Continuous enzymatic process for preparing peptides

Details Continuous enzymatic process for preparing peptides Continuous enzymatic process for preparing peptides

1988-09-30

1991-10-15

Naff, David M.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Enzymatic production of a protein or polypeptide

435180, 435288, 435803, 435813, 530335, 530338, 530339, 530344, C12P 2100, C12N 1108, C07K 0106, C07K 0114

Patent

active

050574159

ABSTRACT:
Peptide preparation is carried out by a continuous process by supplying a serine protease or peptidase enzyme retained in a reaction vessel with an alkyl ester of an N-protected amino acid or oligopeptide and a recycle stream containing an amino acid or oligopeptide to form an N-protected chain extended peptide, separating the N-protected chain extended peptide by adsorption on a hydrophobic absorbent, and eluting and recovering the adsorbed N-protected chain extended peptide. Adsorption is carried out without adjusting the pH from that in the reaction vessel, and adsorber effluent is recycled to the recycle stream. The protease or peptidase enzyme may be immobilized, and there is substantial exclusion of organic solubilizers in the reaction vessel. A preferred N-protecting group is an N-phenacyl group. The N-protecting group can be separated from the N-protected chain extended peptide with a deprotecting enzyme to recover the chain extended peptide. Apparatus is used that provides for continuous operation of the process.

REFERENCES:
patent: 4304858 (1981-12-01), Wandrey et al.
patent: 4326031 (1982-04-01), Wandrey et al.
patent: 4339534 (1982-07-01), Johansen et al.
Cramer et al., "Tandem Use of Carboxypeptidase Y Reactor and Displacement Chromatograph for Peptide Synthesis", Journal of Chromatography, 394 (1987) pp. 305-314.
Widmer et al., "Enzymatic Peptic Synthesis", Peptides 1982, Walter de Gruyter & Co. Berlin New York, 1983, pp. 375-379.
Fuganti et al., "Immobilized Penicillinacylase: Application to the Synthesis of the Dipeptide Aspartame", Tetrahedron Letters, vol. 27, No. 27, 1986, pp. 3191-3194.

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