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US4225672A
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Method for producing maltooligosaccharide glycosides
Assignee: Individual
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Filing: 1979-02-27
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Granted: 1980-09-30
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Expires: 1999-02-27
Abstract
Abstract Disclosed herein is a method for producing maltooligosaccharide glycosides in substantially pure form which includes the steps of incubating a glucosyl donor and a glucosyl acceptor in the presence of a glucanotransferase enzyme under transglycosylating conditions and separating the maltooligosaccharide glycoside from the reaction mixture.
Inventors
- 👤 Leo M. Hall
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-01 23:06
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/10; C12N9/1048; C12N9/1051; C12N9/1074; C07; C07H; C07H3/00; C07H3/06; C12P; C12P19/00; C12P19/44; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/832; Y10S435/837; Y10S435/852
Claims Analysis
Total Claims: 9
Claim 1
Independent Claim
61 words
A method for producing an α or β maltooligosaccharide glycoside derivative in substantially pure form, comprising incubating an aqueous solution of a glucosyl donor and a chromogenic or fluorogenic glycoside in the presence of a glucanotransferase enzyme (E.C.2.4.1.19) under transglycosylating conditions to form a reaction mixture containing the maltooligosaccharide glycoside derivative; and separating the maltooligosaccharide glycoside derivative from the reaction mixture.
Claim 2
Independent Claim
61 words
A method for producing an α or β maltooligosaccharide glycoside derivative in substantially pure form, comprising incubating an aqueous solution of a glucosyl donor and a chromogenic or fluorogenic glycoside in the presence of a glucanotransferase enzyme (E.C.2.4.1.19) under transglycosylating conditions to form a reaction mixture containing the maltooligosaccharide glycoside derivative; and separating the maltooligosaccharide glycoside derivative from the reaction mixture.
Claim 3
Dependent Claim
47 words
The method of claim 1 wherein the glucosyl donor is selected from the group consisting of α-cyclic dextrin, β-cyclic dextrin, linear oligosaccharides, maltotriose, maltotetraose, maltopentaose, maltohexaose, maltoheptaose, starch, glycogen and dextrins; the chromogenic or fluorogenic gylcoside is selected from the group consisting of p-nitrophenyl-α-D-glycoside, p-nitrophenyl-β-D-glycoside, 4-methylumbelliferyl-α-D-glycoside, 4-methylumbelliferyl-β-D-glycoside.
Claim 4
Independent Claim
33 words
The method of claims 1 or 2 wherein the glucanotransferase enzyme is an enzyme produced by an organism selected from the group consisting of Bacillus macerans, Bacillus stearothermophilus, Bacillus megaterium, and Klebsiella pneumoniae.
Claim 5
Independent Claim
45 words
The method of claims 1 or 2 wherein the transglycoslating conditions include a pH of from about 4.9 to about 6.0, a reaction temperature of from about 10° C. to about 60° C., and a reaction time from about 3 minutes to about 5 hours.
Claim 6
Independent Claim
46 words
The method of claims 1 or 2 wherein the transglycosolating conditions include a pH of from about 5.2 to about 5.4, a reaction temperature of from about 38° C. to about 45° C., and a reaction time of from about 25 minutes to about 150 minutes.
Claim 7
Dependent Claim
85 words
The method of claim 4 wherein the glucosyl donor is α-cyclic dextrin, and is employed in a concentration of from about 0.10 to about 0.25 grams per ml, the chromogenic or fluorogenic glycoside is selected from the group consisting of p-nitrophenyl-α-D-glycoside, p-nitrophenyl-β-D-glycoside, 4-methylumbelliferyl-α-D-glycoside, and 4-methylumbelliferyl-β-D-glucoside, and is employed at a concentration of from about 1.5 to about 26 milligrams per ml, and the glucanotransferase enzyme is an enzyme produced by the organism Bacillus macerans and is employed at a concentration sufficient to catalyze the reaction.
Claim 8
Dependent Claim
95 words
The method of claim 5 wherein the glucosyl donor is α-cyclic dextrin, and is employed in a concentration of from about 0.10 to about 0.25 grams per ml, the chromogenic or fluorogenic glycoside is selected from the group consisting of p-nitrophenyl-α-D-glycoside, p-nitrohenyl-β-D-glycoside, 4-methylumbelliferyl-α-D-glycoside, and 4-methylumbelliferyl-β-D-glycoside, and is employed at a concentration of from about 1.5 to about 26 milligrams per ml, and the glucanotransferase enzyme is an enzyme produced by the organism Bacillus macerans and is employed at a concentration of about 0.3 Units to about 3.0 Units of activity per ml of reaction mixture.
Claim 9
Independent Claim
157 words
The method of claims 1 or 2 wherein the glucosyl donor is α-cyclic dextrin and the maltooligosaccharide glycoside derivative is separated from the reaction mixture by the steps of: a. denaturing the glucanotransferase by adjusting the pH of the reaction mixture to about 1.8 to 2.5, heating rapidly to 80° C. to 85° C. and, after about 2 to 3 minutes, chilling rapidly to room temperature; b. adjusting the pH of the reaction mixture to about 6 to 7 and adding about 0.08 to about 0.20 ml of a liquid chlorinated hydrocarbon, then stirring for about 18 hours at about 4° C., to precipitate unreacted α-cyclic dextrin; c. removing the precipitated α-cyclic dextrin from the reaction mixture thereby forming a clarified reaction mixture; d. chromatographically separating the maltooligosaccharide glycoside derivative from the clarified reaction mixture on a microcrystalline cellulose column using ethanol-water eluates; e. collecting the maltooligosaccharide glycoside-containing fractions; and f. evaporating the collected fractions to dryness.
References Cited (Backward Citations)
US4102747A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1983-09-30 | 1984-03-31 | 1984-09-30 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1987-09-30 | 1988-03-31 | 1988-09-30 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1991-09-30 | 1992-03-31 | 1992-09-30 | $7700.00 | Window Closed / Lapsed |