Alkaline protease catalysis of a secondary amine to form a peptide bond.
Procedures for peptide bond formation using esters of L-amino acids as the acyl donor and secondary amines such as derivatives of D- or L-proline, or pyroglutamic acid as the nucleophile in anhydrous 2-methyl-2-propanol catalyzed by alcalase or subtilisin Carlsberg have been developed. Kinetic studies have shown that all secondary amines tested had catalytic efficiencies (kcat/km) in the range 84-423 min-1 M-1. Both enzymes have identical catalytic properties. The selectivity of the enzyme-catalyzed reaction in 2-methyl-2-propanol shows that at the s-1 subsite of the enzyme only L-amino acids are substrates as acyl donors, and at the s-1' subsite both D- and L-amino acids are substrates. Optimal conditions for preparing precursors of the renin inhibitors Z-Ala-Pro-OBzl, Z-Lys(Z)-Pro-OBzl and Z-Lys(TFA)-Pro-OBzl were studied.
Chen
S T
ST
Laboratory of Biocatalysis, Academia Sinica, Taiwan.
Kao
C L
CL
Wang
K T
KT
D000577:Amides / Q000378:metabolism; D000588:Amines / Q000378:metabolism*; D001665:Binding Sites; D002384:Catalysis; D004151:Dipeptides / Q000378:metabolism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D010455:Peptides / Q000378:metabolism*; D011392:Proline / Q000031:analogs & derivatives / Q000378:metabolism; D011761:Pyrrolidonecarboxylic Acid / Q000378:metabolism; D013381:Subtilisins / Q000378:metabolism*
D000577:Amides; D000588:Amines; D004151:Dipeptides; D010455:Peptides; D011392:Proline; D013381:Subtilisins; D011761:Pyrrolidonecarboxylic Acid