CN112137116B - Starch-based dietary fiber for high yield of butyric acid and processing method thereof - Google Patents
Starch-based dietary fiber for high yield of butyric acid and processing method thereof Download PDFInfo
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
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- C—CHEMISTRY; METALLURGY
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Abstract
The invention discloses a starch-based dietary fiber for high yield of butyric acid and a processing method thereof, belonging to the technical field of modern nutritional food processing. The invention uses starch as raw material, and the bonding mode of starch chains is reconstructed by a composite enzyme synergistic catalysis-membrane filtration grading coupling technology composed of alpha-amylase and glycosyltransferase, so that the high-yield butyric acid starch-based dietary fiber is prepared. The synthesized starch-based dietary fiber not only has high solubility and low viscosity, but also is easy to ferment by intestinal microorganisms to produce butyric acid with high yield, thereby having higher nutritional quality. The product solves the problems of complex extraction process, low yield, poor safety, high cost and the like of starch-based dietary fibers, and the application range of the product can relate to the fields of functional foods, medicines and the like.
Description
Technical Field
The invention relates to a starch-based dietary fiber for high yield of butyric acid and a processing method thereof, belonging to the technical field of modern food nutrition food processing.
Background
With the rapid development of the economy in China, the living standard of residents in China is greatly improved. The coarse food grain has a coarse taste, so that the eating habits of residents in China gradually deviate from the traditional eating modes mainly comprising vegetable food, and the eating structure is gradually changed to the western style. Thus, the spectrum of diseases for residents in China is also gradually westernized, obesity, diabetes and cardiovascular diseases are becoming more and more common, and the prevalence of hypertension and type II diabetes has even reached and overtaken the United states. Therefore, the research and development of the novel starch-based dietary fiber has important effects on improving the physical health level of people, enhancing the chronic disease prevention and treatment work, reducing the disease burden and improving the health of residents.
Starch is used as the most abundant carbohydrate resource in China, and has wide sources and low price. The starch is used as a raw material, and the synthesis of the starch-based dietary fiber with certain digestion resistance has important significance for the development of the food industry. Compared with starch, the starch-based dietary fiber still maintains a large amount of undigested parts under the action of various digestive enzymes of the gastrointestinal tract, has the effect of lowering the blood sugar response level, and is beneficial to preventing the prevalence rate of various chronic diseases. In addition, undigested fractions are fermented to produce large amounts of short chain fatty acids under the action of intestinal microorganisms after entering the large intestine, wherein butyric acid can regulate the growth of intestinal beneficial bacteria, maintain the integrity of colonic mucosa, reduce the prevalence of diseases such as cancer and ulcerative colitis, and can effectively maintain the intestinal health of the body. And because starch-based dietary fibers are easier to add to foods, there is a great deal of interest. However, the existing starch-based dietary fiber is mainly produced by extracting from bean dregs, wheat bran, jerusalem artichoke and other crops and processing byproducts thereof through a mechanical method, a chemical method, an enzymatic method and a mixing method. For example, the existing enzyme method mainly adopts protease, amylase, cellulase and the like to extract from crops and processing byproducts, and the starch-based dietary fiber extracted by the method has low yield, low purity and poor safety, and the preparation process is complex and has high cost, so that the method cannot meet the increasingly-growing social demands. Based on the reasons, the preparation method of the high-yield butyric acid starch-based dietary fiber is convenient, efficient and safe, and has important significance for the development of the food industry.
Disclosure of Invention
In order to solve the problems, the invention provides a convenient, efficient and safe preparation method of the high-yield butyric acid starch-based dietary fiber. The method solves the problems of complicated process, low yield, poor safety, high cost and the like of the existing starch-based dietary fiber extraction, and the application range of the method can relate to the fields of functional foods, medicines and the like, for example, epidemiological researches show that the starch-based dietary fiber can be used alone or in combination with probiotics to play a plurality of health care functions, and can reduce diarrhea in intestinal tracts, constipation, obesity and diabetes; meanwhile, the starch-based dietary fiber can prevent colon cancer, treat chronic diseases to a certain extent, and has the reputation of 'intestinal tract cleaning'.
Specifically, the invention is realized by the following technical scheme: a processing method of high-yield butyric acid starch-based dietary fiber takes starch as a substrate, and specifically comprises the following steps:
(1) Weighing starch with a certain mass to prepare starch milk with the concentration of 10-30wt%, and heating to above 70 ℃ until the starch milk is completely gelatinized;
(2) Transferring the starch paste solution to an enzyme-membrane coupling reactor after the temperature is reduced to 30-55 ℃, adding a complex enzyme solution consisting of alpha-amylase and glycosyltransferase according to the enzyme adding amount of 1-2.5U/g substrate, and reacting for 12-36h in a heat preservation way;
(3) And collecting membrane filtrate, inactivating enzyme, centrifuging, and drying the obtained supernatant to obtain the high-yield butyric acid starch-based dietary fiber.
In one embodiment of the invention, the heating of step (1) is preferably boiling water bath heating.
In one embodiment of the invention, the alpha-amylase is a medium temperature commercial alpha-amylase; the glycosyltransferase is transglycosylase derived from a microorganism of GH70 family, and the enzyme activity ratio of the alpha-amylase to the glycosyltransferase is 5:1-1:10.
In one embodiment of the invention, the GH70 family microorganisms comprise GH70 family fungi and bacteria.
In one embodiment of the present invention, the glycosyltransferase is a multifunctional transglycosylase derived from GH70 family such as Lactobacillus reuteri, lactobacillus fermentum, streptococcus thermophilus, azotobacter chroococcus etc.
In one embodiment of the invention, the glycosyltransferase is capable of hydrolyzing the product of the enzymatic hydrolysis of starch by an alpha-amylase to produce alpha-1, 4/6 and alpha-1, 6-glucose-alpha-1, 4 structural dietary fiber.
In one embodiment of the present invention, the starch is any one of corn starch, potato starch, tapioca starch, rice starch, wheat starch, etc., or any one of ordinary starch, waxy starch, high amylose starch, starch dextrin of the above-mentioned starches.
In one embodiment of the invention, in step (1), a certain mass of starch is weighed to prepare starch milk with the concentration of 10-30wt%, and the starch is dispersed in a buffer salt solution (pH 6.5-7.0) to obtain a starch suspension.
In one embodiment of the invention, the membrane cutoff molecular weight of the enzyme membrane reactor is 10-60kDa.
The second purpose of the invention is to provide the high-yield butyric acid starch-based dietary fiber prepared by the processing method.
In one embodiment of the present invention, the high-yield butyrate-based dietary fiber has a molecular weight of 0.5 to 9.5X10 7 Da。
In one embodiment of the invention, the alpha-1, 6 glycosidic linkage content in the high yield butyrate starch-based dietary fiber is greater than 20%.
A third object of the present invention is to provide a food product and a food additive comprising the above-mentioned high-yield butyrate starch-based dietary fiber.
The fourth object of the invention is to provide a processing method of the high-yield starch butyrate-based dietary fiber and application of the high-yield starch butyrate-based dietary fiber in the fields of food and medicine.
The invention has the following advantages:
(1) The invention utilizes the multifunctional carbohydrase synergistic catalysis-membrane filtration grading coupling technology to obtain the starch-based dietary fiber of high-yield butyric acid, has simple and convenient steps, mild reaction conditions and high safety, realizes continuous and low-cost green production, fully utilizes rich starch resources in China, and improves the quality of modern foods.
(2) The product of the invention has outstanding performance of producing butyric acid by intestinal fermentation and more remarkable health care function, and truly meets the requirements of vast consumers on modern functional foods. The starch-based dietary fiber obtained by the invention is not easy to digest in the stomach and the small intestine of a human body, short-chain fatty acid, especially butyric acid, is easy to be produced by microbial fermentation in the large intestine, and compared with the original substrate, the butyric acid produced by intestinal fermentation is improved by more than 2 times, so that the intestinal health of the human body can be regulated.
(3) The starch-based dietary fiber prepared by the invention can be used as a food additive for processing various foods, so that the value of the product is greatly improved, and the starch-based dietary fiber has important significance for improving the health level of people, thereby having higher social benefit and economic effect.
Drawings
FIG. 1 starch-based dietary fiber prepared in example 1 1 H NMR chart.
Detailed Description
The testing method comprises the following steps:
molecular weight measurement: the starch-based dietary fiber was formulated as a 5mg/mL aqueous solution and its molecular weight distribution was determined using the HPSEC-MALL-RI system.
1 H NMR test: 25mg of starch-based dietary fiber was weighed into a clean 1.5mL centrifuge tube, dissolved in 0.5mL of heavy water, and lyophilized for 24h. Repeating the steps2 times, heavy water exchange is carried out. Dissolving the sample with 0.5mL heavy water, and performing nuclear magnetic resonance spectrometer 1 H NMR analysis.
Measurement of starch-based dietary fiber structure: the starch-based dietary fiber was accurately weighed and dissolved in 25mM acetic acid-sodium acetate buffer (pH 4.0) to prepare a reaction solution of 2.5mg/mL, to which 0.5U of enzyme combinations (I: isoamylase, isopropenylase and beta-amylase; II: isoamylase; III: isoamylase and beta-amylase) were added, respectively, and water-bath was conducted at 40℃for 24 hours. The polymerization degree of the hydrolysis product in the above reaction solution was then measured by HPAEC-PAD.
The method for measuring the butyric acid produced by in vitro anaerobic fermentation comprises the following steps: preparing anaerobic fermentation culture medium with starch-based dietary fiber or starch as carbon source, adding feces diluent according to 10% of the additive amount, placing into an incubator at 37 ℃ for anaerobic fermentation, sampling at different fermentation time, and measuring the butyric acid content in the fermentation liquid by gas chromatography GC.
Corn starch, corn starch dextrins, high amylose corn starch, potato starch, wheat starch were purchased from Shanghai national starch company.
Alpha-amylase was purchased from Sigma; glycosyltransferases are described in literature Microal Starch-Converting Enzymes: recent Insights and Perspectives, comprehensive Reviews in Food Science and Food Safety,2018,17 (5): 1238-1260 comprises the steps of enzyme gene excavation, engineering bacterium construction, fermentation enzyme production and the like.
The present invention is further described below with reference to examples, but embodiments of the present invention are not limited thereto.
Example 1
200g of corn starch was dispersed in acetic acid-sodium acetate buffer (25 mM, pH 6.5) to give a 20wt% starch suspension which was boiled in water until complete gelatinization. And (3) when the temperature is reduced to 50 ℃, transferring the mixture into an enzyme membrane reactor (membrane interception size is 10 kDa), simultaneously adding 400U of alpha-amylase and glycosyltransferase (the enzyme activity ratio of the alpha-amylase to glycosyltransferase is 5:1) compound enzyme liquid, carrying out heat preservation reaction for 36h, collecting membrane filtrate, heating to deactivate the enzyme activity, carrying out centrifugal treatment, and drying the obtained supernatant to obtain the starch-based dietary fiber.
The purity of the starch-based dietary fiber of the embodiment is up to98.3% and a molecular weight of 1.1X10 7 Da, the content of alpha-1, 6 glycosidic bonds is 33%; the butyric acid content produced by in vitro simulated anaerobic fermentation is 2.7 times that of the original substrate corn starch.
Example 2
300g of corn starch dextrin was dispersed in acetic acid-sodium acetate buffer (25 mM, pH 6.5) to give a 30wt% starch suspension which was gelatinized in a boiling water bath. And (3) when the temperature is reduced to 35 ℃, transferring into an enzyme membrane reactor (membrane interception size is 10 kDa), simultaneously adding 300U of alpha-amylase and glycosyltransferase (the enzyme activity ratio of the alpha-amylase to glycosyltransferase is 1:1) into the enzyme membrane reactor, carrying out heat preservation reaction for 24 hours, collecting membrane filtrate, heating to inactivate enzyme, carrying out centrifugal treatment, and drying the obtained supernatant to obtain the starch-based dietary fiber.
The purity of the starch-based dietary fiber of the embodiment reaches 96.5%, and the molecular weight is 0.6X10 7 Da; the content of alpha-1, 6 glycosidic bond is 24%; the butyric acid content produced by in vitro simulated anaerobic fermentation is 3.1 times that of the original substrate starch dextrin.
Example 3
200g of high amylose corn starch was dispersed in acetic acid-sodium acetate buffer (25 mM, pH 6.5) to give a 30wt% starch suspension which was gelatinized in a boiling water bath. And (3) when the temperature is reduced to 40 ℃, transferring into an enzyme membrane reactor (membrane interception size is 10 kDa), simultaneously adding 500U of alpha-amylase and glycosyltransferase (the enzyme activity ratio of the alpha-amylase to glycosyltransferase is 5:1) compound enzyme liquid, carrying out heat preservation reaction for 12 hours, collecting membrane filtrate, heating to inactivate enzyme, carrying out centrifugal treatment, and drying the obtained supernatant to obtain the starch-based dietary fiber.
The purity of the starch-based dietary fiber of this example was 99.1% and the molecular weight was determined to be 2.7X10 7 Da; the content of alpha-1, 6 glycosidic bond is 30%; the butyric acid content produced by in vitro simulated anaerobic fermentation is 2.7 times that of the high amylose starch of the original substrate.
Example 4
200g of wheat starch was dispersed in acetic acid-sodium acetate buffer (25 mM, pH 6.5) to give a 20wt% starch suspension which was boiled in water until complete gelatinization. And (3) when the temperature is reduced to 50 ℃, transferring the mixture into an enzyme membrane reactor (membrane interception size is 50 kDa), simultaneously adding 400U of alpha-amylase and glycosyltransferase (the enzyme activity ratio of the alpha-amylase to glycosyltransferase is 5:1) compound enzyme liquid, carrying out heat preservation reaction for 36h, collecting membrane filtrate, heating to deactivate the enzyme activity, carrying out centrifugal treatment, and drying the obtained supernatant to obtain the starch-based dietary fiber.
The purity of the starch-based dietary fiber of the embodiment reaches 97.2%, and the molecular weight is 4.6X10 7 Da; the content of alpha-1, 6 glycosidic bond is 25%; the butyric acid content produced by in vitro simulated anaerobic fermentation is 2.5 times that of the original substrate wheat starch.
Example 5
200g of potato starch was dispersed in acetic acid-sodium acetate buffer (25 mM, pH 6.5) to give a 20wt% starch suspension which was boiled in water until complete gelatinization. And (3) when the temperature is reduced to 50 ℃, transferring the mixture into an enzyme membrane reactor (membrane interception size is 50 kDa), simultaneously adding 400U of alpha-amylase and glycosyltransferase (the enzyme activity ratio of the alpha-amylase to glycosyltransferase is 5:1) compound enzyme liquid, carrying out heat preservation reaction for 36h, collecting membrane filtrate, heating to deactivate the enzyme activity, carrying out centrifugal treatment, and drying the obtained supernatant to obtain the starch-based dietary fiber.
The purity of the starch-based dietary fiber of this example was 97.8%, and the molecular weight was measured to be 4.0X10 7 Da; the content of alpha-1, 6 glycosidic bond is 29%; the butyric acid content produced by in vitro simulated anaerobic fermentation is 3.0 times that of the original substrate potato starch.
The method of the invention can treat common starch, high amylose starch and starch dextrin of tapioca starch, rice starch and various corn starch to obtain the starch with molecular weight of 0.5-9.5X10 7 Da. The alpha-1, 6 glycosidic bond content is higher than 20%, and compared with the original substrate, the butyric acid produced by intestinal fermentation is improved by more than 2.0 times.
Comparative example 1
200g of corn starch was dispersed in acetic acid-sodium acetate buffer (25 mM, pH 6.5) to give a 20wt% starch suspension which was boiled in water until complete gelatinization. And (3) when the temperature is reduced to 50 ℃, transferring the mixture into an enzyme membrane reactor (membrane interception size is 10 kDa), simultaneously adding 400U of glycosyltransferase enzyme solution, carrying out heat preservation reaction for 36h, collecting membrane filtrate, heating to deactivate enzyme activity, carrying out centrifugal treatment, and drying the obtained supernatant to obtain the starch-based dietary fiber.
The purity of the prepared starch-based dietary fiber is 92.2 percent and the molecular weight is 9.0 multiplied by 10 6 Da; the content of alpha-1, 6 glycosidic bond is 18%; at the position ofThe butyric acid content produced by in vitro simulated anaerobic fermentation is 1.6 times of that of the original substrate corn starch.
Comparative example 2
200g of wheat starch was dispersed in acetic acid-sodium acetate buffer (25 mM, pH 6.5) to give a 20wt% starch suspension which was boiled in water until complete gelatinization. And (3) when the temperature is reduced to 50 ℃, transferring the mixture into an enzyme membrane reactor (membrane interception size is 10 kDa), simultaneously adding 400U of glycosyltransferase enzyme solution, carrying out heat preservation reaction for 36h, collecting membrane filtrate, heating to deactivate enzyme activity, carrying out centrifugal treatment, and drying the obtained supernatant to obtain the starch-based dietary fiber.
The purity of the obtained starch-based dietary fiber is 90.5 percent and the molecular weight is 1.5X10 8 Da; the content of alpha-1, 6 glycosidic bond is 14%; the butyric acid content produced by in vitro simulated anaerobic fermentation is 1.1 times that of the original substrate wheat starch.
Comparative example 3
200g of potato starch was dispersed in acetic acid-sodium acetate buffer (25 mM, pH 6.5) to give a 20wt% starch suspension which was boiled in water until complete gelatinization. And (3) when the temperature is reduced to 50 ℃, transferring the mixture into an enzyme membrane reactor (membrane interception size is 50 kDa), simultaneously adding 400U of glycosyltransferase enzyme solution, carrying out heat preservation reaction for 36h, collecting membrane filtrate, heating to deactivate enzyme activity, carrying out centrifugal treatment, and drying the obtained supernatant to obtain the starch-based dietary fiber.
The purity of the prepared starch-based dietary fiber is 85.7 percent and the molecular weight is 2.9X10 8 Da; the content of alpha-1, 6 glycosidic bond is 11%; the butyric acid content produced by in vitro simulated anaerobic fermentation is 0.9 times that of the original substrate potato starch.
Comparative example 4
Referring to example 1, the amounts of alpha-amylase and glycosyltransferase were replaced with 80U and 900U from 400U, respectively, to produce the corresponding functional sugar products. The performance results of the resulting functional sugar product are shown in Table 1.
TABLE 1 results of dietary fiber efficacy obtained with different amounts of multifunctional enzyme systems
| Dosage (U/g) | Increase in production of butyric acid by fermentation |
| 80 | 0.7 |
| 900 | 1.1 |
While the invention has been described with reference to the preferred embodiments, it is not limited thereto, and various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (6)
1. The processing method of the high-yield butyric acid starch-based dietary fiber is characterized by comprising the following steps of:
(1) Weighing starch with a certain mass to prepare starch milk with the concentration of 10-30wt%, and heating to above 70 ℃ until the starch milk is completely gelatinized;
(2) Transferring the starch paste solution to an enzyme-membrane coupling reactor after the temperature is reduced to 30-55 ℃, adding a complex enzyme solution consisting of alpha-amylase and glycosyltransferase according to the enzyme adding amount of 1-2.5U/g substrate, and carrying out heat preservation reaction for 12-36h; the alpha-amylase is a medium temperature alpha-amylase; the glycosyltransferase is transglycosylase derived from lactobacillus reuteri, lactobacillus fermentum, streptococcus thermophilus and azotobacter chroococcus GH70 family microorganisms, wherein the enzyme activity ratio of the alpha-amylase to the glycosyltransferase is 5:1-1:1, a step of;
(3) Collecting membrane filtrate, inactivating enzyme, centrifuging, and drying the obtained supernatant to obtain high-yield butyric acid starch-based dietary fiber; the molecular weight of the high-yield butyric acid starch-based dietary fiber is 0.5-9.5X10 7 Da, alpha-1, 6 glycosidic bond content is higher than 20%.
2. The method for processing high-yield butyrate starch-based dietary fiber according to claim 1, wherein the starch is any one of corn starch, potato starch, tapioca starch, rice starch, and wheat starch.
3. The method for processing high-yield butyrate-based dietary fiber according to claim 2, wherein the starch is any one of common starch, waxy starch, high-amylose starch, and starch dextrin.
4. The high-yield starch butyrate-based dietary fiber prepared by the processing method of the high-yield starch butyrate-based dietary fiber according to any one of claims 1-3.
5. A food product or food additive comprising the high yield of starch butyrate-based dietary fiber according to claim 4.
6. A processing method of the high-yield starch butyrate-based dietary fiber according to any one of claims 1-3 or application of the high-yield starch butyrate-based dietary fiber according to claim 4 in the field of food.
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| WO2019153611A1 (en) * | 2018-02-06 | 2019-08-15 | 江南大学 | Method for preparing highly branched dextrin product |
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| WO2019153611A1 (en) * | 2018-02-06 | 2019-08-15 | 江南大学 | Method for preparing highly branched dextrin product |
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