title: Immobilization of lysozyme-CLEA onto electrospun chitosan nanofiber for effective antibacterial applications.
abstract: Chitosan (CS) nanofibers with a diameter of 150-200nm were fabricated from a mixed chitosan/poly (vinyl alcohol) (PVA) solution by the electrospinning method. The nascent CS/PVA nanofibers were treated with 0.5M NaOH solution to make stable CS nanofibers by removing PVA under aqueous conditions. Hen egg-white lysozyme was immobilized on electrospun CS nanofibers via cross-linked enzyme aggregates (CLEAs) and used for effective and continuous antibacterial applications. The maximum amount of lysozyme immobilized on the CS nanofibers was determined to be 62.3mg/g of nanofibers under the optimum conditions. The immobilized lysozyme-CLEA retained more than 75.4% of its initial activity after 80 days of storage at room temperature, while the free lysozyme lost all of its activity under the same conditions. In addition, the immobilized lysozyme-CLEA retained more than 76% of its activity after 100 consecutive uses. Finally, the durability of the lysozyme-CLEA immobilized CS nanofibers showed bacteriostasis ratios of 82.4%, 79.8%, 83.4%, and 84.1% after 10 cycles against 4 pathogenic bacteria, viz. Staphylococcus aureus, Bacillus subtilis, Shigella flexneri, and Psedomonas aeruginosa, respectively. These results demonstrated that lysozyme-CLEA immobilized CS nanofibers could be used as a promising material for enhanced and continuous antibacterial applications.
authors.affiliation: Department of Microbiology, Chungbuk National University, 52 Naesudong-Ro, Heungduk-Gu, Cheongju 361-763, South Korea.
authors.forename: Jae-Min
authors.initials: JM
authors.lastname: Park
authors.forename: Mina
authors.initials: M
authors.lastname: Kim
authors.forename: Hyun-Sung
authors.initials: HS
authors.lastname: Park
authors.forename: Am
authors.initials: A
authors.lastname: Jang
authors.forename: Jiho
authors.initials: J
authors.lastname: Min
authors.forename: Yang-Hoon
authors.initials: YH
authors.lastname: Kim
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001419:Bacteria / Q000187:drug effects / Q000254:growth & development; D002645:Chickens; D048271:Chitosan / Q000494:pharmacology*; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008826:Microbial Sensitivity Tests; D009113:Muramidase / Q000378:metabolism*; D057139:Nanofibers / Q000737:chemistry* / Q000648:ultrastructure; D011142:Polyvinyl Alcohol / Q000737:chemistry; D020836:Protein Structure, Quaternary; D059027:Recycling; D013696:Temperature; D013997:Time Factors; D023822:Tissue Engineering
chemical_list: D000900:Anti-Bacterial Agents; D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D011142:Polyvinyl Alcohol; D048271:Chitosan; C421864:hen egg lysozyme; D009113:Muramidase