| Autor: |
Olshannikova SS; Biophysics and Biotechnology Department, Voronezh State University, 394018 Voronezh, Russia., Malykhina NV; Biophysics and Biotechnology Department, Voronezh State University, 394018 Voronezh, Russia., Lavlinskaya MS; Biophysics and Biotechnology Department, Voronezh State University, 394018 Voronezh, Russia.; Metagenomics and Food Biotechnologies Laboratory, Voronezh State University of Engineering Technologies, 394036 Voronezh, Russia., Sorokin AV; Biophysics and Biotechnology Department, Voronezh State University, 394018 Voronezh, Russia.; Metagenomics and Food Biotechnologies Laboratory, Voronezh State University of Engineering Technologies, 394036 Voronezh, Russia.; Polymer Science and Colloid Chemistry Department, Voronezh State University, 394018 Voronezh, Russia., Yudin NE; Polymer Science and Colloid Chemistry Department, Voronezh State University, 394018 Voronezh, Russia., Vyshkvorkina YM; Phystech School of Biological and Medical Physics, Moscow Institute of Physics and Technology, 141701 Dolgoprudnyi, Russia., Lukin AN; Research Core Center of Voronezh State University, Voronezh State University, 394018 Voronezh, Russia., Holyavka MG; Biophysics and Biotechnology Department, Voronezh State University, 394018 Voronezh, Russia.; Physics Department, Sevastopol State University, 299053 Sevastopol, Russia., Artyukhov VG; Biophysics and Biotechnology Department, Voronezh State University, 394018 Voronezh, Russia. |
| Abstrakt: |
Briefly, 2-(4-Acetamido-2-sulfanilamide) chitosan, which is a chitosan water-soluble derivative, with molecular weights of 200, 350, and 600 kDa, was successfully synthesized. The immobilization of ficin, papain, and bromelain was carried out by complexation with these polymers. The interaction mechanism of 2-(4-acetamido-2-sulfanilamide) chitosan with bromelain, ficin, and papain was studied using FTIR spectroscopy. It was found that the hydroxy, thionyl, and amino groups of 2-(4-acetamido-2-sulfanilamide) chitosan were involved in the complexation process. Molecular docking research showed that all amino acid residues of the active site of papain formed hydrogen bonds with the immobilization matrix, while only two catalytically valuable amino acid residues took part in the H-bond formation for bromelain and ficin. The spectral and in silico data were in good agreement with the catalytic activity evaluation data. Immobilized papain was more active compared to the other immobilized proteases. Moreover, the total and specific proteolytic activity of papain immobilized on the carrier with a molecular weight of 350 kDa were higher compared to the native one due to the hyperactivation. The optimal ratio of protein content (mg × g -1 of carrier), total activity (U × mL -1 of solution), and specific activity (U × mg -1 of protein) was determined for the enzymes immobilized on 2-(4-acetamido-2-sulfanilamide) chitosan with a molecular weight of 350 kDa. |
