US3869349A
Expired / Lapsed
US
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Method for the enzymatic hydrolysis of cholesterol esters
Assignee: Eastman Kodak Co
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Filing: 1974-03-25
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Granted: 1975-03-04
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Expires: 1994-03-25
Abstract
Abstract A totally enzymatic method for the hydrolysis of cholesterol esters using a lipase having cholesterol esterase activity and a protease and a method for quantitative determination of total cholesterol in compositions containing both free and esterified cholesterol comprising enzymatically hydrolyzing the cholesterol esters with a lipase having cholesterol esterase activity and a protease and determining total cholesterol by gas-liquid chromatography or some other suitable technique are described.
Inventors
- 👤 Charles T Goodhue
- 👤 Hugh A Risley
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 06:44
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/60; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/832; Y10S435/839; Y10S435/886; Y10S435/897; Y10S435/911; Y10S435/913; Y10S435/918; Y10S435/921
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
59 words
IN A PROCESS OF ASSAYING AN AQUEOUS PROTEINACEOUS LIQUID CONTAINING CHOLESTEROL ESTERS FOR TOTAL CHOLESTEROL CONTENT IN WHICH SAID ESTER IS HYDROLYZED TO LIBERATE ALL OF SAID CHOLESTER OL FOLLOWED BY DETERMINING THE AMOUNT OF CHOLESTEROL, THE IMPROVEMENT COMPRISING EFFECTING SAID HYDROLYSIS BY TREATING SAID AQUEOUS PROTEINACEOUS LIQUID WITH BOTH A LIPASE HAVING CHOLESTEROL ESTRASE ACTIVITY AND A PROTEASE.
Claim 2
Dependent Claim
59 words
IN A PROCESS OF ASSAYING AN AQUEOUS PROTEINACEOUS LIQUID CONTAINING CHOLESTEROL ESTERS FOR TOTAL CHOLESTEROL CONTENT IN WHICH SAID ESTER IS HYDROLYZED TO LIBERATE ALL OF SAID CHOLESTER OL FOLLOWED BY DETERMINING THE AMOUNT OF CHOLESTEROL, THE IMPROVEMENT COMPRISING EFFECTING SAID HYDROLYSIS BY TREATING SAID AQUEOUS PROTEINACEOUS LIQUID WITH BOTH A LIPASE HAVING CHOLESTEROL ESTRASE ACTIVITY AND A PROTEASE.
Claim 3
Dependent Claim
77 words
The improved method of claim 1 wherein said lipase having cholesterol esterase activity releases at least 25 mg% cholesterol (in 2 hours at 37*C under nitrogen) when 50 mg of a preparation of said lipase in 5 ml 0.1 M phosphate buffer, pH 7.0, is used to treat a dispersion of cholesteryl linoleate prepared by dispersing 200 mg cholesteryl linoleate in 5 ml of ethyl ether and 100 ml boiling water containing 430 mg of sodium cholate.
Claim 4
Dependent Claim
51 words
The improved method of claim 1 wherein said aqueous liquid is serum and said treating step is accomplished with a mixture comprising from about 600 to about 1500 units of said lipase having cholesterol esterase activity and from about 50 to about 500 units of said protease per ml of serum.
Claim 5
Independent Claim
31 words
The method of clain 1 wherein said treatment is carried out at a temperature of between about 25* and 55*C and at a pH of between about 6.5 and about 9.5.
Claim 6
Independent Claim
31 words
The method of clain 1 wherein said treatment is carried out at a temperature of between about 25* and 55*C and at a pH of between about 6.5 and about 9.5.
Claim 7
Dependent Claim
16 words
The improved method of claim 1 wherein said lipase having esterase activity is a microbial lipase.
Claim 8
Dependent Claim
16 words
The improved method of claim 5 wherein said microbial lipase is the lipase from Candida cylindracca.
Claim 9
Dependent Claim
27 words
The improved method of claim 1 wherein said lipase is selected from the group consisting of wheat germ lipase, pancreatic lipases and the lipase from Candida cylindracca.
Claim 10
Dependent Claim
32 words
The improved method of claim 1 wherein said protease is selected from the group consisting of Alpha -chymotrypsin, papain, bromelain, Bacillus subtilis protease, Aspergillus oryzae protease, Streptomyces griseus protease and mixtures thereof.
Claim 11
Dependent Claim
27 words
The improved method of claim 8 wherein said lipase is selected from the group consisting of wheat germ lipase, pancreatic lipases and the lipase from Candida cylindracca.
Claim 12
Dependent Claim
31 words
The method of claim 9 wherein said treatment is Carried out at a temperature of between about 25 and 55*C and at a pH of between about 6.5 and about 9.5.
References Cited (Backward Citations)
DE2224132A1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1978-03-03 | 1978-09-02 | 1979-03-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1982-03-03 | 1982-09-02 | 1983-03-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1986-03-03 | 1986-09-02 | 1987-03-04 | $7700.00 | Window Closed / Lapsed |