US4092219A
Expired / Lapsed
US
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Preparation of enzyme-membrane complexes
Assignee: Individual
•
Filing: 1975-08-13
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Granted: 1978-05-30
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Expires: 1995-08-13
Abstract
Abstract The amount of enzyme complexed to a protein membrane is increased by treating the protein before or after forming the membrane with a proteolytic enzyme. Preferred proteolytic enzymes are pepsin, trypsin and pronase. Treatment is carried out at a temperature between 15° and 25° C for a period of 1 to 12 hours. Enzymes are complexed to the protein membrane, after treatment, by swelling and washing the membrane and contacting the membrane with an enzyme.
Inventors
- 👤 Po-Min Lin
- 👤 Jack R. Giacin
- 👤 Seymour G. Gilbert
- 👤 Joseph G. Leeder
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 05:15
CPC Classifications
B; B01; B01D; B01D69/00; B01D69/14; B01D69/141; B01D69/142; B01D69/144; C; C12; C12N; C12N11/00; C12N11/02
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
131 words
A method for preparing an enzyme-membrane complex which comprises providing a membrane of collagen, said collagen including a well-ordered crystalline region and a telopeptide region, treating said membrane with a proteolytic enzyme selected from the group consisting of pepsin, trypsin, pronase, and mixtures thereof at a temperature between about 15° and 25° C., and for a period of from about 1 to 12 hours, swelling said membrane at a pH of between about 2 and 12, washing said swollen membrane, and complexing an enzyme to said washed membrane by contacting said membrane with an aqueous solution of an enzyme, said contacting of said membrane with said enzyme being carried out for a period of between about 12 and 48 hours and at a temperature of between about 4° and 15° C.
Claim 2
Independent Claim
131 words
A method for preparing an enzyme-membrane complex which comprises providing a membrane of collagen, said collagen including a well-ordered crystalline region and a telopeptide region, treating said membrane with a proteolytic enzyme selected from the group consisting of pepsin, trypsin, pronase, and mixtures thereof at a temperature between about 15° and 25° C., and for a period of from about 1 to 12 hours, swelling said membrane at a pH of between about 2 and 12, washing said swollen membrane, and complexing an enzyme to said washed membrane by contacting said membrane with an aqueous solution of an enzyme, said contacting of said membrane with said enzyme being carried out for a period of between about 12 and 48 hours and at a temperature of between about 4° and 15° C.
Claim 3
Dependent Claim
32 words
The method of claim 1 wherein said proteolytic enzyme comprises pepsin, and is maintained in an aqueous solution at a pH from 2 to 4 for from about 1 to 12 hours.
Claim 4
Dependent Claim
33 words
The method of claim 1 wherein said proteolytic enzyme comprises pronase, and is maintained in an aqueous solution at a pH of from 6 to 8 for from about 1 to 6 hours.
Claim 5
Dependent Claim
13 words
The method of claim 3 wherein said pH is maintained at about 7.2.
Claim 6
Dependent Claim
20 words
The method of claim 1 wherein said membrane is prepared by casting a film from a dispersion of said collagen.
Claim 7
Dependent Claim
27 words
The method of claim 5 wherein said film is aged for a period of from 3 to 200 days prior to said treatment with said proteolytic enzyme.
Claim 8
Dependent Claim
11 words
The method of claim 5 wherein said proteolytic enzyme comprises pepsin.
Claim 9
Dependent Claim
24 words
An enzyme-membrane complex containing greater than about 100 mg of said enzyme per gram of said complex prepared by the process of claim 1.
Claim 10
Dependent Claim
32 words
The enzyme-membrane complex of claim 8 wherein said enzyme complexed to said membrane is selected from the group consisting of lysozyme, urease, amylase, invertase, cellulase, glucose isomerase, β-galactosidase, and compatible mixtures thereof.
Claim 11
Independent Claim
140 words
A method for preparing an enzyme-membrane complex which comprises preparing a dispersion of collagen, said collagen including a well-ordered crystalline region and a telopeptide region, treating said collagen in the dispersion with a proteolytic enzyme selected from the group consisting of pepsin, trypsin, pronase, and mixtures thereof at a temperature of between about 15° and 25° C. for a period of from about 1 to 12 hours, casting a membrane of said collagen, swelling said membrane at a pH of between about 2 and 12, washing said swollen membrane and complexing an enzyme to said washed membrane by contacting said membrane with an aqueous solution of an enzyme, said contacting of said membrane with said enzyme being carried out for a period of between about 12 and 48 hours and at a temperature of between about 4° and 15° C.
Claim 12
Independent Claim
140 words
A method for preparing an enzyme-membrane complex which comprises preparing a dispersion of collagen, said collagen including a well-ordered crystalline region and a telopeptide region, treating said collagen in the dispersion with a proteolytic enzyme selected from the group consisting of pepsin, trypsin, pronase, and mixtures thereof at a temperature of between about 15° and 25° C. for a period of from about 1 to 12 hours, casting a membrane of said collagen, swelling said membrane at a pH of between about 2 and 12, washing said swollen membrane and complexing an enzyme to said washed membrane by contacting said membrane with an aqueous solution of an enzyme, said contacting of said membrane with said enzyme being carried out for a period of between about 12 and 48 hours and at a temperature of between about 4° and 15° C.
Claim 13
Dependent Claim
29 words
The method of claim 10 wherein said enzyme to be complexed with said membrane is selected from the group consisting of oxidoreductases, transferases, hydrolases, isomerases, and compatible mixtures thereof.
Claim 14
Dependent Claim
32 words
The method of claim 10 wherein said enzyme to be complexed with said membrane is selected from the group consisting of lysozyme, urease, amylase, invertase, cellulase, glucose-isomerase, B-galactosidase and compatible mixtures thereof.
Claim 15
Dependent Claim
10 words
The method of claim 10 including drying said enzyme-membrane complex.
Claim 16
Dependent Claim
24 words
An enzyme-membrane complex containing greater than about 100 mg of said enzyme per gram of said complex prepared by the process of claim 10.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1981-05-29 | 1981-11-28 | 1982-05-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-05-29 | 1985-11-28 | 1986-05-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-05-29 | 1989-11-28 | 1990-05-30 | $7700.00 | Window Closed / Lapsed |