Claims
- 1. A process for producing a peptide having the formula:
- X--A--B--Y
- wherein A and B are the same or different and each represents an amino acid residue or a peptide residue, X represents an amino acid protective group, Y represents a carboxyl protective group, which comprises:
- reacting an acid component of an amino acid or peptide having an N-terminal protective group or a salt thereof which has the formula
- X--A--OH
- with an amine component of an amino acid or peptide having a C-terminal protective group or a salt thereof which has the formula
- H--B--Y
- in the presence of metalloproteinase enzyme produced from microorganisms in an aqueous solution having a pH necessary to maintain the activity of said metalloproteinase enzyme.
- 2. The process of claim 1, wherein the microorganism is selected from the group consisting of Bacillus subtilis, Bacillus thermoproteoliticus, Streptomyces caespitosus, Bacillus megaterium, Bacillus polymyxa, Streptomyces griseus, Streptomyces naraensis, Streptomyces fradiae, Tseudomonas aeruginosa, Aspergillus oryzae, Clostridium histolyticum and Proteus aeruginosa.
- 3. The process of claim 1 wherein the pH of said solution is maintained at the desired level sufficient to sustain enzyme activity by reacting the amino acid or peptide reactants in a buffer solution having a pH of 6 to 8.
- 4. The process of claim 1 wherein the pH is maintained at the desired level by adding an acid or a base to the reaction solution depending upon the detected pH of the reaction mixture.
- 5. The process of claim 1 wherein the reaction is conducted by using the starting materials in a ratio of 0.8 to 2 moles of said acid component, X--A--OH per one mole of said amine component H--B--Y.
- 6. The process of claim 1, wherein the reaction is conducted by adding 10 to 500 mg of metalloproteinase per one mmole of the amine component, H--B--Y.
- 7. The process of claim 1, wherein the N-terminal protective group of the acid component X--A--OH is tertiary alkoxycarbonyl, benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, p-toluenesulfonyl, or o-nitrosulfenyl; and the C-terminal protective group of the amine component; H--B--Y is non-substituted amino, primary alkoxy, tertiary alkoxy, benzyloxy, p-nitrobenzyloxy, benzhydryloxy, benzylamino, 2,4-dimethoxybenzylamino or benzhydrylamino.
- 8. The process of claim 1, wherein A and B are the same or different and represent an amino acid residue or peptide residue wherein the amino acid is an aliphatic amino acid, an oxyamino acid, a sulfur-containing amino acid, a monoamino dicarboxylic acid, a diamino monocarboxylic acid, an aromatic amino acid or an heterocyclic amino acid.
- 9. The process of claim 1, wherein an inhibitor for the proteolytic enzyme present as an impurity to said metalloproteinase enzyme is added to said aqueous solution and wherein said inhibitor is extracted from oats, fava, kidney beans or potato.
- 10. The process of claim 7, wherein said tertiary alkoxycarbonyl group is t-butyloxycarbonyl or t-amyloxycarbonyl; said primary alkoxy group is methoxy or ethoxy; and said tertiary alkoxy group is t-butoxy.
- 11. The process of claim 8, wherein said aliphatic amino acid is glycine, alanine, valine, norvaline, leucine, isoleucine or norleucine; said oxyamino acid is serine, threonine or homoserine; said sulfur-containing amino acid is methionine, cystine or cysteine; said monoamino dicarboxylic acid is aspartic acid or glutamic acid; said diamino monocarboxylic acid is ornithine, lysine, arginine; said aromatic amino acid is phenylalanine or tyrosine; and said heterocyclic amino acid is histidine or tryptophan.
Priority Claims (2)
Number |
Date |
Country |
Kind |
50-126877 |
Oct 1975 |
JPX |
|
51-43687 |
Apr 1976 |
JPX |
|
Parent Case Info
This is a continuation of application Ser. No. 734,010, filed Oct. 19, 1976, now abandoned.
US Referenced Citations (3)
Number |
Name |
Date |
Kind |
3803327 |
Fujimaki et al. |
Apr 1974 |
|
3888837 |
Tsumita et al. |
Jun 1975 |
|
3972773 |
Isowa et al. |
Aug 1976 |
|
Non-Patent Literature Citations (4)
Entry |
Journal of Biological Chemistry, vol. 204, pp. 891 to 902 (1953). |
Journal of Biological Chemistry, vol. 185, pp. 629 to 641 (1950). |
Biochemical & Biophysical Research Communications, vol. 21, No. 3, pp. 242 to 256 (1965). |
Methods in Enzymology, vol. 19, pp. 883 to 889. |
Continuations (1)
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Number |
Date |
Country |
Parent |
734010 |
Oct 1976 |
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