EP0188342A2
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EP
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Method and apparatus for enzymatic synthesis
Assignee: Genentech Inc
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Filing: 1986-01-13
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Granted: —
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Expires: 2006-01-13
Abstract
Abstract Enzyme-catalyzed synthetic reactions having otherwise unfavourable equilibria are facilitated by recovery of product by electrodialysis. Methods and apparatus are provided that can be operated continuously to produce high yields of enriched product without recourse to post-synthetic product manipulations heretofore employed that arte expensive and present a risk of residual toxic by-products. The invention is applied to advantage in the preparation of alpha-L-aspartyl-L-phenylalanine lower alkyl esters.
Inventors
- 👤 Bradley Richard Snedecor
- 👤 Chung Chih Hsu
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-28 23:39
CPC Classifications
C; C12; C12P; C12P21/00; C07; C07K; C07K5/00; C07K5/04; C07K5/06; C07K5/06104; C07K5/06113; C07K5/06121; C07K5/0613
Claims Analysis
Total Claims: 26
Claim 1
Independent Claim
110 words
L. A process comprising (a) contacting one or more reactants with an enzyme to produce a composition comprising a mixture of said reactants and one or more products, wherein either all of the products or all of the reactants bear substantially zero net ionic charge, provided that if all reactants bear substantially zero net ionic charge then the products bear a substantial net ionic charge, or if all products bear substantially zero net ionic charge then the reactants bear a substantial net ionic charge; (b) electrodialyzing the composition of step a) whereby product is separated from reactant; (c) recovering product; and (d) recycling separated reactant into contact with the enzyme.
Claim 2
Dependent Claim
11 words
The process of claim 1 wherein the product is a polypeptide.
Claim 3
Dependent Claim
11 words
The process of claim 1 wherein the product is a dipeptide.
Claim 4
Dependent Claim
24 words
The process of claim 3 wherein the dipeptide is APM and the reactants are aspartic acid and phenylalanine methyl ester or the salts thereof.
Claim 5
Dependent Claim
18 words
The process of claim 1 wherein the enzyme is capable of acting on substrates to form peptide bonds.
Claim 6
Dependent Claim
11 words
The process of claim 5 wherein the enzyme is a protease.
Claim 7
Dependent Claim
30 words
The process of claim 6 wherein the protease is an exoprotease capable of catalyzing the formation of a peptide bond between aspartic acid, or its salts, and phenylalanine methyl ester.
Claim 8
Dependent Claim
16 words
The process of claim 1 wherein the enzyme is present within a living or killed cell.
Claim 9
Dependent Claim
23 words
The process of claim 1 wherein the enzyme is immobilized by covalent bonding to a support, by physical entrapment or by ionic adsorption.
Claim 10
Dependent Claim
16 words
The process of claim 1 wherein the enzyme is covalently immobilized on an ion exchange membrane.
Claim 11
Dependent Claim
14 words
The process of claim 9 wherein the enzyme is ionically adsorbed onto a membrane.
Claim 12
Dependent Claim
13 words
The process of claim 1 wherein the composition further contains an organic solvent.
Claim 13
Dependent Claim
19 words
The process of claim 1 wherein the enzyme is not separated from a reactant solution by a permeable membrane.
Claim 14
Independent Claim
35 words
An apparatus comprising an immobilized enzyme in fluid communication with a means for electrodialysis, means for recirculating reactant from the electrodialysis means to the immobilized enzyme, and means for recovering product from the electrodialysis means.
Claim 15
Independent Claim
77 words
An apparatus comprising an electrodialysis salt depleting chamber having limits defined at least in part by an ion and water permeable ion exchange membrane, means for applying a voltage across the chamber, an electrodialysis salt accumulating chamber situated on the opposite side of the membrane from the salt depleting chamber, an enzyme in fluid communication with the salt depleting chamber, and means for recycling fluid to the enzyme from either the salt depleting or salt accumulating chamber.
Claim 16
Dependent Claim
20 words
L6. The apparatus of claim 15 wherein the enzyme is capable of acting on substrates to form a peptide bond.
Claim 17
Dependent Claim
11 words
The apparatus of claim 16 wherein the enzyme is a protease.
Claim 18
Dependent Claim
15 words
The apparatus of claim 15 additionally comprising a means for supplying reactants to the enzyme.
Claim 19
Dependent Claim
23 words
The apparatus of claim 15 wherein the enzyme is immobilized by covalent bonding to a support, by physical entrapment or by ionic adsorption.
Claim 20
Dependent Claim
15 words
The apparatus of claim 19 wherein the enzyme is immobilized on the ion exchange membrane.
Claim 21
Dependent Claim
22 words
The apparatus of claim 19 wherein the enzyme is present in a reactor chamber in fluid communication with the salt depleting chamber.
Claim 22
Dependent Claim
21 words
The apparatus of claim 21 including means for removing inorganic salts and water from fluid recycled to the enzyme reactor chamber.
Claim 23
Dependent Claim
18 words
The apparatus of claim 15 including means for precipitating product which is removed from the salt depleting chamber.
Claim 24
Dependent Claim
21 words
The apparatus of claim 15 wherein the means for recycling fluid conducts fluid from the salt accumulating chamber to the enzyme.
Claim 25
Dependent Claim
62 words
An electrodialysis apparatus having salt accumulating and depleting cells, means for introducing charged reagents into the salt accumulating cell, means for removing product from the salt depleting cells, an enzyme immobilized in the salt accumulating cell in the region of highest concentration of charged ions and non-electrically conductive means for recycling fluid from the salt accumulating cell into the salt depleting cell.
Claim 26
Dependent Claim
14 words
The device of claim 25 wherein the means for introducing and removing are pumps.
References Cited (Backward Citations)
No citation records indexed.
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