3% at 72 h of reaction, respectively. The latter biocatalyst exhibited similar performance compared to Novozym® 435. Purified biodiesel was characterized by different techniques. Transesterification reaction carried out under microwave irradiation exhibited higher yield and productivity than conventional heating. The operational stability of immobilized lipase PS was determined in repeated batch runs under conventional and microwave heating systems, revealing half-life times of 430.4 h and 23.5 h, respectively.

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“…Human bone morphogenetic protein, for example, covalently bound to titanium using anchor molecules, significantly increases osteoinduction . PIII treatment of PEEK can produce this bioactivity without the need for anchor molecules and at the same time prevent adverse stress shielding associated with the use of metallic implants that result from a mismatch of material moduli . Despite the advantages, this process has not been optimised and fully characterised for PEEK.…”

Section: Introductionmentioning

confidence: 99%

Bio-Activation of Polyether Ether Ketone Using Plasma Immersion Ion Implantation: A Kinetic Model

Wakelin

Kondyurin

Wise

et al. 2014

Plasma Process. Polym.

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Plasma immersion ion implantation (PIII) is used in this study to improve the surface bioactivity of polyether ether ketone (PEEK) by modifying the chemical structure and introducing radicals. The kinetic properties of the treated surface are characterised here, allowing for tuning of the PIII process in order to produce PEEK with optimised bioactivity. The optimum PIII ion fluence for PEEK is determined to be 1 × 1016 ions/cm2. A model is proposed that accounts for the decay of radicals, nitrogen loss, oxygen incorporation and changes in surface energy within the structure. The model is based on the premise that there is a distribution of local chemical environments produced by PIII. The distribution of chemical environments results in a bioactive surface that has a long shelf life, compatible with clinical needs.

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Effect of Low Molecular Weight Additives on Immobilization Strength, Activity, and Conformation of Protein Immobilized on PVC and UHMWPE

Cited by 30 publications

References 67 publications

Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa

Mechanism of Action of Surface Immobilized Antimicrobial Peptides Against Pseudomonas aeruginosa

Selection of Lipases for the Synthesis of Biodiesel from Jatropha Oil and the Potential of Microwave Irradiation to Enhance the Reaction Rate

Bio-Activation of Polyether Ether Ketone Using Plasma Immersion Ion Implantation: A Kinetic Model

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