US2017314004A1PendingUtilityA1
Enzymes
Est. expirySep 14, 2031(~5.1 yrs left)· nominal 20-yr term from priority
C12C 12/002C12Y 302/01014C12P 7/06C12C 11/003A23L 5/25C12N 9/2408C12Y 302/01006C12N 9/24C12Y 302/01008D21H 17/005C12N 9/2482Y02E50/17C12N 9/244C12N 9/242C12H 1/14Y02E50/10
50
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Claims
Abstract
The present invention relates to new enzymes with improved properties and to compositions comprising these enzymes suitable for use in the production of a food, feed, or malt beverage product, such as in a brewing process.
Claims
exact text as granted — not AI-modified1 . An enzyme exhibiting endo-1,4-β-xylanase activity, which enzyme comprises an amino acid sequence having at least 80% identity with any one selected from SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:17, and SEQ ID NO:18, or any functional fragment thereof.
2 . The enzyme according to claim 1 , which enzyme has a ratio in activity on soluble arabinoxylan substrate (WE-AX) to insoluble arabinoxylan substrate (WU-AX) arabinoxylan substrate of less than about 7.0, such as less than about 6.5, such as less than about 6.0, such as less than about 5.5, such as less than about 5.0, such as less than about 4.5.
3 . An enzyme exhibiting endo-1,3(4)-β-glucanase activity, which enzyme comprises an amino acid sequence having at least 80% identity with any one selected from SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, or any functional fragment thereof.
4 . The enzyme according to claim 1 , which enzyme has a temperature optimum in the range of 40-70° C., such as in the range of 45-65° C., such as in the range of 50-65° C., such as in the range of 55-65° C.
5 . The enzyme according to claim 4 , wherein said enzyme has at least 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99% identity with any one amino acid sequence selected from SEQ ID NO: 1-18, or any functional fragment thereof.
6 . The enzyme according to claim 5 having a total number of amino acids of less than 350, such as less than 340, such as less than 330, such as less than 320, such as less than 310, such as less than 300 amino acids, such as in the range of 200 to 350, such as in the range of 220 to 345 amino acids.
7 . The enzyme according to claim 6 , wherein the amino acid sequence of said enzyme has at least one, two, three, four, five, six, seven, eight, nine or ten amino acid substitutions as compared to any one amino acid sequence selected from SEQ ID NO: 1-18, or any functional fragment thereof.
8 . The enzyme according to claim 7 , wherein the amino acid sequence of said enzyme has a maximum of one, two, three, four, five, six, seven, eight, nine or ten amino acid substitutions compared to any one amino acid sequence selected from SEQ ID NO: 1-18, or any functional fragment thereof.
9 . The enzyme according to claim 8 , which enzyme comprises the amino acid sequence identified by any one of SEQ ID NO: 1-18, or any functional fragment thereof.
10 . The enzyme according to claim 3 , which enzyme consists of the amino acid sequence identified by any one of SEQ ID NO: 1-18, or any functional fragment thereof.
11 . A DNA construct comprising a DNA sequence encoding an enzyme according to claim 1 .
12 . A recombinant expression vector comprising a DNA construct according to claim 11 .
13 . A cell that has been transformed with a DNA construct of claim 11 .
14 . A preparation comprising an enzyme according to claim 10 .
15 . A composition comprising an enzyme exhibiting endo-1,4-β-xylanase activity according to claim 1 in combination with any one or more β-glucanase.
16 . The composition according to claim 15 , wherein said one or more β-glucanase is an enzyme exhibiting endo-1,3(4)-β-glucanase activity according to any one of claims 3 - 10 .
17 . A composition comprising an enzyme exhibiting endo-1,3(4)-β-glucanase activity according to claim 3 in combination with any one or more xylanase.
18 . The composition according to claim 17 , wherein said one or more xylanase is an enzyme exhibiting endo-1,4-β-xylanase activity.
19 . The composition according to claim 18 , wherein said endo-1,3(4)-β-glucanase activity and said endo-1,4-β-xylanase activity are derived from at least two different enzymes, such as at least two different enzymes from two different species.
20 . The composition according to claim 19 , comprising a combination of at least two enzymes, said two enzymes, or two enzymes with an amino acid sequence having at least 80% sequence identity with the respective SEQ ID, or any functional fragment thereof, being selected from the list consisting of
SEQ ID NO:1 and SEQ ID NO:7; SEQ ID NO:2 and SEQ ID NO:7; SEQ ID NO:3 and SEQ ID NO:7; SEQ ID NO:4 and SEQ ID NO:7; SEQ ID NO:5 and SEQ ID NO:7; SEQ ID NO:6 and SEQ ID NO:7; SEQ ID NO:17 and SEQ ID NO:7; SEQ ID NO:18 and SEQ ID NO:7; SEQ ID NO:1 and SEQ ID NO:8; SEQ ID NO:2 and SEQ ID NO:8; SEQ ID NO:3 and SEQ ID NO:8; SEQ ID NO:4 and SEQ ID NO:8; SEQ ID NO:5 and SEQ ID NO:8; SEQ ID NO:6 and SEQ ID NO:8; SEQ ID NO:17 and SEQ ID NO:8; SEQ ID NO:18 and SEQ ID NO:8; SEQ ID NO:1 and SEQ ID NO:9; SEQ ID NO:2 and SEQ ID NO:9; SEQ ID NO:3 and SEQ ID NO:9; SEQ ID NO:4 and SEQ ID NO:9; SEQ ID NO:5 and SEQ ID NO:9; SEQ ID NO:6 and SEQ ID NO:9; SEQ ID NO:17 and SEQ ID NO:9; SEQ ID NO:18 and SEQ ID NO:9; SEQ ID NO:1 and SEQ ID NO:10; SEQ ID NO:2 and SEQ ID NO:10; SEQ ID NO:3 and SEQ ID NO:10; SEQ ID NO:4 and SEQ ID NO:10; SEQ ID NO:5 and SEQ ID NO:10; SEQ ID NO:6 and SEQ ID NO:10; SEQ ID NO:17 and SEQ ID NO:10; SEQ ID NO:18 and SEQ ID NO:10; SEQ ID NO:1 and SEQ ID NO:11; SEQ ID NO:2 and SEQ ID NO:11; SEQ ID NO:3 and SEQ ID NO:11; SEQ ID NO:4 and SEQ ID NO:11; SEQ ID NO:5 and SEQ ID NO:11; SEQ ID NO:6 and SEQ ID NO:11; SEQ ID NO:17 and SEQ ID NO:11; SEQ ID NO:18 and SEQ ID NO:11; SEQ ID NO:1 and SEQ ID NO:12; SEQ ID NO:2 and SEQ ID NO:12; SEQ ID NO:3 and SEQ ID NO:12; SEQ ID NO:4 and SEQ ID NO:12; SEQ ID NO:5 and SEQ ID NO:12; SEQ ID NO:6 and SEQ ID NO:12; SEQ ID NO:17 and SEQ ID NO:12; SEQ ID NO:18 and SEQ ID NO:12; SEQ ID NO:1 and SEQ ID NO:13; SEQ ID NO:2 and SEQ ID NO:13; SEQ ID NO:3 and SEQ ID NO:13; SEQ ID NO:4 and SEQ ID NO:13; SEQ ID NO:5 and SEQ ID NO:13; SEQ ID NO:6 and SEQ ID NO:13; SEQ ID NO:17 and SEQ ID NO:13; SEQ ID NO:18 and SEQ ID NO:13; SEQ ID NO:1 and SEQ ID NO:14; SEQ ID NO:2 and SEQ ID NO:14; SEQ ID NO:3 and SEQ ID NO:14; SEQ ID NO:4 and SEQ ID NO:14; SEQ ID NO:5 and SEQ ID NO:14; SEQ ID NO:6 and SEQ ID NO:14; SEQ ID NO:17 and SEQ ID NO:14; SEQ ID NO:18 and SEQ ID NO:14; SEQ ID NO:1 and SEQ ID NO:15; SEQ ID NO:2 and SEQ ID NO:15; SEQ ID NO:3 and SEQ ID NO:15; SEQ ID NO:4 and SEQ ID NO:15; SEQ ID NO:5 and SEQ ID NO:15; SEQ ID NO:6 and SEQ ID NO:15; SEQ ID NO:17 and SEQ ID NO:15; SEQ ID NO:18 and SEQ ID NO:15; SEQ ID NO:1 and SEQ ID NO:16; SEQ ID NO:2 and SEQ ID NO:16; SEQ ID NO:3 and SEQ ID NO:16; SEQ ID NO:4 and SEQ ID NO:16; SEQ ID NO:5 and SEQ ID NO:16; SEQ ID NO:6 and SEQ ID NO:16; SEQ ID NO:17 and SEQ ID NO:16; and SEQ ID NO:18 and SEQ ID NO:16.
21 . The composition according to claim 20 , wherein when used prior to the lautering in a brewing application the total pressure built up is reduced to a value of less than 470 mm WC, such as less than 450 mm WC, such as less than 430 mm WC, such as less than 410 mm WC, such as less than 390 mm WC, such as less than 370 mm WC, such as less than 350 mm WC, such as less than 330 mm WC, such as less than 310 mm WC, such as less than 300 mm WC, such as less than 290 mm WC.
22 . The composition according to claim 21 , wherein when used prior to the lautering in a brewing application the total pressure built up is reduced by at least 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93 or 95% compared to the use of a negative control without said composition.
23 . The composition according to claim 22 , wherein when used in a brewing application prior to the wort separation, the wort filterability as measured by volume wort collected after 5 min of filtration relative to a control without enzymes is increased to above 1.5, such as above 1.6, such as above 1.7, such as above 1.8, such as above 1.9, such as above 2.0, such as above 2.1, such as above 2.2, such as above 2.3, such as above 2.4, such as above 2.5.
24 . The composition according to claim 23 , when used in a brewing application prior to the wort separation, the wort filterability as measured by volume wort collected after 5 min of filtration is increased by at least 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, or 300% as compared to the use of a negative control without said composition.
25 . The composition according to claim 24 , comprising any one or more further enzyme.
26 . The composition according to claim 25 , wherein said one or more further enzyme is selected from list consisting of a xylanase classified in EC 3.2.1.32, EC 3.2.1.136, or EC 3.2.1.156, a cellulase, a laminarinase, an endo-1,5-α-L-arabinanase, a beta-D-glucoside glucohydrolase, a β-Xylosidase, a cellobiohydrolase, a glucan 1,4-beta-glucosidase, a xyloglucan-specific exo-beta-1,4-glucanase and an α-N-Arabinofuranosidase.
27 . Use of an enzyme according to claim 1 in the production of a food, feed, or malt beverage product.
28 . Use of an enzyme according to claim 1 in the production of dough or baked products.
29 . Use of an enzyme according to claim 1 in the preparation of pulp or paper.
30 . Use of an enzyme according to claim 1 for the preparation of cereal components.
31 . The use according to claim 30 , in which the cereal is rye, wheat, or barley.
32 . Use of an enzyme according to claim 1 in the production of beer or modification of by-products from a brewing process.
33 . Use of an enzyme according to claim 1 in the production of wine or juice.
34 . Use of an enzyme according to claim 1 in the production of a first- or second-generation biofuel, such as bioethanol.
35 . Method of altering filterability of a starch comprising material, said method comprising the step of treating said starch comprising material with enzyme according to claim 1 .
36 . Method of reducing pressure built up during lautering in a brewing application, said method comprising the step of treating a brewing mash with enzyme according to claim 1 .
37 . Method for the production of a food, feed, or beverage product, such as an alcoholic or non-alcoholic beverage, such as a cereal- or malt-based beverage like beer or whiskey, said method comprising the step of treating a starch comprising material with enzyme according to claim 1 .
38 . Method for the production of a brewing mash, said method comprising the step of treating a starch comprising material with enzyme according to claim 1 .
39 . Method for the production of a first- or second-generation biofuel, such as bioethanol, said method comprising the step of treating a starch comprising material with an enzyme according to claim 1 .
40 . Product obtained by the method according to claim 39 .
41 . A composition comprising the product according to claim 40 , such as wherein the product is in a range of 0.1%-99.9%.Join the waitlist — get patent alerts
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