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Title: Carbohydrate protease conjugates: Stabilized proteases for peptide synthesis

Abstract

The synthesis of oligopeptides using stable carbohydrate protease conjugates (CPCs) was examined in acetonitrile solvent systems. CPC[{alpha}-chymotrypsin] was used for the preparation of peptides containing histidine, phenylalanine, tryptophan in the P{sub 1} position in 60-93% yield. The CPC[{alpha}-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 peptides containing both hydrophylic and hydrophobic amino acids. The P{sub 2} specificity of papain for aromatic residues was utilized for the 2 + 3 coupling of Z-Tyr-Gly-OMe to H{sub 2}N-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-methacryl- amido-2-deoxy-D-glucose), poly(2-methacrylamido-2-deoxy-D-galactose), and poly(5-methacryl-amido-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 {alpha}-chymotrypsin gave a CPC with catalytic activity in potassium phosphate buffer atmore » 90{degrees}C for 2 h. 1 fig., 1 tab., 40 refs.« less

Authors:
; ; ;  [1]
  1. Ohio State Univ., Columbus, OH (United States)|[Lawrence Berkeley Lab., CA (United States)
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
588654
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Organic Chemistry; Journal Volume: 60; Other Information: PBD: 1995
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; PROTEINS; STRUCTURE-ACTIVITY RELATIONSHIPS; ACETONITRILE; AMINO ACIDS; CHYMOTRYPSIN; ESTERS; GALACTOSE; GLUCOSE; HYDROLYSIS; LEUCINE; OXIDATION; PAPAIN; PEPTIDES; POTASSIUM; RIBOSE; SPECIFICITY; STABILITY; SYNTHESIS; CARBOHYDRATES; ALDEHYDES; CATALYSTS

Citation Formats

Wartchow, C.A., Wang, Peng, Bednarski, M.D., and Callstrom, M.R. Carbohydrate protease conjugates: Stabilized proteases for peptide synthesis. United States: N. p., 1995. Web. doi:10.1021/jo00112a049.
Wartchow, C.A., Wang, Peng, Bednarski, M.D., & Callstrom, M.R. Carbohydrate protease conjugates: Stabilized proteases for peptide synthesis. United States. doi:10.1021/jo00112a049.
Wartchow, C.A., Wang, Peng, Bednarski, M.D., and Callstrom, M.R. Sun . "Carbohydrate protease conjugates: Stabilized proteases for peptide synthesis". United States. doi:10.1021/jo00112a049.
@article{osti_588654,
title = {Carbohydrate protease conjugates: Stabilized proteases for peptide synthesis},
author = {Wartchow, C.A. and Wang, Peng and Bednarski, M.D. and Callstrom, M.R.},
abstractNote = {The synthesis of oligopeptides using stable carbohydrate protease conjugates (CPCs) was examined in acetonitrile solvent systems. CPC[{alpha}-chymotrypsin] was used for the preparation of peptides containing histidine, phenylalanine, tryptophan in the P{sub 1} position in 60-93% yield. The CPC[{alpha}-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 peptides containing both hydrophylic and hydrophobic amino acids. The P{sub 2} specificity of papain for aromatic residues was utilized for the 2 + 3 coupling of Z-Tyr-Gly-OMe to H{sub 2}N-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-methacryl- amido-2-deoxy-D-glucose), poly(2-methacrylamido-2-deoxy-D-galactose), and poly(5-methacryl-amido-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 {alpha}-chymotrypsin gave a CPC with catalytic activity in potassium phosphate buffer at 90{degrees}C for 2 h. 1 fig., 1 tab., 40 refs.},
doi = {10.1021/jo00112a049},
journal = {Journal of Organic Chemistry},
number = ,
volume = 60,
place = {United States},
year = {Sun Dec 31 00:00:00 EST 1995},
month = {Sun Dec 31 00:00:00 EST 1995}
}