title: T4 lysozyme fused with cellulose-binding module for antimicrobial cellulosic wound dressing materials.
abstract: OBJECTIVE
  Bacterial infection is a major challenge in wound care. Antimicrobial wound dressings are of great value for treating wound infections. Endolysins are evolving as a new class of antimicrobials with multiple applications. This study describes the production and evaluation of T4 lysozyme (T4Lyz), product of gene e of the T4 bacteriophage, fused with Cellulose Binding Module (CBM) for facile immobilization to cellulosic wound dressing.
  
  
  RESULTS
  Genes encoding T4Lyz-CBM and T4Lyz were cloned and expressed in Escherichia coli and the enzymes were purified by cation exchange chromatography. While the CBM tag did not alter the optimum pH and stability features of T4Lyz, the lytic activity of the fusion protein was lowered. The bactericidal activity of T4Lyz-CBM, determined by viable count plating assay after 1 h incubation with Micrococcus lysodeikticus was 97·5% with 10 μg ml(-1) , and 99·96% and 95% for E. coli and Pseudomonas mendocina, respectively, with 200 μg ml(-1) enzyme. T4Lyz-CBM was immobilized to wound dressing gauze with a capacity of 5·5 μg mg(-1) matrix, whereas the unmodified T4Lyz did not exhibit any binding. The immobilized protein retained its bactericidal activity against Gram-positive and Gram-negative bacteria. Both free and immobilized T4Lyz-CBM, after heat denaturation, retained their bactericidal activities against Gram-negative bacteria only. The immobilized enzyme exhibited higher stability than the free enzyme when stored in dry form or in the presence of polyol stabilizers.
  
  
  CONCLUSIONS
  Tagging T4Lyz with CBM provides a facile, irreversible binding to cellulosic wound dressing while retaining its activity. This approach may be suitable even for other antimicrobial enzymes and -peptides.
  
  
  CONCLUSIONS
  The spread of antibiotic resistance requires innovative strategies for discovery and development of effective antimicrobial alternatives. This report presents a novel strategy for producing antimicrobial wound dressing materials.
authors.affiliation: Division of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, Lund, Sweden.|Department of Microbiology and Immunology, Faculty of Pharmacy, Al-Azhar University, Assiut, Egypt.
authors.forename: A
authors.initials: A
authors.lastname: Abouhmad
authors.affiliation: Division of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, Lund, Sweden.
authors.forename: G
authors.initials: G
authors.lastname: Mamo
authors.affiliation: Division of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, Lund, Sweden.|Department of Microbiology and Immunology, Faculty of Pharmacy, Beni-Suef University, Beni-Suef, Egypt.
authors.forename: T
authors.initials: T
authors.lastname: Dishisha
authors.affiliation: Department of Microbiology and Immunology, Faculty of Pharmacy, Cairo University, Cairo, Egypt.
authors.forename: M A
authors.initials: MA
authors.lastname: Amin
authors.affiliation: Division of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, Lund, Sweden.
authors.forename: R
authors.initials: R
authors.lastname: Hatti-Kaul
mesh_terms: D000890:Anti-Infective Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D017122:Bacteriophage T4 / Q000201:enzymology*; D001458:Bandages; D002482:Cellulose / Q000737:chemistry*; D004352:Drug Resistance, Microbial; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000494:pharmacology*; D004926:Escherichia coli / Q000235:genetics; D006090:Gram-Negative Bacteria / Q000187:drug effects; D006094:Gram-Positive Bacteria / Q000187:drug effects; D008826:Microbial Sensitivity Tests; D009113:Muramidase / Q000737:chemistry / Q000494:pharmacology*; D014946:Wound Infection / Q000382:microbiology / Q000517:prevention & control
chemical_list: D000890:Anti-Infective Agents; D004800:Enzymes, Immobilized; D002482:Cellulose; D009113:Muramidase
keywords: T4 lysozyme; antimicrobial activity; cellulose binding module; fusion protein; immobilization; storage stability; wound dressing