Process for the enzymatic preparation of protected and unprotected di- and oligopeptides in aqueous solutions

US5304470A · kind A · utility

Assignees

• Forschungszentrum Jülich GmbH · DE
• Degussa AG · DE

Inventors

• Andreas Fischer · Heppenheim, DE
• Alexander Schwarz · Soest, DE
• Christian Wandrey · Jülich, DE
• Guenter Knaup · Bruchköbel, DE
• Andreas Bommarius · Frankfurt, DE
• Karl-Heinz Drauz · Alzenau, DE

Key dates

• Filing date: Jan 23, 1992
• Grant date: Apr 19, 1994
• Expiry date: Jan 23, 2012

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12P21/02
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Protected and unprotected di- or oligopeptides are synthesized by reacting an N-terminally protected .alpha.-amino acid alkyl ester or peptide alkyl ester of the formula X--E--R.sup.1 with an amino acid or a di- or oligopeptide or a derivative thereof of the formula H.sub.2 N--Q--R.sup.2 in aqueous solution in the presence of a hydrolase, and, removing protective groups from the reaction product separated from the reaction mixture, where E is the residue of an .alpha.-amino acid or of a di- or oligopeptide, R.sup.1 is lower alkyl and X is a group which carries a charge or is polar at the pH values used for the reaction and which increases the solubility by a factor >5 compared with compounds wherein X=H, Q is the residue of an amino acid or of a di- or oligopeptide, and R.sup.2 is an optionally esterified or amidated acid group. In a preferred embodiment, the peptide or (.alpha.-)amino acid ester concentration is greater than 50 mM, wherein in a particularly preferred embodiment, the concentration ranges from 100 to 1000 mM with approximately comparable nucleophile and electrophile concentrations. The preferred substrate/enzyme ratios are >10.sup.5 M/M or >10.sup.3 M/M when papain …