US4576913A
Expired / Lapsed
US
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NAD(P)-Independent glycerol dehydrogenase, a process for the preparation thereof and the use thereof for the determination of glycerol and triglycerides
Assignee: Roche Diagnostics GmbH
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Filing: 1984-03-23
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Granted: 1986-03-18
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Expires: 2004-03-23
Abstract
Abstract The present invention provides a new NAD(P)-independent glycerol dehydrogenase and a process for obtaining it, wherein a micro-organism of the species Pseudomonas, Serratia, Klebsiella, Erwinia, Aspergillus, Penicillium, Rhizopus, Acetobacter or Gluconobacter which is capable of forming this enzyme is cultured in a nutrient medium and the enzyme isolated from the culture broth. The present invention also provides processes for the determination of glycerol and of triglycerides which make use of this new enzyme.
Inventors
- 👤 Osao Adachi
- 👤 Minoru Ameyama
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 21:40
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0006; C12Q; C12Q1/00; C12Q1/26; C12Q1/32; C12Q1/61; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/823
Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
18 words
Glycerol dehydrogenase which catalyzes the reaction of glycerol to dihydroxyacetone characterized in that said glycerol dehydrogenase is NAD(P)-independent.
Claim 2
Dependent Claim
18 words
Glycerol dehydrogenase which catalyzes the reaction of glycerol to dihydroxyacetone characterized in that said glycerol dehydrogenase is NAD(P)-independent.
Claim 3
Dependent Claim
42 words
Process for obtaining an NAD(P)-independent glycerol dehydrogenase comprising culturing, in a nutrient medium, a micro-organism of the species Pseudomonas, Serratia, Klebsiella, Erwinia, Aspergillus, Penicillium, Rhizopus, Acetobacter or Gluconobacter which is capable of forming this enzyme and isolating enzyme from the culture broth.
Claim 4
Dependent Claim
42 words
Process for obtaining an NAD(P)-independent glycerol dehydrogenase comprising culturing, in a nutrient medium, a micro-organism of the species Pseudomonas, Serratia, Klebsiella, Erwinia, Aspergillus, Penicillium, Rhizopus, Acetobacter or Gluconobacter which is capable of forming this enzyme and isolating enzyme from the culture broth.
Claim 5
Dependent Claim
23 words
Process according to claim 2, wherein the micro-organism used is Acetobacter xylinus DSM 2623, Gluconobacter cerinus DSM 2622 or Gluconobacter industrius DSM 2621.
Claim 6
Dependent Claim
10 words
NAD(P)-independent glycerol dehydrogenase, obtained by the process of claim 2.
Claim 7
Dependent Claim
55 words
Process for the determination of glycerol, comprising converting glycerol into dihydroxyacetone with an NAD(P)-independent glycerol dehydrogenase according to claim 1 in the presence of a proton and electron acceptor, and measuring the decrease of the oxidised acceptor or the increase of the reduced acceptor or the increase of dihydroxyacetone as a measure of the glycerol.
Claim 8
Dependent Claim
19 words
Process according to claim 5, wherein oxygen is used as the acceptor and the consumption of oxygen is measured.
Claim 9
Dependent Claim
18 words
Process according to claim 5, wherein a first acceptor redox system is coupled with a further redox indicator.
Claim 10
Dependent Claim
29 words
Process according to claim 7, wherein the first acceptor is phenazine methosulphate and the further redox indicator is 2,6-dichlorophenol-indophenol, the absorption decrease of 2,6-dichlorophenol-indophenol being measured at 600 nm.
Claim 11
Dependent Claim
27 words
Process according to claim 7, wherein the first acceptor is phenazine methosulphate and the further redox indicator is a tetrazolium salt, the formazane formation being measured photometrically.
Claim 12
Dependent Claim
66 words
Process for the determination of triglycerides, comprising saponifying the triglycerides to glycerol and free fatty acids converting the free glycerol into dihydroxyacetone with an NAD(P)-independent glycerol dehydrogenase according to claim 1 in the presence of a proton and electron acceptor, and measuring the decrease of the oxidised acceptor or the increase of the reduced acceptor or the increase of dihydroxyacetone as a measure of the glycerol.
Claim 13
Dependent Claim
87 words
Agent for the determination of glycerol, by means of which glycerol is reacted with the help of a glycerol dehydrogenase in the presence of an appropriate acceptor to give dihydroxyacetone, the decrease of the oxidised acceptor or the increase of the reduced acceptor or the increase of dihydroxyacetone being measurable, said agent comprising an NADP-independent glycerol dehydrogenase according to claim 1 and a means for the determination of the decrease of the oxidised acceptor or the increase of the reduced acceptor or the increase of the dihydroxyacetone.
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GB1441642A
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US4223090A
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JPS5533603A
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1989-03-17 | 1989-09-16 | 1990-03-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-03-17 | 1993-09-16 | 1994-03-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-03-17 | 1997-09-16 | 1998-03-18 | $7700.00 | Window Closed / Lapsed |