Enzymatic synthesis of chromogenic substrates for Glu,Asp-specific proteinases.
Glu,Asp-specific endopeptidases represent a new subfamily of chymotrypsin-like proteolytic enzymes. These enzymes prefer Glu or Asp residues in the P1 position of the substrates. p-Nitroanilides of N-acylated di-, tri- and tetrapeptides with C-terminal glutamic or aspartic acid residues have been obtained. Acyl peptide p-nitroanilides were synthesized via acylation of glutamic or aspartic acid p-nitroanilides using methyl esters of the respective N-acylated peptides, generally with good yields. The reactions were performed in organic solvents using subtilisin 72 sorbed on silica as a catalyst. The kinetic parameters for the hydrolysis of these p-nitroanilides with proteinases from Bacillus intermedius and Bacillus licheniformis were determined.
Milgotina
E I
EI
V.M. Stepanov Laboratory of Protein Chemistry, Institute of Genetics and Selection of Industrial Microorganisms, Moscow, Russia.
Shcheglov
A S
AS
Lapa
G B
GB
Chestukhina
G G
GG
Voyushina
T L
TL
D000813:Anilides / Q000737:chemistry; D001224:Aspartic Acid / Q000378:metabolism; D001407:Bacillus / Q000201:enzymology; D001671:Biochemistry / Q000379:methods*; D002863:Chromogenic Compounds / Q000138:chemical synthesis* / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism; D007700:Kinetics; D012697:Serine Endopeptidases / Q000378:metabolism*; D013379:Substrate Specificity; D013381:Subtilisins / Q000378:metabolism
D000813:Anilides; D002863:Chromogenic Compounds; D001224:Aspartic Acid; D018698:Glutamic Acid; D012697:Serine Endopeptidases; D013381:Subtilisins; C028344:glutamyl endopeptidase