Carbohydrate protease conjugates

Stabilized proteases for peptide synthesis

Charles A. Wartchow, Peng Wang, Mark D. Bednarski, Matthew R Callstrom

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The synthesis of oligopeptides using stable carbohydrate protease conjugates (CPCs) was examined in acetonitrile solvent systems. CPC[α-chymotrypsin] was used for the preparation of peptides containing histidine, phenylalanine, tyrosine, and tryptophan in the P1 position in 60-93% yield. The CPC[α-chymotrypsin]-catalyzed synthesis of octamer Z-Gly-Gly-Phe-Gly-Gly-Phe-Gly-Gly-OEt from Z-Gly-Gly-Phe-Gly-Gly-Phe-OMe was achieved in 71% yield demonstrating that synthesis was the dominant kinetic process relative to amide hydrolysis. CPC[papain] was used to synthesize peptides containing both hydrophilic and hydrophobic amino acids. The P2 specificity of papain for aromatic residues was utilized for the 2 + 3 coupling of Z-Tyr-Gly-OMe to H2N-Gly-Phe-Leu-OH to generate the leucine enkephalin derivative in 79% yield. Although papain is nonspecific for the hydrolysis of N-benzyloxycarbonyl amino acid methyl esters in aqueous solution, the rates of synthesis for these derivitives with nucleophile leucine tert-butyl ester differed by nearly 2 orders of magnitude. CPC[thermolysin] was used to prepare the aspartame precursor Z-Asp-Phe-OMe in 90% yield. The increased stability of CPCs prepared from periodate-modified poly(2-methacrylamido-2-deoxy-D-glucose), poly(2-methacrylamido-2-deoxy-D-galactcose), and poly(5-methacrylamido-5-deoxy-D-ribose), carbohydrate materials designed to increase the aldehyde concentration in aqueous solution, suggests that the stability of CPCs is directly related to the aldehyde concentration of the carbohydrate material. Periodate oxidation of poly(2-methacrylamido-2-deoxy-D-glucose) followed by covalent attachment to α-chymotrypsin gave a CPC with catalytic activity in potassium phosphate buffer at 90°C for 2 h.

Original languageEnglish (US)
Pages (from-to)2216-2226
Number of pages11
JournalJournal of Organic Chemistry
Volume60
Issue number7
StatePublished - 1995
Externally publishedYes

Fingerprint

Peptide Hydrolases
Carbohydrates
Peptides
Papain
Chymotrypsin
Deoxyglucose
Aldehydes
Hydrolysis
Esters
Aspartame
Thermolysin
Leucine Enkephalin
Amino Acids
Oligopeptides
Nucleophiles
Ribose
Phenylalanine
Histidine
Amides
Tryptophan

ASJC Scopus subject areas

  • Organic Chemistry

Cite this

Carbohydrate protease conjugates : Stabilized proteases for peptide synthesis. / Wartchow, Charles A.; Wang, Peng; Bednarski, Mark D.; Callstrom, Matthew R.

In: Journal of Organic Chemistry, Vol. 60, No. 7, 1995, p. 2216-2226.

Research output: Contribution to journalArticle

Wartchow, Charles A. ; Wang, Peng ; Bednarski, Mark D. ; Callstrom, Matthew R. / Carbohydrate protease conjugates : Stabilized proteases for peptide synthesis. In: Journal of Organic Chemistry. 1995 ; Vol. 60, No. 7. pp. 2216-2226.
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abstract = "The synthesis of oligopeptides using stable carbohydrate protease conjugates (CPCs) was examined in acetonitrile solvent systems. CPC[α-chymotrypsin] was used for the preparation of peptides containing histidine, phenylalanine, tyrosine, and tryptophan in the P1 position in 60-93{\%} yield. The CPC[α-chymotrypsin]-catalyzed synthesis of octamer Z-Gly-Gly-Phe-Gly-Gly-Phe-Gly-Gly-OEt from Z-Gly-Gly-Phe-Gly-Gly-Phe-OMe was achieved in 71{\%} yield demonstrating that synthesis was the dominant kinetic process relative to amide hydrolysis. CPC[papain] was used to synthesize peptides containing both hydrophilic and hydrophobic amino acids. The P2 specificity of papain for aromatic residues was utilized for the 2 + 3 coupling of Z-Tyr-Gly-OMe to H2N-Gly-Phe-Leu-OH to generate the leucine enkephalin derivative in 79{\%} yield. Although papain is nonspecific for the hydrolysis of N-benzyloxycarbonyl amino acid methyl esters in aqueous solution, the rates of synthesis for these derivitives with nucleophile leucine tert-butyl ester differed by nearly 2 orders of magnitude. CPC[thermolysin] was used to prepare the aspartame precursor Z-Asp-Phe-OMe in 90{\%} yield. The increased stability of CPCs prepared from periodate-modified poly(2-methacrylamido-2-deoxy-D-glucose), poly(2-methacrylamido-2-deoxy-D-galactcose), and poly(5-methacrylamido-5-deoxy-D-ribose), carbohydrate materials designed to increase the aldehyde concentration in aqueous solution, suggests that the stability of CPCs is directly related to the aldehyde concentration of the carbohydrate material. Periodate oxidation of poly(2-methacrylamido-2-deoxy-D-glucose) followed by covalent attachment to α-chymotrypsin gave a CPC with catalytic activity in potassium phosphate buffer at 90°C for 2 h.",
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