Native Subtilisin Karlsberg and Modified Subtilisin 72 As Effective Catalysts of Peptide Bond Formation in Organic Media

O. M. Anikina ^a , T A. Semashko ^b , E. S. Oksenoit ^b , E. N. Lysogorskaya ^b , and I. Yu. Filippova ^{b #}

^# Phone: +7 (495) 939-5529; e-mail: irfilipp@genebee.msu.su

^a Faculty of Bioengineering and Bioinformatics, Moscow State University , Vorob'evy gory, Moscow , 119992 Russia

^b Faculty of Chemistry, Moscow State University , Vorob'evy gory, Moscow , 119992 Russia

Received July 6, 2005; in final form, July 13, 2005

Abstract: The activity and stability of native subtilisin Karlsberg and subtilisin 72 and their complexes with sodium dodecyl sulfate (SDS) in organic solvents were studied. The kinetic constants of the hydrolysis of specific chromogenic peptide substrates Z-Ala-Ala-Leu-pNA and Glp-Ala-Ala-Leu-pNA by the subtilisins were determined. It was found that the subtilisin Karlsberg complex with SDS in anhydrous organic solvents is an effective catalyst of peptide synthesis with multifunctional amino acids in positions P@1 and P'@1 (Glu, Arg, and Asp) containing unprotected side ionogenic groups.

Key words: enzymatic peptide synthesis, organic solvents, SDS–subtilisin Karlsberg complex

Russian Journal of Bioorganic Chemistry 2006, 32 (2):116-121