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US4430427A
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Red absorbing combination of alcohol oxidase and an azide compound
Assignee: MOLDOWAN LABORATORIES Inc
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Filing: 1980-11-04
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Granted: 1984-02-07
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Expires: 2000-11-04
Abstract
Abstract Combining active alcohol oxidase with sufficient amount of an azide compound has been found to form a red absorbing combination. Formation of the red absorbing combination enables determining the presence of active alcohol oxidase by adding the azide compound to a preparation and observing the resultant color. Additionally, alcohol oxidase may be purified by adding the azide compound to a preparation containing active alcohol oxidase in an amount effective to produce a red absorbing complex and separating the red absorbing complex from the preparation.
Inventors
- 👤 Thomas R. Hopkins
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 17:09
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0006; C12Q; C12Q1/00; C12Q1/26; Y; Y10; Y10S; Y10S435/00; Y10S435/814; Y10S435/815; Y10S435/816
Claims Analysis
Total Claims: 31
Claim 1
Independent Claim
64 words
A composition of matter comprising active alcohol oxidase; and an azide compound selected from the group of compounds having the formula R"(N 3 ) x wherein R" is a metal atom, a hydrogen atom, or the ammonium radical, and N 3 is the moiety N═N═N, the azide compound being present in an amount effective to form a red absorbing combination with active alcohol oxidase.
Claim 2
Dependent Claim
64 words
A composition of matter comprising active alcohol oxidase; and an azide compound selected from the group of compounds having the formula R"(N 3 ) x wherein R" is a metal atom, a hydrogen atom, or the ammonium radical, and N 3 is the moiety N═N═N, the azide compound being present in an amount effective to form a red absorbing combination with active alcohol oxidase.
Claim 3
Dependent Claim
21 words
A composition as in claim 1 wherein: the azide is selected from the group consisting of azide salts of electropositive metals.
Claim 4
Dependent Claim
32 words
A composition as in claim 1 wherein: the azide is present in an amount in the range of about 1 mole azide/mole alcohol oxidase to about 5.0×10 7 moles azide/mole alcohol oxidase.
Claim 5
Dependent Claim
32 words
A composition as in claim 1 wherein: the azide is present in an amount in the range of about 8 moles azide/mole alcohol oxidase to about 5.0×10 7 moles azide/mole alcohol oxidase.
Claim 6
Dependent Claim
31 words
A composition as in claim 1 wherein: the azide is present in an amount in the range of about 10 moles azide/mole alcohol oxidase to about 2000 moles azide/mole alcohol oxidase.
Claim 7
Dependent Claim
40 words
A composition as in claim 1 wherein: the alcohol oxidase is isolated from genus Pichia; and the azide is present in an amount in the range of about 0.07 mg azide/gram alcohol oxidase to about 80,000 mg azide/gram alcohol oxidase.
Claim 8
Dependent Claim
40 words
A composition as in claim 1 wherein: the alcohol oxidase is isolated from genus Pichia; and the azide is present in an amount in the range of about 0.1 mg azide/gram alcohol oxidase to about 80,000 mg azide/gram alcohol oxidase.
Claim 9
Dependent Claim
40 words
A composition as in claim 1 wherein: the alcohol oxidase is isolated from genus Pichia; and the azide is present in an amount in the range of about 0.7 mg azide/gram alcohol oxidase to about 135 mg azide/gram alcohol oxidase.
Claim 10
Dependent Claim
56 words
A composition as in claim 1 wherein: the alcohol oxidase has a specific activity in the range of about 10-12 Enzyme Units per mg protein; and the azide is present in an amount in the range of about 7×10 6 - mg azide (N 3 )/Enzyme Unit to about 8 mg azide (N 3 )/Enzyme Unit.
Claim 11
Dependent Claim
55 words
A composition as in claim 1 wherein: the alcohol oxidase has a specific activity in the range of about 10-12 Enzyme Units per mg protein; and the azide is present in an amount in the range of about 7×10 -5 mg azide (N 3 )/Enzyme Unit to about 8 mg azide (N 3 )/Enzyme Unit.
Claim 12
Dependent Claim
56 words
A composition as in claim 1 wherein: the alcohol oxidase has a specific activity in the range of about 10-12 Enzyme Units per mg protein; and the azide is present in an amount in the range of about 7×10 -4 mg azide (N 3 )/Enzyme Unit to about 1.35×10 -2 mg azide (N 3 )/Enzyme Unit.
Claim 13
Independent Claim
73 words
A method of purifying an active alcohol oxidase comprising: adding an azide compound selected from the group of compounds having the formula R"(N 3 ) x wherein R" is a metal atom, a hydrogen atom, or the ammonium radical, and N 3 is the moiety N═N═N to a preparation comprising the active alcohol oxidase in an amount effective for producing a red absorbing complex; and separating the red absorbing complex from the preparation.
Claim 14
Independent Claim
73 words
A method of purifying an active alcohol oxidase comprising: adding an azide compound selected from the group of compounds having the formula R"(N 3 ) x wherein R" is a metal atom, a hydrogen atom, or the ammonium radical, and N 3 is the moiety N═N═N to a preparation comprising the active alcohol oxidase in an amount effective for producing a red absorbing complex; and separating the red absorbing complex from the preparation.
Claim 15
Dependent Claim
35 words
A method as in claim 12 wherein adding an azide comprises: adding an amount of sodium azide in the range of about 1 mole azide/mole alcohol oxidase to about 5.0×10 7 moles azide/mole alcohol oxidase.
Claim 16
Dependent Claim
35 words
A method as in claim 12 wherein adding an azide comprises: adding an amount of sodium azide in the range of about 8 moles azide/mole alcohol oxidase to about 5.0×10 7 moles azide/mole alcohol oxidase.
Claim 17
Dependent Claim
34 words
A method as in claim 12 wherein adding an azide comprises: adding an amount of sodium azide in the range of about 10 moles azide/mole alcohol oxidase to about 2000 moles azide/mole alcohol oxidase.
Claim 18
Dependent Claim
44 words
A method as in claim 12 wherein adding an azide comprises: adding an amount of sodium azide in the range of about 0.07 mg azide/gram alcohol oxidase to about 80,000 mg azide/gram alcohol oxidase; and wherein the alcohol oxidase is isolated from genus Pichia.
Claim 19
Dependent Claim
44 words
A method as in claim 12 wherein adding an azide comprises: adding an amount of sodium azide in the range of about 0.1 mg azide/gram alcohol oxidase to about 80,000 mg azide/gram alcohol oxidase; and wherein the alcohol oxidase is isolated from genus Pichia.
Claim 20
Dependent Claim
44 words
A method as in claim 12 wherein adding an azide comprises: adding an amount of sodium azide in the range of about 0.7 mg azide/gram alcohol oxidase to about 135 mg azide/gram alcohol oxidase; and wherein the alcohol oxidase is isolated from genus Pichia.
Claim 21
Dependent Claim
59 words
A method as in claim 12 wherein adding an azide comprises: adding an amount of sodium azide in the range of about 7×10 -6 mg azide (N 3 )/Enzyme Unit to about 8 mg azide (N 3 )/Enzyme Unit; and wherein the alcohol oxidase has a specific activity in the range of about 10-12 Enzyme Units per mg protein.
Claim 22
Dependent Claim
59 words
A method as in claim 12 wherein adding an azide comprises: adding an amount of sodium azide in the range of about 7×10 -5 mg azide (N 3 )/Enzyme Unit to about 8 mg azide (N 3 )/Enzyme Unit; and wherein the alcohol oxidase has a specific activity in the range of about 10-12 Enzyme Units per mg protein.
Claim 23
Dependent Claim
60 words
A method as in claim 12 wherein adding an azide comprises: adding an amount of sodium azide in the range of about 7×10 -4 mg azide (N 3 )/Enzyme Unit to about 1.35×10 -2 mg azide (N 3 )/Enzyme Unit; and wherein the alcohol oxidase has a specific activity in the range of about 10-12 Enzyme Units per mg protein.
Claim 24
Dependent Claim
28 words
The method of claim 12 in which the red absorbing complex is separated from the preparation by the use of differential solubility using ammonium sulfate or polyethylene glycol.
Claim 25
Dependent Claim
19 words
The method of claim 12 in which the red absorbing complex is separated from the preparation by specific precipitation.
Claim 26
Dependent Claim
19 words
The method of claim 12 in which the red absorbing complex is separated from the preparation by column chromatography.
Claim 27
Dependent Claim
19 words
The method of claim 12 in which the red absorbing complex is separated from the preparation by preparative electrophoresis.
Claim 28
Dependent Claim
19 words
The method of claim 12 in which the red absorbing complex is separated from the preparation by preparative ultracentrifugation.
Claim 29
Dependent Claim
20 words
The method of claim 12 in which the red absorbing complex is separated from the preparation by dialysis to crystallization.
Claim 30
Independent Claim
79 words
A method of determining the presence of active alcohol oxidase comprising: observing the color of a composition of matter comprising an alcohol oxidase and an azide compound selected from the group of compounds having the formula R"(N 3 ) x wherein R" is a metal atom, a hydrogen atom, or the ammonium radical, and N 3 is the moiety N═N═N the azide compound being present in an amount effective to form a red absorbing combination with active alcohol oxidase.
Claim 31
Independent Claim
79 words
A method of determining the presence of active alcohol oxidase comprising: observing the color of a composition of matter comprising an alcohol oxidase and an azide compound selected from the group of compounds having the formula R"(N 3 ) x wherein R" is a metal atom, a hydrogen atom, or the ammonium radical, and N 3 is the moiety N═N═N the azide compound being present in an amount effective to form a red absorbing combination with active alcohol oxidase.
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) | 1987-02-06 | 1987-08-08 | 1988-02-07 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-02-06 | 1991-08-08 | 1992-02-07 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-02-06 | 1995-08-08 | 1996-02-07 | $7700.00 | Window Closed / Lapsed |