CN107287263B - Preparation method for high-purity maltose and co-production of beta-limit dextrin - Google Patents

Preparation method for high-purity maltose and co-production of beta-limit dextrin Download PDF

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CN107287263B
CN107287263B CN201710668516.7A CN201710668516A CN107287263B CN 107287263 B CN107287263 B CN 107287263B CN 201710668516 A CN201710668516 A CN 201710668516A CN 107287263 B CN107287263 B CN 107287263B
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maltose
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CN107287263A (en
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邵先豹
李方华
干昭波
窦光朋
杨腾腾
张明站
刘双双
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Shandong Bailong Chuangyuan Bio Tech Co Ltd
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    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/22Preparation of compounds containing saccharide radicals produced by the action of a beta-amylase, e.g. maltose
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    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/04Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/12Disaccharides

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Abstract

The invention relates to a preparation method of high-purity maltose and beta-limit dextrin, which comprises the following steps: (1) preparing starch milk; (2) gelatinizing, adding beta-amylase to carry out saccharification reaction to prepare a saccharified product; (3) decolorizing and deproteinizing the saccharified product, desalting and concentrating to obtain a concentrated product; (4) separating the concentrated product by chromatography to respectively prepare maltose and beta-limit dextrin; (5) carrying out decoloration ion exchange, concentration, drying and refining on maltose to prepare high-purity maltose; (6) the beta-limit dextrin is subjected to decoloration ion, ion exchange, concentration, drying and refining to prepare the beta-limit dextrin with the low DE value. The invention generates maltose and beta-limit dextrin by the action of beta-amylase after gelatinization, which is beneficial to separation, thereby obtaining high-purity maltose and low DE value beta-limit dextrin.

Description

Preparation method for high-purity maltose and co-production of beta-limit dextrin
Technical Field
The invention relates to a preparation method of high-purity maltose and co-production of beta-limit dextrin, belonging to the technical field of biological engineering.
Background
The maltose syrup is prepared by taking high-quality starch as a raw material and carrying out liquefaction, saccharification, decoloration, filtration and refinement concentration, and takes maltose as a main component product. Maltose (maltose) is a disaccharide with two glucose units connected by an alpha-1, 4 glycosidic linkage, also known as maltobiose. Because the position of the hydroxyl of C1 is different, two isomers are provided, namely alpha-isomer and beta-isomer. The maltose syrup may be classified into general maltose syrup, high maltose syrup and ultra high maltose syrup according to the quality fraction of maltose. The maltose syrup with the maltose mass fraction below 60 percent is common maltose syrup, the maltose mass fraction between 60 percent and 70 percent is called high maltose syrup, and the maltose mass fraction above 70 percent is called ultrahigh maltose syrup.
Because of the characteristics of low moisture absorption, high moisture retention, moderate sweetness, good crystallization resistance, oxidation resistance, moderate viscosity, good chemical stability, low freezing point and the like, the maltose syrup is widely applied to the industries of candies, cold drink products, dairy products, beer, jelly, baked food, seasonings, enzyme preparations, instant food, meat products and the like.
At present, high-temperature amylase is mostly adopted for hydrolysis and liquefaction in maltose production, the components of a final product are complex, a certain amount of glucose and maltotriose are contained, subsequent separation and purification are not facilitated, the separation difficulty of the product is increased, and the application of the product in food is limited due to low purity.
Chinese patent document CN103194509A (application number 201310143750.X) discloses a method for preparing high-purity beta-limit dextrin, comprising the following steps: a) saccharifying with biological enzyme: saccharifying by using beta-amylase to generate saccharified liquid containing beta-limit dextrin; b) chromatographic separation and purification: removing micromolecular sugar in the saccharified liquid by utilizing a chromatographic separation technology to obtain high-purity beta-limit dextrin sugar liquid: c) refining: and (3) carrying out activated carbon decoloration filtration, ion exchange desalination and vacuum concentration on the obtained high-purity beta-limit dextrin sugar solution to obtain a high-purity beta-limit dextrin product. However, the product has poor quality due to more micromolecule saccharides with different polymerization degrees, and the market demand cannot be met.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a preparation method for high-purity maltose and beta-limit dextrin coproduction.
The technical scheme of the invention is as follows:
a preparation method of high-purity maltose and beta-limit dextrin co-production comprises the following steps:
(1) mixing waxy corn starch and reverse osmosis water to prepare starch milk with the mass percentage concentration of 10-30% and the pH value of 4-6;
(2) heating the starch milk prepared in the step (1) to 70-95 ℃, gelatinizing for 60-120 min, cooling to 50-65 ℃, adding beta-amylase to perform saccharification reaction, and deactivating enzyme to prepare a saccharification product;
(3) carrying out decoloration and protein removal treatment on the saccharified product prepared in the step (2), desalting and concentrating to prepare a concentrated product with the fixed mass percentage of 40-60%;
(4) carrying out chromatographic separation on the concentrated product prepared in the step (3) to respectively prepare maltose and beta-limit dextrin;
(5) carrying out decoloration ion exchange, concentration, drying and refining on the maltose prepared in the step (4) to prepare high-purity maltose;
(6) and (4) carrying out decoloration ion exchange, concentration, drying and refining on the beta-limit dextrin prepared in the step (4) to prepare the beta-limit dextrin with the low DE value.
Preferably, in the step (2), the beta-amylase is added in an amount of 7X 104-7×105 0DP/t dry base starch.
According to the present invention, in the step (2), the saccharification reaction time is preferably 30 to 50 hours.
According to the invention, in the step (2), the enzyme inactivation condition is that the temperature is kept at 85-100 ℃ for 20-60 min.
According to the preferable selection of the invention, in the step (3), the decolorization and deproteinization treatment is carried out by adding active carbon with the mass percent of 0.5-2% of dry starch, treating for 20-60 min at the temperature of 50-85 ℃, and filtering to remove impurities.
Further preferably, the filtration is plate-and-frame filter-pressing filtration.
Preferably, in step (3), the desalting is desalting by cation-anion exchange.
According to the invention, in the step (4), the chromatographic separation is carried out at the temperature of 70-85 ℃.
According to the invention, in the step (4), the purity of maltose is more than 98%, and the beta-limit dextrin DE is 1-4%.
The decolorization, ion exchange, concentration, drying and refining in the step (5) and the step (6) are all conventional techniques in the field.
The invention has the advantages of
1. The inventor finds that after the conventional liquefaction treatment, small molecular saccharides with different polymerization degrees are generated, and the generation of the small molecular saccharides can cause difficulty in subsequent purification and increase of heterosaccharide, so that the product quality is seriously influenced; therefore, the inventor generates maltose and beta-limit dextrin through the action of beta-amylase after gelatinization, and is beneficial to separation, thereby obtaining high-purity maltose and low DE value beta-limit dextrin;
2. the waxy corn starch containing 100% of amylopectin is used as the raw material, so that the content of glucose, maltotriose, maltotetraose and other components is reduced, the subsequent purification is facilitated, and the prepared product has high purity and good quality;
3. according to the invention, after the saccharification step, a high-temperature system at 70-85 ℃ is adopted for separation, so that the viscosity of the feed liquid is prevented from being increased due to a large amount of beta-limit dextrin, and the problems of high column pressure and low separation efficiency in the conventional chromatographic separation at 60 ℃ are avoided, thereby being beneficial to reducing the viscosity of the product and ensuring the smooth separation and purification.
Detailed Description
In order to better illustrate the technical solutions of the present invention, the technical solutions of the present invention are further described below by way of examples, but the scope of the present invention is not limited thereto.
Source of raw materials
Waxy corn starch from Changchun Dacheng group
beta-Amylase available from Jenenaceae with enzyme Activity 14000DP/g (1)0DP units refer to the amount of enzyme required to reduce 5ml of a Fehlin reagent in a reaction of 0.1ml of an 5% strength enzyme sample solution with 100ml of a particular substrate at 20 ℃ for 1 hour
Example 1
A preparation method of high-purity maltose and beta-limit dextrin co-production comprises the following steps:
(1) mixing waxy corn starch and reverse osmosis water to prepare starch milk with the mass percentage concentration of 10 percent and the pH value of 4;
(2) heating the starch milk prepared in the step (1) to 70 ℃, gelatinizing for 60min, cooling to 50 ℃, and adding 7 x 10 beta-amylase4 0Carrying out saccharification reaction on DP/t dry base starch for 30 hours, and inactivating enzyme at 85 ℃ for 20min to obtain a saccharification product;
(3) carrying out decoloration and deproteinization treatment on the saccharified product prepared in the step (2), desalting and concentrating through cation and anion exchange to prepare a concentrated product with the fixed mass percentage of 40%;
the decolorization and deproteinization treatment comprises the steps of adding active carbon with the mass percent of 0.5% of dry starch, treating for 20min at the temperature of 50 ℃, and filtering to remove impurities through plate-and-frame filter pressing;
(4) carrying out chromatographic separation on the concentrated product prepared in the step (3) at the temperature of 70 ℃ to respectively prepare maltose and beta-limit dextrin;
the purity of maltose was determined to be 96.5%, the other 3.5%, and the DE value of beta-limit dextrin was 1.2%.
(5) Carrying out decoloration ion exchange, concentration, drying and refining on the maltose prepared in the step (4) to prepare high-purity maltose;
(6) and (4) carrying out decoloration ion exchange, concentration, drying and refining on the beta-limit dextrin prepared in the step (4) to prepare the beta-limit dextrin with the low DE value.
The purity of maltose is 99.5% and the DE value of beta-limit dextrin is 1% through detection.
Example 2
A preparation method of high-purity maltose and beta-limit dextrin co-production comprises the following steps:
(1) mixing waxy corn starch and reverse osmosis water to prepare starch milk with the mass percentage concentration of 20 percent and the pH value of 5;
(2) heating the starch milk prepared in the step (1) to 85 ℃, gelatinizing for 100min, cooling to 60 ℃, and adding beta3.5X 10 Amylase5 0DP/t dry base starch.
Performing saccharification reaction for 40 hours, and inactivating enzyme at 90 ℃ for 40min to obtain a saccharification product;
(3) carrying out decoloration and deproteinization treatment on the saccharified product prepared in the step (2), desalting and concentrating through cation and anion exchange to prepare a concentrated product with the fixed mass percentage of 50%;
the decolorization and deproteinization treatment comprises the steps of adding active carbon with the mass percent of 1% of dry starch, treating for 40min at the temperature of 70 ℃, and filtering to remove impurities through plate-and-frame filter pressing;
(4) carrying out chromatographic separation on the concentrated product prepared in the step (3) at the temperature of 75 ℃ to respectively prepare beta-limit dextrin;
the purity of maltose was tested to be 96.1%, the other 3.9%, and the DE value of beta-limit dextrin was 2.3%.
(5) Carrying out decoloration ion exchange, concentration, drying and refining on the maltose prepared in the step (4) to prepare high-purity maltose;
(6) and (4) carrying out decoloration ion exchange, concentration, drying and refining on the beta-limit dextrin prepared in the step (4) to prepare the beta-limit dextrin with the low DE value.
The purity of maltose is 99.3% and the DE value of beta-limit dextrin is 2% by detection.
Example 3
A preparation method of high-purity maltose and beta-limit dextrin co-production comprises the following steps:
(1) mixing waxy corn starch and reverse osmosis water to prepare starch milk with the mass percentage concentration of 30 percent and the pH value of 6;
(2) heating the starch milk prepared in the step (1) to 95 ℃, gelatinizing for 120min, cooling to 65 ℃, and adding 7 x 10 beta-amylase5 0DP/t dry base starch.
Carrying out saccharification reaction for 50 hours, and inactivating enzyme at 100 ℃ for 60min to obtain a saccharification product;
(3) carrying out decoloration and deproteinization treatment on the saccharified product prepared in the step (2), desalting and concentrating through cation and anion exchange to prepare a concentrated product with the fixed mass percentage of 60%;
the decolorization and deproteinization treatment comprises the steps of adding active carbon with the mass percent of 2% of dry starch, treating for 60min at the temperature of 85 ℃, and filtering to remove impurities through plate-and-frame filter pressing;
(4) carrying out chromatographic separation on the concentrated product prepared in the step (3) at the temperature of 85 ℃ to respectively prepare maltose and beta-limit dextrin;
the purity of maltose was found to be 95.7%, the other 4.3%, and the DE value of beta-limiting dextrin was 2.5%.
(5) Carrying out decoloration ion exchange, concentration, drying and refining on the maltose prepared in the step (4) to prepare high-purity maltose;
(6) and (4) carrying out decoloration ion exchange, concentration, drying and refining on the beta-limit dextrin prepared in the step (4) to prepare the beta-limit dextrin with the low DE value.
The purity of maltose is 98.7% and the DE value of beta-limit dextrin is 2.3%.
Comparative example 1
The process as described in example 2, except that the starting material was ordinary corn starch.
Common corn starch is available from Shandong West king corn Co.
A preparation method of high-purity maltose and beta-limit dextrin co-production comprises the following steps:
(1) mixing common corn starch with reverse osmosis water to prepare starch milk with the mass percentage concentration of 20 percent and the pH value of 5;
(2) heating the starch milk prepared in the step (1) to 85 ℃, gelatinizing for 100min, cooling to 60 ℃, and adding beta-amylase 3.5 multiplied by 105 0Carrying out saccharification reaction on DP/t dry base starch for 40 hours, and inactivating enzyme at 90 ℃ for 40min to obtain a saccharification product;
(3) carrying out decoloration and deproteinization treatment on the saccharified product prepared in the step (2), desalting and concentrating through cation and anion exchange to prepare a concentrated product with the fixed mass percentage of 50%;
the decolorization and deproteinization treatment comprises the steps of adding active carbon with the mass percent of 1% of dry starch, treating for 40min at the temperature of 70 ℃, and filtering to remove impurities through plate-and-frame filter pressing;
(4) carrying out chromatographic separation on the concentrated product prepared in the step (3) at the temperature of 75 ℃ to respectively prepare maltose and beta-limit dextrin;
the detection proves that the purity of the maltose is 90.5 percent, the content of the heterosaccharide such as glucose, maltotriose, maltotetraose and the like is 7.2 percent, other impurities are 2.3 percent, and the DE value of the beta-limit dextrin is 12.2 percent.
(5) Performing decoloration ion exchange, concentration, drying and refining on the maltose prepared in the step (4) to prepare maltose;
(6) and (4) carrying out decoloration ion exchange, concentration, drying and refining on the beta-limit dextrin prepared in the step (4) to prepare the beta-limit dextrin with the low DE value.
The detection proves that the purity of the maltose is 91.4 percent, the content of glucose, maltotriose, maltotetraose and other heterosugars is 8.6 percent, and the DE value of the beta-limit dextrin is 10.6 percent.
Comparative example 2
The preparation as described in example 2, except that a liquefaction step was used.
A preparation method of high-purity maltose and beta-limit dextrin co-production comprises the following steps:
(1) mixing waxy corn starch and reverse osmosis water to prepare starch milk with the mass percentage concentration of 20 percent and the pH value of 5;
(2) adding alpha high-temperature amylase into the starch milk prepared in the step (1), liquefying at 95-110 ℃, cooling to 60 ℃, and adding 3.5 multiplied by 10 beta-amylase5 0Carrying out saccharification reaction on DP/t dry base starch for 40 hours, and inactivating enzyme at 90 ℃ for 40min to obtain a saccharification product;
(3) carrying out decoloration and deproteinization treatment on the saccharified product prepared in the step (2), desalting and concentrating through cation and anion exchange to prepare a concentrated product with the fixed mass percentage of 50%;
the decolorization and deproteinization treatment comprises the steps of adding active carbon with the mass percent of 1% of dry starch, treating for 40min at the temperature of 70 ℃, and filtering to remove impurities through plate-and-frame filter pressing;
(4) carrying out chromatographic separation on the concentrated product prepared in the step (3) at the temperature of 75 ℃ to respectively prepare maltose and beta-limit dextrin;
the detection proves that the purity of the maltose is 80.1%, the small molecular sugar is 15.3%, other impurities are 4.6%, and the DE value of the beta-limit dextrin is 23%.
(5) Performing decoloration ion exchange, concentration, drying and refining on the maltose prepared in the step (4) to prepare maltose;
(6) and (4) carrying out decoloration ion exchange, concentration, drying and refining on the beta-limit dextrin prepared in the step (4) to prepare the beta-limit dextrin with the low DE value.
The purity of maltose is 82.7%, the purity of small molecular sugar is 17.3%, and the DE value of beta-limit dextrin is 20.7%.
The comparison of the data shows that compared with comparative examples 1 and 2, the product obtained in the embodiment of the invention reduces the content of components such as glucose, maltotriose, maltotetraose and the like, is beneficial to subsequent purification, and the prepared product has high purity and good quality and is beneficial to application in food.

Claims (5)

1. A preparation method for high-purity maltose and beta-limit dextrin co-production is characterized by comprising the following steps:
(1) mixing waxy corn starch and reverse osmosis water to prepare starch milk with the mass percentage concentration of 10-30% and the pH value of 4-6;
(2) heating the starch milk prepared in the step (1) to 70-95 ℃, gelatinizing for 60-120 min, cooling to 50-65 ℃, adding beta-amylase to perform saccharification reaction, and deactivating enzyme to prepare a saccharification product;
the addition amount of the beta-amylase is 7 multiplied by 104-7×1050DP/t dry base starch, and the saccharification reaction time is 30-50 hours;
(3) carrying out decoloration and protein removal treatment on the saccharified product prepared in the step (2), desalting and concentrating to prepare a concentrated product with the fixed mass percentage of 40-60%;
(4) carrying out chromatographic separation on the concentrated product prepared in the step (3) at the temperature of 70-85 ℃ to respectively prepare maltose and beta-limit dextrin;
(5) carrying out decoloration ion exchange, concentration, drying and refining on the maltose prepared in the step (4) to prepare high-purity maltose;
(6) and (4) carrying out decoloration ion exchange, concentration, drying and refining on the beta-limit dextrin prepared in the step (4) to prepare the beta-limit dextrin with the low DE value.
2. The preparation method according to claim 1, wherein in the step (2), the enzyme inactivation condition is that the temperature is kept at 85-100 ℃ for 20-60 min.
3. The preparation method according to claim 1, wherein in the step (3), the decolorization and deproteinization treatment is carried out by adding 0.5-2% by mass of activated carbon to dry starch, treating at 50-85 ℃ for 20-60 min, and filtering to remove impurities.
4. The method of claim 3, wherein the filtration is a plate and frame filter press filtration.
5. The method according to claim 1, wherein in the step (3), the desalting is desalting by cation-anion exchange.
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CN109234329A (en) * 2018-09-25 2019-01-18 山东百龙创园生物科技股份有限公司 A kind of preparation method of high-purity maltotetraose coproduction limit dextrin
CN112111542B (en) * 2020-09-01 2022-11-15 山东百龙创园生物科技股份有限公司 Preparation method of high-purity isomaltooligosaccharide co-produced resistant dextrin

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