CN105623937A - Saccharifying method - Google Patents
Saccharifying method Download PDFInfo
- Publication number
- CN105623937A CN105623937A CN201610097716.7A CN201610097716A CN105623937A CN 105623937 A CN105623937 A CN 105623937A CN 201610097716 A CN201610097716 A CN 201610097716A CN 105623937 A CN105623937 A CN 105623937A
- Authority
- CN
- China
- Prior art keywords
- beer
- amylopectase
- aforementioned
- beerwort
- amylase
- 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.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 78
- 235000013405 beer Nutrition 0.000 claims abstract description 64
- 102000004190 Enzymes Human genes 0.000 claims description 49
- 108090000790 Enzymes Proteins 0.000 claims description 49
- 229940088598 enzyme Drugs 0.000 claims description 49
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 38
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 claims description 33
- 102100022624 Glucoamylase Human genes 0.000 claims description 32
- 239000004382 Amylase Substances 0.000 claims description 26
- 240000008042 Zea mays Species 0.000 claims description 26
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 26
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 26
- 235000005822 corn Nutrition 0.000 claims description 26
- 239000002773 nucleotide Substances 0.000 claims description 23
- 125000003729 nucleotide group Chemical group 0.000 claims description 21
- 235000013339 cereals Nutrition 0.000 claims description 20
- 240000005979 Hordeum vulgare Species 0.000 claims description 18
- 235000007340 Hordeum vulgare Nutrition 0.000 claims description 18
- 239000000203 mixture Substances 0.000 claims description 18
- 244000082988 Secale cereale Species 0.000 claims description 16
- 235000007238 Secale cereale Nutrition 0.000 claims description 16
- 210000000582 semen Anatomy 0.000 claims description 16
- 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 claims description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- 108010028688 Isoamylase Proteins 0.000 claims description 12
- 108091005804 Peptidases Proteins 0.000 claims description 12
- 239000004365 Protease Substances 0.000 claims description 12
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 12
- 241000228245 Aspergillus niger Species 0.000 claims description 11
- 108010059892 Cellulase Proteins 0.000 claims description 11
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 11
- 240000006394 Sorghum bicolor Species 0.000 claims description 10
- 238000009396 hybridization Methods 0.000 claims description 10
- 241000894006 Bacteria Species 0.000 claims description 9
- 240000007594 Oryza sativa Species 0.000 claims description 9
- 235000007164 Oryza sativa Nutrition 0.000 claims description 9
- 239000004373 Pullulan Substances 0.000 claims description 9
- 229920001218 Pullulan Polymers 0.000 claims description 9
- 235000007230 Sorghum bicolor Nutrition 0.000 claims description 9
- 235000019423 pullulan Nutrition 0.000 claims description 9
- 235000009566 rice Nutrition 0.000 claims description 9
- 210000000544 articulatio talocruralis Anatomy 0.000 claims description 8
- 235000007199 Panicum miliaceum Nutrition 0.000 claims description 7
- 239000002253 acid Substances 0.000 claims description 7
- 244000022185 broomcorn panic Species 0.000 claims description 7
- 229940106157 cellulase Drugs 0.000 claims description 7
- 230000000295 complement effect Effects 0.000 claims description 7
- 150000007523 nucleic acids Chemical group 0.000 claims description 7
- 235000013334 alcoholic beverage Nutrition 0.000 claims description 6
- 230000035784 germination Effects 0.000 claims description 6
- 241000193755 Bacillus cereus Species 0.000 claims description 5
- 235000021440 light beer Nutrition 0.000 claims description 5
- 238000003860 storage Methods 0.000 claims description 4
- 241001134780 Bacillus acidopullulyticus Species 0.000 claims description 3
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 claims description 3
- 241000959173 Rasamsonia emersonii Species 0.000 claims description 3
- 108090000637 alpha-Amylases Proteins 0.000 abstract description 16
- 230000008569 process Effects 0.000 abstract description 7
- 238000005360 mashing Methods 0.000 abstract description 2
- 239000000523 sample Substances 0.000 description 28
- 230000000694 effects Effects 0.000 description 24
- 229920002472 Starch Polymers 0.000 description 21
- 235000019698 starch Nutrition 0.000 description 21
- 239000008107 starch Substances 0.000 description 21
- 235000000346 sugar Nutrition 0.000 description 16
- 239000002671 adjuvant Substances 0.000 description 13
- 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 12
- 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 12
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 12
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 12
- 238000000855 fermentation Methods 0.000 description 11
- 230000004151 fermentation Effects 0.000 description 11
- 239000000758 substrate Substances 0.000 description 11
- 229920000945 Amylopectin Polymers 0.000 description 10
- 102000004139 alpha-Amylases Human genes 0.000 description 10
- 229940024171 alpha-amylase Drugs 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 229920001353 Dextrin Polymers 0.000 description 9
- 239000004375 Dextrin Substances 0.000 description 9
- 235000001014 amino acid Nutrition 0.000 description 9
- 150000001413 amino acids Chemical class 0.000 description 9
- 235000019425 dextrin Nutrition 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 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 7
- 239000008103 glucose Substances 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 150000008163 sugars Chemical class 0.000 description 7
- 108010019077 beta-Amylase Proteins 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 239000007979 citrate buffer Substances 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 241000193830 Bacillus <bacterium> Species 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 230000002538 fungal effect Effects 0.000 description 5
- 241000228212 Aspergillus Species 0.000 description 4
- IMQLKJBTEOYOSI-UHFFFAOYSA-N Diphosphoinositol tetrakisphosphate Chemical compound OP(O)(=O)OC1C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C1OP(O)(O)=O IMQLKJBTEOYOSI-UHFFFAOYSA-N 0.000 description 4
- 241000233866 Fungi Species 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 230000002255 enzymatic effect Effects 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 239000002002 slurry Substances 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000008399 tap water Substances 0.000 description 4
- 235000020679 tap water Nutrition 0.000 description 4
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 3
- 101710146708 Acid alpha-amylase Proteins 0.000 description 3
- 240000006439 Aspergillus oryzae Species 0.000 description 3
- 101710130006 Beta-glucanase Proteins 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 241001480714 Humicola insolens Species 0.000 description 3
- 240000003183 Manihot esculenta Species 0.000 description 3
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 238000006460 hydrolysis reaction Methods 0.000 description 3
- 229910052740 iodine Inorganic materials 0.000 description 3
- 239000011630 iodine Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000004890 malting Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 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 2
- 102000013142 Amylases Human genes 0.000 description 2
- 108010065511 Amylases Proteins 0.000 description 2
- 241001513093 Aspergillus awamori Species 0.000 description 2
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 2
- 241000193744 Bacillus amyloliquefaciens Species 0.000 description 2
- 241000194108 Bacillus licheniformis Species 0.000 description 2
- 241000193403 Clostridium Species 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 241000628997 Flos Species 0.000 description 2
- 241000223218 Fusarium Species 0.000 description 2
- 241000223198 Humicola Species 0.000 description 2
- 241000582914 Saccharomyces uvarum Species 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000228341 Talaromyces Species 0.000 description 2
- 241000193447 Thermoanaerobacter thermohydrosulfuricus Species 0.000 description 2
- 241000223259 Trichoderma Species 0.000 description 2
- 239000008186 active pharmaceutical agent Substances 0.000 description 2
- 235000019418 amylase Nutrition 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- GDEBSAWXIHEMNF-UHFFFAOYSA-O cupferron Chemical compound [NH4+].O=NN([O-])C1=CC=CC=C1 GDEBSAWXIHEMNF-UHFFFAOYSA-O 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000007599 discharging Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000006260 foam Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000002386 leaching Methods 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 229920001592 potato starch Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000017854 proteolysis Effects 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- 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 1
- 229910019626 (NH4)6Mo7O24 Inorganic materials 0.000 description 1
- ZIIUUSVHCHPIQD-UHFFFAOYSA-N 2,4,6-trimethyl-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide Chemical compound CC1=CC(C)=CC(C)=C1S(=O)(=O)NC1=CC=CC(C(F)(F)F)=C1 ZIIUUSVHCHPIQD-UHFFFAOYSA-N 0.000 description 1
- 108010011619 6-Phytase Proteins 0.000 description 1
- 108091005508 Acid proteases Proteins 0.000 description 1
- 229920000856 Amylose Polymers 0.000 description 1
- 101000757144 Aspergillus niger Glucoamylase Proteins 0.000 description 1
- 241000972773 Aulopiformes Species 0.000 description 1
- 241000680658 Bacillus deramificans Species 0.000 description 1
- 241000219310 Beta vulgaris subsp. vulgaris Species 0.000 description 1
- 229920002498 Beta-glucan Polymers 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 241001509321 Clostridium thermoamylolyticum Species 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 101100381997 Danio rerio tbc1d32 gene Proteins 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 241000605909 Fusobacterium Species 0.000 description 1
- 241000193385 Geobacillus stearothermophilus Species 0.000 description 1
- 229920001503 Glucan Polymers 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 108010029541 Laccase Proteins 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 101710117655 Maltogenic alpha-amylase Proteins 0.000 description 1
- 101100381999 Mus musculus Tbc1d32 gene Proteins 0.000 description 1
- 108020005187 Oligonucleotide Probes Proteins 0.000 description 1
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 108010064785 Phospholipases Proteins 0.000 description 1
- 102000015439 Phospholipases Human genes 0.000 description 1
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 241000124033 Salix Species 0.000 description 1
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 1
- 235000021536 Sugar beet Nutrition 0.000 description 1
- 241001484137 Talaromyces leycettanus Species 0.000 description 1
- 241001136490 Thermomyces dupontii Species 0.000 description 1
- 235000009430 Thespesia populnea Nutrition 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 235000015107 ale Nutrition 0.000 description 1
- 210000003423 ankle Anatomy 0.000 description 1
- 230000008901 benefit Effects 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
- 235000013361 beverage Nutrition 0.000 description 1
- 235000015496 breakfast cereal Nutrition 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000004464 cereal grain Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000013256 coordination polymer Substances 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000002478 diastatic effect Effects 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- IHZDYHDJAVUIBH-UHFFFAOYSA-L disodium hydrogenarsenate Chemical compound [Na+].[Na+].O[As]([O-])([O-])=O IHZDYHDJAVUIBH-UHFFFAOYSA-L 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- -1 for instance Proteins 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000010903 husk Substances 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 235000021430 malt vinegar Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000005497 microtitration Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000002751 oligonucleotide probe Substances 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 229940085127 phytase Drugs 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
- VZOPRCCTKLAGPN-ZFJVMAEJSA-L potassium;sodium;(2r,3r)-2,3-dihydroxybutanedioate;tetrahydrate Chemical compound O.O.O.O.[Na+].[K+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O VZOPRCCTKLAGPN-ZFJVMAEJSA-L 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000000197 pyrolysis Methods 0.000 description 1
- 239000012925 reference material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 235000019515 salmon Nutrition 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 229940074446 sodium potassium tartrate tetrahydrate Drugs 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 230000003019 stabilising effect Effects 0.000 description 1
- 239000013589 supplement Substances 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
- 239000005418 vegetable material Substances 0.000 description 1
- 238000005406 washing Methods 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
- C12C7/00—Preparation of wort
- C12C7/04—Preparation or treatment of the mash
- C12C7/047—Preparation or treatment of the mash part of the mash being unmalted cereal mash
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2451—Glucanases acting on alpha-1,6-glucosidic bonds
- C12N9/2457—Pullulanase (3.2.1.41)
-
- 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)
- C12Y302/01041—Pullulanase (3.2.1.41)
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
本发明涉及糖化方法，具体地提供用于产生啤酒用麦芽汁的方法，其中包括从谷物形成醪，和将所述醪与支链淀粉酶接触。The present invention relates to a mashing process and in particular provides a process for producing beer wort comprising forming mash from grain and contacting said mash with pullulanase.
Description
The present patent application is based on the applying date and is December in 2007 12, and application number is the divisional application that 200780101920.0 (international application no is PCT/US2007/087209), name are called the application for a patent for invention of " method for saccharifying ".
Quoting sequence table
The present invention comprises the sequence table of computer-reader form. By reference computer-reader form is expressly incorporated herein.
Invention field
The present invention relates to for producing medicated beer beerwort and for producing the method for saccharifying of the improvement of medicated beer.
Background of invention
In modern method for saccharifying, when the diastatic Fructus Hordei Germinatus amount in enzyme is relatively low or allows to use all auxiliary corn (adjunctgrist), often enzyme agent as a supplement is added. Also in the saccharifying carrying out the fully Fructus Hordei Germinatus of modification with high-load enzyme, enzyme is used, to increase the amount of extraction recovery and fermentable sugars. Therefore known application debranching enzyme, for instance, isoamylase or amylopectase, to increase the yield of fermentable sugars. Application debranching enzyme in the method for low-heat beer can be produced being used for. This method is the theme of (MBAATechnicalQuarterly, 1977,14:105), U.S. Patent No. 4,528,198,4,666,718 and 4,318,927 and GB2056484 and the GB2069527 such as Willow.
Summary of the invention
The present inventor it has now surprisingly been found that by use certain amylopectase, it is possible to use lesser amount of pheron and realize saccharifying.
Therefore, in first aspect, the method that the present invention is provided to produce medicated beer beerwort, wine with dregs is formed including by corn, and described wine with dregs is contacted with amylopectase (E.C.3.2.1.41), wherein said amylopectase has aminoacid, aminoacid sequence at least 50% shown in described aminoacid sequence a) and SEQIDNO:4 is identical, or b) by under low stringent condition with the nucleic acid sequence encoding of following hybridization: i) encode the complementary strand of the nucleotide sequence of aminoacid sequence shown in SEQIDNO:4, or ii) subsequence of at least 100 nucleotide of (i).
In second aspect, the present invention provides according to the beerwort produced by the method for first aspect.
In the third aspect, beerwort that is that the present invention provides the concentration produced by the method for first aspect and/or that dry.
In fourth aspect, the present invention provides the medicated beer produced by the beerwort of second and third aspect.
In the 5th, the present invention provides the compositions suitable in the method for first aspect, described compositions includes amylopectase (E.C.3.2.1.41), glucoamylase and optional ��-amylase, wherein amylopectase has aminoacid sequence, aminoacid sequence at least 50% shown in described aminoacid sequence a) and SEQIDNO:4 is identical, or b) by under low stringent condition with the nucleic acid sequence encoding of following hybridization: i) encode the complementary strand of the nucleotide sequence of aminoacid sequence shown in SEQIDNO:4, or ii) subsequence of at least 100 nucleotide of (i).
In particular it relates to every as follows:
1. the method being used for producing medicated beer beerwort (Brewer ' swort), forms wine with dregs including from corn, contacts with amylopectase with by described wine with dregs, and wherein said amylopectase has following aminoacid sequence:
A) identical with aminoacid sequence at least 86% shown in SEQIDNO:4; Or
B) by nucleic acid sequence encoding, described nucleotide sequence under low stringent condition with following hybridization:
I) complementary strand of the nucleotide sequence of aminoacid sequence shown in coding SEQIDNO:4; Or
Ii) subsequence of at least 100 nucleotide of (i).
2. the method according to item 1, wherein said amylopectase is derived from addicted to acid Pullulan bacillus cereus (Bacillusacidopullulyticus).
3. the method according to item 1 or 2, it farther includes to contact described wine with dregs with glucoamylase and/or ��-amylase.
4. the method any one of aforementioned item, wherein glucoamylase and/or ��-amylase are derived from aspergillus niger (Aspergillusniger) or Ai Mosen ankle joint bacterium (Talaromycesemersonii).
5. the method any one of aforementioned item, it farther includes to contact described wine with dregs with the enzyme selected from lower group: cellulase, isoamylase, xylanase and protease.
6. the method any one of aforementioned item, wherein said corn includes the grain germinateing and/or not germinateing.
7. the method any one of aforementioned item, the grain wherein not germinateed and/or the grain of germination are selected from following: Fructus Hordei Vulgaris, Semen Tritici aestivi, rye (Secale cereale L.), Sorghum vulgare Pers., broomcorn millet, Semen Maydis and rice.
8. the method any one of aforementioned item, the grain wherein germinateed includes the germination grain being selected from the Fructus Hordei Vulgaris, Semen Tritici aestivi, rye (Secale cereale L.), Sorghum vulgare Pers., broomcorn millet, Semen Maydis and the rice that germinate.
9. the method any one of aforementioned item, wherein said beerwort is concentration and/or dries.
10. the method any one of aforementioned item, it also includes fermenting wort to obtain alcoholic beverage.
11. the method according to item 10, wherein said alcoholic beverage is medicated beer.
12. the method according to item 11, wherein said medicated beer is to like youngster's medicated beer, strong love youngster's medicated beer, Bitter, stou, alms bowl that dark beer, old storage medicated beer, export-oriented medicated beer, malt liquor, barley broo, low malt medicated beer, high alcohol beer, low alcohol beer, low-heat beer or light beer thoroughly.
13. the beerwort produced by the method any one of aforementioned item.
14. the concentration any one of aforementioned item and/or dry beerwort.
15. the medicated beer produced by the beerwort any one of aforementioned item.
16. the compositions suitable in the method any one of aforementioned item, described compositions includes amylopectase, glucoamylase and optional ��-amylase, and wherein said amylopectase has following aminoacid sequence, described aminoacid sequence
A) identical with aminoacid sequence at least 50% shown in SEQIDNO:4; Or
B) by under low stringent condition with the nucleic acid sequence encoding of following hybridization:
I) coding SEQIDNO:4 shown in aminoacid sequence nucleotide sequence complementary strand or
Ii) subsequence of at least 100 nucleotide of (i).
17. the compositions according to item 16, wherein glucoamylase and/or ��-amylase are derived from aspergillus niger or Ai Mosen ankle joint bacterium.
Detailed Description Of The Invention
Brewing method is it is well known in the art that and generally include malting (malting), saccharifying (mashing) and the step fermented. Saccharifying is that autoclase barley malt becomes to ferment with the Starch Conversion of solid adjuvant material and not fermentable sugar in the future, thus the method producing there is the beerwort of desirable composition. Traditional saccharifying includes broken barley malt and adjuvant and water at design temperature with to set volume mixture, so that the biochemical change started in malting technique proceeds. Method for saccharifying carries out one period in various temperature, to activate responsible protein degradation matter and the endogenous enzymes of sugar. Up to now, the most important change brought in saccharifying is that starch molecule is changed into fermentable sugars. The main enzyme being responsible for Starch Conversion in traditional method for saccharifying is ��-and beta amylase. ��-amylase by resolving into the shorter chain and the soluble and solvable starch that quickly reduces that much can be subject to beta amylase attack by starch molecule. The disaccharide produced is maltose. Except the maltose formed in saccharifying, also produce short branched glucose oligomer. Short branched glucose oligomer is not fermentable sugar, it is possible to makes finished beer taste better, and increases heat.
After saccharifying, when all starch all decompose, it is necessary to separate liquid extract (beerwort) from solid (distiller grains). The separation of beerwort and filter critically important, because solid comprises substantial amounts of protein, (poorlymodified) starch of basic unmodified, fatty material, silicate and polyphenol (tannin). After distiller grains separation of wort, it is possible to ferment distiller grains to produce medicated beer with brewer's yeast.
Details about conventional brewing method can at ResearchandTeachingInstituteofBrewing, " TechnologyBrewingandMalting " of the WolfgangKunze of Berlin (VLB), second edition revised edition in 1999, finds in ISBN3-921690-39-0.
The short branched glucose oligomer formed in saccharifying can be hydrolyzed further by adding exogenous enzyme (enzyme added except Fructus Hordei Germinatus). Debranching enzyme such as amylopectase (pullulanase) and isoamylase (isoamylase) are hydrolyzed the side chain ��-1-6 glycosidic bond in these oligomers, thus discharging glucose or maltose and straight chain oligomer, it is subject to endogenous enzymes (being derived from Fructus Hordei Germinatus) and/or exogenous enzyme, such as, the effect of ��-amylase, beta amylase and glucoamylase.
The present invention provides and is suitable to produce the new method of the not beerwort that fermentable sugars content is low. The method uses regioselective Pullulanase activity.
Definition
In content of this disclosure, employ multiple terms that those of ordinary skill in the art are generally understood that. But, several terms employ specific implication, and implication is limited as follows.
As it is used herein, term " corn (grist) " is interpreted as the material containing starch or sugar, it is the basis of Beer Production, for instance, barley malt and adjuvant. Generally, corn does not comprise the water of any interpolation.
Term " Fructus Hordei Germinatus " is interpreted as cereal grains (maltedcerealgrain), particularly Fructus Hordei Vulgaris of any germination.
It is interpreted as in corn by term " adjuvant " not being the part of barley malt. Adjuvant can include the vegetable material of any rich in starch, and e.g., the corn not germinateed, such as Fructus Hordei Vulgaris, rice, Semen Maydis, Semen Tritici aestivi, rye (Secale cereale L.), Sorghum vulgare Pers. and the sugar and/or the syrup that are prone to fermentation.
Being interpreted as wrapping amyloid slurry by term " wine with dregs (mash) ", this slurry comprises the corn being dipped in water.
Term " beerwort " is interpreted as in saccharifying and extracts the unfermentable liquid efflunent (liquorrun-off) after corn.
Term " distiller grains " is interpreted as (drained) solid drained remaining after extracting corn and separation of wort.
Term " medicated beer " is understood herein to the beerwort after fermentation, i.e. from barley malt, the pick-me-up (alcoholicbeverage) that optional adjuvant and Flos lupuli (Flos Humuli Lupuli) are brewageed.
Term " homologous sequence " is used for being characterized as below sequence, it has and known array at least 70%, preferably at least 75%, or at least 80%, or at least 85%, or 90%, or at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or even at least 100% identical aminoacid sequence. the relevant portion of the aminoacid sequence determined for homology is mature polypeptide, i.e. without signal peptide. term " homologous sequence " is also used for being characterized in the DNA sequence of low stringency, middle stringency, medium/high stringency, high stringency, or even very high stringency and known array hybridization. for determining low, in or the suitable experiment condition of intermolecular hybrid of high stringency nucleotide probe and homologous dna or RNA sequence include: by comprise DNA fragmentation or RNA filter membrane preimpregnation with in 5xSSC (sodium chloride/sodium citrate, Sambrook etc., 1989) hybridization 10 minutes in, and at 5xSSC, 5xDenhardt solution (Sambrook etc., 1989), by filter membrane prehybridization (Sambrook etc. in the solution of 0.5%SDS and 100 microgram/ml degeneration supersound process salmon sperm DNA, 1989), then random (Feinberg and the Vogelstein caused that concentration is 10ng/ml is being comprised, 1983, Anal.Biochem.132:6-13), hybridize 12 hours at about 45 DEG C with in the same solution of (specific activity > 1x109cpm/ microgram) probe of 32P-dCTP labelling. then by filter membrane at 2xSSC, in 0.5%SDS, at about 55 DEG C (low stringencies), more preferably from about 60 DEG C (middle stringency), more preferably from about 65 DEG C (medium/high stringency), even more preferably about 70 DEG C (high stringency), and even more preferably about 75 DEG C (very high stringencies) wash twice totally 30 minutes. use x-ray sheet detection under these conditions with the molecule of oligonucleotide probe hybridization.
By the disclosure is interpreted as the homogeneity degree between two sequences about polypeptide or term " homogeneity " that DNA sequence used and related to, represent the difference (derivation) between first sequence and second sequence. Homogeneity can be passed through computer program known in the art and suitably determine, GAP (ProgramManualfortheWisconsinPackage, the Version8 provided in GCG program package, August1994, GeneticsComputerGroup, 575ScienceDrive, Madison, Wisconsin, USA53711) (Needleman, and Wunsch S.B., C.D., (1970), JournalofMolecularBiology, 48,443-453). Peptide sequence is compared, uses following setting: gap opening penalty (GAPcreationpenalty) be 3.0 and gap extension penalty (GAPextensionpenalty) be 0.1. Homogeneity degree between aminoacid sequence and the different aminoacid sequences (" exogenous array ") of the present invention is calculated as below: the number mated completely in two sequence alignments, divided by the shortest person in the length of " invention sequence " or the length of " exogenous array ". Result is expressed as percentage identities.
Beerwort produces
According to first aspect, the method that the present invention is provided to produce medicated beer beerwort, wine with dregs is formed including by corn, and described wine with dregs is contacted with amylopectase (E.C.3.2.1.41), wherein said amylopectase has following aminoacid sequence, aminoacid sequence at least 50% shown in described aminoacid sequence a) and SEQIDNO:4, at least 60%, at least 70%, at least 75%, at least 80%, or at least 85%, or 90%, or at least 95%, at least 96%, at least 97%, at least 98% or even at least 99% is identical or b) by nucleic acid sequence encoding, described nucleotide sequence is at low stringency, middle stringency, medium/high stringency, high stringency, or under even very high stringent conditions with following hybridization: i) encode the complementary strand of the nucleotide sequence of aminoacid sequence shown in SEQIDNO:4, or ii) at least 100 nucleotide of (i), at least 200 nucleotide, at least 300 nucleotide, at least 500 nucleotide, at least 1000 nucleotide, or the subsequence of even at least 1500 nucleotide. in a preferred embodiment, amylopectase has aminoacid sequence, the aminoacid sequence of described aminoacid sequence and SEQIDNO:3 is more or less the same in 100 aminoacid, it is preferably no more than 80 aminoacid, it is more preferably no more than 50 aminoacid, it is more preferably no more than 30 aminoacid, even more preferably from no more than 20 aminoacid, and most preferably no more than 10 aminoacid.
The corn of first aspect includes amyloid germinated ceral and/or adjuvant. Corn can preferably include 0%-100%, it is preferable that 20%-100%, it is preferable that 30%-100%, more preferably 40%-100%, even more preferably from 50%-100%, more preferably 60%-100%, such as the adjuvant of 80%-100% or even most preferably 90%-100%, non-germinated ceral and/or non-germinating barley. In a specific embodiment, adjuvant is made up of 100% non-germinating barley. Additionally, corn preferably comprises 0%-100%, it is preferable that 20%-100%, preferred 30%-100%, more preferably 40%-100%, even more preferably from 50%-100%, more preferably 60%-100%, or most preferably 70%-100%, or even most preferably 90%-100% germinated ceral and/or non-germinating barley. In a specific embodiment, corn comprises about 50% germinated ceral, for instance, germinating barley, and about 50% adjuvant, for instance non-germinated ceral, such as non-germinating barley.
The germinated ceral used in the method for first aspect includes the corn of any germination, and is preferably selected from the germinated ceral of the Fructus Hordei Vulgaris, Semen Tritici aestivi, rye (Secale cereale L.), Sorghum vulgare Pers., broomcorn millet, Semen Maydis and the rice that germinate, and most preferably germinating barley.
The adjuvant used in the method for first aspect can obtain from tuber, root, stem, leaf, beans, frumentum and/or complete grain. Adjuvant can comprise raw and/or purified starch and/or sugary material, it is derived from plant, such as Semen Tritici aestivi, rye (Secale cereale L.), Herba bromi japonici, Semen Maydis, rice, buys sieve Sorghum vulgare Pers. (milo), broomcorn millet, Sorghum vulgare Pers., Rhizoma Solani tuber osi, Rhizoma Dioscoreae esculentae, Maninot esculenta crantz. (cassava), tapioca (tapioca), sago (sago), Fructus Musae, sugar beet and/or Caulis Sacchari sinensis. Preferably, accessory package is containing non-germinated ceral, for instance, the non-germinated ceral selected from following: Fructus Hordei Vulgaris, Semen Tritici aestivi, rye (Secale cereale L.), Sorghum vulgare Pers., broomcorn millet, Semen Maydis and rice, and most preferably non-germinating barley. Before the method for saccharifying of the present invention, in process or afterwards, it is possible to add in barley malt wine with dregs and comprise the carbohydrate that can be easy to fermentation such as the adjuvant of sugar or syrup, but add preferably in after method for saccharifying.
According to the present invention, amylopectase (E.C.3.2.1.41) enzymatic activity is provided by external source and is present in wine with dregs. Can formed before wine with dregs, in process or rear in wine with dregs composition such as water and/or corn, add amylopectase. In particularly preferred embodiments, ��-amylase (E.C.3.2.1.1) and/or glucoamylase (E.C.3.2.1.3) add together with amylopectase, and are present in wine with dregs.
In another preferred embodiment, adding other enzyme in wine with dregs, described enzyme is selected from lower group: isoamylase, protease, laccase, xylanase, lipase, phospholipase, phytase, phytin (phytin) and esterase.
In method for saccharifying, the starch extracted from corn is hydrolyzed into fermentable sugared and less dextrin gradually. Preferably, before extracting beerwort, iodine test is starch negative by wine with dregs.
The temperature of method for saccharifying is generally stepped up under control, and in wherein often walking, the effect of a kind of enzyme surpasses other enzyme, final protein degradation matter, cell wall and starch. Saccharification temperature curve is generally known in the art. In the present invention, saccharifying (starch degradation) step in method for saccharifying is preferably in 60 DEG C-66 DEG C, more preferably in 61 DEG C-65 DEG C, even more preferably in 62 DEG C-64 DEG C, and carries out most preferably at 63 DEG C-64 DEG C. In one embodiment of the invention, saccharification temperature is 64 DEG C.
Obtaining beerwort from wine with dregs and generally include the beerwort elimination (strain) distiller grains, namely described distiller grains form insoluble grain and the rice husk material of the part of corn. Hot water can be gone out the extract of any remnants by distiller grains with flushing or drip washing (sparge) from corn. Application heat-stable cellulase causes that beta glucan level effectively declines to promote wort filtration in the method for the invention, so that it is guaranteed that circulation time reduces and high extraction recovery. Preferably, extraction recovery is at least 80%, it is preferable that at least 81%, more preferably at 82%, and even more desirably at least 83%, such as at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, and most preferably at least 91%.
After the corn distiller grains separation of wort of any of above embodiment of first aspect, it is possible to be used as is beerwort or by its dehydration to provide concentration and/or dry beerwort. Concentration and/or dry beerwort can serve as wine brewing extract, and Fructus Hordei Germinatus extract flavoring agent (flavoring), for nonalcoholic Fructus Hordei Germinatus beverage, malt vinegar, breakfast cereals, for confection etc.
In a preferred embodiment, fermenting wort is to produce alcoholic beverage, preferably beer, such as, like youngster's medicated beer (ale), strong love youngster's medicated beer (strongale), Bitter (bitter), stou (stout), alms bowl that dark beer (porter) thoroughly, old storage medicated beer (lager), export-oriented medicated beer (exportbeer), malt liquor (maltliquor), barley broo, low malt medicated beer (happoushu), high alcohol beer (high-alcoholbeer), low alcohol beer (low-alcoholbeer), low-heat beer (low-caloriebeer) or light beer (lightbeer). the fermentation of beerwort can include input (pitchwith) yeast slurry in beerwort, and this slurry comprises fresh yeast (being namely previously not used for the yeast of the present invention) or described yeast can be the yeast reclaimed. the yeast of application can be adapted for any yeast of brewing, it is especially selected from the yeast of Saccharomyces sp (Saccharomycesspp.), such as saccharomyces cerevisiae and Saccharomyces uvarum (S.uvarum), it includes the natural or artificially generated variant of these organisms. it is known by those skilled in the art for producing the fermentation process of the beerwort of medicated beer.
The method of the present invention can include adding silica hydrogel (silicahydrogel) in fermenting wort, to increase the colloidal stability (colloidalstability) of medicated beer. These methods can also include adding kieselguhr (kieselguhr) in the beerwort of fermentation and filtering so that medicated beer vivid (bright).
According to an aspect of the present invention, it is provided that the medicated beer produced by the beerwort of second or third aspect, the medicated beer as produced by beerwort fermentation is produced medicated beer. Medicated beer can be any type beer, such as, youngster's medicated beer, strong love youngster's medicated beer, stou, alms bowl that dark beer, old storage medicated beer, Bitter, export-oriented medicated beer, malt liquor, low malt medicated beer, high alcohol beer, low alcohol beer, low-heat beer or light beer thoroughly are liked.
Enzyme
To be applied to the exogenous enzyme in the present invention should according to they treatment temperatures in the methods of the invention and their pH in wine with dregs when keep the ability of fully activity to select, and should add with effective dose. Enzyme can be derived from any source, it is preferable that from plant or algae, more preferably from microorganism, as from antibacterial or fungus.
Amylopectase (E.C.3.2.1.41)
The preferred amylopectase used in the method and/or compositions of the present invention is the amylose enzyme with following aminoacid sequence, described aminoacid sequence and the sequence at least 50% shown in SEQIDNO:4, such as at least 55%, such as at least 60%, such as at least 65%, such as at least 66%, such as at least 70%, such as at least 75%, such as at least 80%, such as at least 85%, such as at least 86%, such as at least 87%, such as at least 88%, such as at least 89%, such as at least 90%, such as at least 95%, as at least 98% or even 100% is identical; Particularly when using following setting with Needle program comparison: matrix is BLOSUM62, and breach initiates point penalty 10.0, gap extension penalty 0.5, non-notch homogeneity matrix.
Most preferably, amylopectase is derived from addicted to acid Pullulan bacillus cereus (Bacillusacidopullulyticus). Amylopectase can have by Kelly etc., aminoacid sequence or homologous sequence disclosed in 1994 (FEMSMicrobiol.Letters115:97-106) (SEQIDNO:6).
Isoamylase (E.C.3.2.1.68)
The another kind of enzyme used in the method for the present invention and/or compositions can be interchangeable debranching enzyme, such as isoamylase (E.C.3.2.1.68). ��-1 in Isoamylase hydrolyses amylopectin (amylopectin) and ��-limit dextrin, 6-D-glucosides branch key, and can not attack amylopectin by isoamylase and be different from amylopectase by the limited action to ��-limit dextrin. Isoamylase can add by effective dose well known to those skilled in the art. Isoamylase can be individually added into or add together with amylopectase.
��-amylase (EC3.2.1.1)
The specific ��-amylase used in the method for the present invention and/or compositions can be bacillus (Bacillus) ��-amylase. Known Bacillus alpha-amylase includes the ��-amylase being derived from the bacterial strain of Bacillus licheniformis (B.licheniformis), bacillus amyloliquefaciens (Bacillusamyloliquefaciens) and bacstearothermophilus (B.stearothermophilus). In the context of the present invention, it is desirable to Bacillus alpha-amylase be that WO99/19467 page 3 the 18th walks to ��-amylase defined in the 6th page of the 27th row. Preferred ��-amylase has aminoacid sequence, and it has at least 90% homogeneity with SEQIDNO:4 (disclosed herein for SEQIDNO:7) in WO99/19467, such as at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or particularly at least 99%. Most preferred product maltogenic alpha-amylase enzyme is SEQIDNO:9 or includes its variant disclosed in WO99/43794. Desired variant and heterozygote are described in WO96/23874, WO97/41213 and WO99/19467. The ��-amylase (E.C.3.2.1.1) originating from bacstearothermophilus being especially desired to, it has in WO99/19467 the aminoacid sequence (disclosed herein for SEQIDNO:10) disclosed in SEQIDNO:3, and it has sudden change: I181*+G182*+N193F.
Bacillus alpha-amylase can with 1.0-1000NU/kgDS, it is preferable that 2.0-500NU/kgDS, it is preferable that the amount of 10-200NU/kgDS adds.
The another kind of specific ��-amylase that the method for the present invention uses can be any fungal alpha-amylase. Such as, it is derived from the ��-amylase of aspergillus (Aspergillus) interior strain, and is preferred from the bacterial strain of aspergillus niger (Aspergillusniger). What be especially desired to is show high homogeneity with the aminoacid sequence (disclosed herein for SEQIDNO:11) shown in SEQIDNO:1 in WO2002/038787, namely at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, the fungal alpha-amylase of at least 80%, at least 85% or even at least 90% homogeneity. Fungal alpha-amylase can with 1-1000AFAU/kgDS, it is preferable that 2-500AFAU/kgDS, it is preferable that the amount of 20-100AFAU/kgDS adds.
Glucoamylase (E.C.3.2.1.3)
The another kind of specific enzyme used in the method for the present invention and/or compositions can be derived from the glucoamylase (E.C.3.2.1.3) of microorganism or plant. Preferably one or more the glucoamylase in lower group of fungus or bacterial origin: Aspergillus glucoamylase, particularly aspergillus niger G1 or G2 glucoamylase (Boel etc., 1984, EMBOJ.3 (5): 1097-1102), or their variant, as disclosed in WO92/00381 and WO00/04136; Aspergillus awamori (A.awamori) glucoamylase (WO84/02921), aspergillus oryzae (A.oryzae) (Agric.Biol.Chem., 1991,55 (4): 941-949), or its variant or fragment.
Other desired glucoamylase includes Talaromyces (Talaromyces) glucoamylase, particularly it is derived from Ai Mosen ankle joint bacterium (Talaromycesemersonii) (WO99/28448), Talaromycesleycettanus (U.S. Patent number Re.32,153), Du Pont's ankle joint bacterium (Talaromycesduponti) and thermophilic ankle joint bacterium (Talaromycesthermophilus) (U.S. Patent number 4,587,215) glucoamylase. Preferred glucoamylase includes the glucoamylase being derived from aspergillus oryzae, the aminoacid sequence as shown in the SEQIDNO:2 in WO00/04136 has at least 90%, at least 92%, at least 95%, at least 96%, the glucoamylase of at least 97%, at least 98% or particularly at least 99% or even at least 90% homogeneity. Other preferred glucoamylase includes the glucoamylase being derived from Ai Mosen ankle joint bacterium, aminoacid sequence (WO99/28448) as saved bacterium with Ai Mosen ankle has at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, or particularly at least 99%, or the glucoamylase of even at least 90% homogeneity.
Desired antibacterial glucoamylase includes the glucoamylase from fusobacterium (Clostridium), particularly pyrolysis clostridium amylobacter (C.thermoamylolyticum) (EP135,138) and Clostridium thermohydrosulfuricum (C.thermohydrosulfuricum) (WO86/01831).
It is also contemplated that commercial product AMG200L; AMG300L; SANTMSUPER and AMGTME (from Novozymes); OPTIDEXTM300 (from GenencorInt.); AMIGASETMAnd AMIGASETMPLUS (from DSM); G-ZYMETMG900 (from EnzymeBio-Systems); G-ZYMETMG990ZR (aspergillus niger glucoamylase and low protease content). Glucoamylase can be added with effective dose well-known to those skilled in the art.
Protease
Suitable protease includes microbial protease, such as fungus and bacterialprotease. Preferred protease is acid protease, i.e. by the protease of the capability representation at the hydrolyzed under acidic conditions protein lower than pH7.
Protease is responsible in wine with dregs the high-molecular-weight protein of total length is degraded into low molecular weight protein (LMWP). Low molecular weight protein (LMWP) is required for the nutrition of yeast, and high-molecular-weight protein ensures foam stability. Therefore technical staff knows protease and should add with aequum, provides slowly (amble) free amino acid and retain enough high-molecular-weight proteins so that foam stabilising for yeast simultaneously. Can with 0.1-1000AU/kgDS, it is preferable that the amount of 1-100AU/kgDS and most preferably 5-25AU/kgDS adds protease.
Cellulase (E.C.3.2.1.4)
Cellulase can be microbe-derived, as be derived from filamentous fungi (such as, aspergillus, trichoderma (Trichoderma), Humicola (Humicola), Fusarium (Fusarium)) bacterial strain. The instantiation of cellulase includes from Humicola insolens (H.insolens) obtainable endoglucanase (endoglucanase), it is also defined by the aminoacid sequence of Figure 14 in WO91/17244, and the endoglucanase of 43kD Humicola insolens described in WO91/17243.
The special fiber element enzyme used in the method for the present invention can be endoglucanase, such as inscribe-Isosorbide-5-Nitrae-1,4 beta-glucanase. What be especially desired to is the 1,4 beta-glucanase (disclosed herein for SEQIDNO:14) shown in SEQIDNO:2 and homologous sequence in WO2003/062409. Operable commercially available cellulase preparation includesWith(can obtain from NovozymesA/S), LAMINEXTMWithCP (can obtain from GenencorInt.) and7069W (can be fromGermany obtains).
Can with 1.0-10000BGU/kgDS, it is preferable that 10-5000BGU/kgDS, it is preferable that the amount of 50-1000BGU/kgDS and most preferably 100-500BGU/kgDS adds 1,4 beta-glucanase.
Materials and methods
Enzyme:
Amylopectase 1, it is derived from addicted to acid Pullulan bacillus cereus, and has sequence shown in SEQIDNO:1. Amylopectase 1 can be obtained as Promozyme400L by Novozymes.
Amylopectase 2, it is derived from Bacillusderamificans (U.S. Patent No. 5,736,375), and has sequence shown in SEQIDNO:2. Amylopectase 2 can be obtained as PromozymeD2 by Novozymes.
Amylopectase 3, it is derived from addicted to acid Pullulan bacillus cereus, and has sequence shown in SEQIDNO:4.
Acid Fungal Alpha-amylase, it is derived from aspergillus niger, and has sequence shown in SEQIDNO:11.
Glucoamylase G1, it is derived from aspergillus niger (boel etc. see above).
Method:
Alpha-amylase activity (NU)
Potato starch is used to may determine that alpha-amylase activity as substrate. This method is based on the decomposition by enzyme of modified (modified) potato starch, and follows the tracks of this reaction by being mixed with iodine solution by the sample of starch/enzymatic solution. Originally, form black-and-blue (blackishbluecolor), but in amylolysis process, blueness is more and more shallow, and gradually become bronzing (reddish-brown), by it compared with colored glass standard.
1000 Novo ��-amylase unit (KNU) are equal to 1000NU. One KNU is defined as at the standard conditions (that is, 37 DEG C of +/-0.05; 0.0003MCa2+; And pH5.6) enzyme amount of 5.26g starch dry matter (MerckAmylumsolubile) that degrade.
Acid alpha-amylase activity (AFAU)
Acid alpha-amylase activity can measure with AFAU (Acid Fungal Alpha-amylase unit), and it is determined relative to enzyme reference material. 1FAU is defined as the enzyme amount of 5.260mg starch dry matter of degrading per hour under following standard conditions.
Acid alpha-amylase is inscribe-��-amylase (Isosorbide-5-Nitrae-��-D-glucosan-glucan hydrolase, E.C.3.2.1.1), the ��-Isosorbide-5-Nitrae-glycosidic bond in catalytic starch intramolecule region, forms dextrin and the oligosaccharide of different chain length. Iodine is directly proportional to starch concentration to the blue intensity of Starch formation. Use reverse colorimetry, measure when particular analysis the minimizing of starch concentration as amylase activity.
Standard conditions/reaction condition:
Glucoamylase activity (AGU)
Novo glucoamylase unit (AGU) is defined as the enzyme amount at 37 DEG C He pH4.3 hydrolysis 1 per minute micromole maltose.
Use from BoehringerMannheim, the GlucoseGOD-Perid test kit of 124036, determine activity by the method revised according to (AEL-SM-0131 can require to obtain from Novozymes) with AGU/ml. Standard substance: AMG-standard substance, lot number 7-1195,195AGU/ml. By 375 �� L substrates (50mM sodium acetate, 1% maltose in pH4.3) 37 DEG C of incubations 5 minutes. Add the 25 �� L enzymes being diluted in sodium acetate. After 10 minutes, by adding 100 �� L0.25MNaOH and stopped reaction. 20 �� L are transferred to 96 hole microtitration plates and add 200 �� LGOD-Perid solution (124036, BoehringerMannheim). After 30 minutes, at the absorbance at room temperature measuring 650nm place, and by AMG-standard substance, with AGU/ml calculated activity. The detailed description of analysis method (AEL-SM-0131) can require to obtain from Novozymes.
Pullulanase activity (PUN):
One Pullulanase Unit (PUN) is defined as in the citrate buffer of pH5, can from the enzyme amount of pullulan substrate formation 1 per minute micromoles glucose at 50 DEG C.
Amylopectase sample is incubation together with substrate (red amylopectin). ��-1 in inscribe amylopectin enzyme hydrolysis redness amylopectin, 6-glycosidic bond, discharge red substrate degradation product. Ethanol is used to be precipitated by undegradable substrate. Discharging the amount of color at 510nm spectrophotometry, it is proportional to the inscribe-Pullulanase activity in sample. Compared with the color that the sample that the color of sample is formed with Pullulanase activity is known produces is formed.
Amylopectase is amylopectin 6-glucosan-hydrolytic enzyme, and enzyme classification number is E.C.3.2.1.41.
Reaction condition
Reagent/substrate
Klorvess Liquid 0.5M
Red pullulan substrate 2%. Supplier is Megazyme, Australia
Citrate buffer 0.05MpH5.0
Citrate buffer 0.05MpH5.0 including 25mM cysteine
Ethanol 99.8%
The amylopectase standard substance prepared product of 904PUN/g is diluted in citrate buffer 0.05M the standard dilutions series of 0.05-0.20PUN/ml.
Blank citrate buffer 0.05M
Enzyme sample is diluted in citrate buffer 0.05M activity for 0.06-0.20PUN/ml, and with standard dilutions series of comparisons.
Embodiment 1
In this embodiment, different amylopectases is analyzed to reduce the ability of the not amount of fermentable sugars (dextrin/DP4/4+) in beerwort.
Using following saccharification temperature curve by 100% malt amylase fully modified, described saccharification temperature curve includes 46 DEG C 26 minutes, then increases to 64 DEG C with 1 DEG C/min, makes temperature remained constant afterwards. Sample is collected at 98,128 and 158 minutes.
Enzyme is added at 0 minute. In all process, glucoamylase and ��-amylase is added respectively with 1000AGU/kgDS and 250AFAU/kgDS. Amylopectase is added according to table 1.
Sample boiled 10 minutes and filter (0.20 micron of aperture). Analyze sample by HPLC, and calculate the percentage ratio of not fermentable sugars (DP4/4+).
Use the data in table 1, by returning, calculate with the 2.74mg pheron/kg same effect of amylopectase 3 needed for amylopectase 1 and the enzyme dosage (see table 2) of amylopectase 2.
By these results it can be seen that amylopectase 3 is maximally effective enzyme. Therefore, amylopectase 3 pheron of the amount needs reducing not fermentable sugars (dextrin/DP4/4+) is less, thereby increases the fermentation (attenuation) of beerwort.
Embodiment 2
Analyze pH curve and the temperature curve of different amylopectase in the present embodiment.
Carry out the research of pH and temperature curve according to alive analysis of relative enzyme by following condition.
Analyze Method And Principle:
By ��-1 in amylopectase hydrolyzing amylopectin, 6-glycosidic bond, and the reducing sugar amount increased by the Somogyi-Nelson method detection revised.
In this experiment, evaluating activity with relative activity, wherein the strongest sample of activity is appointed as 100%. Testing conditions is as follows:
Buffer: citrate 0.1M+0.2M phosphate (regulates, be pH5 in temperature curve) in pH curve
Substrate: 0.2% amylopectin Sigma (p-4516)
Temperature: be 60 DEG C in pH curve, regulates in temperature curve
Response time: 30 minutes
According to Nelson, 1944, the principle detection described in J.Biol.Chem.153:375-380 and Somogyi, 1945, J.Biol.Chem.160:61-68 is by the reducing sugar of amylopectin enzyme r e lease. In short, by add SomogyiShi copper (cobber) reagent corresponding to the volume of sample volume (such as, the sample of 2ml adds 2ml) and to terminate hydrolysis. Sample is boiled 20 minutes, and cool down before color reaction. Reacted corresponding to the Nelson reagent (such as, adding 2ml during sample+Somogyi cupferron 4ml altogether) of sample 1/2 volume by addition. Sample mix 2 minutes, is subsequently adding and the water of Nelson reagent equivalent. In the dark incubated samples 30 minutes, and measure at 540nm in spectrophotometer.
Reagent can be made by:
Somogyi cupferron
At 1000mlH2O dissolves 70.2gNa2HPO4x2H2O and 80.0gKNAC4H4O6x4H2O (kaliumsodiumtartrat (sodium potassium tartrate tetrahydrate)) (slightly heating). Add 60gNaOH; 16.0gCuSO4x5H2O and 360.0gNa2SO4And add water to 2000ml. PH to 10.8 is adjusted with NaOH
Nelson reagent:
At 1200mlH2O dissolves 100.0g (NH4)6Mo7O24x7H2O. It is carefully added into 84.0mlH2SO4. Additionally, at 100mlH2O dissolves 12.00gNa2HAsO4x7H2O (disodium hydrogen arsenate), and this solution is slowly added in the first solution and adds water to 2000ml.
Table 3 below and 4 gives pH and the temperature curve of three kinds of amylopectases.
These are it is shown that compare with other two kinds of amylopectases, and amylopectase 3 has wide pH curve and activity when high temperature. These character make amylopectase 3 become enzyme very sane in wine brewing (saccharifying condition), especially for when saccharification temperature is 62 DEG C-65 DEG C.
Embodiment 3
Carry out leaching saccharifying (infusionmashing) test with amylopectase 1 and 3 (SEQIDNO:1 and 4). 6 saccharifying samples are prepared for substrate (corn) with 100% Fructus Hordei Vulgaris.
Fructus Hordei Vulgaris (DS%:86.73) is ground, and for each sample, by the 50.0g sample (total DS43.34g) the 200g tap water with 50 DEG C and 3.0ml1MH3PO4Mixing.
All samples all adds the identical enzymatic mixture of pullulan free enzyme.
Then amylopectase is added according to following table to sample 1-6:
Enzyme dose active/g
Then test sample in the automatization's sacchariferous equipment running following program.
| Time (minute) | Temperature DEG C |
| 0-30 | 50 |
| 30-44 | Rise to 64 |
| 44-104 | 64 |
| 104-120 | Rise to 80 |
| 120-140 | 80 |
| 140-155 | It is down to 20 |
After saccharifying, all samples all adds tap water and to 300g altogether and filters. Then the sample of filtration is boiled 10 minutes and dilute with deionized water 1:1. By centrifugal for 50ml subsample column criterion density analysis of going forward side by side, calculate RDF% (true horizon of fermentation). Result provides as RDF% with the % beerwort represented sugar (DP2 and DP4+).
| PUN/g | Amylopectase 1 | Amylopectase 3 |
| 0 | 0 | 0 |
| 0.1 | 64 | 64.6 |
| 0.2 | 65 | 66.1 |
| 0.5 | 67.3 | 69.5 |
It is shown that for obtaining high %RDF, amylopectase 3 is substantially better than amylopectase 1, and amylopectase 3 can produce the maltose level (DP2 level) even better than amylopectase 1.
Embodiment 4
Carrying out leaching saccharifying test with amylopectase 3 (SEQIDNO:4), the maltose to evaluate amylopectase 3 produces character. 6 saccharifying samples are prepared for substrate (corn) with 100% Fructus Hordei Vulgaris.
Fructus Hordei Vulgaris (DS%:86.73) is ground, and for each sample, by the 50.0g sample (total DS43.34g) the 200g tap water with 50 DEG C and 5%Na2SO3And 1MH3PO4Mixing.
All samples all adds the identical enzymatic mixture of pullulan free enzyme.
Then amylopectase 3 is added according to following table to sample 1-6:
Then following program test sample in automatization's sacchariferous equipment is run.
After saccharifying, in all samples, add tap water to total amount be 300g and filter. Then filtered sample is boiled 10 minutes, and dilute with deionized water 1:1. 50ml subsample is centrifuged and is analyzed. Result is as follows:
| PUN/g | % glucose | % maltose | % dextrin | RDF% |
| 0 | 3.8 | 47.5 | 34 | 61.2 |
| 0.1 | 3.8 | 48.2 | 32 | 63.2 |
| 0.3 | 3.8 | 49.8 | 28.8 | 65.7 |
| 0.5 | 3.7 | 51.2 | 26.4 | 68.6 |
| 1 | 3.7 | 52.6 | 23.9 | 71 |
| 2 | 3.6 | 55.6 | 20.1 | 74.3 |
Result shows that maltose concentration increases along with the dosage of amylopectase 3 and increases, and the after fermentation of the increase of maltose % (RDF%) increases. Dextrin part (HPLC analyzes DP4/4+) reduces at the same time.
Only barley beta-amylase can produce maltose in this reaction, and amylopectase 3 can promote the effect of beta amylase of Fructus Hordei Vulgaris.
Concentration of glucose is low, and not by the impact of amylopectase 3 effect, advantage when this is to ferment the beerwort produced. By barley beta-amylase, and being increased by the fermentation (RDF%) of beerwort, amylopectase 3 helps degraded dextrin, and promotes the formation of maltose.
Claims (17)
1. the method being used for producing medicated beer beerwort (Brewer ' swort), forms wine with dregs including from corn, contacts with amylopectase with by described wine with dregs, and wherein said amylopectase has following aminoacid sequence:
A) identical with aminoacid sequence at least 86% shown in SEQIDNO:4; Or
B) by nucleic acid sequence encoding, described nucleotide sequence under low stringent condition with following hybridization:
I) complementary strand of the nucleotide sequence of aminoacid sequence shown in coding SEQIDNO:4; Or
Ii) subsequence of at least 100 nucleotide of (i).
2. method according to claim 1, wherein said amylopectase is derived from addicted to acid Pullulan bacillus cereus (Bacillusacidopullulyticus).
3. the method according to claim 1 or 2, it farther includes to contact described wine with dregs with glucoamylase and/or ��-amylase.
4. the method any one of aforementioned claim, wherein glucoamylase and/or ��-amylase are derived from aspergillus niger (Aspergillusniger) or Ai Mosen ankle joint bacterium (Talaromycesemersonii).
5. the method any one of aforementioned claim, it farther includes to contact described wine with dregs with the enzyme selected from lower group: cellulase, isoamylase, xylanase and protease.
6. the method any one of aforementioned claim, wherein said corn includes the grain germinateing and/or not germinateing.
7. the method any one of aforementioned claim, the grain wherein not germinateed and/or the grain of germination are selected from following: Fructus Hordei Vulgaris, Semen Tritici aestivi, rye (Secale cereale L.), Sorghum vulgare Pers., broomcorn millet, Semen Maydis and rice.
8. the method any one of aforementioned claim, the grain wherein germinateed includes the germination grain being selected from the Fructus Hordei Vulgaris, Semen Tritici aestivi, rye (Secale cereale L.), Sorghum vulgare Pers., broomcorn millet, Semen Maydis and the rice that germinate.
9. the method any one of aforementioned claim, wherein said beerwort is concentration and/or dries.
10. the method any one of aforementioned claim, it also includes fermenting wort to obtain alcoholic beverage.
11. method according to claim 10, wherein said alcoholic beverage is medicated beer.
12. method according to claim 11, wherein said medicated beer is to like youngster's medicated beer, strong love youngster's medicated beer, Bitter, stou, alms bowl that dark beer, old storage medicated beer, export-oriented medicated beer, malt liquor, barley broo, low malt medicated beer, high alcohol beer, low alcohol beer, low-heat beer or light beer thoroughly.
13. the beerwort produced by the method any one of aforementioned claim.
14. the concentration any one of aforementioned claim and/or dry beerwort.
15. the medicated beer produced by the beerwort any one of aforementioned claim.
16. the compositions suitable in the method any one of aforementioned claim, described compositions includes amylopectase, glucoamylase and optional ��-amylase, and wherein said amylopectase has following aminoacid sequence, described aminoacid sequence
A) identical with aminoacid sequence at least 50% shown in SEQIDNO:4; Or
B) by under low stringent condition with the nucleic acid sequence encoding of following hybridization:
I) coding SEQIDNO:4 shown in aminoacid sequence nucleotide sequence complementary strand or
Ii) subsequence of at least 100 nucleotide of (i).
17. compositions according to claim 16, wherein glucoamylase and/or ��-amylase are derived from aspergillus niger or Ai Mosen ankle joint bacterium.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007801019200A CN101918525A (en) | 2007-12-12 | 2007-12-12 | Saccharification method |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007801019200A Division CN101918525A (en) | 2007-12-12 | 2007-12-12 | Saccharification method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105623937A true CN105623937A (en) | 2016-06-01 |
Family
ID=56044447
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610097716.7A Pending CN105623937A (en) | 2007-12-12 | 2007-12-12 | Saccharifying method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105623937A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113999740A (en) * | 2021-11-19 | 2022-02-01 | 广州南沙珠江啤酒有限公司 | Strong beer and preparation method thereof |
| WO2023225459A2 (en) | 2022-05-14 | 2023-11-23 | Novozymes A/S | Compositions and methods for preventing, treating, supressing and/or eliminating phytopathogenic infestations and infections |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1090325A (en) * | 1992-12-28 | 1994-08-03 | 索尔维公司 | Pullulanase, microorganism producing it, its preparation method and use |
| CN1309701A (en) * | 1998-07-02 | 2001-08-22 | 诺沃奇梅兹有限公司 | Starch debranching enzymes |
| CN1671833A (en) * | 2002-07-25 | 2005-09-21 | 诺维信公司 | Mashing process |
| WO2005121305A1 (en) * | 2004-06-08 | 2005-12-22 | Novozymes A/S | Mashing process |
-
2007
- 2007-12-12 CN CN201610097716.7A patent/CN105623937A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1090325A (en) * | 1992-12-28 | 1994-08-03 | 索尔维公司 | Pullulanase, microorganism producing it, its preparation method and use |
| CN1309701A (en) * | 1998-07-02 | 2001-08-22 | 诺沃奇梅兹有限公司 | Starch debranching enzymes |
| CN1671833A (en) * | 2002-07-25 | 2005-09-21 | 诺维信公司 | Mashing process |
| WO2005121305A1 (en) * | 2004-06-08 | 2005-12-22 | Novozymes A/S | Mashing process |
Non-Patent Citations (1)
| Title |
|---|
| 杨兵: "运用不同质量的麦芽进行双醪浸出糖化工艺的研究", 《食品与发酵工业》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113999740A (en) * | 2021-11-19 | 2022-02-01 | 广州南沙珠江啤酒有限公司 | Strong beer and preparation method thereof |
| CN113999740B (en) * | 2021-11-19 | 2023-12-01 | 广州南沙珠江啤酒有限公司 | Strong beer and preparation method thereof |
| WO2023225459A2 (en) | 2022-05-14 | 2023-11-23 | Novozymes A/S | Compositions and methods for preventing, treating, supressing and/or eliminating phytopathogenic infestations and infections |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP2222830B1 (en) | Mashing process | |
| CN102277341B (en) | Alcohol product processes | |
| CN101952409B (en) | Brewing process | |
| CN102220191B (en) | Saccharification process | |
| RU2426775C2 (en) | Wort production method | |
| CN103502420B (en) | Method for producing beer wort | |
| EP2499227B1 (en) | A brewing method | |
| US20060057684A1 (en) | Mashing process | |
| WO2007144393A1 (en) | Mashing process | |
| CN104718289A (en) | Method for producing beer wort | |
| WO2005121305A1 (en) | Mashing process | |
| CN102186965B (en) | Brewing process | |
| CN109923201A (en) | The method for preparing grain extract | |
| EP2222831B1 (en) | Brewing process | |
| US8765199B2 (en) | Mashing process | |
| CN105623937A (en) | Saccharifying method | |
| JP2025063767A (en) | Wort production method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20160601 |