US4329429A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Lactase preparation
Assignee: Danisco Finland Oy
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Filing: 1980-03-24
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Granted: 1982-05-11
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Expires: 2000-03-24
Abstract
Abstract A process for the release of lactase from yeast cells comprising contacting the cells with an alkyl alcohol or a dialkyl ketone, followed by extraction of the lactase in an aqueous solution at a pH in the range 5.5 to 8.0 is disclosed. Also disclosed is a process for the selective reduction of protease activity in lactase preparations comprising heating an aqueous solution of lactase containing protease impurities in the presence of glycerol. Lactase stability during the heating process can optionally be enhanced by addition of manganous ion.
Inventors
- 👤 Dennis M. Fenton
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 11:03
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/24; C12N9/2402; C12N9/2468; C12N9/2471; C12N9/78; C12N9/86; C12Y; C12Y302/00; C12Y302/01; C12Y302/01023; C12Y305/00; C12Y305/02; C12Y305/02006; Y; Y10; Y10S; Y10S435/00; Y10S435/814; Y10S435/8215; Y10S435/911; Y10S435/921
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
113 words
A process for the release of lactase from lactase-containing yeast cells comprising the steps of (a) contacting said cells with from about 20 weight % to 95 weight % of a compound selected from alkyl alcohols of 1 to 4 carbon atoms and dialkyl ketones having from 1 to 4 carbon atoms in each alkyl group, based on the total weight of said cells and said compound, at a temperature from 5° C. to 35° C., and then (b) contacting said cells with an aqueous solution having a pH from about 5.5 to 8.0 at a temperature from about 5° C. to 35° C. and separating the cells from the resulting lactase-containing solution.
Claim 2
Dependent Claim
113 words
A process for the release of lactase from lactase-containing yeast cells comprising the steps of (a) contacting said cells with from about 20 weight % to 95 weight % of a compound selected from alkyl alcohols of 1 to 4 carbon atoms and dialkyl ketones having from 1 to 4 carbon atoms in each alkyl group, based on the total weight of said cells and said compound, at a temperature from 5° C. to 35° C., and then (b) contacting said cells with an aqueous solution having a pH from about 5.5 to 8.0 at a temperature from about 5° C. to 35° C. and separating the cells from the resulting lactase-containing solution.
Claim 3
Dependent Claim
17 words
A process according to claim 1 wherein said cells are of the genera Kluyveromyces, Candida, or Torula.
Claim 4
Dependent Claim
24 words
A process according to claim 2 wherein the temperature of each of steps (a) and (b) is from about 20° C. to 30° C.
Claim 5
Dependent Claim
18 words
A process according to claim 2 wherein the pH of said aqueous solution is from 6.4 to 7.0.
Claim 6
Dependent Claim
19 words
A process according to claim 2 wherein the compound of step (a) is methanol, ethanol, isopropanol or mixtures thereof.
Claim 7
Dependent Claim
11 words
A process according to claim 5 wherein said compound is ethanol.
Claim 8
Dependent Claim
23 words
A process according to claim 5 wherein the amount of the compound of step (a) is 60 weight % to 90 weight %.
Claim 9
Dependent Claim
17 words
A process according to claim 2 wherein said cells are separated from said compound after step (a).
Claim 10
Dependent Claim
46 words
A process according to claim 6 wherein the temperature of each of steps (a) and (b) is from 20° C. to 30° C. and the aqueous solution of step (b) is selected from about 0.1 M potassium phosphate buffer and about 0.1 M sodium phosphate buffer.
Claim 11
Dependent Claim
15 words
A process according to claim 9 wherein said cells are of the species Kluyveromyces fragilis.
Claim 12
Dependent Claim
15 words
A process according to claim 9 wherein said cells are of the species Kluyveromyces lactis.
Claim 13
Dependent Claim
15 words
A process according to claim 9 wherein said cells are of the species Torula cremoris.
Claim 14
Dependent Claim
65 words
A process according to claim 1 comprising the additional step of heating an aqueous solution of said released lactase and from about 30 to 90 weight % glycerol, based on the total weight of said solution, at a temperature from about 35° C. to about 60° C. at a pH from about 5.5 to 8.0 until the protease activity in said solution is substantially reduced.
Claim 15
Dependent Claim
63 words
A process for selectively reducing protease activity from a protease-containing lactase preparation comprising heating an aqueous solution containing said preparation and from about 30 to 90 weight percent glycerol, based on the total weight of said solution, at a temperature from about 35° C. to about 60° C. at a pH from about 5.5 to 8 until said protease activity is substantially reduced.
Claim 16
Dependent Claim
63 words
A process for selectively reducing protease activity from a protease-containing lactase preparation comprising heating an aqueous solution containing said preparation and from about 30 to 90 weight percent glycerol, based on the total weight of said solution, at a temperature from about 35° C. to about 60° C. at a pH from about 5.5 to 8 until said protease activity is substantially reduced.
Claim 17
Dependent Claim
18 words
A process according to claim 14 wherein the temperature is from about 45° C. to about 55° C.
Claim 18
Dependent Claim
21 words
A process according to claim 14 wherein the glycerol concentration in said solution is from about 40 to 60 weight percent.
Claim 19
Dependent Claim
15 words
A process according to claim 14 wherein the pH is from about 6.4 to 7.0.
Claim 20
Dependent Claim
23 words
A process according to claim 14 wherein said solution contains manganous ion in a concentration from about 10 -4 to 10 -2 Molar.
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) | 1985-05-10 | 1985-11-09 | 1986-05-11 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1989-05-10 | 1989-11-09 | 1990-05-11 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1993-05-10 | 1993-11-09 | 1994-05-11 | $7700.00 | Window Closed / Lapsed |