Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Enzymatic production of a protein or polypeptide
Patent
1993-01-19
1994-11-29
Robinson, Douglas W.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Enzymatic production of a protein or polypeptide
435227, C12P 2100, C12N 978
Patent
active
053690168
ABSTRACT:
A peptide amidase isolated from the flavedo of citrus fruits, preferably oranges, which is capable of catalyzing the selective hydrolytic elimination of the free amino group on the C-terminal end of peptide amides but which does not cleave peptide bonds. The enzyme accepts D-amino acid residues in the C-terminal position, although the hydrolysis rate is much slower than with L-amino acid residues. The enzyme is weakly inhibited by serine protease inhibitors; has an optimal pH of 7.5.+-.1.5, an optimum temperature of 30.degree. C. at pH 7.5 and has an isoelectric point of pH 9.5. The peptide amidase is stable at pH 6.0-9.0. The molecular weight of the purified enzyme is 32,000.+-.3000 daltons. A peptide amidase according to the present invention is particularly useful in the production of peptides by continuous enzymatic reaction of N-protected amino acid or peptide alkyl esters with amides of amino acids. In the continuous reaction, the synthesized peptide amide is hydrolyzed by the peptide amidase and separated by anion exchange from the amide of the amino acid which can be recycled.
REFERENCES:
patent: 4339534 (1982-07-01), Johansen et al.
patent: 5057415 (1991-10-01), Schwartz et al.
Breddam, K., Carlsberg Res. Commun. 49:535-54 (1984), p. 2.
Zuber, H., Hoppe Seyler's Z. Phys. Chem., 349: 1337-1352 (1968).
Steinke et al., Enz. Microb. Technol., 12:836-840 (1990).
Kula Maria-Regina
Schwarz Alexander
Steinke Doerte
Wandrey Christian
Degussa - Aktiengesellschaft
Forschungszentrum Juelich GmbH
Robinson Douglas W.
Weber Jon P.
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