CN115028748B - Method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification - Google Patents
Method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification Download PDFInfo
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Abstract
The invention discloses a method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification. The method comprises the following steps: preparing waxy corn starch into 10-20% starch emulsion, debranching the starch emulsion by using pullulanase, and preparing debranched starch. Preparing debranched starch into a 10-20% solution, adding a certain amount of malic acid (the mass ratio of malic acid to starch is 0.5-0.7), regulating the pH value to 1.5-2.5 by NaOH, standing for 11 h, drying at 50 ℃ until the water content is lower than 10%, crushing, and reacting at 130-150 ℃ for 4-6 h. Washing, drying, crushing and sieving to obtain the malic acid esterified debranched starch. The substitution degree of the product is up to 0.866, the content of resistant starch is up to 95.78%, and the product can be used in low GI value foods.
Description
Technical Field
The invention belongs to the technical field of modified starch production, and particularly relates to a method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification.
Background
Starch is a major source of energy in daily life, but has the disadvantages of cold water intolerance, high viscosity, low transparency, intolerance to enzymes and the like, and is greatly limited in application. Therefore, the starch modification is always a hot spot in the field of starch, the modified starch has a changed structure, the performance is greatly improved, and the modified starch is widely applied to the fields of food and the like. Resistant Starch (RS) refers to starch that cannot be enzymatically hydrolyzed in the small intestine but can react with volatile fatty acids in the human gastrointestinal colon. The resistant starch can effectively improve postprandial blood sugar load, thereby regulating blood sugar of a human body and preventing diabetes. In addition, the resistant starch can produce short chain fatty acid, has a regulating effect on intestinal flora and reduces the incidence rate of intestinal diseases. Therefore, the development of novel resistant starches has been a research hotspot in the low GI food field.
Esterified starch is a resistant starch (RS 4) obtained by reacting an organic or inorganic acid with starch, such as starch phosphate, starch sulfate, starch acetate, starch citrate, starch alkenyl succinate, etc. The digestion resistance and the thermal stability of the esterified starch are remarkably high, and the improvement degree is proportional to the substitution degree. However, it has been found that the degree of substitution of the esterified starch is still relatively low and that the content of resistant starch is to be increased.
Pullulanase is a starch debranching enzyme capable of specifically cutting alpha-1, 6 glycosidic bonds in amylopectin, and cutting off the whole branch structure to produce short amylose. Therefore, the invention mainly uses pullulanase to debranch waxy corn starch, thereby generating more hydroxyl groups to promote the esterification reaction of malic acid and starch, and further improving the substitution degree and the resistant starch content of the malic acid esterified starch.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification, which mainly uses pullulanase to debranch waxy corn starch so as to generate more hydroxyl sites to promote the malic acid esterification reaction, thereby improving the substitution degree to improve the content of the resistant starch.
In order to solve the technical problems, the invention adopts the following technical scheme:
the method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification is characterized by comprising the following steps:
(1) Boiling 10-20% of waxy corn starch emulsion in a boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and oscillating in a constant-temperature water bath shaking table at 50 ℃ to debranch 24 and h to obtain debranched starch solution;
(2) Boiling debranched starch solution, inactivating enzyme, adding absolute ethanol for precipitation, centrifuging to obtain precipitate, freeze-drying 24 h, grinding and sieving to obtain debranched starch;
(3) Preparing debranched starch into a 10-20% solution, adding malic acid, regulating the pH value of the solution to 1.5-2.5 by NaOH, standing at room temperature for 11-h, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining a debranched starch-malic acid premix;
(4) And (3) crushing the debranched starch-malic acid premix, placing the crushed debranched starch-malic acid premix in a glass vessel, reacting for 4-6 hours at 130-150 ℃, washing the crushed debranched starch-malic acid premix with water for multiple times until the pH value of the washing solution is neutral, and drying, crushing and sieving the crushed debranched starch-malic acid premix to obtain the malic acid esterified debranched starch.
In the step (2), the mass ratio of the debranched starch solution to the absolute ethyl alcohol is 1:2-1:4.
In the step (3), the ratio of the added malic acid to the starch is 0.5-0.7.
In the step (3), naOH is added to adjust the pH value to be 1.5-2.5.
In the step (4), the reaction temperature is 130-150 ℃.
The product prepared by the invention is odorless and white powder, the substitution degree is up to 0.866, and the content of resistant starch is up to 95.78%.
The cooling, drying, pulverizing, etc. are all common techniques in the current food processing field.
Compared with the prior art, the invention has the following advantages:
(1) The enzyme modification can react with specific chemical bonds, and has few byproducts and high safety.
(2) Malic acid is one of the food additives and is naturally present in fruits. The malic acid is oxidized to obtain oxaloacetic acid, which is an important intermediate product of internal circulation metabolism of human body. Harmless to human body, and the obtained product can be directly applied to food production.
(3) The substitution degree and the resistant starch content of the product prepared by the invention are high, and the product can be used as a novel dietary fiber and also can be used in low GI functional foods.
Detailed Description
The invention will be further illustrated with reference to specific examples. It is to be understood that the following examples are intended to illustrate the present invention and are not to be construed as limiting the scope of the invention, and that numerous insubstantial modifications and adaptations can be made by those skilled in the art in light of the foregoing disclosure.
Example 1
The method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification comprises the following steps:
(1) Boiling 10% waxy corn starch emulsion in boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and shaking in a 50 ℃ constant temperature water bath shaking table to remove branches 24 and h to obtain debranched starch solution;
(2) Boiling debranched starch solution, inactivating enzyme, adding absolute ethanol according to a ratio of 1:2 for precipitation, centrifuging to obtain precipitate, freeze-drying 24 h, grinding and sieving to obtain debranched starch;
(3) Preparing debranched starch into a 10% solution, adding malic acid (the mass ratio of malic acid to starch is 0.5), regulating the pH value of the solution to 2 by NaOH, standing at room temperature for 11 hours, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining a debranched starch-malic acid premix;
(4) Pulverizing debranched starch-malic acid premix, placing in a glass vessel, reacting at 140 deg.C for 4 h, washing with water for several times until the pH of the washing solution is neutral, drying, pulverizing, and sieving to obtain malic acid esterified debranched starch.
The product is white powder, and has no foreign odor. The degree of substitution was 0.482 and the resistant starch content was 79.97%.
Example 2
The method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification comprises the following steps:
(1) Boiling 15% waxy corn starch emulsion in boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and shaking in a 50 ℃ constant temperature water bath shaking table to remove branches 24 and h to obtain debranched starch solution;
(2) Boiling debranched starch solution, inactivating enzyme, adding absolute ethanol according to a ratio of 1:3 for precipitation, centrifuging to obtain precipitate, freeze-drying 24 h, grinding and sieving to obtain debranched starch;
(3) Preparing debranched starch into a 15% solution, adding malic acid (the mass ratio of malic acid to starch is 0.6), regulating the pH value of the solution to 1.5 by NaOH, standing at room temperature for 11 hours, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining debranched starch-malic acid premix;
(4) Pulverizing debranched starch-malic acid premix, placing in a glass vessel, reacting at 140 deg.C for 5 h, washing with water for several times until the pH of the washing solution is neutral, drying, pulverizing, and sieving to obtain malic acid esterified debranched starch.
The product is white powder, and has no foreign odor. The degree of substitution was 0.745 and the resistant starch content was 95.01%.
Example 3
The method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification comprises the following steps:
(1) Boiling 20% waxy corn starch emulsion in boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and shaking in a 50 ℃ constant temperature water bath shaking table to remove branches 24 and h to obtain debranched starch solution;
(2) Boiling debranched starch solution, inactivating enzyme, adding absolute ethanol according to a ratio of 1:4 for precipitation, centrifuging to obtain precipitate, freeze-drying 24 h, grinding and sieving to obtain debranched starch;
(3) Preparing debranched starch into a 20% solution, adding malic acid (the mass ratio of malic acid to starch is 0.6), regulating the pH value of the solution to 2.5 by NaOH, standing at room temperature for 11 h, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining debranched starch-malic acid premix;
(4) Pulverizing debranched starch-malic acid premix, placing in a glass vessel, reacting at 130 ℃ for 6 h, washing with water for many times until the pH of the washing solution is neutral, drying, pulverizing, and sieving to obtain malic acid esterified debranched starch.
The product is white powder, and has no foreign odor. The degree of substitution was determined to be 0.613 and the resistant starch content was 92.80%.
Example 4
The method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification comprises the following steps:
(1) Boiling 10% waxy corn starch emulsion in boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and shaking in a 50 ℃ constant temperature water bath shaking table to remove branches 24 and h to obtain debranched starch solution;
(2) Boiling debranched starch solution, inactivating enzyme, adding absolute ethanol according to a ratio of 1:2 for precipitation, centrifuging to obtain precipitate, freeze-drying 24 h, grinding and sieving to obtain debranched starch;
(3) Preparing debranched starch into a 10% solution, adding malic acid (the mass ratio of malic acid to starch is 0.7), regulating the pH value of the solution to 1.5 by NaOH, standing at room temperature for 11 h, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining debranched starch-malic acid premix;
(4) Pulverizing debranched starch-malic acid premix, placing in a glass vessel, reacting at 150deg.C for 6 h, washing with water for several times until pH of the washing solution is neutral, drying, pulverizing, and sieving to obtain malic acid esterified debranched starch.
The product is white powder, and has no foreign odor. The degree of substitution was determined to be 0.603 and the resistant starch content was determined to be 92.69%.
Example 5
The method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification comprises the following steps:
(1) Boiling 10% waxy corn starch emulsion in boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and shaking in a 50 ℃ constant temperature water bath shaking table to remove branches 24 and h to obtain debranched starch solution;
(2) Boiling debranched starch solution, inactivating enzyme, adding absolute ethanol according to a ratio of 1:2 for precipitation, centrifuging to obtain precipitate, freeze-drying 24 h, grinding and sieving to obtain debranched starch;
(3) Preparing debranched starch into a 10% solution, adding malic acid (the mass ratio of malic acid to starch is 0.7), regulating the pH value of the solution to 2 by NaOH, standing at room temperature for 11 h, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining a debranched starch-malic acid premix;
(4) Pulverizing debranched starch-malic acid premix, placing in a glass vessel, reacting at 130 ℃ for 6 h, washing with water for many times until the pH of the washing solution is neutral, drying, pulverizing, and sieving to obtain malic acid esterified debranched starch.
The product is white powder, and has no foreign odor. The degree of substitution was determined to be 0.866 and the resistant starch content was determined to be 95.78%.
Example 6
The method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification comprises the following steps:
(1) Boiling 10% waxy corn starch emulsion in boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and shaking in a 50 ℃ constant temperature water bath shaking table to remove branches 24 and h to obtain debranched starch solution;
(2) Boiling debranched starch solution, inactivating enzyme, adding absolute ethanol according to a ratio of 1:2 for precipitation, centrifuging to obtain precipitate, freeze-drying 24 h, grinding and sieving to obtain debranched starch;
(3) Preparing debranched starch into a 10% solution, adding malic acid (the mass ratio of malic acid to starch is 0.7), regulating the pH value of the solution to 2.5 by NaOH, standing at room temperature for 11 h, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining debranched starch-malic acid premix;
(4) Pulverizing debranched starch-malic acid premix, placing in a glass vessel, reacting at 140 deg.C for 6 h, washing with water for many times until the pH of the washing solution is neutral, drying, pulverizing, and sieving to obtain malic acid esterified debranched starch.
The product is white powder, and has no foreign odor. The degree of substitution was determined to be 0.644 and the resistant starch content was determined to be 93.54%.
Example 7
The method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification comprises the following steps:
(1) Boiling 10% waxy corn starch emulsion in boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and shaking in a 50 ℃ constant temperature water bath shaking table to remove branches 24 and h to obtain debranched starch solution;
(2) Boiling debranched starch solution, inactivating enzyme, adding absolute ethanol according to a ratio of 1:2 for precipitation, centrifuging to obtain precipitate, freeze-drying 24 h, grinding and sieving to obtain debranched starch;
(3) Preparing debranched starch into a 10% solution, adding malic acid (the mass ratio of malic acid to starch is 0.5), regulating the pH value of the solution to 1.5 by NaOH, standing at room temperature for 11 h, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining debranched starch-malic acid premix;
(4) Pulverizing debranched starch-malic acid premix, placing in a glass vessel, reacting at 130 ℃ for 6 h, washing with water for many times until the pH of the washing solution is neutral, drying, pulverizing, and sieving to obtain malic acid esterified debranched starch.
The product is white powder, and has no foreign odor. The degree of substitution was determined to be 0.42 and the resistant starch content was determined to be 70.23%.
Example 8
The method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification comprises the following steps:
(1) Boiling 10% waxy corn starch emulsion in boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and shaking in a 50 ℃ constant temperature water bath shaking table to remove branches 24 and h to obtain debranched starch solution;
(2) Boiling debranched starch solution, inactivating enzyme, adding absolute ethanol according to a ratio of 1:2 for precipitation, centrifuging to obtain precipitate, freeze-drying 24 h, grinding and sieving to obtain debranched starch;
(3) Preparing debranched starch into a 10% solution, adding malic acid (the mass ratio of malic acid to starch is 0.5), regulating the pH value of the solution to 2.5 by NaOH, standing at room temperature for 11 h, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining debranched starch-malic acid premix;
(4) Pulverizing debranched starch-malic acid premix, placing in a glass vessel, reacting at 150deg.C for 6 h, washing with water for several times until pH of the washing solution is neutral, drying, pulverizing, and sieving to obtain malic acid esterified debranched starch.
The product is white powder, and has no foreign odor. The degree of substitution was determined to be 0.456 and the resistant starch content was 76.58%.
Example 9
The method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification comprises the following steps:
(1) Boiling 10% waxy corn starch emulsion in boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and shaking in a 50 ℃ constant temperature water bath shaking table to remove branches 24 and h to obtain debranched starch solution;
(2) Boiling debranched starch solution, inactivating enzyme, adding absolute ethanol according to a ratio of 1:2 for precipitation, centrifuging to obtain precipitate, freeze-drying 24 h, grinding and sieving to obtain debranched starch;
(3) Preparing debranched starch into a 10% solution, adding malic acid (the mass ratio of malic acid to starch is 0.6), regulating the pH value of the solution to 2 by NaOH, standing at room temperature for 11 h, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining a debranched starch-malic acid premix;
(4) Pulverizing debranched starch-malic acid premix, placing in a glass vessel, reacting at 150deg.C for 6 h, washing with water for several times until pH of the washing solution is neutral, drying, pulverizing, and sieving to obtain malic acid esterified debranched starch.
The product is white powder, and has no foreign odor. The degree of substitution was determined to be 0.546 and the resistant starch content was determined to be 86.75%.
The present invention is not limited to the above-described embodiments, and the above-described embodiments and descriptions are merely illustrative of the principles of the present invention, and various changes and modifications may be made therein without departing from the spirit and scope of the invention, which is defined in the appended claims. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (1)
1. The method for preparing high-content resistant starch by enzymolysis debranching and malic acid esterification is characterized by comprising the following steps:
(1) Boiling 10-20% of waxy corn starch emulsion in a boiling water bath, sterilizing at high temperature, completely gelatinizing, cooling, adding pullulanase, and oscillating in a constant-temperature water bath shaking table at 50 ℃ to debranch 24 and h to obtain debranched starch solution;
(2) Boiling a debranching starch solution, inactivating enzyme, adding absolute ethyl alcohol for precipitation, wherein the mass ratio of the debranching starch solution to the absolute ethyl alcohol is 1:2-1:4, centrifuging to obtain a precipitate, freeze-drying 24 h, grinding and sieving to obtain debranching starch;
(3) Preparing debranched starch into a 10-20% solution, adding malic acid, wherein the ratio of the added malic acid to the starch is 0.6-0.7, regulating the pH value of the solution to 1.5-2.5 by NaOH, standing at room temperature for 11 h, and drying at 50 ℃ until the water content is lower than 10%, thus obtaining a debranched starch-malic acid premix;
(4) And (3) crushing the debranched starch-malic acid premix, placing the crushed debranched starch-malic acid premix in a glass vessel, reacting for 5-6 hours at 130-150 ℃, washing the crushed debranched starch-malic acid premix with water for multiple times until the pH value of the washing solution is neutral, and drying, crushing and sieving the crushed debranched starch-malic acid premix to obtain the malic acid esterified debranched starch.
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