Proteinases immobilized on poly(vinyl alcohol) cryogel: novel biocatalysts for peptide synthesis in organic mediaстатья
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Дата последнего поиска статьи во внешних источниках: 8 сентября 2021 г.
Аннотация:Covalent immobilization of subtilisin and thermolysin on cryogel of poly(vinyl alcohol) was carried out. The biocatalysts obtained are characterized by high stability in water and in DMF-MeCN mixtures of various compositions. The synthetic efficiency of immobilized subtilisin in the multiple iterative synthesis of the peptide Z-Ala-Ala-Leu-Phe-pNA was examined in organic mixtures of different solvent compositions, Immobilized subtilisin exhibits high synthetic activity in organic media. A series of N-acylated p-nitroanilides of tetrapeptides of the general formula Z-Ala-Ala-Xaa-Yaa-pNA (Z is benzyloxycarbonyl, Xaa = Leu, Lys, or Glu, Yaa = Phe or Asp; pNA = 4-NO2-C6H4NH-) were synthesized in 70-98% yields using immobilized subtilisin as a biocatalyst without activation and protection of the ionogenic groups of polyfunctional amino acids. Immobilized thermolysin in a DMF-MeCN mixture catalyzed the formation of the peptide Z-Ala-Ala-Leu-pNA, which was obtained in 90% yield (during 1 h). It was demonstrated that the biocatalyst can be used repeatedly and that it retained activity after storage in an aqueous buffer during 6 months.