EP1812547A1 - Maischverfahren und dazu geeignete enzymzusammensetzung - Google Patents
Maischverfahren und dazu geeignete enzymzusammensetzungInfo
- Publication number
- EP1812547A1 EP1812547A1 EP05746301A EP05746301A EP1812547A1 EP 1812547 A1 EP1812547 A1 EP 1812547A1 EP 05746301 A EP05746301 A EP 05746301A EP 05746301 A EP05746301 A EP 05746301A EP 1812547 A1 EP1812547 A1 EP 1812547A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- xylanase
- mash
- proceeding
- wort
- mashing
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims description 51
- 102000004190 Enzymes Human genes 0.000 title claims description 43
- 108090000790 Enzymes Proteins 0.000 title claims description 43
- 230000008569 process Effects 0.000 title claims description 39
- 238000005360 mashing Methods 0.000 title abstract description 35
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 claims abstract description 37
- 108010059892 Cellulase Proteins 0.000 claims abstract description 20
- 238000001914 filtration Methods 0.000 claims abstract description 16
- 241000499912 Trichoderma reesei Species 0.000 claims abstract description 12
- 229920002472 Starch Polymers 0.000 claims description 31
- 239000008107 starch Substances 0.000 claims description 31
- 235000019698 starch Nutrition 0.000 claims description 31
- 235000013405 beer Nutrition 0.000 claims description 24
- 230000000694 effects Effects 0.000 claims description 21
- 239000007864 aqueous solution Substances 0.000 claims description 13
- 239000000413 hydrolysate Substances 0.000 claims description 12
- 241000228215 Aspergillus aculeatus Species 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 229920001221 xylan Polymers 0.000 claims description 5
- 241000228257 Aspergillus sp. Species 0.000 claims description 4
- 241000233866 Fungi Species 0.000 claims description 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 3
- 150000004823 xylans Chemical class 0.000 claims description 2
- 108010013043 Acetylesterase Proteins 0.000 claims 1
- 102100036617 Monoacylglycerol lipase ABHD2 Human genes 0.000 claims 1
- 108010041969 feruloyl esterase Proteins 0.000 claims 1
- 238000013124 brewing process Methods 0.000 abstract description 6
- 241000228212 Aspergillus Species 0.000 abstract description 2
- 229940088598 enzyme Drugs 0.000 description 38
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 22
- 235000006085 Vigna mungo var mungo Nutrition 0.000 description 21
- 240000005616 Vigna mungo var. mungo Species 0.000 description 21
- 241000209219 Hordeum Species 0.000 description 16
- 235000007340 Hordeum vulgare Nutrition 0.000 description 16
- 229920002498 Beta-glucan Polymers 0.000 description 11
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 11
- 239000000758 substrate Substances 0.000 description 11
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 description 10
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 10
- 235000000346 sugar Nutrition 0.000 description 8
- 235000013339 cereals Nutrition 0.000 description 7
- 238000011084 recovery Methods 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 108090000637 alpha-Amylases Proteins 0.000 description 6
- 150000001720 carbohydrates Chemical class 0.000 description 6
- 235000014633 carbohydrates Nutrition 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 238000000926 separation method Methods 0.000 description 6
- UGXQOOQUZRUVSS-ZZXKWVIFSA-N [5-[3,5-dihydroxy-2-(1,3,4-trihydroxy-5-oxopentan-2-yl)oxyoxan-4-yl]oxy-3,4-dihydroxyoxolan-2-yl]methyl (e)-3-(4-hydroxyphenyl)prop-2-enoate Chemical compound OC1C(OC(CO)C(O)C(O)C=O)OCC(O)C1OC1C(O)C(O)C(COC(=O)\C=C\C=2C=CC(O)=CC=2)O1 UGXQOOQUZRUVSS-ZZXKWVIFSA-N 0.000 description 5
- 229920000617 arabinoxylan Polymers 0.000 description 5
- 229940106157 cellulase Drugs 0.000 description 5
- 238000004890 malting Methods 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000005096 rolling process Methods 0.000 description 5
- 150000008163 sugars Chemical class 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 230000015556 catabolic process Effects 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000006731 degradation reaction Methods 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 230000002538 fungal effect Effects 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000004458 spent grain Substances 0.000 description 4
- 108010001817 Endo-1,4-beta Xylanases Proteins 0.000 description 3
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 3
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 102000004139 alpha-Amylases Human genes 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 108010019077 beta-Amylase Proteins 0.000 description 3
- 230000001461 cytolytic effect Effects 0.000 description 3
- 230000002255 enzymatic effect Effects 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 235000015095 lager Nutrition 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 239000005017 polysaccharide Substances 0.000 description 3
- 150000004804 polysaccharides Chemical class 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
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- 235000015041 whisky Nutrition 0.000 description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 2
- 229920001353 Dextrin Polymers 0.000 description 2
- 239000004375 Dextrin Substances 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- 108010009736 Protein Hydrolysates Proteins 0.000 description 2
- 235000007238 Secale cereale Nutrition 0.000 description 2
- 240000006394 Sorghum bicolor Species 0.000 description 2
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 2
- 229910000831 Steel Inorganic materials 0.000 description 2
- 235000013334 alcoholic beverage Nutrition 0.000 description 2
- 235000015107 ale Nutrition 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 239000004464 cereal grain Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000000593 degrading effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 235000019425 dextrin Nutrition 0.000 description 2
- YERABYSOHUZTPQ-UHFFFAOYSA-P endo-1,4-beta-Xylanase Chemical group C=1C=CC=CC=1C[N+](CC)(CC)CCCNC(C(C=1)=O)=CC(=O)C=1NCCC[N+](CC)(CC)CC1=CC=CC=C1 YERABYSOHUZTPQ-UHFFFAOYSA-P 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
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- 238000002156 mixing Methods 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
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- 239000010959 steel Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- LUEWUZLMQUOBSB-FSKGGBMCSA-N (2s,3s,4s,5s,6r)-2-[(2r,3s,4r,5r,6s)-6-[(2r,3s,4r,5s,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5s,6r)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-4,5-dihydroxy-2-(hydroxymethyl)oxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](O[C@@H](OC3[C@H](O[C@@H](O)[C@@H](O)[C@H]3O)CO)[C@@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-FSKGGBMCSA-N 0.000 description 1
- DBTMGCOVALSLOR-UHFFFAOYSA-N 32-alpha-galactosyl-3-alpha-galactosyl-galactose Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(OC2C(C(CO)OC(O)C2O)O)OC(CO)C1O DBTMGCOVALSLOR-UHFFFAOYSA-N 0.000 description 1
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 241001513093 Aspergillus awamori Species 0.000 description 1
- 241000228245 Aspergillus niger Species 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000007319 Avena orientalis Nutrition 0.000 description 1
- 241000209763 Avena sativa Species 0.000 description 1
- 235000007558 Avena sp Nutrition 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000194103 Bacillus pumilus Species 0.000 description 1
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 1
- 101710104295 Beta-1,4-xylanase Proteins 0.000 description 1
- 108010084185 Cellulases Proteins 0.000 description 1
- 102000005575 Cellulases Human genes 0.000 description 1
- RXVWSYJTUUKTEA-UHFFFAOYSA-N D-maltotriose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(O)C(CO)O1 RXVWSYJTUUKTEA-UHFFFAOYSA-N 0.000 description 1
- -1 F. oxysporum Chemical compound 0.000 description 1
- 241000223218 Fusarium Species 0.000 description 1
- 241000223221 Fusarium oxysporum Species 0.000 description 1
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 description 1
- 102100022624 Glucoamylase Human genes 0.000 description 1
- 108050008938 Glucoamylases Proteins 0.000 description 1
- 229920002581 Glucomannan Polymers 0.000 description 1
- 108700023372 Glycosyltransferases Proteins 0.000 description 1
- 241000223198 Humicola Species 0.000 description 1
- 241001480714 Humicola insolens Species 0.000 description 1
- 241001373560 Humicola sp. Species 0.000 description 1
- 235000008694 Humulus lupulus Nutrition 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- 239000004368 Modified starch Substances 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 241001451060 Poitrasia Species 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 241000135252 Rhizomucor sp. Species 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 241000582914 Saccharomyces uvarum Species 0.000 description 1
- 241000209056 Secale Species 0.000 description 1
- 244000062793 Sorghum vulgare Species 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 241000223258 Thermomyces lanuginosus Species 0.000 description 1
- 235000009430 Thespesia populnea Nutrition 0.000 description 1
- 241000223259 Trichoderma Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 244000098338 Triticum aestivum Species 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
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- 230000004913 activation Effects 0.000 description 1
- 229940024171 alpha-amylase Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 150000004783 arabinoxylans Chemical class 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 235000019998 barley wine Nutrition 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
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- 235000020087 canadian whiskey Nutrition 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
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- 238000009795 derivation Methods 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 108010091371 endoglucanase 1 Proteins 0.000 description 1
- 108010091384 endoglucanase 2 Proteins 0.000 description 1
- 108010092450 endoglucanase Z Proteins 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 1
- 108010038658 exo-1,4-beta-D-xylosidase Proteins 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
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- 239000012530 fluid Substances 0.000 description 1
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- 239000011521 glass Substances 0.000 description 1
- 229940046240 glucomannan Drugs 0.000 description 1
- 229930182478 glucoside Natural products 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 102000045442 glycosyltransferase activity proteins Human genes 0.000 description 1
- 108700014210 glycosyltransferase activity proteins Proteins 0.000 description 1
- 239000010903 husk Substances 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 235000020088 irish whiskey Nutrition 0.000 description 1
- 235000021440 light beer Nutrition 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- FYGDTMLNYKFZSV-UHFFFAOYSA-N mannotriose Natural products OC1C(O)C(O)C(CO)OC1OC1C(CO)OC(OC2C(OC(O)C(O)C2O)CO)C(O)C1O FYGDTMLNYKFZSV-UHFFFAOYSA-N 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 235000019713 millet Nutrition 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000019426 modified starch Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 235000009048 phenolic acids Nutrition 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 235000020004 porter Nutrition 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- KUIXZSYWBHSYCN-UHFFFAOYSA-L remazol brilliant blue r Chemical compound [Na+].[Na+].C1=C(S([O-])(=O)=O)C(N)=C2C(=O)C3=CC=CC=C3C(=O)C2=C1NC1=CC=CC(S(=O)(=O)CCOS([O-])(=O)=O)=C1 KUIXZSYWBHSYCN-UHFFFAOYSA-L 0.000 description 1
- 238000011160 research Methods 0.000 description 1
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- 239000005720 sucrose Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 235000018553 tannin Nutrition 0.000 description 1
- 229920001864 tannin Polymers 0.000 description 1
- 239000001648 tannin Substances 0.000 description 1
- 125000000969 xylosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)CO1)* 0.000 description 1
- FYGDTMLNYKFZSV-BYLHFPJWSA-N β-1,4-galactotrioside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@H](CO)O[C@@H](O[C@@H]2[C@@H](O[C@@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-BYLHFPJWSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
- C12C5/00—Other raw materials for the preparation of beer
- C12C5/004—Enzymes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
- C12C7/00—Preparation of wort
- C12C7/04—Preparation or treatment of the mash
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12C—BEER; PREPARATION OF BEER BY FERMENTATION; PREPARATION OF MALT FOR MAKING BEER; PREPARATION OF HOPS FOR MAKING BEER
- C12C7/00—Preparation of wort
- C12C7/14—Lautering, i.e. clarifying wort
- C12C7/16—Lautering, i.e. clarifying wort by straining
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01004—Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01008—Endo-1,4-beta-xylanase (3.2.1.8)
Definitions
- the present invention relates, inter alia, to a mashing and filtration step in a process for the production of an alcoholic beverage, and to a composition useful in the mashing and filtration step in such a process.
- the invention provides in a first aspect a process for production of a mash having enhanced filterability and/or improved extract yield after filtration, which comprises; preparing a mash in the presence of enzyme activities and filtering the mash to obtain a wort, wherein the enzyme activities comprise endoglucanase activities derived from Trichoderma Reesei and a xylanase of GH family 10.
- the invention provides a composition
- a composition comprising; an endoglucanase derived from Trichoderma reesei; and, a xylanase GH 10 derived from Aspergillus sp., preferably from Aspergillus aculeatus (SEQ ID NO:1).
- the invention provides uses of a composition according to the second aspect in a process comprising reducing the viscosity of an aqueous solution comprising a starch hydrolysate, in a process comprising filtering of an aqueous solution comprising a starch hydrolysate, in a process wherein the aqueous solution comprising a starch hydrolysate is a mash for beer making and/or in a process wherein the aqueous solution comprising a starch hydrolysate is a feed composition.
- the final step is to dry the green malt in the kiln.
- the temperature regime in the kiln determines the colour of the barley malt and the amount of enzymes which survive for use in the mashing process.
- Low temperature kilning is more appropriate for malts when it is essential to preserve enzymatic activity.
- Malts kilned at high temperatures have very little or no enzyme activity but are very high in colouring such as caramelized sugars as well as in flavouring compounds.
- Mashing is the process of converting starch from the milled barley malt and solid adjuncts into fermentable and unfermentable sugars to produce wort of the desired composition.
- Traditional mashing involves mixing milled barley malt and adjuncts with water at a set temperature and volume to continue the biochemical changes initiated during the malting process.
- the mashing process is conducted over a period of time at various temperatures in order to activate the endogenous malt enzymes responsible for the degradation of proteins and carbohydrates.
- the principal enzymes responsible for starch conversion in a traditional mashing process are alpha- and beta-amylases.
- Alpha-amylase very rapidly reduces insoluble and soluble starch by splitting starch molecules into many shorter chains that can be attacked by beta-amylase.
- the disaccharide produced is maltose.
- step- infusion This mashing procedure involves a series of rests at various temperatures, each favouring one of the necessary endogenous enzyme activities.
- step- infusion To day the double-mash infusion system is the most widely used system for industrial production of beer, especially lager type beer. This system prepares two separate mashes. It utilizes a cereal cooker for boiling adjuncts and a mash tun for well-modified, highly enzymatically active malts.
- mashing may be performed in the presence of added enzyme compositions comprising the enzymes necessary for the hydrolysis of the grist starch.
- These enzymes may comprise alpha- amylases, pullulanases, beta-amylases and glucoamylases.
- the liquid extract the wort
- the solids spent grains i.e. the insoluble grain and husk material forming part of grist.
- Wort separation is important because the solids contain large amounts of non-starch polysaccharides, protein, poorly modified starch, fatty material, silicates, and polyphenols (tannins).
- Important non-starch polysaccharides present in cereal grains are beta-glucan and arabinoxylan.
- the endosperm cell wall of barley comprises 75% beta-glucan, 20% arabinoxylan, and 5% remaining protein with small amount of cellulose, glucomannan and phenolic acids.
- Long chains of barley arabinoxylans, and to a lesser degree beta-glucan, which have not been modified due to enzymatic hydrolysis may cause formation of gels when solubilised in water, these gels will strongly increase wort viscosity and reduce filterability.
- beta-glucan has been reduced to smaller oligomers, as unmodified beta-glucans later on will give rise to haze stability problems in the final beer.
- enzymatic compositions comprising endoglucanases and xylanases, such as Ultraflo® or Viscozyme®, are often used in the mashing step to improve wort separation.
- the objectives of wort separation include the following: • to obtain good extract recovery, • to obtain good filterability, and • to produce clear wort. Extraction recovery and filterability are important for the economy in the brewing process, while the wort clarity is a must in order to produce a beer which does not develop haze. Extraction recovery, filterability and wort clarity is greatly affected by the standard of the grist, e.g. the barley malt and the types of adjunct, as well as the applied mashing procedure.
- the wort may be fermented with brewers yeast to produce a beer.
- grist is understood as the starch or sugar containing material that's the basis for beer production, e.g. the barley malt and the adjunct.
- malt is understood as any malted cereal grain, in particular barley.
- adjunct is understood as the part of the grist which is not barley malt.
- the adjunct may be any carbohydrate rich material.
- mash is understood as a aqueous starch slurry, e.g. comprising crushed barley malt, crushed barley, and/or other adjunct or a combination hereof, steeped in water to make wort.
- wort is understood as the unfermented liquor run-off following extracting the grist during mashing.
- spent grains is understood as the drained solids remaining when the grist has been extracted and the wort separated from the mash.
- beer is here understood as fermented wort, e.g. an alcoholic beverage brewed from barley malt, optionally adjunct and hops.
- extract recovery in the wort is defined as the sum of soluble substances extracted from the grist (malt and adjuncts) expressed in percentage based on dry matter.
- a thermostable enzyme is understood as an enzyme that under the temperature regime and the incubation period applied in the processes of the present invention in the amounts added is capable of sufficient degradation of the substrate in question.
- homology when used about polypeptide or DNA sequences and referred to in this disclosure is understood as the degree of homology between two sequences indicating a derivation of the first sequence from the second.
- the homology may suitably be determined by means of computer programs known in the art such as GAP provided in the GCG program package (Program Manual for the Wisconsin Package, Version 8, August 1994, Genetics Computer Group, 575 Science Drive, Madison, Wisconsin, USA 53711 ) (Needleman, S.B. and Wunsch, CD., (1970), Journal of Molecular Biology, 48, 443-453.
- GAP GAP creation penalty of 3.0
- GAP extension penalty 0.1.
- DP is the degree of polymerisation, herein used for average number of glucose units in polymers in a polysaccharide hydrolysate.
- GH Glycoside Hydrolase Families
- Binding Modules applied in this disclosure follows the concept of Coutinho, P.M. & Henrissat, B. (1999) CAZy - Carbohydrate-Active Enzymes server at URL: http://afmb.cnrs- mrs.fr/ ⁇ cazy/CAZY/index.html or alternatively Coutinho, P.M. & Henrissat, B. 1999;
- Genetics, Biochemistry and Ecology of Cellulose Degradation K. Ohmiya, K. Hayashi, K. Sakka, Y. Kobayashi, S. Karita and T.
- composition according to the proceeding aspect may be used in a process comprising reducing the viscosity of an aqueous solution comprising a starch hydrolysate.
- the composition may even be used in a process comprising filtering of an aqueous solution comprising a starch hydrolysate.
- the aqueous solution comprising a starch hydrolysate is a mash for beer making, and in another preferred embodiment the aqueous solution comprising a starch hydrolysate is a feed composition.
- the process of the invention may be applied in the mashing of any grist.
- the grist may comprise any starch and/or sugar containing plant material derivable from any plant and plant part, including tubers, roots, stems, leaves and seeds.
- the grist comprises grain, such as grain from barley, wheat, rye, oat, corn, rice, milo, millet and sorghum, and more preferably, at least 10%, or more preferably at least 15%, even more preferably at least 25%, or most preferably at least 35%, such as at least 50%, at least 75%, at least 90% or even 100% (w/w) of the grist of the wort is derived from grain.
- the grist comprises malted grain, such as barley malt.
- At least 10%, or more preferably at least 15%, even more preferably at least 25%, or most preferably at least 35%, such as at least 50%, at least 75%, at least 90% or even 100% (w/w) of the grist of the wort is derived from malted grain.
- the mashing enzymes may be exogenously supplied.
- the enzymes mostly used as starch degrading enzymes include pullulanases, alpha- amylases and amyloglucosidases. The use of starch degrading enzymes in mashing is well- known to the skilled person.
- Adjunct comprising readily fermentable carbohydrates such as sugars or syrups may be added to the malt mash before, during or after the mashing process of the invention but is preferably added after the mashing process.
- a part of the adjunct may be treated with a protease and/or an endoglucanase, and/or heat treated before being added to the mash of the invention.
- starch extracted from the grist is gradually hydrolyzed into fermentable sugars and smaller dextrins.
- the mash is starch negative to iodine testing, before wort separation.
- the application of the appropriate xylanase and endoglucanase activities in the process of the present invention results in efficient reduction of beta-glucan and arabino- xylan level facilitating wort separation, thus ensuring reduced cycle time, high extract recovery and clear wort.
- the wort produced by the process of the first aspect of the invention may be fermented to produce a beer. Fermentation of the wort may include pitching the wort with a yeast slurry comprising fresh yeast, i.e. yeast not previously used for the invention or the yeast may be recycled yeast.
- the yeast applied may be any yeast suitable for beer brewing, especially yeasts selected from Saccharomyces spp. such as S. cerevisiae and S.
- the processes for fermentation of wort for production of beer are well known to the person skilled in the arts.
- the process of the invention may include adding silica hydrogel to the fermented wort to increase the colloidal stability of the beer.
- the processes may further include adding kieselguhr to the fermented wort and filtering to render the beer bright.
- the beer produced by fermenting the wort of the invention may be any type of beer, e.g. ale, strong ale, stout, porter, lager, pilsner, bitter, export beer, malt liquor, happoushu, Iambic, barley wine, high- alcohol beer, low-alcohol beer, low-calorie beer or light beer.
- the beer produced by the process of the invention may be distilled to recover ethanol, e.g. for whisky production.
- Contemplated are any kind of whisky (spelled "whiskey” in US and Ireland) include bourbon, Canadian whisky, Irish whiskey, rye, and scotch.
- xylanase For the present purposes a xylanase is an enzyme classified as EC 3.2.1.8. The official name is endo-1 ,4-beta-xylanase. The systematic name is 1 ,4-beta-D-xylan xylanohydrolase.
- endo-(1-4)-beta-xylanase (1-4)-beta- xylan 4-xylanohydrolase; endo-1, 4-xylanase; xylanase; beta-1 ,4-xylanase; endo-1 ,4- xylanase; endo-beta-1 ,4-xylanase; endo-1, 4-beta-D-xylanase; 1 ,4-beta-xylan xylanohydrolase; beta-xylanase; beta- 1 ,4-xy Ian xylanohydrolase; endo-1 ,4-beta-xylanase; beta-D-xylanase.
- the reaction catalysed is the endohydrolysis of 1 ,4-beta-D-xylosidic linkages in xylans.
- the xylanase to be used for the present invention may be of any origin including mammalian, plant or animal origin it is presently preferred that the xylanase is of microbial origin.
- the xylanase may be one derivable from a filamentous fungus or a yeast.
- Xylanases have been found in a number of fungal species, in particular species of Aspergillus, such as A. niger, A. awamori, A. aculeatus and A. oryzae, Trichoderma, such as T.
- xylanases have also been found in bacterial species, e.g. within the genus Bacillus, such as B. pumilus.
- the xylanase is derived from a filamentous fungus such as from Aspergillus sp., Bacillus sp., Humicola sp., Myceliophotora sp., Poitrasia sp.
- the xylanase to be used in the present invention is a Glycoside Hydrolase Family 10 (GH10) xylanase.
- GH10 Glycoside Hydrolase Family 10
- the xylanase to be used according to the process of the invention the xylanase from Aspergillus aculeatus shown as SEQ ID NO:1 (XYL II) in the present disclosure and/or described in WO9421785A1 and comprising the partial amino acid sequence shown as SEQ ID NO:5 therein.
- any xylanase having and or comprising a sequence having at least 50%, at least 60%, at least 70%, at least 80%, or even at least 90% homology to any of the aforementioned sequences.
- Endoglucanases For the present purposes an endoglucanase is an enzyme classified as EC 3.2.1.4.
- the endoglucanase activity to be used for the present invention may be any composition derived from Trichoderma reesei, preferably the composition is obtained by submerged fermentation, such as the composition Celluclast® available from Novozymes A S. Celluclast has a pronounced viscosity-reducing effect on soluble cellulosic substrates.
- the cellulase composition Laminex BG® a commercial cellulase preparation produced by Trichoderma reesei and available from Genencor International Inc.
- the xylanolytic activity can be expressed in FXU(S)-units, determined at pH 6.0 with
- Azo-Wheat arabinoxylan (arabinoxylan dyed with remazol brilliant blue, Megazyme) as substrate.
- a xylanase sample is incubated with the remazol-xylan substrate.
- the background of non-degraded dyed substrate is precipitated by ethanol.
- the remaining blue colour in the supernatant (as determined spectrophotometrically at 585 nm) is proportional to the xylanase activity, and the xylanase units are then determined relatively to an enzyme standard at standard reaction conditions, i.e.
- Endoclucanase activity The cellulytic activity may be measured in fungal endoglucanase units (FBG), determined on a 0.5% beta-glucan substrate at 30°C, pH 5.0 and reaction time 30 min.
- Fungal endoglucanase reacts with beta-glucan releases glucose or reducing carbohydrate which is determined as reducing sugar according to the Somogyi-Nelson method.
- 1 fungal endoglucanase unit (FBG) is the amount of enzyme which according to the above outlined standard conditions, releases glucose or reducing carbohydrate with a reduction capacity equivalent to 1 micromol glucose per minute.
- the cellulytic activity may be measured in endo-glucanase units (EGU), determined at pH 6.0 with carboxymethyl cellulose (CMC) as substrate.
- EGU endo-glucanase units
- a substrate solution is prepared, containing 34.0 g/l CMC (Hercules 7 LFD) in 0.1 M phosphate buffer at pH 6.0.
- the enzyme sample to be analyzed is dissolved in the same buffer. 5 ml substrate solution and 0.15 ml enzyme solution are mixed and transferred to a vibration viscosimeter (e.g. MIVI 3000 from Sofraser, France), thermostated at 40DC for 30 minutes.
- One EGU is defined as the amount of enzyme that reduces the viscosity to one half under these conditions.
- the amount of enzyme sample should be adjusted to provide 0.01-0.02 EGU/ml in the reaction mixture.
- the arch standard is defined as 880 EGU/g.
- Enzymes A Celluclast ® 1.5 L, a commercial preparation produced by Trichoderma reesei comprising cellulase activities 700 EGU/g and xylanase activities 200 FXU(S)/g.
- B composition of the invention comprising the cellulase and xylanase activities, respectively 700 EGU/g and 200 FXU(S)/g, from Trichoderma reesei and the GH10 xylanase from Aspergillus aculeatus shown in SEQ ID NO:1 250 FXU(S)/g.
- C Laminex BG®, a commercial preparation produced by Trichoderma reesei comprising cellulase activities 1370 EGU/g and xylanase activities.
- the mash was prepared according to EBC: 4.5.1 using malt grounded according to EBC: 1.1. Mashing trials were performed in 500 ml lidded vessels incubated in water bath with stirring and each containing a mash with 44 g malt and adjusted to a total weight of 300 ⁇ 0.2 g with water preheated to 50°C. The wort produced was app. 12% Plato.
- Mashing temperature profile Unless otherwise stated mashing was carried out using an initial incubation temperature at 50°C for 30 minutes, followed by an 17 min increasing step with 1°C/min to 67°C remaining here for 40 min. The profile is continued with an 8 min increasing step with
- E, - 100 b)E 2 100 - M
- E-, the extract content of sample
- E 2 the extract content of dry grist
- P the extract content in wort
- Plato M the moisture content of the grist
- in % (m/m) 800 the amount of destilled water added into the mash to 100 g of grist
- Viscosity Automated Microviscometer (AMVn) is based on the rolling ball principle.
- the sample to be measured is introduced into a glass capillary in which a steel ball rolls.
- the viscous properties of the test fluid can be determined by measuring the rolling time of the steel ball.
- the rolling time t 0 of a ball over a defined measuring distance in a capillary is measured.
- the dynamic viscosity ⁇ of the sample is calculated from the calibration constant K ⁇ ( ⁇ ) of the measuring system, the rolling time to and the difference of density ⁇ p between the ball and the sample.
- K ⁇ ( ⁇ ) of the measuring system K ⁇ ( ⁇ ) of the measuring system
- ⁇ 1 K ⁇ ⁇ a) • f Q • (p k - p s ),
- ⁇ Dynamic vis cos ity of the sample
- Example 1 A mash was prepared from a well modified malt (172 mg ⁇ -glucan/l). Four treatments according to table 1 were applied. The results are presented in table 2 The treatment B wherein the enzyme composition comprised a GH10 xylanase had a significantly better performance on viscosity reduction than blank, as well as than the treatments A and C consisting only of a T. reesei preparation. The differences are significant an at least the 99.9% level.
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PCT/DK2005/000367 WO2005118769A1 (en) | 2004-06-03 | 2005-06-02 | Mashing process and enzyme composition useful therein |
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GB0702844D0 (en) * | 2007-02-14 | 2007-03-28 | Leuven K U Res & Dev | Improving taste of beer |
WO2010118257A2 (en) * | 2009-04-08 | 2010-10-14 | Danisco Us Inc. | Endoglucanase for reducing the viscosity of a plant material slurry |
US9012186B2 (en) | 2009-04-27 | 2015-04-21 | The Board Of Trustees Of The University Of Illinois | Hemicellulose-degrading enzymes |
BRPI1012166B1 (pt) * | 2009-05-07 | 2020-10-27 | Dupont Nutrition Biosciences Aps | complexo de enzimas com atividade de beta 1,4-endoglucano hidrolase, seu uso e seu processo de produção |
MX2012003091A (es) | 2009-09-23 | 2012-04-30 | Danisco Us Inc | Enzimas de glicosil hidrolasa novedosas y usos de las mismas. |
CN102639697B (zh) | 2009-11-06 | 2015-03-25 | 诺维信股份有限公司 | 具有木聚糖酶活性的多肽和编码该多肽的多核苷酸 |
EP2516663B1 (de) | 2009-12-23 | 2020-02-12 | Danisco US Inc. | Verfahren zur verbesserung der effizienz von simultanen verzuckerungs- und gärungsreaktionen |
DK2686434T3 (da) | 2011-03-17 | 2021-10-11 | Danisco Us Inc | Fremgangsmåde til reduktion af viskositet i saccharificeringsproces |
HUE037237T2 (hu) | 2011-09-14 | 2018-08-28 | Dupont Nutrition Biosci Aps | Kompozíciók, amelyek tartalmaznak enzimeket endo-1,4-béta-xilanáz aktivitással és enzimeket endo-1,3(4)-béta glukanáz aktivitással |
CN104736702A (zh) | 2012-08-03 | 2015-06-24 | 杜邦营养生物科学有限公司 | 酶 |
RU2695461C2 (ru) * | 2013-09-05 | 2019-07-23 | Новозимс А/С | Способ снижения вязкости в способе пивоварения |
EP3017706A1 (de) | 2014-11-05 | 2016-05-11 | Dupont Nutrition Biosciences ApS | Enzyme zum Mälzen |
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KR20000010743A (ko) * | 1996-05-03 | 2000-02-25 | 한스 발터 라벤 | 향상된 여과성 및/또는 증대된 수율을 갖는 맥아즙을 제조하는방법 |
AU720139B2 (en) * | 1996-08-05 | 2000-05-25 | Syngenta Mogen Bv | Improved process for the production of alcoholic beverages using maltseed |
AU2002210409A1 (en) * | 2000-11-10 | 2002-05-21 | Novozymes A/S | Ethanol process |
EP1471799A2 (de) * | 2002-01-25 | 2004-11-03 | DSM IP Assets B.V. | Hitzestabile enzympräparate |
EP1570041A1 (de) * | 2002-12-05 | 2005-09-07 | Novozymes A/S | Maischeverfahren für die bierherstellung |
CN1788083B (zh) * | 2003-03-10 | 2011-10-05 | 诺维信公司 | 酒精产品生产方法 |
EP2258829A3 (de) * | 2003-12-19 | 2013-03-27 | Novozymes A/S | Maischeverfahren |
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