CN108315374B - Preparation method of high-branch modified starch granules - Google Patents
Preparation method of high-branch modified starch granules Download PDFInfo
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- CN108315374B CN108315374B CN201810048745.3A CN201810048745A CN108315374B CN 108315374 B CN108315374 B CN 108315374B CN 201810048745 A CN201810048745 A CN 201810048745A CN 108315374 B CN108315374 B CN 108315374B
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- 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
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- 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
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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- 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
- C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins
Abstract
The invention relates to the technical field of starch production, and particularly discloses a green preparation method of ultrahigh-branch modified starch granules. The green preparation method of the ultrahigh-branch modified starch granules is characterized by comprising the following steps: the method comprises the steps of taking sweet potato starch as an acceptor and malto-oligosaccharide as a donor, and sequentially carrying out enzymatic hydrolysis and transglycosylation on the sweet potato starch by using branching enzyme, beta-amylase and glucoside transferase to obtain the ultrahigh-branch modified starch granules with high branch short-chain proportion. The method has the advantages of simple process, short preparation period, no need of adding special equipment, low production cost, suitability for industrial production, no need of any additive during production, safety and no pollution.
Description
Technical Field
The invention relates to the technical field of starch production, in particular to a preparation method of high-branch modified starch granules.
Background
Starch is widely used in the food and industrial fields as an important industrial raw material to enhance various processing properties of the product. Starch is also an important component of the human diet, which is digested by the human body and converted to glucose to provide the energy necessary for human life activities. However, with the advancement of food processing technology and the intensive research on starch science, it has been found that raw starch has great limitations in application: the stability is poor, and the aging is easy; the solubility is poor; in the starch gel, partial segments of starch molecules can displace to generate viscous flow, so that the elasticity of the starch gel is changed, and the starch gel has high hardness, rough feeling and the like. Meanwhile, the native starch has a high digestion rate in vivo, so that the postprandial blood sugar is easily and rapidly increased, and diseases such as hyperglycemia, obesity, cardiovascular diseases and the like are caused.
In order to solve the problems of the original starch, the modification of the original starch by adopting physical, chemical, genetic or enzymatic methods and composite modification methods is currently studied, so as to improve the physicochemical property and the digestion property of the original starch. The preparation method of the enzymatic modified starch has the advantages of environmental protection, simple and mild action conditions, easy digestion and decomposition by human bodies and the like.
Disclosure of Invention
The invention provides a preparation method of high-branch modified starch granules, which has the advantages of short preparation period, low production cost, safety and no pollution, and is used for making up the defects of the prior art.
The invention is realized by the following technical scheme:
a preparation method of high-branch modified starch granules is characterized by comprising the following steps: the method comprises the steps of taking sweet potato starch as an acceptor and malto-oligosaccharide as a donor, and sequentially carrying out enzymatic hydrolysis and transglycosylation on the sweet potato starch by using branching enzyme, beta-amylase and glucoside transferase to obtain the ultrahigh-branch modified starch granules with high branch short-chain proportion.
The invention adopts a compound enzyme method to modify sweet potato starch, firstly, when the sweet potato starch is acted by branching enzyme, alpha-1, 4-glycosidic bond in amylose and amylopectin linear region in the starch is hydrolyzed to generate cluster and some linear glucose residues, and simultaneously, the glucose residues are catalytically transferred to be connected to other chains to promote the formation of the alpha-1, 6-glycosidic bond, generate new branches, and increase the branch density of starch molecules. Then, beta-amylase is used for further hydrolysis to shorten the external chain length, finally, free glucose residues and malto-oligosaccharide are connected to other chains in a-1,6 glycosidic bond mode by using transglucosylation effect of glucosyltransferase to form more new short chain side branches, the branch density is further increased, a compact structure is formed, the ultrahigh branch modified starch granules containing higher branch short chain proportion number are obtained, the branch degree of the sweet potato starch is improved, and no chemical residue is left.
The method specifically comprises the following steps:
(1) preparing 5 (w/v)% starch solution from sweet potato starch, and adjusting pH of the starch solution to 6.5;
(2) adding branching enzyme into the starch solution, oscillating in water bath for hydrolysis, inactivating enzyme, and cooling to obtain enzymatic hydrolysate;
(3) adjusting the pH value of the enzymatic hydrolysate to 5.2, adding beta-amylase, oscillating and hydrolyzing in a water bath, then inactivating enzyme activity, and cooling to obtain enzymatic hydrolysate;
(4) adding malto-oligosaccharide into the enzymatic hydrolysate, uniformly mixing, adjusting the pH to 5.0, adding glucoside transferase, placing in a water bath for oscillating enzymolysis, then inactivating enzyme, and cooling to obtain a sample solution;
(5) and adding absolute ethyl alcohol into the sample solution, centrifuging, and freeze-drying the obtained precipitate to obtain the product.
The more preferable technical scheme of the invention is as follows:
in the step (1), 10g of sweet potato starch is weighed, and 0.02M sodium acetate buffer solution with pH of 6.5 is added until the volume of starch slurry is 200 mL.
In the step (2), 308U/g of branching enzyme is added into the starch solution, the starch solution is placed in a water bath at 60 ℃ for oscillation for 20-24h, and then 2.5mL of 1mol/L NaOH solution is added for inactivating the enzyme activity.
In the step (3), 10u/g of beta-amylase is added into the enzymatic hydrolysate, the enzymatic hydrolysate is placed in a water bath at 37 ℃ for oscillation for 4-6h, and then 2.5mL of 1mol/L NaOH solution is added for inactivating the enzyme.
In the step (4), adding malto-oligosaccharide accounting for 5% of the weight of the enzymatic hydrolysate, uniformly mixing, adding 9088U/g of glucoside transferase, placing in a water bath at 55 ℃ for shaking enzymolysis for 10-12h, and then adding 2.5mL of 1mol/L NaOH solution for inactivating enzyme; wherein the malto-oligosaccharide is formed by mixing 20% of maltotetraose, 30% of maltopentaose, 40% of maltohexaose and 10% of maltoheptaose in weight ratio.
In the step (5), 3 times of volume of absolute ethyl alcohol is added into the sample solution, and the mixture is centrifuged for 10min under the condition of 3000r/min to obtain a precipitate.
The method has the advantages of simple process, short preparation period, no need of adding special equipment, low production cost, suitability for industrial production, no need of any additive during production, safety and no pollution.
Drawings
The invention will be further described with reference to the accompanying drawings.
FIG. 1 is SEM electron micrograph of raw sweet potato starch;
FIG. 2 is an SEM electron micrograph of a modified starch granule product of the present invention;
FIG. 3 is an image of the original sweet potato starch under a polarizing microscope by black cross observation;
FIG. 4 is an image of a modified starch granule of the present invention obtained by observing it with a polarizing microscope under a black cross.
Detailed Description
Example (b): preparation method of high-branch modified starch granules
The method specifically comprises the following steps:
(1) preparation of sweet potato starch solution: accurately weighing 10g of sweet potato starch sample to prepare 5% (w/v) starch solution, and adjusting the pH value of the starch solution to 6.5;
(2) and (3) enzymolysis by using branching enzyme: branching enzyme (308U/g), placing in water bath at 60 ℃ for oscillation for 24h, then adding 2.5mL of 1mol/L NaOH solution to inactivate the enzyme, and cooling;
(3) beta-amylase hydrolysis: adjusting the pH value of the solution to 5.2, adding 10U/g of beta-amylase, carrying out enzymolysis for 6h at the reaction temperature of 37 ℃, adding 2.5mL of 1mol/L NaOH solution to inactivate the enzyme, and cooling;
(4) addition of malto-oligosaccharide: weighing 5g of malto-oligosaccharide mixture comprising 20% of maltotetraose, 30% of maltopentaose, 40% of maltohexaose and 10% of maltoheptaose, adding into the enzymatic hydrolysate, and mixing uniformly;
(5) and (3) carrying out enzymolysis on glucoside transferase: adjusting the pH value of the solution to 5.0, adding 9088U/g of glucoside transferase, placing in a water bath at 55 ℃ for shaking and enzymolysis for 12h, and then adding 2.5mL of 1mol/L NaOH solution to inactivate the enzyme;
(6) and cooling the sample liquid to room temperature, adding 3 times of volume of absolute ethyl alcohol, centrifuging at 3000r/min for 10min, and freeze-drying the obtained precipitate to obtain the ultrahigh-branch modified starch granules.
The physicochemical and digestive characteristics of the modified starch granules obtained in the example are detected, and the detection proves that compared with the original starch, the solubility of the modified starch granules is remarkably improved from 9.86% to 47.65%, and the viscosity of the starch paste is increased by 43.24%; the gelatinization peak temperature is reduced from 68.98 ℃ to 56.94 ℃; the aging degree of the starch paste is obviously reduced from 58.94% to 36.57% when the starch paste is stored for one week at 4 ℃; the slowly digested and resistant starch contents increased significantly from 24.65% and 8.68% to 48.34% and 26.87%, respectively.
The modified starch granules obtained in this example were subjected to an application test:
(1) the additive is added into flour products to delay aging and improve the taste, viscosity stability, elasticity and the like of the flour products;
(2) can be used as additive for foundation make-up in cosmetic industry;
(3) the starch is used as a multifunctional food additive in food processing, can facilitate the processing technology, provide excellent texture for food, and improve the thickening, suspending, water retaining and stabilizing capabilities of starch;
(4) used as a functional ingredient in various foods, remarkably reduces the digestion rate and thus lowers the release amount of blood sugar.
Claims (6)
1. A preparation method of high-branch modified starch granules is characterized by comprising the following steps: taking sweet potato starch as an acceptor and malto-oligosaccharide as a donor, sequentially carrying out enzymatic hydrolysis and enzyme deactivation cooling on the sweet potato starch by using branching enzyme and beta-amylase to obtain an enzymatic hydrolysate, adding the malto-oligosaccharide into the enzymatic hydrolysate, uniformly mixing, adjusting the pH to 5.0, adding glucoside transferase, placing in a water bath for oscillating enzymolysis, then carrying out enzyme deactivation, and cooling to obtain high-branch modified starch granules containing high branch short chain proportion number;
the method specifically comprises the following steps: (1) accurately weighing 10g of sweet potato starch sample to prepare 5% (w/v) starch solution, and adjusting the pH value of the starch solution to 6.5; (2) adding branching enzyme into the starch solution, oscillating in water bath for hydrolysis, inactivating enzyme, and cooling to obtain enzymatic hydrolysate; (3) adjusting the pH value of the enzymatic hydrolysate to 5.2, adding beta-amylase, oscillating and hydrolyzing in a water bath, then inactivating enzyme activity, and cooling to obtain enzymatic hydrolysate; (4) adding 5g of malto-oligosaccharide into the enzymatic hydrolysate, uniformly mixing, adjusting the pH to 5.0, adding glucoside transferase, placing in a water bath for oscillating enzymolysis, then inactivating enzyme, and cooling to obtain a sample solution; the malto-oligosaccharide is formed by mixing 20% of maltotetraose, 30% of maltopentaose, 40% of maltohexaose and 10% of maltoheptaose in weight ratio; (5) and adding absolute ethyl alcohol into the sample solution, centrifuging, and freeze-drying the obtained precipitate to obtain the product.
2. The method for preparing highly branched modified starch granules according to claim 1, wherein: in the step (1), the sweet potato starch is added into 0.02M sodium acetate buffer solution with pH of 6.5 until the volume of starch slurry is 200 mL.
3. The method for preparing highly branched modified starch granules according to claim 1, wherein: in the step (2), 308U/g of branching enzyme is added into the starch solution, the starch solution is placed in a water bath at 60 ℃ for oscillation for 20-24h, and then 2.5mL of 1mol/L NaOH solution is added for inactivating the enzyme activity.
4. The method for preparing highly branched modified starch granules according to claim 1, wherein: in the step (3), 10U/g beta-amylase is added into the enzymatic hydrolysate, the enzymatic hydrolysate is placed in a water bath at 37 ℃ for oscillation for 4-6h, and then 2.5mL of 1mol/L NaOH solution is added for inactivating the enzyme.
5. The method for preparing highly branched modified starch granules according to claim 1, wherein: in the step (4), 9088U/g of glucoside transferase is added, the mixture is placed in a water bath at 55 ℃ for shaking enzymolysis for 10-12 hours, and then 2.5mL of 1mol/L NaOH solution is added for inactivating the enzyme activity.
6. The method for preparing highly branched modified starch granules according to claim 1, wherein: in the step (5), 3 times of volume of absolute ethyl alcohol is added into the sample solution, and the mixture is centrifuged for 10min under the condition of 3000r/min to obtain a precipitate.
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| CN109988798A (en) * | 2019-04-16 | 2019-07-09 | 齐鲁工业大学 | A kind of environment-friendly preparation method thereof of rice resistant starch |
| CN110438183A (en) * | 2019-07-25 | 2019-11-12 | 哈尔滨商业大学 | Combined-enzyme method modified starch prepares the application of the method and low DE value maltodextrin of low DE value maltodextrin in food |
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| FR2840612B1 (en) * | 2002-06-06 | 2005-05-06 | Roquette Freres | HIGHLY BRANCHED SOLUBLE GLUCOSE POLYMERS AND PROCESS FOR OBTAINING THEM |
| KR100868329B1 (en) * | 2007-02-01 | 2008-11-12 | 씨제이제일제당 (주) | A method for preparing enzymatically highly branched-amylose and amylopectin cluster |
| CN101715911A (en) * | 2009-12-10 | 2010-06-02 | 上海市农业科学院 | Method for preparing digestion starch containing slow digestion starch by enzymolysis |
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