Abstract
Carboxypeptidase Y catalyzed reactions between a common acyl component (Bz-Ala-OMe) and a variety of amine compounds are described. The α-amino acid amides tested were—with the exception of isoglutamine— incorporated in high yields (70–95%). For the free α-amino acids, the yields were below 60% and fluctuated widely. For the cases examined, the enzyme only reacted with thel-enatiomers of these compounds. Dipeptides were not accepted as amine components. Glycinonitril, glycine-n-methyl amide and threonine-n-methyl amide reacted in low yields (<20%) only. None of the secondary amines tested (sarcosine, sarcosine methyl ester,n-methyl-alanine, proline and proline amide) was incorporated to form an amide bond. However, several structurally interesting primary amines (e.g.,l-alaninol, cyclopropylamine, β-alanine amide, taurine etc.) reacted in good yields (40–60%), indicating that carboxypeptidase Y may also become useful for the synthesis of certain peptide analogues.
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Abbreviations
- Bz:
-
benzoyl
- CPD-Y:
-
carboxypeptidase Y
- HPLC:
-
high pressure liquid chromatography
- Sar:
-
sarcosine
- Taurine:
-
2-aminoethanesulfonic acid
- TEAF:
-
triethylammonium formate buffer
- TEAP:
-
triethylammonium phosphate buffer
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Amino acids abbreviated in upper case refer to thel-enantiomer and those in lower case refer to thed-enantiomer. Other abbreviations of amino acids, amino acid derivatives and peptides are according to the guidelines of the IUPAC-IUB Commission on Biochemical Nomenclature. The binding notation for the enzyme and the substrates is that ofSchechter andBerger (16). Accordingly, amino acid residues in the substrate are referred to as P1, P2…Pi in the amino-terminal direction away from the scissible bond and Pi for the carboxy-terminal residue that is hydrolyzed off. Enzyme subsites are denoted S1, S2…Si and Si in correspondence with the substrate.
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Widmer, F., Breddam, K. & Johansen, J.T. Influence of the structure of amine components on carboxypeptidase Y catalyzed amide bond formation. Carlsberg Res. Commun. 46, 97–106 (1981). https://doi.org/10.1007/BF02906202
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DOI: https://doi.org/10.1007/BF02906202