Recombinant spider silk coatings functionalized with enzymes targeting bacteria and biofilms

• Published in: MicrobiologyOpen (Weinheim) 2020-04, Vol.9 (4), p.e993-n/a
• Main Authors: Seijsing, Fredrik, Nilebäck, Linnea, Öhman, Oskar, Pasupuleti, Rajeev, Ståhl, Camilla, Seijsing, Johan, Hedhammar, My
• Format: Article
• Language: English
• Subjects: absorption; Aggregatibacter actinomycetemcomitans; Anti-Bacterial Agents - pharmacology; antibacterial; antibacterial activity; Antibacterial materials; antibiofilm; antibiofilm activity; Antibiotics; antiinfective agent; Antiinfectives and antibacterials; Bacteria; Bacteria - drug effects; Bacteria - growth & development; Bacterial Adhesion - drug effects; bacterial colonization; Bacterial Proteins - genetics; Bacterial Proteins - pharmacology; bacteriolysis; Bacteriophages - metabolism; Biodegradation; biofilm degradation; Biofilms; Biofilms - drug effects; Biofilms - growth & development; catalysis; chemical modification; coating; Coatings; Colonization; controlled study; Degradation; dispersin B; endolysin; Enzymatic activity; Enzyme activity; enzyme degradation; enzyme immobilization; Enzymes; Fluorescence; Fusion protein; gene fusion; Glycerol; Glycoside Hydrolases - genetics; Glycoside Hydrolases - pharmacology; Immobilization; material coating; Medical equipment; Microbalances; Mutation; nonhuman; Original; Peptides; peptidoglycan; Peptidoglycans; Phages; priority journal; protein protein interaction; proteinase; Proteins; quantitative analysis; Quartz crystal microbalance; Quartz crystals; Recombinant Proteins - genetics; Recombinant Proteins - pharmacology; recombinant spider silk; SAL 1 protein; SAP 1 protein; Sensors; Silk; Silk - genetics; Silk - pharmacology; Sortase; sortase A; Staphylococcus; Substrates; Surgical implants; Turbidity; turbidity reduction; unclassified drug; Viral Proteins - genetics
• ISSN: 2045-8827; 2045-8827
• DOI: 10.1002/mbo3.993

Bacteria forming biofilms on surgical implants is a problem that might be alleviated by the use of antibacterial coatings. In this article, recombinant spider silk was functionalized with the peptidoglycan degrading endolysin SAL‐1 from the staphylococcal bacteriophage SAP‐1 and the biofilm‐matrix‐degrading enzyme Dispersin B from Aggregatibacter actinomycetemcomitans using direct genetic fusion and/or covalent protein–protein fusion catalyzed by Sortase A. Spider silk assembly and enzyme immobilization was monitored using quartz crystal microbalance analysis. Enzyme activity was investigated both with a biochemical assay using cleavage of fluorescent substrate analogues and bacterial assays for biofilm degradation and turbidity reduction. Spider silk coatings functionalized with SAL‐1 and Disperin B were found to exhibit bacteriolytic effect and inhibit biofilm formation, respectively. The strategy to immobilize antibacterial enzymes to spider silk presented herein show potential to be used as surface coatings of surgical implants and other medical equipment to avoid bacterial colonization. Surgical implants could potentially be protected from colonization of bacteria by having a protective coating with antibacterial properties. Herein, we investigate strategies to use assembly of silk functionalized with antibacterial enzymes to prepare such coatings. Both the peptidoglycan degrading enzyme SAL‐1 and the biofilm‐matrix degrading enzyme Dispersin B were shown effective when immobilized using a silk coating.