US4194067A
Expired / Lapsed
US
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Process for the purification of carbohydrate containing enzymes
Assignee: Bayer Corp
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Filing: 1978-07-31
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Granted: 1980-03-18
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Expires: 1998-07-31
Abstract
Abstract This invention relates to a method for purifying a carbohydrate containing enzyme which is a desired enzyme preferred to be separated from a mixture. The method comprises the steps of mixing a solution containing the carbohydrate containing enzyme with a carbohydrate modifying reagent. The carbohydrate modifying reagent reacts with the carbohydrate attached to the enzyme, thereby modifying its chemical structure. The modified enzyme is then separated from the other undesirable enzymes or proteins in the mixture by a suitable chemical separation method, for example, gel filtration chromatography. The method was used in the separation of glucose oxidase from catalase, a separation which by previous methods was very inefficient.
Inventors
- 👤 Melvin H. Keyes
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 22:15
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0065; C12N9/0006; Y; Y10; Y10S; Y10S435/00; Y10S435/814
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
52 words
A method for separating a carbohydrate containing enzyme from the enzyme catalase, comprising: a. mixing a solution containing said carbohydrate containing enzyme with a carbohydrate modifying reagent, b. allowing said carbohydrate containing enzyme and said modifying reagent to react to modify said carbohydrate, c. and separating said enzyme from said catalase enzyme.
Claim 2
Independent Claim
52 words
A method for separating a carbohydrate containing enzyme from the enzyme catalase, comprising: a. mixing a solution containing said carbohydrate containing enzyme with a carbohydrate modifying reagent, b. allowing said carbohydrate containing enzyme and said modifying reagent to react to modify said carbohydrate, c. and separating said enzyme from said catalase enzyme.
Claim 3
Dependent Claim
17 words
The method of claim 1 wherein said modifying agent oxidizes said carbohydrates on said carbohydrate containing enzyme.
Claim 4
Dependent Claim
14 words
The method of claim 2 wherein said modifying reagent is an inorganic oxidizing agent.
Claim 5
Dependent Claim
12 words
The method of claim 3 wherein said oxidizing agent is sodium periodate.
Claim 6
Dependent Claim
12 words
The method of claim 1 wherein said modifying reagent is an enzyme.
Claim 7
Dependent Claim
11 words
The method of claim 5 wherein said enzyme is a dextranase.
Claim 8
Dependent Claim
11 words
The method of claim 5 wherein said enzyme is an amylase.
Claim 9
Dependent Claim
11 words
The method of claim 5 wherein said enzyme is a cellulase.
Claim 10
Dependent Claim
11 words
The method of claim 5 wherein said enzyme is a glucoamylase.
Claim 11
Dependent Claim
13 words
The method of claim 1 wherein said carbohydrate containing enzyme is an oxidase.
Claim 12
Dependent Claim
11 words
The method of claim 10 wherein said oxidase is glucose oxidase.
Claim 13
Dependent Claim
13 words
The method of claim 1 wherein said carbohydrate containing enzyme is a dehydrogenase.
Claim 14
Dependent Claim
22 words
The method of claim 1 wherein said separation of said enzyme from said catalase enzyme is conducted by gel filtration chromatography procedures.
Claim 15
Independent Claim
51 words
A method for separating glucose oxidase contained in an enzyme mixture with catalase comprising: a. adding to said mixture an aqueous solution of dextranase, b. allowing said mixture and dextranase to react thereby removing the carbohydrate from the glucose oxidase, and c. separating said carbohydrate-free glucose oxidase from said reacted mixture.
Claim 16
Independent Claim
51 words
A method for separating glucose oxidase contained in an enzyme mixture with catalase comprising: a. adding to said mixture an aqueous solution of dextranase, b. allowing said mixture and dextranase to react thereby removing the carbohydrate from the glucose oxidase, and c. separating said carbohydrate-free glucose oxidase from said reacted mixture.
Claim 17
Dependent Claim
18 words
The method of claim 14 wherein said dextranase is immobilized on a solid support in said aqueous solution.
Claim 18
Dependent Claim
30 words
The method of claim 15 wherein said dextranase is separated from said reacted mixture by filtration, and said carbohydrate-free glucose oxidase and catalase are separated by gel filtration chromatography procedures.
Claim 19
Independent Claim
54 words
A method for separating glucose oxidase contained in an enzyme mixture with catalase comprising adding to said mixture an aqueous solution of sodium periodate in an amount sufficient to react and partially oxidize the carbohydrate of said glucose oxidase, and separating said oxidized carbohydrate-containing glucose oxidase from said reacted mixture by gel filtration chromatography.
Claim 20
Independent Claim
54 words
A method for separating glucose oxidase contained in an enzyme mixture with catalase comprising adding to said mixture an aqueous solution of sodium periodate in an amount sufficient to react and partially oxidize the carbohydrate of said glucose oxidase, and separating said oxidized carbohydrate-containing glucose oxidase from said reacted mixture by gel filtration chromatography.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-03-18 | 1983-09-17 | 1984-03-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-03-18 | 1987-09-17 | 1988-03-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-03-18 | 1991-09-17 | 1992-03-18 | $7700.00 | Window Closed / Lapsed |