title: Solid-phase synthesis of a peptide-ligand affinity matrix for isolation of chymosin.
abstract: Aminopropyl Perloza beaded cellulose was used as the support for solid-phase synthesis of resin-bound Val-dLeu-Pro-Phe-Phe-Val-dLeu, an inhibitor of aspartic proteases. Both Boc and Fmoc SPPS methodologies were employed in separate syntheses. The peptide-resins were characterized by amino acid analysis. The peptide-resin from the Fmoc synthesis gave the better amino acid analysis of the two syntheses and was used for further studies. Following modification of the peptide N-terminus by succinylation, the peptide-resin was able to bind chymosin (E.C. 3.5.21.4). The peptide resin was used for isolation of chymosin from a crude recombinant broth.
authors.affiliation: Separation Science Unit, Massey University, New Zealand.
authors.forename: D R
authors.initials: DR
authors.lastname: Englebretsen
authors.forename: D R
authors.initials: DR
authors.lastname: Harding
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors; D002482:Cellulose; D002846:Chromatography, Affinity; D012085:Chymosin / Q000302:isolation & purification* / Q000378:metabolism; D005449:Fluorenes; D005563:Formic Acid Esters; D008024:Ligands; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000378:metabolism; D011480:Protease Inhibitors / Q000138:chemical synthesis* / Q000737:chemistry; D011994:Recombinant Proteins / Q000302:isolation & purification; D012116:Resins, Plant
chemical_list: D000596:Amino Acids; D005449:Fluorenes; D005563:Formic Acid Esters; D008024:Ligands; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D009842:Oligopeptides; D011480:Protease Inhibitors; D011994:Recombinant Proteins; D012116:Resins, Plant; C000345:t-butyloxycarbonyl group; C085596:succinyl-valyl-leucyl-prolyl-phenylalanyl-phenylalanyl-valyl-leucinamide; D002482:Cellulose; D016282:Aspartic Acid Endopeptidases; D012085:Chymosin