US4135980A
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Choline oxidase
Assignee: Asahi Kasei Corp
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Filing: 1977-04-22
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Granted: 1979-01-23
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Expires: 1997-04-22
Abstract
Abstract A novel enzyme choline oxidase is produced by culturing a microorganism belonging to the genus Arthrobacter, and particularly the species Arthrobacter globiformis B-0577 FERM-P No. 3518, NRRL B-11097, and isolating the choline oxidase thus produced from the culture medium. Choline oxidase is useful for the determination of choline or betaine aldehyde in a sample by mixing the new product with the sample and then measuring the generated hydrogen peroxide, betaine or consumed oxygen.
Inventors
- 👤 Shigeru Ikuta
- 👤 Yoshifumi Horiuchi
- 👤 Hideo Misaki
- 👤 Kazuo Matsuura
- 👤 Shigeyuki Imamura
- 👤 Naoki Muto
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-28 05:46
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0006; C12N1/00; C12N1/20; C12N1/205; C12N9/14; C12N9/16; C12Q; C12Q1/00; C12Q1/26; C12Q1/32; C12Q1/34; C12Q1/44; C12Q1/46; C12R; C12R2001/00; C12R2001/01; C12R2001/06; C12R2001/465; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/83
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
67 words
An enzyme choline oxidase which has the following properties: (1) substrate specificity: a specific activity for a compound of the formula ##STR3## wherein R 1 is --CH 2 OH or --CHO; (2) optimum pH: pH 8 (tris-HCl buffer); (3) isoelectric point: 4.6 (electrophoresis using carrier-type ampholyte); (4) molecular weight: 84,000 ± 2,100 [gel filtration using Sephadex G-150]; (5) enzyme action: catalyzes the reaction [I] and/or [II]: ##STR4##
Claim 2
Dependent Claim
67 words
An enzyme choline oxidase which has the following properties: (1) substrate specificity: a specific activity for a compound of the formula ##STR3## wherein R 1 is --CH 2 OH or --CHO; (2) optimum pH: pH 8 (tris-HCl buffer); (3) isoelectric point: 4.6 (electrophoresis using carrier-type ampholyte); (4) molecular weight: 84,000 ± 2,100 [gel filtration using Sephadex G-150]; (5) enzyme action: catalyzes the reaction [I] and/or [II]: ##STR4##
Claim 3
Dependent Claim
35 words
Choline oxidase as claimed in claim 1 wherein the optimum temperature is about 40° C. (tris-HCl buffer), stable pH is about pH 8 (tris-HCl buffer) and heat stability is below about 40° C. (tris-HCl buffer).
Claim 4
Dependent Claim
35 words
A process for production of choline oxidase which comprises culturing a choline oxidase-producing microorganism belonging to genus Arthrobacter in a nutrient culture medium and isolating the choline oxidase thus produced from the cultured mass thereof.
Claim 5
Dependent Claim
35 words
A process for production of choline oxidase which comprises culturing a choline oxidase-producing microorganism belonging to genus Arthrobacter in a nutrient culture medium and isolating the choline oxidase thus produced from the cultured mass thereof.
Claim 6
Dependent Claim
14 words
A process as claimed in claim 3 wherein the culture medium contains free choline.
Claim 7
Dependent Claim
19 words
A process as claimed in claim 3 wherein the choline oxidase-producing microorganism is Arthrobacter globiformis B-0577 FERM-P No. 3518.
Claim 8
Dependent Claim
30 words
A process as claimed in claim 3 wherein the cultured microorganism belonging to genus Arthrobacter is subjected to lysozyme treatment for the isolation of choline oxidase from the cultured mass.
Claim 9
Dependent Claim
34 words
Method for determination of choline or betaine aldehyde, which method comprises treating a sample containing choline or betaine aldehyde with a choline oxidase as claimed in claim 1 and measuring the generated hydrogen peroxide.
Claim 10
Dependent Claim
32 words
Method as claimed in claim 7 wherein the said sample comprises a liquid containing choline or betaine aldehyde, or a liquid containing choline derivative or betaine aldehyde derivative and hydrolyzing reagent therefor.
Claim 11
Dependent Claim
13 words
Method as claimed in claim 8 wherein the choline derivative is a phospholipid.
Claim 12
Dependent Claim
15 words
Method as claimed in claim 9 wherein the phospholipid is lecithin, lysolecithin or a sphingomyelin.
Claim 13
Dependent Claim
13 words
Method as claimed in claim 8 wherein the hydrolyzing reagent is phospholipase D.
Claim 14
Dependent Claim
22 words
Method as claimed in claim 11 wherein the said phospholipase D is an enzyme obtained from the culture of Streptomyces chromofuscus A-0848.
Claim 15
Dependent Claim
12 words
Method as claimed in claim 8 wherein the choline derivative is benzoylcholine.
Claim 16
Dependent Claim
13 words
Method as claimed in claim 8 wherein the hydrolyzing reagent is choline esterase.
Claim 17
Dependent Claim
33 words
Method as claimed in claim 7 wherein the measurement of hydrogen peroxide is carried out by the system comprising one or more chromogens for color development depending on the concentration of hydrogen peroxide.
Claim 18
Dependent Claim
18 words
Method as claimed in claim 15 wherein the measurement of hydrogen peroxide is carried out by using peroxidase.
Claim 19
Dependent Claim
19 words
Method as claimed in claim 16 wherein the peroxidase is used in an amount of 0.2 to 0.5 unit.
Claim 20
Dependent Claim
25 words
Method as claimed in claim 15 wherein the coloring reagent is used in an amount of two molar excess per mole of hydrogen peroxide formed.
Claim 21
Dependent Claim
16 words
Method as claimed in claim 15 wherein the said system consists of 4-aminoantipyrine, phenol and peroxidase.
Claim 22
Dependent Claim
20 words
Method as claimed in claim 7 wherein the choline oxidase is used in an amount of 1.5 to 5 units.
Claim 23
Dependent Claim
20 words
Method as claimed in claim 7 wherein the said sample contains phospholipid, phospholipase D, choline oxidase, 4-aminoantipyrine, phenol and peroxidase.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1982-01-22 | 1982-07-24 | 1983-01-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1986-01-22 | 1986-07-24 | 1987-01-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1990-01-22 | 1990-07-24 | 1991-01-23 | $7700.00 | Window Closed / Lapsed |