Influence of the substrate structure on carboxypeptidase Y catalyzed peptide bond formation

  title={Influence of the substrate structure on carboxypeptidase Y catalyzed peptide bond formation},
  author={Klaus Breddam and Fred Widmer and Jack T. Johansen},
  journal={Carlsberg Research Communications},
It is shown that for carboxypeptidase Y catalyzed peptide synthesis the coupling yields using peptide esters as substrates and amino acids as nucleophiles are strongly dependent on the C-terminal amino acid residue. A dependence on the length of the acyl donating peptide chain is also observed, indicating the influence of residues other than the one directly involved in acylation of the enzyme. Using amino acid amides as nucleophiles, the yields are generally high (80%–100%). No peptide ester… 
Carboxypeptidase Y catalyzed C-terminal modifications of peptides
It is demonstrated that carboxypeptidase Y catalyzes the exchange of C-terminal amino acid residues in peptides for various other groups, and the coupling yields for all the transacylation reactions are dependent on the hydrophobicity of the amino acid leaving the active site.
Malt carboxypeptidase catalyzed aminolysis reactions
The influence of the nucleophile H-Val-NH2 on the kinetic parameters for the cleavage of Nα-CBZ-Lys-p-nitrophenylester indicates that the acylation step is rate-limiting, and that H- Val-NH 2 has two different modes of binding to the enzyme.
Influence of the structure of amine components on carboxypeptidase Y catalyzed amide bond formation
Several structurally interesting primary amines reacted in good yields, indicating that carboxypeptidase Y may also become useful for the synthesis of certain peptide analogues.
Carboxypeptidase Y catalyzed peptide synthesis using amino acid alkyl esters as amine components
Carboxypeptidase Y catalyzed transacylation reactions between N-protected amino acid methyl esters or peptide methyl esters as initial acyl components and methyl, ethyl, isopropyl or tert.-butyl
Enzymatic properties of malt carboxypeptidase II in hydrolysis and aminolysis reactions
The enzyme is easily saturated with amino acid amides indicating the formation of a complex between nucleophile and acyl-enzyme intermediate prior to deacylation, which renders it suitable for synthesis of peptide bonds where the acyl and/or the imine portion is donated by a basic amino acid residue.
Influence of guanidine derivatives on the specificity of malt carboxypeptidase
Kinetic data for the hydrolysis of various substrates indicate that phenylguanidine binds to the S1′ binding site of the enzyme in such a manner that the rate of cleavage of ester or amide substrates with large groups in the P1′ position is reduced while substrateswith small groups in this position are hydrolyzed with increased rates.
Amino acid methyl esters as amine components in CPD-Y catalyzed peptide synthesis: Control of side reactions
It is demonstrated that knowledge of the kinetic constants for the CPD-Y catalyzed hydrolysis of ester substrates can be exploited in such a way that amino acid esters may be used as amine components without oligomerization.
Carboxypeptidase S-1 from Penicillium janthinellum: Enzymatic properties in hydrolysis and aminolysis reactions
  • K. Breddam
  • Biology, Chemistry
    Carlsberg research communications
  • 1988
Using amino acids and amino acid amides as nucleophiles carboxypeptidase S-1 efficiently catalyses the formation of peptide bonds by aminolysis of peptides and peptide esters provided that the substrate contains a basic amino acid residue at the P1 position.


Enzymatic peptide synthesis. Carboxypeptidase Y catalyzed formation of peptide bonds
The present study suggests that carboxypeptidase Y, having a broad specificity for amino acid side chains, may become a general catalyst for enzymatic peptide synthesis in the homogeneous phase.
Carboxypeptidase Y catalyzed transpeptidations and enzymatic peptide synthesis
The data presented in this paper indicate that for peptides of certain unique amino acid sequences the carboxypeptidase Y catalyzed transpeptidations may be applicable in the field of enzymatic peptide synthesis.
Kinetic studies of carboxypeptidase Y. I. Kinetic parameters for the hydrolysis of synthetic substrates.
Kinetic parameters for carboxypeptidase Y confirmed that the enzyme released COOH-terminal proline and beta-alanine at an appreciable rate, as well as neutral amino acids with aromatic and aliphatic side chains at a very high speed.
Kinetic studies of carboxypeptidase Y. II. Effects of substrate and product analogs on peptidase and esterase activities.
Reversible inhibition of the peptidase and esterase activities of CPase Y [EC] was investigated with substrate and product analogs known to be inhibitors or effectors of pancreatic
On the size of the active site in proteases. II. Carboxypeptidase-A.
Kinetic studies of carboxypeptidase Y. III. Action on ester, amide, and anilide substrates and the effects of some environmental factors.
Hydroxylaminolysis and the kinetic isotope effects of deuterium oxide indicated, with some reservations, a reaction mechanism which proceeds via the formation of an acyl intermediate.
Demonstration of the acyl-enzyme mechanism for the hydrolysis of peptides and anilides by chymotrypsin.
The ultimate proof for nonspecific substrates is that the structures of the crystalline indolylacryloyl- chymo- trypsin and carbamyl-chymotrypsin have been solved by X-ray diffraction methods.
Does carboxypeptidase Y have intrinsic endopeptidase activity?