Influence of Soil Humic and Fulvic Acid on the Activity and Stability of Lysozyme and Urease

Humic substances (HS), including humic acids (HA) and fulvic acids (FA), are important components of soil systems. HS form strong complexes with oppositely charged proteins, which will lead to changes in the enzyme activity. The effect of soil HS on the activity and stability of two enzymes was investigated as a function of pH, ionic strength, and mass ratio HS/enzyme. Humic acid (JGHA) and fulvic acid (JGFA) are negatively charged, lysozyme is net positive at pH values below 10.4, and urease is net positive below pH 5.2 or net negative above pH 5.2. The enzyme activities in the HS-enzyme complexes were suppressed when the enzymes were oppositely charged to the HS. The largest activity suppression was observed around the mass ratio HS/enzyme where the HS-protein complex was at its isoelectric point (IEP). At the IEP strong aggregation of the complexes led to encapsulation of the enzyme. The ionic strength was important; an increase decreased complex formation, but increased aggregation. Due to the larger hydrophobicity of JGHA than JGFA, the reduction in enzyme activity was stronger for JGHA. The enzyme stability also decreased maximally at mass ratio around the IEP of the complex when HS and protein were oppositely charged. When urease and HS were both negatively charged no complexes were formed, but the presence of JGHA or JGFA improved the activity and stability of the enzyme

Formation and Characterization of Light-Responsive TEMPO-Oxidized Konjac Glucomannan Microspheres

A light-responsive delivery system has been developed. It consists of gelly micro­spheres made of TEMPO-oxidized Konjac gluco­mannan (OKGM) polymers where the carboxyl (COO^–) groups are cross-linked via ferric ions (Fe³⁺) and in which functional ingredients may be incorporated. By irradiation with (simulated) sunlight, the microspheres degrade, thereby releasing the encapsulated component(s). The degree of oxidation (DO) of the OKGM polymers could be well-controlled between 15 and 80%, as confirmed by proton titrations and FT-IR spectroscopy. OKGM of DO 80% was selected to prepare the microspheres because the high COO^– content leads to a high density of cross-links, yielding a strong gel. The electro­kinetic potential of the OKGM particles increases with increasing pH and decreasing salt concentration. Mössbauer and FT-IR spectroscopy revealed that the cross-links are formed through two modes of COO^––Fe³⁺ coordination, that is, 68.4% by bridging and 31.6% by unidentate binding. Thus, the unique properties of the OKGM microspheres make them potentially applicable as light-controlled biocompatible delivery systems

Antiadhesive Polymer Brush Coating Functionalized with Antimicrobial and RGD Peptides to Reduce Biofilm Formation and Enhance Tissue Integration

This paper describes the synthesis and characterization of polymer–peptide conjugates to be used as infection-resistant coating for biomaterial implants and devices. Antiadhesive polymer brushes composed of block copolymer Pluronic F-127 (PF127) were functionalized with antimicrobial peptides (AMP), able to kill bacteria on contact, and arginine–glycine–aspartate (RGD) peptides to promote the adhesion and spreading of host tissue cells. The antiadhesive and antibacterial properties of the coating were investigated with three bacterial strains: Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa. The ability of the coating to support mammalian cell growth was determined using human fibroblast cells. Coatings composed of the appropriate ratio of the functional components: PF127, PF127 modified with AMP, and PF127 modified with RGD showed good antiadhesive and bactericidal properties without hampering tissue compatibility