Subtilisin and alpha-chymotrypsin catalyzed synthesis of peptides containing arginine and lysine p-nitroanilides as C-terminal moieties.
p-Nitroanilides of N-acylated di-, tri- and tetrapeptides with C-terminal arginine or lysine residues have been obtained, as a rule with good yields, via acylation of arginine or lysine p-nitroanilides by methyl esters of respective N-acylated peptides, catalyzed by subtilisin or alpha-chymotrypsin. The synthesis might be performed by two routes--by reaction in water-organic solvent mixtures, catalyzed by dissolved enzyme, or by condensation of the components in organic solvents with low water content in the presence of any enzyme distributed over a silica support surface. The second approach seems to be preferable due to suppression of hydrolytic side reactions and improved stability of an enzyme. Subtilisin 72 is especially effective as a catalyst for the acylation of p-nitroanilides by N-protected tripeptide methyl esters--the derivatives capable of occupying the S1, S2 and S3 subsites of its extended binding site. Even dipeptide esters with D-amino acids in P2 position can be applied for p-nitroanilide acylation. The efficiency of alpha-chymotrypsin as a catalyst for peptide synthesis is more limited due to restricted specificity of this enzyme.
Stepanov
V M
VM
Laboratory of Protein Chemistry, Institute of Genetics and Selection of Industrial Microorganisms, Moscow, Russia.
Terent'eva EYu
Voyushina
T L
TL
Gololobov MYu
D000595:Amino Acid Sequence; D000813:Anilides / Q000138:chemical synthesis; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000737:chemistry; D002417:Cattle; D002918:Chymotrypsin / Q000378:metabolism*; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry; D008969:Molecular Sequence Data; D015394:Molecular Structure; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D012997:Solvents; D013381:Subtilisins / Q000378:metabolism*
D000813:Anilides; D010455:Peptides; D012997:Solvents; C072567:lysine 4-nitroanilide; C018833:arginine-4-nitroanilide; D001120:Arginine; D013381:Subtilisins; D002918:Chymotrypsin; D008239:Lysine