CN112029809A - Method for producing high-purity maltose by multi-enzyme synergistic saccharification - Google Patents
Method for producing high-purity maltose by multi-enzyme synergistic saccharification Download PDFInfo
- Publication number
- CN112029809A CN112029809A CN202010959754.5A CN202010959754A CN112029809A CN 112029809 A CN112029809 A CN 112029809A CN 202010959754 A CN202010959754 A CN 202010959754A CN 112029809 A CN112029809 A CN 112029809A
- Authority
- CN
- China
- Prior art keywords
- saccharification
- enzyme
- maltose
- temperature
- purity
- 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
- 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 title claims abstract description 116
- 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 title claims abstract description 116
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 30
- 230000002195 synergetic effect Effects 0.000 title claims abstract description 20
- 108090000790 Enzymes Proteins 0.000 claims abstract description 49
- 102000004190 Enzymes Human genes 0.000 claims abstract description 49
- 229920002472 Starch Polymers 0.000 claims abstract description 35
- 239000008107 starch Substances 0.000 claims abstract description 35
- 235000019698 starch Nutrition 0.000 claims abstract description 35
- 238000001914 filtration Methods 0.000 claims abstract description 26
- 239000006228 supernatant Substances 0.000 claims abstract description 23
- 235000013336 milk Nutrition 0.000 claims abstract description 22
- 239000008267 milk Substances 0.000 claims abstract description 22
- 210000004080 milk Anatomy 0.000 claims abstract description 22
- 239000000758 substrate Substances 0.000 claims abstract description 22
- 238000002156 mixing Methods 0.000 claims abstract description 21
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 19
- 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 claims abstract description 15
- 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 claims abstract description 15
- 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 claims abstract description 15
- 239000008103 glucose Substances 0.000 claims abstract description 15
- 239000012535 impurity Substances 0.000 claims abstract description 15
- 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 claims abstract description 15
- 238000001694 spray drying Methods 0.000 claims abstract description 15
- 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 claims abstract description 15
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims abstract description 14
- 108090000637 alpha-Amylases Proteins 0.000 claims abstract description 14
- 108010019077 beta-Amylase Proteins 0.000 claims abstract description 14
- 238000013375 chromatographic separation Methods 0.000 claims abstract description 14
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000003456 ion exchange resin Substances 0.000 claims abstract description 13
- 229920003303 ion-exchange polymer Polymers 0.000 claims abstract description 13
- 239000007788 liquid Substances 0.000 claims abstract description 13
- 239000012528 membrane Substances 0.000 claims abstract description 13
- 238000011033 desalting Methods 0.000 claims abstract description 12
- 238000001179 sorption measurement Methods 0.000 claims abstract description 11
- LUEWUZLMQUOBSB-UHFFFAOYSA-N UNPD55895 Natural products OC1C(O)C(O)C(CO)OC1OC1C(CO)OC(OC2C(OC(OC3C(OC(O)C(O)C3O)CO)C(O)C2O)CO)C(O)C1O LUEWUZLMQUOBSB-UHFFFAOYSA-N 0.000 claims abstract description 9
- UYQJCPNSAVWAFU-UHFFFAOYSA-N malto-tetraose Natural products OC1C(O)C(OC(C(O)CO)C(O)C(O)C=O)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(O)C(CO)O2)O)C(CO)O1 UYQJCPNSAVWAFU-UHFFFAOYSA-N 0.000 claims abstract description 9
- LUEWUZLMQUOBSB-OUBHKODOSA-N maltotetraose 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]3[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-OUBHKODOSA-N 0.000 claims abstract description 9
- 238000001704 evaporation Methods 0.000 claims abstract description 8
- 230000008020 evaporation Effects 0.000 claims abstract description 7
- 238000005119 centrifugation Methods 0.000 claims abstract description 5
- 238000000926 separation method Methods 0.000 claims abstract description 5
- 229940088598 enzyme Drugs 0.000 claims description 70
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 31
- 239000008367 deionised water Substances 0.000 claims description 17
- 229910021641 deionized water Inorganic materials 0.000 claims description 17
- 239000004382 Amylase Substances 0.000 claims description 12
- 108010065511 Amylases Proteins 0.000 claims description 12
- 102000013142 Amylases Human genes 0.000 claims description 12
- 235000019418 amylase Nutrition 0.000 claims description 12
- 238000010438 heat treatment Methods 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 9
- 239000011552 falling film Substances 0.000 claims description 9
- 229910021645 metal ion Inorganic materials 0.000 claims description 3
- 229920005654 Sephadex Polymers 0.000 claims description 2
- 239000012507 Sephadex™ Substances 0.000 claims description 2
- 229940079919 digestives enzyme preparation Drugs 0.000 claims description 2
- 238000004042 decolorization Methods 0.000 claims 1
- 235000020357 syrup Nutrition 0.000 abstract description 28
- 239000006188 syrup Substances 0.000 abstract description 28
- 229920001353 Dextrin Polymers 0.000 abstract description 6
- 239000004375 Dextrin Substances 0.000 abstract description 6
- 235000019425 dextrin Nutrition 0.000 abstract description 6
- 238000000034 method Methods 0.000 abstract description 6
- 229920000945 Amylopectin Polymers 0.000 abstract description 4
- 238000007670 refining Methods 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 abstract description 3
- 102000004169 proteins and genes Human genes 0.000 abstract description 3
- 230000009849 deactivation Effects 0.000 abstract description 2
- 239000012860 organic pigment Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 12
- 239000003480 eluent Substances 0.000 description 8
- 229910052799 carbon Inorganic materials 0.000 description 7
- 230000000052 comparative effect Effects 0.000 description 7
- 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 3
- 229920001542 oligosaccharide Polymers 0.000 description 3
- 150000002482 oligosaccharides Chemical class 0.000 description 3
- 235000009508 confectionery Nutrition 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 235000020429 malt syrup Nutrition 0.000 description 2
- MIDXCONKKJTLDX-UHFFFAOYSA-N 3,5-dimethylcyclopentane-1,2-dione Chemical compound CC1CC(C)C(=O)C1=O MIDXCONKKJTLDX-UHFFFAOYSA-N 0.000 description 1
- 229920000856 Amylose Polymers 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 230000009286 beneficial effect 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
- 235000013361 beverage Nutrition 0.000 description 1
- 235000013736 caramel Nutrition 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000015203 fruit juice Nutrition 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
- 238000010253 intravenous injection Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/22—Preparation of compounds containing saccharide radicals produced by the action of a beta-amylase, e.g. maltose
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/12—Disaccharides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/16—Preparation of compounds containing saccharide radicals produced by the action of an alpha-1, 6-glucosidase, e.g. amylose, debranched amylopectin
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a method for producing high-purity maltose by multi-enzyme synergistic saccharification, which comprises the steps of starch size mixing, liquefaction, filtration and activated carbon adsorption to obtain a substrate, pullulanase, maltotriose enzyme and maltotetraose enzyme are sequentially added, beta-amylase is finally added for continuous saccharification, the temperature is raised for enzyme deactivation, yeast is added into the obtained saccharified liquid for reaction and then centrifugation is carried out, and the high-purity maltose is prepared by taking supernatant through ion exchange resin, membrane separation, chromatographic separation, evaporation concentration and spray drying. The method comprises the steps of adjusting the concentration of starch milk to 18-25 degrees Be, carrying out primary filtration and decoloration after liquefaction, adding a plurality of enzymes for synergistic saccharification, adding yeast to remove glucose, carrying out centrifugation to remove protein and organic pigment impurities, fully saccharifying dextrin and amylopectin to reduce the content of the dextrin and amylopectin, improving the purity of maltose, desalting the obtained saccharified liquid by ion exchange resin, carrying out membrane separation and refining, further purifying the maltose by chromatographic separation, evaporating and concentrating to produce high maltose syrup, and carrying out spray drying to obtain the high-purity maltose.
Description
Technical Field
The invention relates to the technical field of maltose production, in particular to a method for producing high-purity maltose by multi-enzyme synergistic saccharification.
Background
Maltose is a disaccharide having two glucose units linked via an α -1,4 glycosidic bond. The traditional maltose is prepared from wheat and glutinous rice, is rich in nutrition, is sweet and delicious, and has a plurality of efficacies of expelling toxin and beautifying, invigorating spleen and replenishing qi, moistening lung and relieving cough, and the like. It can be prepared into maltose syrup, which has the advantages of low sweetness, low hygroscopicity, high moisture retention and the like, and has wide application. Can be used in various fields of food industry, and is mainly used for processing caramel paste color, candy and fruit juice beverage; the method can also be used in the pharmaceutical industry, and in the pharmaceutical field, the refined maltose can be used for producing maltose intravenous injection, so that the preparation of high-purity maltose is particularly important.
At present, in the production of malt syrup, a double-enzyme method or an enzyme-enzyme combination method is widely applied, but a single amylase or two enzymes have not ideal saccharification effect on starch, the purity of the produced malt syrup is not high, certain limitation is caused on the field needing high-purity maltose such as medical and pharmaceutical, and the enzyme-enzyme combination method commonly used in industry causes certain chemical pollution and influences the environment.
Disclosure of Invention
The invention aims to provide a method for producing high-purity maltose by utilizing multi-enzyme synergistic saccharification to improve the content of maltose in maltose syrup and by removing glucose and impurities through treatment after liquefaction and refining after saccharification to separate other oligosaccharides.
To achieve the object, the present invention provides a method for producing high-purity maltose by multi-enzyme cooperative saccharification, comprising the steps of:
(1) adding water into the size mixing tank, starting a stirrer, adding starch, uniformly mixing, adjusting the pH value to 5.5-6.2, and controlling the concentration of starch milk to 18-25 degrees Be;
(2) uniformly stirring starch milk and alpha-high temperature amylase, and performing jet liquefaction, wherein the liquefaction temperature is controlled to be 85-105 ℃, and the enzyme adding amount is 5-10U/g;
(3) filtering the liquefied solution to remove redundant impurities, and adding activated carbon for adsorption and decoloration to obtain a substrate A suitable for saccharification;
(4) sequentially adding pullulanase, maltotriose enzyme and maltotetraose enzyme into a substrate A for synergistic saccharification, controlling the saccharification temperature at 50-65 ℃ and the saccharification time at 15-20 h, finally adding beta-amylase for continuous saccharification, controlling the saccharification temperature at 55-65 ℃ and the saccharification time at 5-10 h, heating to inactivate enzyme after the saccharification is finished, adding yeast when the temperature is reduced to 35 ℃, removing glucose, and centrifuging the obtained final saccharified liquid to obtain a supernatant B;
(5) desalting the supernatant B through ion exchange resin to remove metal ions, and then performing membrane separation and chromatographic separation;
(6) the high-purity maltose is prepared by evaporation concentration and spray drying.
Further preferably, the water used in the step (1) is deionized water.
Preferably, the filtering in the step (3) is performed by filtering macromolecular impurities in the solution by using a plate and frame filter press, wherein the filtering pressure is 0.5-0.7 mpa, and the water flow is 60-75L/min.
Further preferably, the adding amount of the activated carbon in the step (3) is 0.1-0.3%, and the decoloring time is 30-40 min.
Further preferably, the enzyme preparations pullulanase, maltotriose, maltotetraose and beta-amylase added in the step (4) for the synergistic saccharification by the multienzyme are added in amounts of 8-15U/ml, 10-15U/ml, 15-20U/ml and 10-20U/ml respectively.
Preferably, in the step (4), the supernatant is obtained after centrifugation is carried out for 30-40 min at the rotating speed of 7500-8500 r/min.
Further preferably, the chromatographic column used in the step (5) is Sephadex G-25.
Preferably, the evaporation and concentration in the step (6) are carried out by adopting a multi-effect falling-film evaporator under the conditions that the vacuum degree is 0.6-1.0 mpa and the temperature is 65-90 ℃, the drying air inlet temperature of spray drying is 180-190 ℃ and the air exhaust temperature is 80-85 ℃.
The invention has the following beneficial effects:
the starch milk concentration is adjusted to be 18-25 degrees Be, the alpha-high temperature amylase is added to enable the starch to be rapidly degraded to be converted into dextrin, amylopectin, oligosaccharide, maltose, glucose and the like, and macromolecular impurities such as protein, organic pigment and the like are removed through primary filtering and decoloring after liquefaction, so that a proper substrate is provided for subsequent saccharification, and the subsequent saccharification effect is improved; adding pullulanase, maltotriose enzyme, maltotetraose enzyme and other multienzymes for synergistic saccharification, firstly converting amylopectin into amylose, and converting dextrin into maltotriose, maltotetraose and maltose, and finally adding beta-amylase for further saccharification to generate more maltose, monosaccharide and limit dextrin; adding yeast into the obtained saccharified solution to hydrolyze glucose, reducing the content of glucose, further improving the purity of maltose, centrifugally removing protein after enzyme deactivation, desalting the saccharified solution after centrifugation by ion exchange resin, removing metal ions, further refining and separating other oligosaccharides and macromolecular limit dextrin by membrane separation and chromatographic separation, improving the purity of maltose, producing high maltose syrup by evaporation concentration, and finally spray drying to obtain the high-purity maltose.
Detailed Description
The embodiments described below are only a part of the embodiments of the present invention, and not all of them. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a method for producing high-purity maltose by multi-enzyme synergistic saccharification, which is characterized by comprising the following steps:
(1) adding water into the size mixing tank, starting a stirrer, adding starch, uniformly mixing, adjusting the pH value to 5.5-6.2, and controlling the concentration of starch milk to 18-25 degrees Be;
(2) uniformly stirring starch milk and alpha-high temperature amylase, and performing jet liquefaction, wherein the liquefaction temperature is controlled to be 85-105 ℃, and the enzyme adding amount is 5-10U/g;
(3) filtering the liquefied solution to remove redundant impurities, and adding activated carbon for adsorption and decoloration to obtain a substrate A suitable for saccharification;
(4) sequentially adding pullulanase, maltotriose enzyme and maltotetraose enzyme into a substrate A for synergistic saccharification, controlling the saccharification temperature at 50-65 ℃ and the saccharification time at 15-20 h, finally adding beta-amylase for continuous saccharification, controlling the saccharification temperature at 55-65 ℃ and the saccharification time at 5-10 h, heating to inactivate enzyme after the saccharification is finished, adding yeast when the temperature is reduced to 35 ℃, removing glucose, and centrifuging the obtained final saccharified liquid to obtain a supernatant B;
(5) desalting the supernatant B through ion exchange resin, separating through a membrane, and performing chromatographic separation by using deionized water as an eluent;
(6) the high-purity maltose is prepared by evaporation concentration and spray drying.
Example 1
This example provides a method for producing high purity maltose by multi-enzyme collaborative saccharification, comprising the steps of:
(1) adding deionized water into the size mixing tank, starting a stirrer, adding starch, uniformly mixing, adjusting the pH value to 5.5-6.2, and controlling the concentration of starch milk to 19 degrees Be;
(2) uniformly stirring starch milk and alpha-high temperature amylase, and performing jet liquefaction, wherein the liquefaction temperature is controlled to be 85-105 ℃, and the enzyme adding amount is 8U/g;
(3) filtering the liquefied solution by using a plate and frame filter press, wherein the filtering pressure is 0.5mpa, the water flow is 60L/min, removing redundant impurities, and adding 0.1% of active carbon for adsorption and decoloration for 30min to obtain a substrate A suitable for saccharification;
(4) sequentially adding 8U/ml pullulanase, 10U/ml maltotriose enzyme and 15U/ml maltotetrase enzyme into a substrate A for synergistic saccharification, controlling the saccharification temperature at 50-65 ℃, the saccharification time at 15h, finally adding 10U/ml beta-amylase for continuous saccharification, controlling the saccharification temperature at 55-65 ℃, the saccharification time at 7h, heating to inactivate the enzymes after the saccharification is finished, adding yeast when the temperature is reduced to 35 ℃, removing glucose, centrifuging the obtained final saccharified liquid at the rotating speed of 7500r/min for 30min, and taking a supernatant B;
(5) desalting the supernatant B through ion exchange resin, separating through a membrane, and performing chromatographic separation by using deionized water as an eluent;
(6) concentrating the high maltose syrup by adopting a multi-effect falling-film evaporator at the vacuum degree of 0.6mpa and the temperature of 65 ℃, preparing the high maltose syrup into high-purity maltose through spray drying, wherein the drying air inlet temperature is 180-190 ℃, and the air exhaust temperature is 80-85 ℃.
The maltose content in the obtained high maltose syrup is 89.9%, and the maltose purity in the high purity maltose is 91.3%.
Example 2
This example provides a method for producing high purity maltose by multi-enzyme collaborative saccharification, comprising the steps of:
(1) adding deionized water into the size mixing tank, starting a stirrer, adding starch, uniformly mixing, adjusting the pH value to 5.5-6.2, and controlling the concentration of starch milk to 19 degrees Be;
(2) uniformly stirring starch milk and alpha-high temperature amylase, and performing jet liquefaction, wherein the liquefaction temperature is controlled to be 85-105 ℃, and the enzyme adding amount is 8U/g;
(3) filtering the liquefied solution by using a plate and frame filter press, wherein the filtering pressure is 0.5mpa, the water flow is 60L/min, removing redundant impurities, and adding 0.1% of active carbon for adsorption and decoloration for 30min to obtain a substrate A suitable for saccharification;
(4) sequentially adding 8U/ml pullulanase, 11U/ml maltotriose enzyme and 17U/ml maltotetrase enzyme into a substrate A for synergistic saccharification, controlling the saccharification temperature at 50-65 ℃, the saccharification time at 15h, finally adding 10U/ml beta-amylase for continuous saccharification, controlling the saccharification temperature at 55-65 ℃, the saccharification time at 7h, heating to inactivate the enzymes after the saccharification is finished, adding yeast when the temperature is reduced to 35 ℃, removing glucose, centrifuging the obtained final saccharified liquid at the rotating speed of 7500r/min for 30min, and taking a supernatant B;
(5) desalting the supernatant B through ion exchange resin, separating through a membrane, and performing chromatographic separation by using deionized water as an eluent;
(6) concentrating the high maltose syrup by adopting a multi-effect falling-film evaporator at the vacuum degree of 0.6mpa and the temperature of 65 ℃, preparing the high maltose syrup into high-purity maltose through spray drying, wherein the drying air inlet temperature is 180-190 ℃, and the air exhaust temperature is 80-85 ℃.
The maltose content in the obtained high maltose syrup is 90.1%, and the maltose purity in the high purity maltose is 92.5%.
Example 3
This example provides a method for producing high purity maltose by multi-enzyme collaborative saccharification, comprising the steps of:
(1) adding deionized water into the size mixing tank, starting a stirrer, adding starch, uniformly mixing, adjusting the pH value to 5.5-6.2, and controlling the concentration of starch milk to 19 degrees Be;
(2) uniformly stirring starch milk and alpha-high temperature amylase, and performing jet liquefaction, wherein the liquefaction temperature is controlled to be 85-105 ℃, and the enzyme adding amount is 8U/g;
(3) filtering the liquefied solution by using a plate and frame filter press, wherein the filtering pressure is 0.5mpa, the water flow is 65L/min, removing redundant impurities, and adding 0.2% of active carbon for adsorption and decoloration for 35min to obtain a substrate A suitable for saccharification;
(4) sequentially adding 9U/ml pullulanase, 11U/ml maltotriose enzyme and 17U/ml maltotetrase enzyme into a substrate A for synergistic saccharification, controlling the saccharification temperature at 50-65 ℃, the saccharification time at 16h, finally adding 12U/ml beta-amylase for continuous saccharification, controlling the saccharification temperature at 55-65 ℃, the saccharification time at 7h, heating to inactivate the enzymes after the saccharification is finished, adding yeast when the temperature is reduced to 35 ℃, removing glucose, centrifuging the obtained final saccharified liquid at the rotating speed of 7500r/min for 30min, and taking a supernatant B;
(5) desalting the supernatant B through ion exchange resin, separating through a membrane, and performing chromatographic separation by using deionized water as an eluent;
(6) concentrating the high maltose syrup by adopting a multi-effect falling-film evaporator at the vacuum degree of 0.8mpa and the temperature of 70 ℃, preparing the high maltose syrup into high-purity maltose through spray drying, wherein the drying air inlet temperature is 180-190 ℃, and the air exhaust temperature is 80-85 ℃.
The maltose content in the obtained high maltose syrup is 90.5%, and the maltose purity in the high purity maltose is 93.2%.
Example 4
This example provides a method for producing high purity maltose by multi-enzyme collaborative saccharification, comprising the steps of:
(1) adding deionized water into the size mixing tank, starting a stirrer, adding starch, uniformly mixing, adjusting the pH value to 5.5-6.2, and controlling the concentration of starch milk to 19 degrees Be;
(2) uniformly stirring starch milk and alpha-high temperature amylase, and performing jet liquefaction, wherein the liquefaction temperature is controlled to be 85-105 ℃, and the enzyme adding amount is 8U/g;
(3) filtering the liquefied solution by using a plate and frame filter press, wherein the filtering pressure is 0.6mpa, the water flow is 70L/min, removing redundant impurities, and adding 0.2% of active carbon for adsorption and decoloration for 35min to obtain a substrate A suitable for saccharification;
(4) sequentially adding 9U/ml pullulanase, 12U/ml maltotriose enzyme and 18U/ml maltotetrase enzyme into a substrate A for synergistic saccharification, controlling the saccharification temperature at 50-65 ℃, the saccharification time at 18h, finally adding 15U/ml beta-amylase for continuous saccharification, controlling the saccharification temperature at 55-65 ℃, the saccharification time at 8h, heating to inactivate the enzymes after the saccharification is finished, adding yeast when the temperature is reduced to 35 ℃, removing glucose, centrifuging the obtained final saccharified liquid at the rotating speed of 7500r/min for 35min, and taking a supernatant B;
(5) desalting the supernatant B through ion exchange resin, separating through a membrane, and performing chromatographic separation by using deionized water as an eluent;
(6) concentrating the high maltose syrup by adopting a multi-effect falling-film evaporator at the vacuum degree of 0.8mpa and the temperature of 75 ℃, preparing the high maltose syrup into high-purity maltose through spray drying, wherein the drying air inlet temperature is 180-190 ℃, and the air exhaust temperature is 80-85 ℃.
The maltose content in the obtained high maltose syrup is 91.4%, and the maltose purity in the high purity maltose is 94%.
Example 5
This example provides a method for producing high purity maltose by multi-enzyme collaborative saccharification, comprising the steps of:
(1) adding deionized water into the size mixing tank, starting a stirrer, adding starch, uniformly mixing, adjusting the pH value to 5.5-6.2, and controlling the concentration of starch milk to 22 degrees Be;
(2) uniformly stirring starch milk and alpha-high temperature amylase, and performing jet liquefaction, wherein the liquefaction temperature is controlled to be 85-105 ℃, and the enzyme adding amount is 10U/g;
(3) filtering the liquefied solution by using a plate and frame filter press, wherein the filtering pressure is 0.7mpa, the water flow is 70L/min, removing redundant impurities, and adding 0.2% of active carbon for adsorption and decoloration for 35min to obtain a substrate A suitable for saccharification;
(4) sequentially adding 12U/ml pullulanase, 12U/ml maltotriose enzyme and 19U/ml maltotetrase enzyme into a substrate A for synergistic saccharification, controlling the saccharification temperature at 50-65 ℃, the saccharification time at 18h, finally adding 15U/ml beta-amylase for continuous saccharification, controlling the saccharification temperature at 55-65 ℃, the saccharification time at 8h, heating to inactivate the enzymes after the saccharification is finished, adding yeast when the temperature is reduced to 35 ℃, removing glucose, centrifuging the obtained final saccharified liquid at the rotating speed of 8000r/min for 30min, and taking a supernatant B;
(5) desalting the supernatant B through ion exchange resin, separating through a membrane, and performing chromatographic separation by using deionized water as an eluent;
(6) concentrating the high maltose syrup by adopting a multi-effect falling-film evaporator at the vacuum degree of 0.8mpa and the temperature of 75 ℃, preparing the high maltose syrup into high-purity maltose through spray drying, wherein the drying air inlet temperature is 180-190 ℃, and the air exhaust temperature is 80-85 ℃.
The maltose content in the obtained high maltose syrup is 92.6%, and the maltose purity in the high purity maltose is 95.3%.
Example 6
This example provides a method for producing high purity maltose by multi-enzyme collaborative saccharification, comprising the steps of:
(1) adding deionized water into the size mixing tank, starting a stirrer, adding starch, uniformly mixing, adjusting the pH value to 5.5-6.2, and controlling the concentration of starch milk to 22 degrees Be;
(2) uniformly stirring starch milk and alpha-high temperature amylase, and performing jet liquefaction, wherein the liquefaction temperature is controlled to be 85-105 ℃, and the enzyme adding amount is 10U/g;
(3) filtering the liquefied solution by using a plate and frame filter press, wherein the filtering pressure is 0.7mpa, the water flow is 75L/min, removing redundant impurities, and adding 0.3% of active carbon for adsorption and decoloration for 40min to obtain a substrate A suitable for saccharification;
(4) sequentially adding 14U/ml pullulanase, 15U/ml maltotriose enzyme and 19U/ml maltotetrase enzyme into a substrate A for synergistic saccharification, controlling the saccharification temperature at 50-65 ℃, the saccharification time at 20h, finally adding 18U/ml beta-amylase for continuous saccharification, controlling the saccharification temperature at 55-65 ℃, the saccharification time at 9h, heating to inactivate the enzymes after the saccharification is finished, adding yeast when the temperature is reduced to 35 ℃, removing glucose, centrifuging the obtained final saccharified liquid at the rotating speed of 8000r/min for 40min, and taking a supernatant B;
(5) desalting the supernatant B through ion exchange resin, separating through a membrane, and performing chromatographic separation by using deionized water as an eluent;
(6) concentrating the high maltose syrup by adopting a multi-effect falling-film evaporator at the vacuum degree of 0.8mpa and the temperature of 80 ℃, preparing the high maltose syrup into high-purity maltose through spray drying, wherein the drying air inlet temperature is 180-190 ℃, and the air exhaust temperature is 80-85 ℃.
The maltose content in the obtained high maltose syrup is 93.3%, and the maltose purity in the high purity maltose is 96.1%.
Example 7
This example provides a method for producing high purity maltose by multi-enzyme collaborative saccharification, comprising the steps of:
(1) adding deionized water into the size mixing tank, starting a stirrer, adding starch, uniformly mixing, adjusting the pH value to 5.5-6.2, and controlling the concentration of starch milk at 23 degrees Be;
(2) uniformly stirring starch milk and alpha-high temperature amylase, and performing jet liquefaction, wherein the liquefaction temperature is controlled to be 85-105 ℃, and the enzyme adding amount is 10U/g;
(3) filtering the liquefied solution by using a plate and frame filter press, wherein the filtering pressure is 0.7mpa, the water flow is 75L/min, removing redundant impurities, and adding 0.3% of active carbon for adsorption and decoloration for 40min to obtain a substrate A suitable for saccharification;
(4) sequentially adding 15U/ml pullulanase, 15U/ml maltotriose enzyme and 20U/ml maltotetrase enzyme into a substrate A for synergistic saccharification, controlling the saccharification temperature at 50-65 ℃, the saccharification time at 20h, finally adding 20U/ml beta-amylase for continuous saccharification, controlling the saccharification temperature at 55-65 ℃, the saccharification time at 10h, heating to inactivate the enzymes after the saccharification is finished, adding yeast when the temperature is reduced to 35 ℃, removing glucose, centrifuging the obtained final saccharified liquid at the rotating speed of 8500r/min for 40min, and taking a supernatant B;
(5) desalting the supernatant B through ion exchange resin, separating through a membrane, and performing chromatographic separation by using deionized water as an eluent;
(6) concentrating the high maltose syrup by adopting a multi-effect falling-film evaporator at the vacuum degree of 1.0mpa and the temperature of 90 ℃, preparing the high maltose syrup into high-purity maltose through spray drying, wherein the drying air inlet temperature is 180-190 ℃, and the air exhaust temperature is 80-85 ℃.
The maltose content in the obtained high maltose syrup is 94.7%, and the maltose purity in the high purity maltose is 97.8%.
Comparative example 1
This comparative example provides a method for producing high-purity maltose by multi-enzyme collaborative saccharification, which is different from the method for producing high-purity maltose provided in example 1 in that maltotriosidase and maltotetraose enzyme are not added at the time of saccharification.
The maltose content in the obtained high maltose syrup is 80.1%, and the maltose purity in the high purity maltose is 81.4%.
Comparative example 2
This comparative example provides a method for producing high-purity maltose by multi-enzyme collaborative saccharification, which is different from the method for producing high-purity maltose provided in example 1 in that yeast is not added after the saccharification is completed.
The maltose content in the obtained high maltose syrup is 84.2%, and the maltose purity in the high purity maltose is 85.6%.
Comparative example 3
This comparative example provides a method for producing high-purity maltose by multi-enzyme cooperative saccharification, which is different from the method for producing high-purity maltose provided in example 1 in that chromatographic separation is not employed in the saccharification liquid refining step.
The maltose content in the obtained high maltose syrup is 84.7%, and the maltose purity in the high purity maltose is 85.9%.
Comparing the results of example 1 with comparative examples 1, 2 and 3, it was found that the process of example 1 greatly improved the purity of maltose.
The above disclosure is intended to cover only the preferred embodiments of the invention, and not to limit the scope of the invention, so that modifications and equivalents may be made within the scope of the invention as claimed.
Claims (8)
1. A method for producing high-purity maltose by multi-enzyme cooperative saccharification is characterized by comprising the following steps:
(1) adding water into the size mixing tank, starting a stirrer, adding starch, uniformly mixing, adjusting the pH value to 5.5-6.2, and controlling the concentration of starch milk to 18-25 degrees Be;
(2) uniformly stirring starch milk and alpha-high temperature amylase, and performing jet liquefaction, wherein the liquefaction temperature is controlled to be 85-105 ℃, and the enzyme adding amount is 5-10U/g;
(3) filtering the liquefied solution to remove redundant impurities, and adding activated carbon for adsorption and decoloration to obtain a substrate A suitable for saccharification;
(4) sequentially adding pullulanase, maltotriose enzyme and maltotetraose enzyme into a substrate A for synergistic saccharification, controlling the saccharification temperature at 50-65 ℃ and the saccharification time at 15-20 h, finally adding beta-amylase for continuous saccharification, controlling the saccharification temperature at 55-65 ℃ and the saccharification time at 5-10 h, heating to inactivate enzyme after the saccharification is finished, adding yeast when the temperature is reduced to 35 ℃, removing glucose, and centrifuging the obtained final saccharified liquid to obtain a supernatant B;
(5) desalting the supernatant B through ion exchange resin to remove metal ions, and then performing membrane separation and chromatographic separation;
(6) the high-purity maltose is prepared by evaporation concentration and spray drying.
2. The method for producing high-purity maltose by multi-enzyme cooperative saccharification according to claim 1, characterized in that the water used in step (1) is deionized water.
3. The method for producing high-purity maltose by multi-enzyme cooperative saccharification according to claim 1, characterized in that the filtering in step (3) is performed by filtering macromolecular impurities in the maltose by using a plate and frame filter press, the filtering pressure is 0.5-0.7 mpa, and the water flow is 60-75L/min.
4. The method for producing high-purity maltose by multi-enzyme cooperative saccharification according to claim 1, characterized in that in step (3), the adding amount of activated carbon is 0.1-0.3%, and the decolorization time is 30-40 min.
5. The method for producing high-purity maltose according to claim 1, wherein the enzyme preparations pullulanase, maltotriose, maltotetraose and β -amylase added in the step (4) are added in amounts of 8-15U/ml, 10-15U/ml, 15-20U/ml and 10-20U/ml, respectively.
6. The method for producing high-purity maltose by multi-enzyme cooperative saccharification according to claim 1, characterized in that in step (4), the supernatant is obtained after centrifugation for 30-40 min at 7500-8500 r/min.
7. The method for producing high-purity maltose by multi-enzyme cooperative saccharification according to claim 1, characterized in that the chromatographic column used in step (5) is Sephadex G-25.
8. The method for producing high-purity maltose through multi-enzyme synergistic saccharification according to claim 1, characterized in that the evaporation and concentration in step (6) are carried out by adopting a multi-effect falling film evaporator under the conditions of a vacuum degree of 0.6-1.0 mpa and a temperature of 65-90 ℃, the drying air inlet temperature of spray drying is 180-190 ℃, and the air exhaust temperature is 80-85 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010959754.5A CN112029809A (en) | 2020-09-14 | 2020-09-14 | Method for producing high-purity maltose by multi-enzyme synergistic saccharification |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010959754.5A CN112029809A (en) | 2020-09-14 | 2020-09-14 | Method for producing high-purity maltose by multi-enzyme synergistic saccharification |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112029809A true CN112029809A (en) | 2020-12-04 |
Family
ID=73589079
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010959754.5A Pending CN112029809A (en) | 2020-09-14 | 2020-09-14 | Method for producing high-purity maltose by multi-enzyme synergistic saccharification |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112029809A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114150028A (en) * | 2021-11-27 | 2022-03-08 | 湖北德安府糖业股份有限公司 | Preparation process of fermented maltose syrup |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1353200A (en) * | 2001-12-06 | 2002-06-12 | 华南理工大学 | Process for preparing high-purity malt sugar by multi-enzyme cooperative saccharification |
| CN103409484A (en) * | 2013-08-23 | 2013-11-27 | 山东福田药业有限公司 | Preparation method of superhigh malt syrup |
| CN107746867A (en) * | 2017-10-31 | 2018-03-02 | 无锡甜丰食品有限公司 | A kind of purification process of malt syrup |
| CN109371078A (en) * | 2018-10-18 | 2019-02-22 | 山东福田药业有限公司 | A kind of high-purity malt sugar preparation process |
-
2020
- 2020-09-14 CN CN202010959754.5A patent/CN112029809A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1353200A (en) * | 2001-12-06 | 2002-06-12 | 华南理工大学 | Process for preparing high-purity malt sugar by multi-enzyme cooperative saccharification |
| CN103409484A (en) * | 2013-08-23 | 2013-11-27 | 山东福田药业有限公司 | Preparation method of superhigh malt syrup |
| CN107746867A (en) * | 2017-10-31 | 2018-03-02 | 无锡甜丰食品有限公司 | A kind of purification process of malt syrup |
| CN109371078A (en) * | 2018-10-18 | 2019-02-22 | 山东福田药业有限公司 | A kind of high-purity malt sugar preparation process |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114150028A (en) * | 2021-11-27 | 2022-03-08 | 湖北德安府糖业股份有限公司 | Preparation process of fermented maltose syrup |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5462864A (en) | Manufacturing method of high purity maltose and its reduced product | |
| CN109207537B (en) | Maltose syrup production process | |
| US3804715A (en) | Process for preparing sugar containing maltose of high purity | |
| CN109371078B (en) | Preparation process of high-purity maltose | |
| CN111100892A (en) | High fructose corn syrup production process | |
| CN112029809A (en) | Method for producing high-purity maltose by multi-enzyme synergistic saccharification | |
| CN111944862B (en) | Trehalose production method | |
| CN110938715B (en) | Maltose crystallization process | |
| CN107287263B (en) | Preparation method for high-purity maltose and co-production of beta-limit dextrin | |
| CN104480160B (en) | A kind of method that coupling sugar is produced using cyclodextrin glycosyltransferase | |
| US7981639B2 (en) | Starch-derived products | |
| CN108774273B (en) | Trehalose crystallization process | |
| CN111705095A (en) | Preparation method of isomaltooligosaccharide | |
| CN114262387B (en) | Preparation method of resistant dextrin | |
| CN113215208A (en) | Preparation method of maltose powder with high maltose content | |
| CN112522346A (en) | Preparation method of high-purity oligomeric maltose | |
| JP3513197B2 (en) | Method for producing high purity maltitol | |
| CN112322680A (en) | Maltose preparation process | |
| JPH0669385B2 (en) | Method for producing sugar with high content of branched oligosaccharide | |
| CN105603022A (en) | Method for preparing high maltose oligosaccharide | |
| CN113430238B (en) | Method for producing resistant dextrin by adding sucrose/fructo-oligosaccharide | |
| KR930000271B1 (en) | Process for the production of low calorie alcoholic beverages | |
| CN1270215A (en) | Starch syrup dedicated for beer and preparing process thereof | |
| JP5961339B2 (en) | Liquid sugar for low sugar beer flavored alcoholic beverage and method for producing the same, and method for producing low sugar beer flavored alcoholic beverage | |
| CN117924381A (en) | Method for preparing isomaltooligosaccharide by utilizing crystallized sugar mother liquor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201204 |
|
| RJ01 | Rejection of invention patent application after publication |