Chymotrypsin-catalysed transpeptidations

Blau, K.; Waley, S. G.

doi:10.1042/bj0570538

Cited by 26 publications

(4 citation statements)

References 6 publications

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“…The demonstration that the liver esterases are all inhibited by DFP, and that a seryl residue in the se-96 quence Gly-Glu-Ser-AIa of horse liver esterase is phos-phorylated by 82P-labeled DFP (Jansz et al, 1959;Blakely et al, 1967), makes it probable that the catalytic action of these enzymes involves the intermediate acylation of a serine residue in the active site, as in the action of chymotrypsin (Hartley, 1960;Bender and Kézdy, 1965) . That chymotrypsin can catalyze acyl transfer from suitable substrates to the -amino group of amino acid residues has long been known (Johnston et at., 1950b; Blau and Waley, 1954). Chymotrypsin, however, is much less effective as a catalyst of such transamidation reactions than papain or ficin (Johnston et al, 1950a;Dowmont and Fruton, 1952;Mycek and Fruton, 1957; Brubacher and Bender, 1966), whose catalytic action involves the intermediate acylation of a cysteine residue (Stockell and Smith, 1957; Lowe and Williams, 1965; Kirsch and Igelstrom, 1966).…”

Section: Resultsmentioning

confidence: 99%

Beef liver esterase as a catalyst of acyl transfer to amino acid esters

Goldberg¹,

Fruton²

1969

Biochemistry

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Section: Resultsmentioning

confidence: 99%

Beef liver esterase as a catalyst of acyl transfer to amino acid esters

Goldberg¹,

Fruton²

1969

Biochemistry

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“…In fact, since Bergmann & Fraenkel-Conrat (1937) first demonstrated papain-catalysed amide (anilide)bond synthesis, many workers have reported enzymic synthesis of peptides, especially with papain (EC 3.4.22.2) (Kimmel & Smith, 1957). Concerning synthesis by chymotrypsin, Bergmann & Fruton (1938) synthesized Bz*-Tyr-Gly-NH2 from Bz-Tyr and Gly-NH2, and Blau & Waley (1954) obtained oligomeric peptides from Tyr-NH2, Phe-NH2 or Tyr-Tyr. Epand (1969) found that Bz-Tyr-Gly-NH2 is produced by chymotrypsin hydrolysis of Bz-Tyr-OEt in the presence of Gly-NH2, although detailed information on the synthesis was not reported.…”

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confidence: 99%

α-Chymotrypsin as the catalyst for peptide synthesis

Morihara¹,

Oka²

1977

Biochemical Journal

126

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alpha-Chymotrypsin (EC 3.4.21.1)-catalysed syntheses of peptides were performed with various N-acylated amino acid or peptide esters as donors, and amino acid derivatives, peptides or their derivatives as acceptors. Under optimal conditions the synthesis was almost quantitative. As acceptor nucleophiles, free amino acids or the ester derivatives were inadequate, but amino acid amides or hydrazides, di- or tri-peptides, or the amides, hydrazides and esters of the peptides were useful. The nucleophile specificity for synthesis was markedly similar to the leaving-group specificity in hydrolysis; hydrophobic or bulky amino acid residues were most effecient at both P1' and P2' positions [notation of Schechter & Berger (1967) Biochem. Biophys. Res. Commun. 27, 157-162], but L-proline as well as D-amino acid residues were the worst choices. The synthesis was further dependent on the solubility of the products synthesized; a higher yield of products was expected with lower solubility. As donor esters, good substrates were all useful. Accordingly, fragment condensation was possible by using N-acylated peptide esters and various peptides. The present study suggested that alpha-chymotrypsin may become a useful tool for peptide synthesis.

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“…Such reactions, well documented for endoproteases (4,13,28), would be of great interest in CPD-Y catalyzed peptide synthesis. In the present paper the results of such studies, employing peptide amides and peptides as substrates, are reported.…”

Section: Introductionmentioning

confidence: 99%

Carboxypeptidase Y catalyzed transpeptidations and enzymatic peptide synthesis

Breddam

Widmer

Johansen

1980

Carlsberg Res. Commun.

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It is shown that carboxypeptidase Y, in addition to its known amidase and peptidase activities, also exhibits peptidyl-amino-acid-amide hydrolase activity, i.e., amino acid amides are released from peptide amides. These three activities were investigated as to their usefulness in catalyzing transpeptidation reactions in the presence of suitable nucleophiles. Using peptides and peptide amides as substrates and amino acids or amino acid amides as nuc[eophiles, it could be shown that transpeptidation products were formed in accordance to each of the above mentioned activities of carboxypeptidase Y. While transpeptidation via the amidase activity results in an elongation of the peptide chain, transpeptidation via peptidase activity results in the exchange of the C-terminal amino acid residue. If the substrate is a peptide amide, transpeptidation via the peptidyl-amino-acid-amide hydrolase activity will result in the formation of a peptide where the C-terminal amino acid amide is replaced by either an amino acid or an amino acid amide.As conditions have not yet been found where a[t three activities are well separated from each other, it is not generally possible to direct these transpeptidations towards a single desired product in high yield. However, 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.Abbreviations: Ac = acetyl: Bz = benzoyl: CBZ = carbobenzoxy: CPD-Y = carboxypeptidase Y; DMF = dimethylformamide; FA = 3-(2-Furylacryloyl)-; HPLC = high pressure liquid chromatography; NaAc = sodium acetate: TEAF = triethylammonium formate; TEAP = triethylammonium phosphate buffer: All other abbreviations of amino acids, amino acid derivatives and peptides are according to the guideline of the IUPAC-IUB Commission on Biochemical Nomenclature.

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Chymotrypsin-catalysed transpeptidations

Cited by 26 publications

References 6 publications

Beef liver esterase as a catalyst of acyl transfer to amino acid esters

Beef liver esterase as a catalyst of acyl transfer to amino acid esters

α-Chymotrypsin as the catalyst for peptide synthesis

Carboxypeptidase Y catalyzed transpeptidations and enzymatic peptide synthesis

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