US3997402A
Expired / Lapsed
US
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Superoxide dismutase and process for production
Assignee: Bpifrance Financement SA
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Filing: 1975-05-06
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Granted: 1976-12-14
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Expires: 1995-05-06
Abstract
Abstract The invention relates to a process for the production of superoxide dismutase and the superoxide dismutase so obtained. The superoxide dismutase obtained is extracted from marine bacterial strains and is characterized in that it comprises non-hematinic iron or copper and zinc, it has a molecular weight of about 33,000 to 40,000 +/- 2500 and a pHi or isoelectric point of about 4 to 8.2 and it has maximum enzyme activity at a pH of about 8.5 to 10 with an optimum at about 9 to 9.5 and the use of said superoxide dismutases for the protection of oxidizable systems from auto-oxidation.
Inventors
- 👤 Adolf Michael Michelson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 18:40
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0089; A; A23; A23B; A23B2/00; A23B2/70; A23B2/725; A23B2/729; A23B2/783; Y; Y10; Y10S; Y10S435/00; Y10S435/814; Y10S435/815; Y10S435/8215; Y10S435/822
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
108 words
A process for the production of superoxide dismutase from cultures of marine bacterial strains comprising dispersing a marine photobacterium bacterial culture in water at about 4° C, adjusting the pH of the dispersion to 6.5 to 8, heating the resulting mixture to 50° to 60° C for a short time, cooling the mixture to about 4° C, removing solids from the mixture, adding neutral salts to the resulting liquid to effect a first fractional precipitation, separating the solids from the mixture, again adding neutrals salts to the resulting liquid to effect a second fractional precipitation and separating the solids from the liquid to recover the solid superoxide dismutase.
Claim 2
Dependent Claim
108 words
A process for the production of superoxide dismutase from cultures of marine bacterial strains comprising dispersing a marine photobacterium bacterial culture in water at about 4° C, adjusting the pH of the dispersion to 6.5 to 8, heating the resulting mixture to 50° to 60° C for a short time, cooling the mixture to about 4° C, removing solids from the mixture, adding neutral salts to the resulting liquid to effect a first fractional precipitation, separating the solids from the mixture, again adding neutrals salts to the resulting liquid to effect a second fractional precipitation and separating the solids from the liquid to recover the solid superoxide dismutase.
Claim 3
Dependent Claim
24 words
The process of claim 1 wherein the marine bacterial strain is selected from the group consisting of Photobacterium phosphoreum, Photobacterium leiognathi and Photobacterium sepia.
Claim 4
Dependent Claim
42 words
The process of claim 1 wherein the superoxide dismutase product is dissolved in a phosphate buffer solution at a pH of 7.8 and dialyzing the resulting solution against the same phosphate buffer solution and recovering the purified superoxide dismutase as inner dialysate.
Claim 5
Dependent Claim
36 words
The process of claim 1 wherein the superoxide dismutase is dissolved in a phosphate buffer solution at a pH of 7.8 and subjecting the said solution to column chromatography and recovering the resulting purified superoxide dismutase.
Claim 6
Dependent Claim
39 words
The process of claim 4 wherein the column chromatography is effected in three stages; the first stage using Sephadex G 200 gel, the second stage using a diethylaminoethyl Sephadex (DEAE-S) and the third stage using Sephadex G 200 gel.
Claim 7
Dependent Claim
34 words
The process of claim 4 wherein the neutral salt for the first fractional crystallization is ammonium sulfate with a final concentration in the liquid of about 30 to 35% of saturation at 4° C.
Claim 8
Dependent Claim
34 words
The process of claim 1 wherein the neutral salt for the second fractional crystallization is ammonium sulfate with a final concentration in the liquid of about 70 to 75% of saturation at 4° C.
Claim 9
Dependent Claim
20 words
The process of claim 1 wherein the said mixture is heated to 50°- 60° C for 3 to 4 minutes.
Claim 10
Dependent Claim
14 words
The process of claim 1 wherein the said neutral salt is neutral ammonium sulfate.
Claim 11
Dependent Claim
54 words
A superoxide dismutase extracted from marine photobacterium bacterial cultures characterized by having a molecular weight of about 40,000 + 2500, a pHi point of about 4 to 7, containing 1 to 2 atoms of non-hematinic iron per mole of superoxide dismutase and having a maximum enzymatic activity at a pH between 8.5 to 10.
Claim 12
Dependent Claim
54 words
A superoxide dismutase extracted from marine photobacterium bacterial cultures characterized by having a molecular weight of about 40,000 + 2500, a pHi point of about 4 to 7, containing 1 to 2 atoms of non-hematinic iron per mole of superoxide dismutase and having a maximum enzymatic activity at a pH between 8.5 to 10.
Claim 13
Dependent Claim
32 words
A superoxide dismutase of claim 10 extracted from Photobacterium leiognathi having a molecular weight of about 42,000, a pHi of about 4.4 and a maximum enzymatic activity at a pH of 9.5.
Claim 14
Dependent Claim
32 words
A superoxide dismutase of claim 10 extracted from Photobacterium sepia having a molecular weight of about 42,500 and a pHi of about 4.1 an optimum enzymatic activity at a pH of 9.5.
Claim 15
Dependent Claim
32 words
A superoxide dismutase of claim 10 extracted from Photobacterium phosphorum having a molecular weight of about 42,000, a pHi of about 4.2 and a maximum enzymatic activity at a pH of 9.5.
Claim 16
Dependent Claim
54 words
A superoxide dismutase extracted from marine photobacterium bacterial cultures characterized by a molecular weight of 33,000 ± 2500, a pHi point of about 8.25, having a maximum enzymatic activity at a pH of 8.5 to 10 and containing about 1 atom of copper and about 2.0 atoms of zinc per mole of superoxide dismutase.
Claim 17
Dependent Claim
54 words
A superoxide dismutase extracted from marine photobacterium bacterial cultures characterized by a molecular weight of 33,000 ± 2500, a pHi point of about 8.25, having a maximum enzymatic activity at a pH of 8.5 to 10 and containing about 1 atom of copper and about 2.0 atoms of zinc per mole of superoxide dismutase.
References Cited (Backward Citations)
US3920521A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1979-12-14 | 1980-06-14 | 1980-12-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1983-12-14 | 1984-06-14 | 1984-12-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1987-12-14 | 1988-06-14 | 1988-12-14 | $7700.00 | Window Closed / Lapsed |