CN113845600A - Preparation method of high-amylose mung bean resistant starch - Google Patents
Preparation method of high-amylose mung bean resistant starch Download PDFInfo
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Abstract
The invention relates to a preparation method of high-amylose mung bean resistant starch, which specifically comprises the following steps: (1) uniformly mixing mung bean powder and water, adjusting pH, carrying out alkali extraction, sequentially sieving and centrifuging to obtain lower-layer crude starch, and sequentially cleaning, centrifuging, drying, crushing and sieving the lower-layer crude starch to obtain mung bean starch; (2) adding water into the mung bean starch obtained in the step (1) to prepare starch paste, cooling after gelatinization, adjusting the pH value, carrying out enzymolysis on the mung bean starch by using pullulanase, carrying out ultrasonic treatment while carrying out enzymolysis, inactivating enzymes after the enzymolysis is finished, carrying out aging, drying, crushing and sieving to obtain the high-amylose mung bean resistant starch. Compared with the prior art, the technology adopts the ultrasonic combined enzyme method to prepare the mung bean resistant starch, and can prepare more mung bean resistant starch and more amylose under the same mung bean dosage.
Description
Technical Field
The invention belongs to the field of functional foods, and particularly relates to a preparation method of high-amylose mung bean resistant starch.
Background
Resistant Starch (RS) is a general name of starch and starch degradation products thereof which can not be absorbed by small intestine of healthy human body, but can be fermented and degraded by microorganisms in intestinal tract, has physiological functions similar to dietary fiber, can directly enter large intestine to be fermented by physiological bacteria, generates a plurality of short chain fatty acids and gases, and has the functions of preventing gastrointestinal diseases and cardiovascular diseases, reducing the risk of conjunctivitis and colon cancer, promoting the growth of beneficial microorganisms in intestinal tract and the absorption of mineral elements, enhancing disease resistance and the like. RS in foods can be broadly divided into five categories: RS1, physical embedded starch; RS2, resistant starch granules; RS3, retrograded starch; RS4, chemically modified starch; RS5, starch-lipid complex. The resistant starch has good processing characteristics, and a proper amount of the resistant starch is added into food, so that the resistant starch not only can be used as a good structure modifier, but also can be used as an excellent dietary fiber enhancer, and can be used for developing high-quality functional health-care food.
Although the starch resources are rich and the varieties are complete in China, the common varieties such as common corn starch, high-amylose corn starch, potato starch and the like are mainly researched by domestic resistant starch, the research on edible beans is less, and a new starch variety and a new process are required to be searched for to improve the content of the resistant starch. The mung beans have a long planting history in China, the variety resources are rich, the total yield is in the top of the world, the mung beans are produced and exported in large countries, and the yield of the mung beans in the whole country in 2019 reaches 2053 ten thousand tons. The mung bean starch in China is excellent in variety, the starch content of the mung bean is as high as about 60%, the amylose content of the mung bean is about 30%, and the coagulation property is good, but at the present stage, the mung bean is mainly used for producing traditional foods such as vermicelli, bean jelly, cakes and the like, the processing level is not high, the added value of the product is low, and the commercialization degree is low, so that the full utilization of mung bean resources is necessary to be enhanced, and related products for fine and deep processing of the mung bean are researched and developed, and the added value of the product is improved.
In the conventional preparation process of the resistant starch, an important factor influencing the yield of the resistant starch is the amylose content in the starch, starch granules swell and break to release the amylose in the process of heating and pasting, then the starch granules are cooled and retrograded, and long-chain polymers are overlapped through double helix (namely, the amylose is recrystallized) to form the resistant starch. The RS3 type resistant starch is prepared by heat treatment, debranching or combination of heat treatment and debranching, and then coagulating and recrystallizing under certain conditions. In the preparation process of the resistant starch, two existing debranching methods are adopted, one is enzyme debranching, and the enzyme method for preparing the resistant starch can greatly reduce the using amount of chemical reagents in the preparation process, improve the quality of the resistant starch and reduce the pollution to the environment, but has the defects of long enzymolysis time and low preparation yield of the resistant starch. The other method is acid hydrolysis, which is to treat starch with acid (hydrochloric acid, sulfuric acid, nitric acid, etc.)), but the debranching effect and safety are not as good as those of the enzyme debranching effect, and the high corrosiveness of acid to equipment is a technical problem which needs to be considered in practical production. The Kim and the Kwak take corn starch as a raw material, and the content of resistant starch in a product obtained by retrogradation through autoclaving under different conditions reaches 9% -12%, so that the effect is more obvious through repeated cold and heat circulation. Gonzalez-Soto et al report that after the autoclaved debranched banana starch is retrograded at different storage temperatures (4-60 ℃) and times (24.0-48.0h), the resistant starch content reaches 13.4% -26.4%. The Weiguyu and the like take cassava starch as a raw material, the concentration of the starch milk is 20%, the starch milk is gelatinized at high temperature, pullulanase is added to act (4u/g dry starch, 8.0h, 55 ℃), and the RS content is 13.36%. The related researches show that the yield and efficiency of the resistant starch prepared by a physical (such as an autoclave method, a wet-heat method and the like) or biochemical method are low, and the problems of long operation time, high energy consumption and safety exist.
At present, few researches on preparation of mung bean resistant starch are carried out at home and abroad, no mung bean resistant starch commodity is sold on the market, mung bean starch which is a characteristic starch variety in China is taken as a raw material, mung bean resistant starch prepared by different methods and researches on application characteristics of the mung bean resistant starch have positive significance for promoting development of deep processing of the mung bean starch in China, abundant mung bean resources in China can be fully developed and utilized, the additional value of the mung bean deep-processed products is improved, and meanwhile, the social benefit and the economic benefit of the resistant starch are also effectively improved.
Disclosure of Invention
The invention aims to provide a preparation method of high-amylose mung bean resistant starch.
The purpose of the invention is realized by the following technical scheme:
a preparation method of high-amylose mung bean resistant starch specifically comprises the following steps:
(1) extraction of mung bean starch
Uniformly mixing mung bean powder and water, adjusting pH, carrying out alkali extraction, sequentially sieving and centrifuging to obtain lower-layer crude starch, and sequentially cleaning, centrifuging, drying, pulverizing and sieving the lower-layer crude starch to obtain mung bean starch (MS), wherein the mung bean starch contains amylose and amylopectin;
(2) preparation of resistant starch (ultrasonic combined enzymolysis method)
Adding water into the mung bean starch obtained in the step (1) to prepare starch paste, cooling after gelatinization, adjusting pH, carrying out enzymolysis on the mung bean starch by pullulanase, carrying out ultrasonic treatment while carrying out enzymolysis, inactivating enzyme after the enzymolysis is finished, aging, drying, crushing and sieving to obtain the high-amylose mung bean resistant starch (MRS), wherein the high-amylose mung bean resistant starch contains amylose and amylopectin, the content of the amylose is higher than that of the amylose in the mung bean starch, and the higher the content of the amylose in the resistant starch is, the better the digestion resistance of the resistant starch is.
In the step (1), the mung beans are ground in a grinding machine to prepare mung bean powder.
In the step (1), the ratio of the mung bean powder to water is 1:6-1:10(w/v), and the water is distilled water.
Preferably, the ratio of mung bean flour to water is 1:8 (w/v).
In step (1), NaOH, which is a commonly used alkali for adjusting pH, is used to adjust the pH of the mung bean flour solution to 9.0 to 11.0, preferably 10.0.
Preferably, the concentration of NaOH is 2 mol/L.
In the step (1), alkali extraction is carried out in water bath at 35-45 ℃ for 2.0-4.0 h.
Preferably, the alkali extraction is carried out in a water bath at 40 ℃ for 3.0 h.
In the step (1), the two sieving processes are both 100-mesh sieving.
In the step (1), the filtrate obtained by sieving is centrifuged at 3500-.
In the step (1), the washing and centrifuging process specifically comprises the following steps: and adding water into the lower-layer crude starch for cleaning, centrifuging again, repeating for multiple times to obtain lower-layer starch, washing the lower-layer starch with absolute ethyl alcohol, centrifuging, and then continuing to wash with water and centrifuging to finally obtain starch precipitate.
Preferably, the first washing with water is repeated 3 times.
Preferably, the water is distilled water.
In the step (1), drying is carried out in a forced air drying oven, the drying temperature is 40 ℃, and the drying time is 24.0-30.0 h.
In the step (1), the total starch content of the obtained mung bean starch is higher than 80%.
In the step (2), the ratio of the mung bean starch to the water is 1:15-1:25(w/v), and preferably 1:20 (w/v).
In the step (2), the gelatinization is carried out in a boiling water bath for 20-30 min.
In step (2), cooling is carried out to 45-55 ℃, preferably 50 ℃.
In step (2), the pH is adjusted to 4.5-5.5, preferably 5.0, using HCl, which is also the acid commonly used to adjust the pH.
Preferably, the concentration of HCl is 0.05 mol/L.
In the step (2), the enzyme activity of the pullulanase is more than or equal to 1000npun/g, and the pullulanase is produced by sigma company.
In the step (2), the addition amount of pullulanase is 20-30npun/g, preferably 27.5npun/g starch (the starch refers to the mung bean starch obtained in the step (1)), the enzyme dosage is related to the enzyme activity, and the enzyme activities produced by different manufacturers are different, but the invention has the advantage of small enzyme dosage in general.
In the step (2), the enzymolysis temperature is 45-50 ℃, and the enzymolysis time is 10.0-14.0 h. The main role of pullulanase is to remove branches and produce amylose molecules, because the higher the amylose content in starch, the more chance there is for crystals to form between amylose molecules when the starch paste is aged, and the corresponding increase in the yield of RS. The enzymolysis time in part of documents reaches 24.0h even 36.0h, while the enzymolysis time in the invention only needs 10.0-14.0h, which is far shorter than the time reported in the documents. The effect is obtained by simultaneously carrying out ultrasonic and enzymolysis reactions, the ultrasonic is beneficial to the diffusion of enzyme, the contact between the enzyme and starch granules is increased, and the efficiency of the enzyme reaction is improved.
Preferably, the enzymolysis temperature is 50 ℃, and the enzymolysis time is 12.0 h.
In the step (2), the power of ultrasonic treatment is 200-300w, the time of ultrasonic treatment is 5-15min, the temperature of ultrasonic treatment is 45-50 ℃, ultrasonic treatment and enzymolysis are carried out simultaneously, and enzymolysis is continued after the ultrasonic treatment is finished.
Preferably, the power of the sonication is 240w, the time of the sonication is 10min and the temperature of the sonication is 50 ℃.
Preferably, a circulating water bath is used to control the temperature during sonication.
In the step (2), after the enzymolysis is finished, the enzyme is inactivated in a boiling water bath for 8-12min, preferably 10 min.
In the step (2), the aging process specifically comprises: aging at 2-6 deg.C for 22.0-26.0 h.
Preferably, the aging process is specifically: aging at 4 deg.C for 24.0 h. The aging of starch is to rearrange and combine starch molecules which are dissolved and expanded in the gelatinization process to form a new compact and crystallized structure, so that the starch molecules are not easy to digest.
In the step (2), drying is carried out in a forced air drying oven, the drying temperature is 40 ℃, the drying time is 24.0-30.0h, and the crushed materials are sieved by a 100-mesh sieve.
The enzyme debranching is most commonly used as pullulanase, the pullulanase can help amylopectin molecules in the mung bean starch to be hydrolyzed to generate amylose molecules, and the higher the content of the amylose is, the stronger the digestion resistance of the generated resistant starch is. The working principle of the pullulanase debranching method is as follows: the amylopectin in the starch is hydrolyzed by alpha-1, 6 glucosidic bonds under the action of enzyme, so that the hydrolysate contains more free amylose molecules, specific parts of the amylose are mutually wound and folded to form a stable double-helix structure, and the double helices can be further polymerized by hydrogen bonds and van der waals force to form an ordered RS crystal with extremely strong resistance to the amylase, so that the resistant starch cannot be digested, decomposed and absorbed in the gastrointestinal tract of a human body. In the enzyme reaction process, the enzyme concentration and the enzymolysis time have great influence on the yield of the final resistant starch, and the excessively high enzyme concentration or the excessively long enzymolysis time can decompose the chain length of amylopectin molecules too short, so that the stable double-helix structure is not formed among amylose molecules, and the yield of RS is influenced. Related studies have shown that the yield and efficiency of resistant starch produced by physical or biochemical methods alone is low.
When the ultrasonic wave is used as a physical method and is combined with an enzyme method to prepare the resistant starch, the yield is improved, the ultrasonic frequency is high, the energy is large, the acoustic effect is generated, the mass transfer rate is increased, the enzymolysis reaction can be accelerated by the ultrasonic wave, the yield of reaction products is increased, the use amount of the enzyme can be reduced, and the operation cost can be saved to a certain extent. The ultrasonic wave method is a relatively new food processing technology and is not widely applied to the industrial production of the resistant starch, and the application of the ultrasonic wave method in the preparation of the mung bean resistant starch is not reported.
When the ultrasonic is applied to the preparation of the mung bean resistant starch, in the ultrasonic process, micro bubbles (cavitation nuclei) in liquid vibrate, grow and continuously gather sound field energy under the action of an ultrasonic field, when the energy reaches a certain threshold value, the cavitation bubbles rapidly collapse and close, namely the bubbles break, and the cavitation bubbles generate local high temperature and high pressure at the moment of rapid collapse, so that corresponding shearing force is generated to destroy the apparent structure and crystal structure of starch particles and better expose enzyme digestion sites on starch molecules, and proper ultrasonic time and ultrasonic power are added, so that the enzymolysis efficiency can be effectively improved, and the enzyme reaction time is reduced. Specifically, the method comprises the following steps: firstly, the mechanical effect generated in the ultrasonic process can cause the starch molecules to vibrate and rotate, and the starch degradation is promoted. Secondly, ultrasonic wave acts on side chain alpha-1, 6 glycosidic bond of the amylopectin molecule to promote the side chain to break, the action can not only reduce the molecular weight of the starch, but also expose the enzyme cutting site of the amylopectin, and the ultrasonic wave and pullulanase generate synergistic action to reduce the content of the amylopectin and improve the content of the amylose. In addition, the cavitation effect formed in the ultrasonic field generates instantaneous high pressure, which affects the starch crystallization area and degrades the amylopectin.
The influence of the ultrasonic power and the ultrasonic time on the yield of the mung bean resistant starch is as follows: along with the increase of ultrasonic power and time, larger pits and pot holes can be formed on the surface of the starch, partial particles are broken, meanwhile, the intensity of mechanical effect and cavitation effect in the ultrasonic process can be influenced by the ultrasonic time and power, the molecular weight of the starch is reduced too much due to overlong ultrasonic time and overhigh ultrasonic power, and a stable double-spiral structure is not easily formed among amylose molecules.
Besides the direct action on the starch, the ultrasonic wave can promote the interaction between the pullulanase and substrate molecules (namely the mung bean starch), strengthen the processes of entering reactants and leaving products from the enzyme activity center and improve the reaction efficiency of the enzyme. The energy released by the ultrasonic wave acting on the enzyme molecules can cause the conformation of the enzyme molecules to be changed to a certain degree, thereby influencing the catalytic activity of the pullulanase.
The invention takes the mung bean starch as the raw material to prepare the high-amylose mung bean resistant starch, and the resistant starch has various physiological functions and good food processing characteristics, thereby being beneficial to the comprehensive utilization of mung beans and improving the added value of mung beans. The invention utilizes the synergy of ultrasonic waves and pullulanase to carry out hydrolysis on mung bean starch molecules, the starch molecules are degraded and enzymolyzed in the treatment process, the ultrasonic waves can promote the enzymolysis reaction while degrading the starch, and the problems of low enzymolysis reaction efficiency, long enzymolysis time, high enzyme consumption and the like in the process of preparing the resistant starch by the enzyme method are solved. Meanwhile, the problems of low yield, long time and high energy consumption of the resistant starch prepared by a single physical preparation method (an autoclave cooling circulation method) are solved, the preparation process of the mung bean resistant starch is optimized, the content of the prepared mung bean resistant starch is 39.42 percent, the highest amylose content is 77.50 percent, the content is far higher than the 6.65 percent resistant starch content and the 30.37 percent amylose content in the mung bean starch raw material, and the yield of the mung bean resistant starch can be improved.
The invention has the beneficial effects that:
(1) in the preparation process of the high-amylose mung bean resistant starch prepared by the pullulanase method, an ultrasonic treatment technology is used for assisting, the enzymolysis of the pullulanase is promoted by ultrasonic, and in order to solve the problems of low enzymolysis reaction efficiency, long enzymolysis time and the like in the process of preparing the resistant starch by the enzymatic method, the higher the amylose content in the resistant starch is, the better the digestion resistance of the resistant starch is.
(2) The physical method used in the invention is ultrasonic treatment, the ultrasonic treatment has excellent effects in food processing and preservation, including higher product yield, shortened processing time, reduced operation and maintenance cost and the like, and the method is a novel physical method for starch modification.
(3) The invention takes the mung bean starch as the raw material to prepare the high-amylose mung bean resistant starch, and the resistant starch has various physiological functions and good food processing characteristics, thereby being beneficial to the comprehensive utilization of mung beans and improving the added value of products.
Drawings
FIG. 1 shows the total starch content in MS, MRS-A, MRS-B, MRS-C;
FIG. 2 shows the content of resistant starch in MS, MRS-A, MRS-B, MRS-C;
FIG. 3 is a scanning electron micrograph of MS (scale: 50 μm);
FIG. 4 is A scanning electron micrograph (scale: 50 μm) of MRS-A;
FIG. 5 is a scanning electron micrograph (scale: 50 μm) of MRS-B;
FIG. 6 is a scanning electron micrograph (scale: 50 μm) of MRS-C;
FIG. 7 is an X-ray diffraction pattern of MS and MRS-A, MRS-B, MRS-C.
Detailed Description
The invention is described in detail below with reference to the figures and specific embodiments. The present invention will be described in further detail with reference to examples, but the scope of the present invention should not be construed as being limited to the examples. The materials and reagents used in this example include: mung bean, sodium hydroxide, absolute ethyl alcohol, hydrochloric acid, pullulanase, a Megazyme total starch determination kit, a Megazyme resistant starch determination kit, and a Megazyme amylose and amylopectin determination kit, which are all commercially available products.
The instrumentation used in this example comprises: the grinding machine, the centrifuge, the blast drying box, the high-pressure steam sterilization pot, the circulating water bath, the ultrasonic cleaner, the refrigerator, the magnetic stirrer, the induction cooker and the scanning electron microscope are all commercial products.
A preparation method of high-amylose mung bean resistant starch specifically comprises the following steps:
(1) extraction of mung bean starch
Uniformly mixing mung bean powder and water in a ratio of 1:6-1:10(w/v), adjusting the pH to 9.0-11.0, carrying out alkali extraction in a water bath at 35-45 ℃ for 2.0-4.0h, sequentially sieving and centrifuging to obtain lower-layer crude starch, sequentially cleaning, centrifuging, drying, crushing and sieving the lower-layer crude starch to obtain mung bean starch, wherein the two sieving processes are 100-mesh sieving, and the cleaning and centrifuging process specifically comprises the following steps: adding water into the lower-layer crude starch for cleaning, centrifuging again, repeating for multiple times to obtain lower-layer starch, washing the lower-layer starch with absolute ethyl alcohol, centrifuging, washing with water and centrifuging continuously to obtain starch precipitate, and drying in a forced air drying oven at 40 ℃ for 24.0-30.0 h;
(2) preparation of resistant starch
Adding water into the mung bean starch obtained in the step (1) to prepare starch paste, wherein the ratio of the mung bean starch to the water is 1:15-1:25(w/v), pasting in a boiling water bath for 20-30min, cooling to 45-55 ℃ after pasting, adjusting the pH to 4.5-5.5, carrying out enzymolysis on the mung bean starch by using pullulanase, carrying out ultrasonic treatment while carrying out enzymolysis, inactivating enzyme in the boiling water bath for 8-12min after the enzymolysis is finished, aging for 22.0-26.0h at the temperature of 2-6 ℃, drying, crushing and sieving to obtain the high-amylose mung bean resistant starch. Wherein the power of ultrasonic treatment is 200-300w, the time of ultrasonic treatment is 5-15min, the temperature of ultrasonic treatment is 45-50 ℃, the temperature in the ultrasonic process is controlled by using a circulating water bath, drying is carried out in a forced air drying box, the temperature of drying is 40 ℃, the drying time is 24.0-30.0h, and the crushed materials are sieved by a 100-mesh sieve.
Example 1
A preparation method of high-amylose mung bean resistant starch comprises the following specific steps:
(1) extraction of mung bean starch
Grinding semen Phaseoli Radiati in mill to obtain semen Phaseoli Radiati powder, mixing 1kg semen Phaseoli Radiati powder with distilled water at a ratio of 1:8(w/v), adjusting pH of semen Phaseoli Radiati powder solution to 10.0 with 2mol/L NaOH, and alkali extracting in 40 deg.C water bath for 3.0 h. Then sieving with 100 mesh sieve, and centrifuging the filtrate at 4000r/min for 10 min. And (3) pouring out the supernatant after centrifugation, scraping green substances on the surface of the precipitate, retaining the lower-layer crude starch, adding distilled water for cleaning, continuing centrifugal separation, and repeating for 3 times to obtain the lower-layer starch. Washing the starch with absolute ethyl alcohol, centrifuging, washing with distilled water, and centrifuging to obtain starch precipitate. Drying the precipitate in a forced air drying oven at 40 deg.C for 30.0 hr, pulverizing, and sieving with 100 mesh sieve to obtain mung bean starch (MS). The total starch content in MS was 85.47% using the Megazyme total starch assay kit, the resistant starch content in MS was 6.65% using the Megazyme resistant starch assay kit, the amylose content in MS was 30.37% using the Megazyme amylose and amylopectin assay kit, and the amylopectin content was 69.63% (this ratio is the ratio of amylose and amylopectin to total starch in the resistant starch sample, the same applies below) as shown in figures 1, 2 and table 1, respectively.
(2) Preparation of resistant starch (ultrasonic combined enzymolysis method)
Adding 200ml of distilled water into 10g of mung bean starch obtained in the step (1), uniformly mixing, gelatinizing in a boiling water bath for 20min, cooling the mung bean starch to 50 ℃, adjusting the pH value to 5.0 by using HCl (0.05mol/L), carrying out enzymolysis on the mung bean starch by using pullulanase (the addition amount of the pullulanase is 27.5npun/g starch, the enzyme activity of the pullulanase is more than 1000npun/g, the production of sigma company), wherein the enzymolysis temperature is 50 ℃, the enzymolysis time is 12.0h, certain ultrasonic treatment is carried out while the mung bean starch is subjected to enzymolysis, a circulating water bath is used for controlling the temperature in the ultrasonic process, the power of the ultrasonic treatment is 240w, the time of the ultrasonic treatment is 10min, the temperature of the ultrasonic treatment is 50 ℃, and the enzyme is inactivated by boiling water bath for 10min after the enzymolysis is finished, aging at 4 deg.C for 24.0h, drying in air drying oven at 40 deg.C, pulverizing, and sieving with 100 mesh sieve to obtain mung bean resistant starch (MRS-B). The total starch content in MRS-B was determined to be 84.20% using the Megazyme total starch assay kit, the resistant starch content in MRS-B was determined to be 39.42% using the Megazyme resistant starch assay kit, the amylose content in MRS-B was determined to be 77.50% using the Megazyme amylose and amylopectin assay kits, and the amylopectin content was determined to be 22.50%, as shown in FIGS. 1, 2, and Table 1, respectively.
Comparative example 1
A method for preparing mung bean resistant starch by adopting an enzymatic hydrolysis method comprises the following specific steps:
(a) extraction of mung bean starch
In accordance with step (1) in example 1. The total starch content in MS was 85.47% using the Megazyme total starch assay kit and 6.65% using the Megazyme resistant starch assay kit, as shown in figures 1, 2 and table 1, respectively.
(b) Preparation of resistant starch
And (b) adding 200ml of distilled water into 10g of mung bean starch obtained in the step (a), uniformly mixing, gelatinizing in a boiling water bath for 20min, cooling to 50 ℃, adjusting the pH to 5.0 by using HCl (0.05mol/L), and carrying out enzymolysis on the mung bean starch by using pullulanase, wherein the enzyme adding amount is 27.5npun/g of starch, the enzymolysis temperature is 50 ℃, and the enzymolysis time is 12.0 h. And (3) after the enzymolysis is finished, inactivating enzyme in A boiling water bath for 10min, aging for 24.0h at 4 ℃, drying in an air drying oven at 40 ℃, crushing, and sieving with A100-mesh sieve to obtain the mung bean resistant starch (MRS-A). The total starch content in MRS-A was 83.48% as determined using Megazyme total starch assay kit, 31.53% as determined using Megazyme resistant starch assay kit, 68.36% as determined using Megazyme amylose and amylopectin assay kit, and 31.64% as determined using Megazyme amylose and amylopectin assay kit, respectively, as shown in FIGS. 1, 2, and Table 1.
Comparative example 2
A method for preparing mung bean resistant starch by adopting a pressure heating cooling circulation method comprises the following specific steps:
(A) extraction of mung bean starch
In accordance with step (1) in example 1. The total starch content in MS was 85.47% using the Megazyme total starch assay kit and 6.65% using the Megazyme resistant starch assay kit, as shown in figures 1, 2 and table 1, respectively.
(B) Preparation of resistant starch
And (2) adding 75ml of distilled water into 15g of mung bean starch obtained in the step (A), uniformly mixing, performing pressure heat treatment at 121 ℃ for 30min, fully gelatinizing, cooling to room temperature, storing at 4 ℃ for 24h, and repeating the steps to obtain starch precipitate. And (3) drying the precipitate in a blast drying oven at 40 ℃, crushing and sieving with a 100-mesh sieve to obtain the mung bean resistant starch (MRS-C). The total starch content in MRS-C was determined to be 84.02% using the Megazyme total starch assay kit, the resistant starch content in MRS-C was determined to be 15.92% using the Megazyme resistant starch assay kit, the amylose content in MRS-C was determined to be 33.87% using the Megazyme amylose and amylopectin assay kit, and the amylopectin content was 66.13%, as shown in FIGS. 1, 2, and Table 1, respectively.
TABLE 1 amylose and amylopectin content in mung bean starch and resistant starch thereof
(Note: abcd letters are different and show significant differences between groups of data)
Performance testing
Testing one: the method for measuring the surface structures of mung bean starch and resistant starch comprises the following specific steps:
taking a proper amount of starch sample to be detected, grinding the starch sample to be detected to ensure that the starch sample is uniformly dispersed, and drying the starch sample in an oven at 105 ℃ for 4.0 h. Fixing the starch sample to be detected on a sample plate by using double-sided conductive adhesive, and carrying out gold plating treatment on the sample plate under a vacuum condition. And then, placing the sample stage in a scanning electron microscope for observation, shooting the appearance of the starch sample particles and analyzing.
FIGS. 3, 4, 5 and 6 are scanning electron micrographs of mung bean starch and its resistant starch. From FIG. 3, mung bean starch (MS), it can be seen that it has an oval shape under scanning electron microscope, part of starch molecules are linked together, and the surface is smooth without wrinkles. FIG. 4 is mung bean resistant starch MRS-A, FIG. 5 is mung bean resistant starch MRS-B, and FIG. 6 is mung bean resistant starch MRS-C. It can be seen from fig. 4, 5 and 6 that the mung bean starch undergoes significant changes in apparent structure after undergoing respective treatments of enzymolysis, ultrasonic combined enzymolysis and autoclaving cooling cycle, wherein the same changes are that starch molecules are changed into irregular fragment shapes, the surfaces of the starch molecules are uneven and have wrinkles, fig. 4 is similar to fig. 5 in a polygonal rhombus shape, but the molecules in fig. 5 are more dispersed than those in fig. 4, which may be due to the effect of ultrasonic treatment, fig. 6 shows that the surface structures of mung bean resistant starch molecules are also irregular fragment shapes, and the wrinkles are more significant due to the treatment of autoclaving cooling cycle. In conclusion, the resistant starch has improved properties compared to native starch.
And (2) testing: the method for measuring the crystal structures and the crystallinities of the mung bean starch and the resistant starch comprises the following specific steps:
and obtaining the X-ray diffraction pattern of the mung bean starch and the resistant starch sample thereof by using an X-ray diffractometer and a step scanning method. The operating conditions were as follows: initial angle: 2 θ is 5 °; end angle: 2 θ is 60 °; step length: 0.05 degree; scanning speed: 4 degree/min; and analyzing the crystallinity and the crystal structure type according to the diffraction pattern.
FIG. 7 is an X-ray diffraction pattern of mung bean starch and resistant starch thereof, from which the crystallinity and the type of crystals are analyzed as shown in Table 2.
TABLE 2 degree of crystallinity and type of crystallinity of mung bean starch and its resistant starch
(Note: abcd letters are different and show significant differences between groups of data)
The mung bean resistant starch MRS-B prepared by the ultrasonic-enzymolysis combined method has the highest crystallinity, the crystallization type is changed from the A + C type of the original starch to a more compact B + C type structure, and the higher crystallinity and the compact crystallization structure are beneficial to improving the digestion resistance of the resistant starch.
In the method for preparing the mung bean resistant starch, the content of the mung bean resistant starch prepared by the ultrasonic combined enzymolysis method is 39.42 percent and the content of amylose is 77.50 percent, so that the content of the resistant starch in the mung bean starch is effectively increased (the content of the resistant starch in the untreated mung bean starch is 6.65 percent and the content of the amylose is 30.37 percent), is higher than the content of the mung bean resistant starch prepared by the enzymolysis method (31.53 percent) and the content of the amylose (68.36 percent), and the content of resistant starch (15.92%) in mung bean prepared by the autoclaving cooling cycle method, the content of amylose (33.87%), and the resistant starch molecules prepared by the ultrasonic-combined enzymolysis method are changed into irregular fragment shapes, the surfaces of the resistant starch molecules are uneven, folds appear, the dispersion degree is larger, and the crystallinity of the resistant starch molecules is higher, and the crystalline structure is more compact, so that the resistant starch molecules show more excellent performance, and in conclusion, the ultrasonic combined enzymolysis method is a better method for preparing the high-amylose mung bean resistant starch.
The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.
Claims (10)
1. The preparation method of the high-amylose mung bean resistant starch is characterized by comprising the following steps of:
(1) extraction of mung bean starch
Uniformly mixing mung bean powder and water, adjusting pH, carrying out alkali extraction, sequentially sieving and centrifuging to obtain lower-layer crude starch, and sequentially cleaning, centrifuging, drying, crushing and sieving the lower-layer crude starch to obtain mung bean starch;
(2) preparation of resistant starch
Adding water into the mung bean starch obtained in the step (1) to prepare starch paste, cooling after gelatinization, adjusting the pH value, carrying out enzymolysis on the mung bean starch by using pullulanase, carrying out ultrasonic treatment while carrying out enzymolysis, inactivating enzymes after the enzymolysis is finished, carrying out aging, drying, crushing and sieving to obtain the high-amylose mung bean resistant starch.
2. The method for preparing high amylose mung bean resistant starch according to claim 1, wherein the ratio of mung bean flour to water in step (1) is 1:6 to 1:10 (w/v).
3. The method for preparing high amylose mung bean resistant starch according to claim 1, wherein in the step (1), NaOH is used to adjust the pH of the mung bean powder solution to 9.0-11.0;
in the step (1), alkali extraction is carried out in water bath at 35-45 ℃ for 2.0-4.0 h.
4. The method for preparing high-amylose mung bean resistant starch according to claim 1, wherein in the step (1), the two siftings are both 100 mesh siftings;
in the step (1), the washing and centrifuging process specifically comprises the following steps: and adding water into the lower-layer crude starch for cleaning, centrifuging again, repeating for multiple times to obtain lower-layer starch, washing the lower-layer starch with absolute ethyl alcohol, centrifuging, and then continuing to wash with water and centrifuging to finally obtain starch precipitate.
5. The method for preparing high amylose mung bean resistant starch according to claim 1, wherein the drying in step (1) is performed in a forced air drying oven at a temperature of 40 ℃ for 24.0 to 30.0 hours.
6. The method for preparing high amylose mung bean resistant starch according to claim 1, wherein the ratio of mung bean starch to water in step (2) is 1:15 to 1:25 (w/v).
7. The method for preparing high-amylose mung bean resistant starch according to claim 1, wherein in the step (2), the gelatinization is carried out in a boiling water bath for 20-30 min;
in the step (2), cooling to 45-55 ℃;
in the step (2), HCl is adopted to adjust the pH value to 4.5-5.5.
8. The method for preparing high-amylose mung bean resistant starch according to claim 1, wherein in the step (2), the enzyme activity of pullulanase is more than or equal to 1000 npun/g;
in the step (2), the addition amount of pullulanase is 20-30npun/g starch;
in the step (2), the enzymolysis temperature is 45-50 ℃, and the enzymolysis time is 10.0-14.0 h.
9. The method for preparing high amylose mung bean resistant starch as claimed in claim 1, wherein in the step (2), the power of the ultrasonic treatment is 200-300w, the time of the ultrasonic treatment is 5-15min, and the temperature of the ultrasonic treatment is 45-50 ℃;
the temperature during sonication was controlled using a circulating water bath.
10. The method for preparing high-amylose mung bean resistant starch according to claim 1, wherein in the step (2), the enzyme is inactivated in a boiling water bath for 8-12min after the enzymolysis is finished;
in the step (2), the aging process specifically comprises: aging at 2-6 deg.C for 22.0-26.0 h;
in the step (2), drying is carried out in a forced air drying oven, the drying temperature is 40 ℃, the drying time is 24.0-30.0h, and the crushed materials are sieved by a 100-mesh sieve.
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| Publication number | Priority date | Publication date | Assignee | Title |
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104356243A (en) * | 2014-10-13 | 2015-02-18 | 河北经贸大学 | New technology for industrial mung bean starch production |
| CN105132492A (en) * | 2015-07-24 | 2015-12-09 | 中国农业科学院农产品加工研究所 | High-resistant-starch-content product prepared by ultrahigh-pressure coordinated enzyme and preparation method of product |
| CN109588695A (en) * | 2018-12-07 | 2019-04-09 | 合肥工业大学 | A kind of preparation method of RS5 type wheat resistant starch |
| CN110144014A (en) * | 2019-06-01 | 2019-08-20 | 江苏昕宇药业有限公司 | A kind of technique of corn production resistant starch |
| CN111041055A (en) * | 2019-12-12 | 2020-04-21 | 江苏大学 | Method for preparing arrowhead resistant starch by using ultrasound in cooperation with pullulanase |
| US20210062236A1 (en) * | 2020-01-17 | 2021-03-04 | Jiangnan University | Preparation Method of Amylodextrin |
| CN113106131A (en) * | 2021-03-31 | 2021-07-13 | 齐鲁工业大学 | Method for preparing slowly digestible starch by using composite modification method |
-
2021
- 2021-09-30 CN CN202111157414.1A patent/CN113845600B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104356243A (en) * | 2014-10-13 | 2015-02-18 | 河北经贸大学 | New technology for industrial mung bean starch production |
| CN105132492A (en) * | 2015-07-24 | 2015-12-09 | 中国农业科学院农产品加工研究所 | High-resistant-starch-content product prepared by ultrahigh-pressure coordinated enzyme and preparation method of product |
| CN109588695A (en) * | 2018-12-07 | 2019-04-09 | 合肥工业大学 | A kind of preparation method of RS5 type wheat resistant starch |
| CN110144014A (en) * | 2019-06-01 | 2019-08-20 | 江苏昕宇药业有限公司 | A kind of technique of corn production resistant starch |
| CN111041055A (en) * | 2019-12-12 | 2020-04-21 | 江苏大学 | Method for preparing arrowhead resistant starch by using ultrasound in cooperation with pullulanase |
| US20210062236A1 (en) * | 2020-01-17 | 2021-03-04 | Jiangnan University | Preparation Method of Amylodextrin |
| CN113106131A (en) * | 2021-03-31 | 2021-07-13 | 齐鲁工业大学 | Method for preparing slowly digestible starch by using composite modification method |
Non-Patent Citations (4)
| Title |
|---|
| 刘淑婷等: "超声-微波协同酶法制备芸豆抗性淀粉工艺优化及结构分析", 《中国食品学报》 * |
| 李伟 等: "鹰嘴豆抗性淀粉提取工艺及其降血脂效应研究", 《现代食品》 * |
| 樊明涛，王银瑞，艾启俊: "绿豆淀粉的提取和绿豆淀粉性质的研究", 《北京农学院学报》 * |
| 赵凯等: "绿豆抗性淀粉制备技术研究", 《食品工业科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115595346A (en) * | 2022-12-15 | 2023-01-13 | 中国农业大学（Cn） | Preparation method of pea III type resistant starch |
| CN115595346B (en) * | 2022-12-15 | 2023-06-23 | 中国农业大学 | Preparation method of pea III-type resistant starch |
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