| Autor: |
Baidamshina DR; Laboratory of Molecular Genetics of Microorganisms, Kazan (Volga Region) Federal University, Kazan 420008, Russia., Koroleva VA; Department of Biophysics and Biotechnology, Voronezh State University, Voronezh 394018, Russia., Olshannikova SS; Department of Biophysics and Biotechnology, Voronezh State University, Voronezh 394018, Russia., Trizna EY; Laboratory of Molecular Genetics of Microorganisms, Kazan (Volga Region) Federal University, Kazan 420008, Russia., Bogachev MI; Biomedical Engineering Research Centre, St. Petersburg Electrotechnical University, St. Petersburg 197376, Russia., Artyukhov VG; Department of Biophysics and Biotechnology, Voronezh State University, Voronezh 394018, Russia., Holyavka MG; Department of Biophysics and Biotechnology, Voronezh State University, Voronezh 394018, Russia., Kayumov AR; Laboratory of Molecular Genetics of Microorganisms, Kazan (Volga Region) Federal University, Kazan 420008, Russia.; Interdepartment Research Laboratory, Kazan State Academy of Veterinary Medicine named after N.E. Bauman, Kazan 420029, Russia. |
| Abstrakt: |
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 Carica papaya 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 -1 ) and specific activity (U mg protein -1 ), leading to the preservation of more than 90% of the initial total activity (U mL -1 ). 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 Staphylococcus aureus and Staphylococcus epidermidis . As a consequence, papain, both soluble and immobilized on medium molecular weight chitosan, is capable of potentiating the efficacy of antimicrobials against biofilm-embedded Staphylococci . Thus, papain immobilized on medium molecular weight chitosan appears a presumably beneficial agent for outer wound treatment for biofilms destruction, increasing antimicrobial treatment effectiveness. |
