US5002872A
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Enzyme mediated coupling reactions
Assignee: Hampshire Chemical Corp
•
Filing: 1989-05-10
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Granted: 1991-03-26
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Expires: 2009-05-10
Abstract
Abstract Disclosed herein is an enzymatic process to bind two amino acids in the presence of a water-miscible solvent. Preferably the amino acids are precursors of aspartame and a preferred solvent is acetonitrile.
Inventors
- 👤 Akiva T. Gross
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 15:02
CPC Classifications
C; C07; C07K; C07K5/00; C07K5/04; C07K5/06; C07K5/06104; C07K5/06113; C07K5/06121; C07K5/0613; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/909
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
95 words
In an enzymatic process to cause peptide bond formation between two substrates selected from the group consisting of N-protected aspartic acid of a salt thereof and a phenylalanine lower alkyl ester wherein the benzylic carbon atom is substituted with hydrogen or one or more labile groups readily replaceable with hydrogen, the improvement comprising carrying out said process in a single liquid phase in the presence of a solvent comprising a mixture of acetonitrile and water wherein the weight ratio of acetonitrile to water is greater than 1:1 and in the presence of a water-containing metalloproteinase.
Claim 2
Dependent Claim
95 words
In an enzymatic process to cause peptide bond formation between two substrates selected from the group consisting of N-protected aspartic acid of a salt thereof and a phenylalanine lower alkyl ester wherein the benzylic carbon atom is substituted with hydrogen or one or more labile groups readily replaceable with hydrogen, the improvement comprising carrying out said process in a single liquid phase in the presence of a solvent comprising a mixture of acetonitrile and water wherein the weight ratio of acetonitrile to water is greater than 1:1 and in the presence of a water-containing metalloproteinase.
Claim 3
Dependent Claim
12 words
A process as in claim 1 wherein the water-containing metalloproteinase is immobilized.
Claim 4
Dependent Claim
11 words
A process as in claim 1 wherein the enzyme is thermolysin.
Claim 5
Dependent Claim
14 words
A process as in claim 1 wherein the N-protective group is formyl or benzyloxycarbonyl.
Claim 6
Dependent Claim
15 words
A process as in claim 1 wherein the phenylalanine benzyl carbon is substituted with hydroxyl.
Claim 7
Dependent Claim
16 words
A process as in claim 1 wherein the lower alkyl substituent of the ester is methyl.
Claim 8
Dependent Claim
19 words
A process as in claim 1 wherein the process is carried out in the presence of an esterase inhibitor.
Claim 9
Dependent Claim
18 words
A process as in claim 1 wherein the N-protective group is tertiary alkoxycarbonyl, benzyloxycarbonyl, p-toluenesulfonyl, o-nitrophenylsulfenyl, or formyl.
Claim 10
Dependent Claim
81 words
A process for manufacturing a dipeptide or polypeptide by reacting an acid component of an amino acid or peptide having an N-terminal protective group or a salt thereof with an amine component of an amino acid or peptide having a C-terminal protective group or a salt thereof in a single liquid phase in the presence of a water-containing metalloproteinase and a solvent comprising a mixture of acetonitrile and water wherein the weight ratio of acetonitrile to water is greater than 1:1.
Claim 11
Dependent Claim
81 words
A process for manufacturing a dipeptide or polypeptide by reacting an acid component of an amino acid or peptide having an N-terminal protective group or a salt thereof with an amine component of an amino acid or peptide having a C-terminal protective group or a salt thereof in a single liquid phase in the presence of a water-containing metalloproteinase and a solvent comprising a mixture of acetonitrile and water wherein the weight ratio of acetonitrile to water is greater than 1:1.
Claim 12
Dependent Claim
30 words
A process as in claim 9 wherein the solvent comprises greater than 50 wt.% acetonitrile and the balance is a mixture of water and one or more water-miscible organic solvents.
Claim 13
Dependent Claim
11 words
A process as in claim 9 wherein the enzyme is thermolysin.
Claim 14
Dependent Claim
18 words
A process as in claim 9 wherein the amino acid having the N-terminal protective group is aspartic acid.
Claim 15
Dependent Claim
22 words
A process as in claim 9 wherein the amino acid having the C-terminal protective group is a lower alkyl ester of phenylalanine.
Claim 16
Dependent Claim
17 words
A process as in claim 13 wherein the protected amino acid is the methyl ester of phenylalanine.
Claim 17
Dependent Claim
14 words
A process as in claim 12 wherein the N-protective group is formyl or benzyloxycarbonyl.
Claim 18
Dependent Claim
16 words
A process as in claim 9 wherein an esterase inhibitor is present in the reaction mixture.
Claim 19
Dependent Claim
18 words
A process as in claim 9 wherein the N-protective group is tertiary alkoxycarbonyl, benzyloxycarbonyl, p-toluenesulfonyl, o-nitrophenylsulfenyl, or formyl.
Claim 20
Dependent Claim
52 words
A process as in claim 1 wherein the phenylalanine lower alkyl ester corresponds to the formula: ##STR2## wherein Ph is phenyl, X is --OH, --SH, --Cl, --Br, --I, --PCOCH 3 , --OCOOCH 3 , --NH 2 or SCH 3 and R is an alkyl group having from 1 to 4 carbon atoms.
Claim 21
Dependent Claim
14 words
A process as in claim 18 wherein X is --OH and R is methyl.
Claim 22
Dependent Claim
17 words
A process as in claim 9 wherein the amino acid having a C-terminal protective group is D-alanine.
Claim 23
Dependent Claim
17 words
A process as in claim 1 wherein the enzyme is the protease of Vibrio proteolyticus ATCC 53559.
Claim 24
Dependent Claim
17 words
A process as in claim 9 wherein the enzyme is the protease of Vibrio proteolyticus ATCC 53559.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1994-03-25 | 1994-09-24 | 1995-03-26 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-03-25 | 1998-09-24 | 1999-03-26 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-03-25 | 2002-09-24 | 2003-03-26 | $7700.00 | Window Closed / Lapsed |