Biochemical Properties and Anti-Biofilm Activity of Chitosan-Immobilized Papain

• Published in: Marine drugs 2021-03, Vol.19 (4), p.197
• Main Authors: Baidamshina, Diana R, Koroleva, Victoria A, Olshannikova, Svetlana S, Trizna, Elena Yu, Bogachev, Mikhail I, Artyukhov, Valeriy G, Holyavka, Marina G, Kayumov, Airat R
• Format: Article
• Language: English
• Subjects: Adsorption; adsorption immobilization; Antiinfectives and antibacterials; Antimicrobial agents; bacterial biofilms; Biofilms; Carica papaya; Chitin; Chitosan; Cysteine proteinase; Cytotoxicity; Deacetylation; Debridement; Enzymes; Glycine; Glycine (amino acid); Immobilization; Latex; Molecular weight; Papain; pH effects; Proteins; Staphylococcus aureus; Staphylococcus epidermidis; Temperature; Thermal stability; Toxicity; Wound healing; Wounds
• ISSN: 1660-3397; 1660-3397
• DOI: 10.3390/md19040197

Chitosan, the product of chitin deacetylation, is an excellent candidate for enzyme immobilization purposes. Here we demonstrate that papain, an endolytic cysteine protease (EC: 3.4.22.2) from latex immobilized on the matrixes of medium molecular (200 kDa) and high molecular (350 kDa) weight chitosans exhibits anti-biofilm activity and increases the antimicrobials efficiency against biofilm-embedded bacteria. Immobilization in glycine buffer (pH 9.0) allowed adsorption up to 30% of the total protein (mg g chitosan ) and specific activity (U mg protein ), leading to the preservation of more than 90% of the initial total activity (U mL ). While optimal pH and temperature of the immobilized papain did not change, the immobilized enzyme exhibited elevated thermal stability and 6-7-fold longer half-life time in comparison with the soluble papain. While one-half of the total enzyme dissociates from both carriers in 24 h, this property could be used for wound-dressing materials design with dosed release of the enzyme to overcome the relatively high cytotoxicity of soluble papain. Our results indicate that both soluble and immobilized papain efficiently destroy biofilms formed by and . As a consequence, papain, both soluble and immobilized on medium molecular weight chitosan, is capable of potentiating the efficacy of antimicrobials against biofilm-embedded . Thus, papain immobilized on medium molecular weight chitosan appears a presumably beneficial agent for outer wound treatment for biofilms destruction, increasing antimicrobial treatment effectiveness.