CN115624186B - High-protein low-GI composite grain composition and preparation method and application thereof - Google Patents

Abstract

The invention provides a high-protein low-GI composite grain composition and a preparation method and application thereof, belonging to the technical field of food, pea protein powder and white kidney bean extract are subjected to secondary enzymolysis, fermented by lactobacillus plantarum, saccharomyces cerevisiae and lactobacillus bulgaricus, freeze-dried to obtain enzymolysis fermented protein peptide powder, and embedded by sodium carboxymethylcellulose, sodium alginate and water-soluble chitosan to obtain slow-release protein particles; grinding oat flour and rice bran powder mixed colloid into fine powder, mixing with wheat amylose, adjusting the moisture content, extruding by a double screw, and spray drying to obtain the low-GI puffed powder; and uniformly mixing the slow-release protein particles, the low-GI puffed powder and the prebiotics sweetener to obtain the high-protein low-GI composite grain composition. The composition disclosed by the invention is fine and smooth in taste, does not contain anti-nutritional factors, has a very low GI value, can play a good role in immunoregulation and blood sugar regulation after being eaten, and has a wide application prospect.

Description

A compound cereal composition with high protein and low GI and its preparation method and application

technical field

The invention relates to the field of food technology, in particular to a compound cereal composition with high protein and low GI and its preparation method and application.

Background technique

With the acceleration of the pace of life, people's diet types and habits have undergone tremendous changes, sub-health and chronic disease populations are gradually increasing, and healthy eating has become the focus of people's attention. The glycemic index (GI) reflects the body's digestion and absorption rate of food and the resulting blood sugar response. The authoritative definition of FAQ/WHO is the ratio of the value-added area under the postprandial blood glucose response curve of a food containing 50 g of available carbohydrates to the value-added area under the postprandial blood glucose response curve of a standard reference food containing the same amount of available carbohydrates. According to the level of GI, food is divided into 3 categories: high GI food, GI>70; medium GI food, 70≥GI>55; low GI food, GI≤55. Low GI foods are digested and absorbed slowly, and the release rate of glucose is slow, which avoids the risk caused by large changes in blood sugar, and is suitable for people who have requirements for blood sugar control. In addition, related studies have shown that low GI foods also have a positive effect on the prevention and treatment of obesity, cardiovascular disease, cancer, Alzheimer's disease and other diseases. Therefore, low GI food has gradually been valued by domestic and foreign enterprises, and has broad prospects for future development.

With the improvement of people's living standards and increasingly superior living conditions, the daily intake of food is greatly over-nutritional, and there are more and more people with high blood pressure, high blood sugar, and high blood fat, and their age is getting younger. People control the three highs and pay more attention to treatment than prevention. Most of them choose to take medicines for reducing the three highs within a certain period of time to reduce the three highs, and pay a certain health price in exchange for temporary results. Therefore, it is particularly important to choose healthy raw materials, not only to ensure the color, aroma and taste of the product, but also to ensure its safety, and to benefit the health of consumers.

Grain composition, processing method and eating state are the main factors affecting food GI. The composition of grains includes starch, protein, fat, dietary fiber, etc. Among them, starch has the greatest impact on GI and plays a decisive role. According to different structures, starch can be divided into amylose and amylopectin; according to different digestion characteristics, starch can be divided into fast digestible starch, slow digestible starch and resistant starch. Cereals high in amylose and resistant starch have a relatively low GI. Different processing methods will affect the microstructure of grain food and the digestion characteristics of starch, and then affect its GI, such as cooking, extrusion puffing, baking, frying, etc. In the actual processing process, resistant starch can be increased through the control of process parameters content, reduce the hydrolysis rate of starch, and change the GI of cereal foods. At the same time, the consumption state of food, including thin consistency and particle size, will also affect GI. Loose and porous foods have a high GI, while hard foods that are hard to digest are relatively low in GI. It can be seen that reducing the content of carbohydrates in food or controlling the decomposition rate of carbohydrates is the basic idea of carrying out low GI food research. Low GI cereals can be produced by using low GI food raw materials, adopting different food formulas, and improving food processing methods.

How to develop a method of maintaining high protein in cereals while reducing the GI value of the cereal composition, and provide a healthy food composition with high protein nutrition and no high sugar increase will be widely loved.

Contents of the invention

The object of the present invention is to propose a compound cereal composition with high protein and low GI and its preparation method and application, which has delicate taste, good dispersibility, no precipitation and agglomeration, and pure taste. amino acids, protein peptides and other easily absorbed protein nutrients. At the same time, it is rich in dietary fiber and has a very low GI value. After eating, it can play a very good role in immune regulation and blood sugar regulation, and has a wide range of applications prospect.

Technical scheme of the present invention is realized like this:

The invention provides a method for preparing a compound cereal composition with high protein and low GI. After secondary enzymolysis of pea protein powder and white kidney bean extract, fermentation by Lactobacillus plantarum, Saccharomyces cerevisiae and Lactobacillus bulgaricus, freezing After drying, the enzymatic fermented protein peptide powder is obtained, and after being embedded in sodium carboxymethyl cellulose, sodium alginate, and water-soluble chitosan, slow-release protein particles are obtained; the mixed colloid of oat flour and rice bran powder is ground into a fine powder , mixed with wheat amylose, adjust the moisture content, through twin-screw extrusion, spray drying, to obtain low GI puffed powder; mix slow-release protein granules, low GI puffed powder and prebiotic sweetener evenly to obtain high protein low GI's complex cereal composition.

As a further improvement of the present invention, comprising the following steps:

S1. First-level enzymatic hydrolysis: mix and disperse pea protein powder and white kidney bean extract in water, add first-level compound protease, heat enzymatic hydrolysis, and inactivate the enzyme to obtain the first-level enzymatic hydrolysis protein peptide product;

S2. Secondary enzymatic hydrolysis: disperse the primary enzymatic protein peptide product prepared in step S1 in water, add secondary compound protease, heat enzymatic hydrolysis, and inactivate the enzyme to obtain the secondary enzymatic protein peptide product;

S3. Activation of fermented probiotics: Inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaulardian medium respectively, activate and cultivate, and obtain strain seed liquid;

S4. Fermentation: Inoculate the seed liquids of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus prepared in step S3 into the secondary enzymatic protein peptide products obtained in step S2, carry out fermentation culture, and freeze-dry to obtain Enzymolysis fermented protein peptide powder;

S5. Preparation of sustained-release protein granules: Dissolve sodium carboxymethyl cellulose, sodium alginate, and water-soluble chitosan in water, add the enzymatic fermented protein peptide powder prepared in step S4, stir and mix evenly, and then add food High-grade emulsifier and edible oil, fast membrane emulsification, dropwise addition of calcium chloride solution, solidification at room temperature, centrifugation, washing, and freeze-drying to obtain slow-release protein particles;

S6. Colloid mill: mix oat flour and rice bran powder evenly, and grind through a colloid mill to obtain dietary fiber fine powder;

S7. Preparation of wheat amylose: the wheat flour is subjected to acid hydrolysis, and the pH value is adjusted to be neutral, then filtered, the filter cake is dried, the supernatant obtained through gelatinization and centrifugation is frozen and crystallized, melted at room temperature into a mixture of ice and water, pumped Filter and dry to obtain wheat amylose;

S8. Preparation of low GI puffed powder: After mixing the dietary fiber fine powder prepared in step S6 and the wheat amylose prepared in step S7 evenly, adjust the moisture content, and obtain low GI puffed powder after twin-screw extrusion;

S9. Preparation of prebiotic sweetener: mix xylitol and steviol glycoside evenly to obtain prebiotic sweetener;

S10. Taste adjustment: uniformly mix the slow-release protein granules prepared in step S5, the low-GI puffed powder prepared in step S8, and the prebiotic sweetener prepared in step S9 to obtain a high-protein and low-GI compound cereal composition.

As a further improvement of the present invention, the mass ratio of pea protein powder and white kidney bean extract described in step S1 is 5-10:3-5, and the primary composite protease is a mixture of alkaline protease and papain, and the mass ratio 5-7:2, the temperature of the enzymolysis is 45-55°C, and the time is 2-4h; the secondary composite protease in step S2 is a mixture of neutral protease and animal protease, and the mass ratio is 3-5 : 1; the temperature of the enzymolysis is 40-60°C, and the time is 1-3h; the method of inactivating the enzyme is 1000-1500W microwave treatment for 3-5min.

As a further improvement of the present invention, the conditions for the activation culture described in step S3 are under micro-anoxic conditions, 35-38°C, 50-70r/min, cultured for 18-24h, and the bacterial content of the bacterial seed solution is 10 ⁸ -10 ⁹ cfu/mL; the inoculum amounts of Lactobacillus plantarum, Saccharomyces cerevisiae and Lactobacillus bulgaricus described in step S4 are 3-5%, 1-2%, 2-3%, and the conditions of the fermentation culture Under micro-anoxic conditions, 36-39°C, 50-70r/min, cultured for 48-72h.

As a further improvement of the present invention, the mass ratio of sodium carboxymethylcellulose, sodium alginate, water-soluble chitosan, enzymatic fermented protein peptide powder, food-grade emulsifier and edible oil in step S5 is 5-7 :2-4:3-7:10-12:1-2:30-50; the food grade emulsifier is selected from sodium stearoyl lactylate, diacetyl tartrate monoglyceride, sucrose fatty ester, distilled monoglyceride At least one of, the edible oil is selected from at least one of peanut oil, soybean oil, corn oil, rapeseed oil, sesame oil, linseed oil, olive oil; the pore size of the fast membrane is between 1000-3000nm, The concentration of the calcium chloride solution is 3-5wt%, and the curing time at normal temperature is 30-50min.

As a further improvement of the present invention, the mass ratio of oat flour and rice bran powder described in step S6 is 5-7:3-5, and the grinding time of the colloid mill is 3-5h; the method of acidolysis described in step S7 is The wheat flour is soaked in 5-10wt% acetic acid solution for 2-4 days, and the solid-to-liquid ratio of the wheat flour and the acetic acid solution is 1:3-5g/mL.

As a further improvement of the present invention, the mass ratio of dietary fiber fine powder and wheat amylose described in step S8 is 3-5:7-12, and the adjusted moisture content is 200-300g/kg, and the twin-screw extruder The barrel temperature of the press is 120-130°C, the screw speed power is 20-40Hz; the mass ratio of xylitol and steviol glycosides in step S9 is 5-7:2-4; the slow-release protein described in step S10 The mass ratio of granule, low GI puffed powder and prebiotic sweetener is 30-50:45-70:5-7.

As a further improvement of the present invention, it specifically includes the following steps:

S1. Primary enzymatic hydrolysis: mix and disperse 5-10 parts by weight of pea protein powder and 3-5 parts by weight of white kidney bean extract in 100 parts by weight of water, add 1-2 parts by weight of primary compound protease, and heat to 45-55 Enzymolysis at ℃ for 2-4 hours, followed by 1000-1500W microwave treatment for 3-5 minutes to obtain the first-grade enzymolysis protein peptide product;

Described primary compound protease is the mixture of alkaline protease and papain, and mass ratio is 5-7:2;

S2. Secondary enzymatic hydrolysis: Disperse 20 parts by weight of the primary enzymatic protein peptide product prepared in step S1 in 50 parts by weight of water, add 1-2 parts by weight of secondary composite protease, heat to 40-60°C for enzymatic hydrolysis 1 -3h, 1000-1500W microwave inactivating enzyme treatment for 3-5min to obtain the secondary enzymatic protein peptide product;

The secondary composite protease is a mixture of neutral protease and animal protease, and the mass ratio is 3-5:1;

S3. Activation of fermented probiotics: Inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaussian medium respectively, under micro-anoxic conditions, 35-38°C, 50-70r/min, activate and cultivate for 18-24h, Obtain the strain seed solution, the bacterial content is 10 ⁸ -10 ⁹ cfu/mL;

S4. Fermentation: Inoculate the seed liquids of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus prepared in step S3 into the secondary enzymatic protein peptide products obtained in step S2 respectively, and the inoculum amounts are 3-5% respectively , 1-2%, 2-3%, under micro-anoxic conditions, 36-39°C, 50-70r/min, fermented for 48-72h, freeze-dried to obtain enzymatic fermented protein peptide powder;

S5. Preparation of sustained-release protein particles: 5-7 parts by weight of sodium carboxymethylcellulose, 2-4 parts by weight of sodium alginate, and 3-7 parts by weight of water-soluble chitosan are dissolved in 30 parts by weight of water, and 10 parts by weight are added. - 12 parts by weight of the enzymatic fermented protein peptide powder prepared in step S4, after stirring and mixing evenly, add 1-2 parts by weight of a food-grade emulsifier and 30-50 parts by weight of edible oil, and quickly emulsify the membrane with a pore size of 1000-3000nm, Add 3-5wt% calcium chloride solution dropwise, solidify at room temperature for 30-50 minutes, centrifuge, wash, and freeze-dry to obtain slow-release protein particles;

S6. Colloid mill: Mix 50-70 parts by weight of oat flour and 30-50 parts by weight of rice bran powder evenly, grind for 3-5 hours in a colloid mill, and pass through a 300-500 mesh screen to obtain fine dietary fiber powder;

S7. Preparation of wheat amylose: soak wheat flour in 5-10wt% acetic acid solution for 2-4 days, the solid-to-liquid ratio of the wheat flour and acetic acid solution is 1:3-5g/mL, adjust the pH to be neutral Finally, filter, dry the filter cake, freeze and crystallize the supernatant obtained by gelatinization and centrifugation, melt at room temperature into an ice-water mixture, filter with suction, and dry to obtain wheat amylose;

S8. Preparation of low GI puffed powder: After mixing the dietary fiber fine powder prepared in 30-50 parts by weight of step S6 and the wheat amylose prepared in 70-120 parts by weight of step S7, the moisture content is adjusted to 200-300g /kg, after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 120-130°C, the screw speed power is 20-40Hz, and low GI puffed powder is obtained;

S9. Preparation of a prebiotic sweetener: uniformly mix 5-7 parts by weight of xylitol and 2-4 parts by weight of steviol glycoside to obtain a prebiotic sweetener;

S10. Taste adjustment: 30-50 parts by weight of the sustained-release protein granules prepared in step S5, 45-70 parts by weight of the low-GI puffed powder prepared in step S8, and 5-7 parts by weight of the prebiotic sweetness prepared in step S9 The ingredients are evenly mixed to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 3-5% CO ₂ , 5-10% O ₂ , the balance is nitrogen, and % is volume percentage.

The present inventors further protect a compound cereal composition with high protein and low GI prepared by the above preparation method.

The present inventors further protect the application of the above-mentioned high-protein and low-GI compound cereal composition in the preparation of low-GI, high-protein nutritional food.

The present invention has following beneficial effects:

There are anti-nutritional factors such as tannin, phytic acid and protease inhibitors in beans. These anti-nutritional factors are regarded as unfavorable factors that hinder the absorption of dietary nutrients and affect the health of the body. Among them, trypsin inhibitors, α-galactooligosaccharides and Anti-nutritional factors such as phytic acid hinder the absorption of nutrients, causing physical discomfort or defects in the sensory and nutritional value of soy milk, and in severe cases, it may lead to risks such as food poisoning. At present, heat treatment is mainly used to inactivate anti-nutritional factors. Excessive heating used to achieve complete inactivation can reduce the solubility of bean protein and destroy nutrients. Moreover, trypsin inhibitors are relatively heat-resistant, and the effect of heat treatment often outweighs the gain.

In the present invention, the enzymolysis method of secondary protease is first adopted to enzymatically hydrolyze legume protein, including pea protein powder and white kidney bean extract. The primary composite protease is a mixture of alkaline protease and papain, Under the enzymatic hydrolysis of protein substrates, the cleavage sites of alkaline protease are all hydrophobic and aromatic amino acid carboxyl amide bonds on the carboxyl side chains, and catalyze the hydrolysis of peptide bonds in protein molecules to produce small molecular weight Peptide papain is a sulfhydryl protease, and papain is complementary to the peptide bond catalyzed by alkaline protease, so that it can hydrolyze proteins to the greatest extent; the secondary composite protease is a mixture of neutral protease and animal protease, and neutral protease is a metal Protease, which preferentially cleaves the peptide bond between leucine and phenylalanine, animal protease can decompose animal protein to macromolecular protein through the control of hydrolysis degree, so as to significantly enhance the nutrition of substrate protein;

After secondary enzymatic hydrolysis, trypsin inhibitors (a type of small molecular protein) can be degraded to the greatest extent, thereby greatly reducing the content of trypsin inhibitors in beans, and at the same time, bean proteins can be better enzymatically hydrolyzed into small molecule peptides to increase the absorption rate of protein;

Furthermore, the secondary enzymatic protein peptide products are fermented by Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus. These probiotics can decompose tannins, α-galactooligosaccharides, and phytic acid through self-proliferation on the one hand. On the other hand, the growth of the three has a synergistic effect. Lactobacillus plantarum can utilize monosaccharides in the early stage of fermentation, and produce formic acid, propionic acid, butyric acid, folic acid, etc. required for the proliferation of Lactobacillus bulgaricus. In the later stage of fermentation, Saccharomyces cerevisiae and Lactobacillus bulgaricus produce a large amount of lactic acid and amino acids, which promote the growth of Lactobacillus plantarum, complement each other, and promote each other, thereby promoting the fermentation of fermentation bacteria and producing a large amount of beneficial nutrients. At the same time, a large number of probiotics Proliferation, the obtained enzymatic fermented protein peptide powder is rich in short-chain fatty acids, live probiotics, amino acids, protein peptides, etc., which consumes almost all anti-nutritional factors and greatly improves the nutritional value of the product.

Further, the present invention prepares a shell material with obvious acid resistance through sodium carboxymethyl cellulose, sodium alginate, and water-soluble chitosan, and wraps the obtained enzymatic fermentation protein peptide in granules, and after entering the human body, Can resist gastric acid erosion, effectively protect live probiotic bacteria against acidic environment, protect protein against pepsin degradation and inactivation, smoothly enter the intestinal tract, absorb nutrients smoothly by the small intestine, probiotic colonization and intestinal wall, play a good role in regulation Immunity and blood sugar, enhance the body's anti-sugar, anti-oxidation, anti-aging functions, and have a certain anti-cancer effect.

Oat flour and rice bran powder are an excellent dietary fiber. After being ground by a colloid mill, a very fine dietary fiber powder is obtained. Wheat flour food refers to food made from refined wheat flour. Common ones include noodles, steamed buns, biscuits, bread, etc. According to different raw materials and processing methods, the GI is 55-85. The invention prepares wheat amylose, on the one hand, after entering the human body, it can form an anti-digestion complex with lipids to reduce the digestibility of starch. On the other hand, amylose can reduce the gelatinization degree of starch, increase crystallinity, and promote starch aging, thereby greatly reducing the GI value of grains. By adding the dietary fiber fine powder prepared by the invention and its suitable particle size to the wheat amylose, the overall GI value can be significantly reduced.

After mixing and extruding oat flour, rice bran flour, and wheat amylose in an appropriate proportion, the vitamins, trace elements, and amino acids in the grain flour can be well preserved. By adjusting the parameters of the appropriate twin-screw extruder, the obtained lower GI puffed powder has a more delicate taste, better dispersibility, no sedimentation and agglomeration, and better sensory quality.

Xylitol and steviol glycosides can not only achieve the effect of low calorie, but also maintain the effect of sweetening. At the same time, they are also very high-quality prebiotics, which can well promote the proliferation of intestinal probiotics, thereby inhibiting the growth of harmful bacteria, and indirectly play a role Good immune regulation and blood sugar regulation.

The compound cereal composition with high protein and low GI prepared by the present invention has delicate taste, good dispersibility, no precipitation and agglomeration, pure taste, and at the same time, does not contain anti-nutritional factors, and is rich in easily absorbed proteins such as amino acids and protein peptides At the same time, it is rich in dietary fiber and has a very low GI value. After eating, it can play a good role in immune regulation and blood sugar regulation, and has broad application prospects.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only These are some embodiments of the present invention. For those skilled in the art, other drawings can also be obtained according to these drawings without any creative effort.

Fig. 1 is the comparative figure of each group's survival rate in test example 1 of the present invention;

Figure 2 is a comparison chart of the release rates of each group in Test Example 1 of the present invention.

Detailed ways

The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some of the embodiments of the present invention, but not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

Pea protein powder, with a content greater than 90%, was purchased from Xi’an Lvteng Biotechnology Co., Ltd.; white kidney bean extract, which is a water extract of white kidney bean, is rich in protein, vitamins, white kidney bean saponins, amino acids, trace elements, etc., and was purchased from Xi'an Prius Biotechnology Co., Ltd.; oat flour, 100-120 mesh, purchased from Zhangjiakou North Oat Food Development Co., Ltd., rice bran powder, 70-100 mesh, purchased from Jiangxi Hehe Industrial Co., Ltd.; wheat flour, moisture quality The fraction is 8%-12%, and the protein mass fraction is ≤0.3%, purchased from Sigma Company of the United States.

Alkaline protease (200000 U/g) and papain (800000 U/g) were purchased from Beijing Suolaibao Technology Co., Ltd.; neutral protease (200000 U/g) and animal protease (100000 U/g) were purchased from From Nanning Donghenghuadao Biotechnology Co., Ltd.;

Lactobacillus plantarum (5 billion cfu/g) was purchased from Weifang Ruichen Biotechnology Co., Ltd.; Saccharomyces cerevisiae (20 billion cfu/g) was purchased from Zhuhai Wenqi Biotechnology Co., Ltd. Lactobacillus bulgaricus was Lactobacillus bulgaricus LB -Z16 (10 billion cfu/g), purchased from Shandong Zhongke Jiayi Bioengineering Co., Ltd.

Example 1

The present embodiment provides a compound cereal composition with high protein and low GI, and its preparation method is as follows:

S1. Primary enzymatic hydrolysis: Mix 5 parts by weight of pea protein powder and 3 parts by weight of white kidney bean extract into 100 parts by weight of water, stir at 500 r/min for 10 minutes, add 1 part by weight of primary compound protease, and heat to 45°C for enzymatic hydrolysis 2h, 1000W microwave treatment for 3min to inactivate the enzyme, and obtain the first-level enzymolysis protein peptide product;

Described primary composite protease is the mixture of alkaline protease and papain, and mass ratio is 5:2;

S2. Secondary enzymatic hydrolysis: Mix 20 parts by weight of the primary enzymatic protein peptide product prepared in step S1 into 50 parts by weight of water, stir at 500r/min for 10min, add 1 part by weight of the secondary compound protease, and heat to 40°C to enzymatically After 1 hour of hydrolysis, 1000W microwave inactivates the enzyme for 3 minutes to obtain the second-level enzymolysis protein peptide product;

The secondary composite protease is a mixture of neutral protease and animal protease, and the mass ratio is 3:1;

S3. Activation of fermented probiotics: Inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaulier medium respectively, and activate and cultivate them for 18 hours at 35°C and 50 r/min under micro-anoxic conditions to obtain the strain seed liquid. The bacteria content is 10 ⁸ cfu/mL;

S4. Fermentation: Inoculate the seed liquids of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus prepared in step S3 into the secondary enzymatic protein peptide products prepared in step S2, respectively, with the inoculation amounts of 3%, 1 %, 2%, under micro-anoxic conditions, 36°C, 50r/min, fermented for 48 hours, freeze-dried to obtain enzymatic fermented protein peptide powder;

S5. Preparation of slow-release protein particles: 5 parts by weight of sodium carboxymethylcellulose, 2 parts by weight of sodium alginate, and 3 parts by weight of water-soluble chitosan are dissolved in 30 parts by weight of water, and 10 parts by weight of step S4 is added to prepare Enzymolysis fermented protein peptide powder, 500r/min stirring and mixing for 15min, adding 1 weight part of sodium stearoyl lactylate and 30 weight parts of soybean oil, rapid membrane emulsification with a pore size of 1000nm, adding 3wt% calcium chloride solution dropwise, and curing at room temperature 30min, centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain sustained-release protein particles;

S6. Colloid mill: Mix 50 parts by weight of oat flour and 30 parts by weight of rice bran powder evenly, grind for 3 hours through a colloid mill, and pass through a 300-mesh sieve to obtain fine dietary fiber powder;

S7. Preparation of wheat amylose: soak the wheat flour in 5wt% acetic acid solution for 2 days, the solid-to-liquid ratio of the wheat flour and the acetic acid solution is 1:3g/mL, after adjusting the pH value to be neutral, filter, filter cake Dry in the sun, gelatinize and centrifuge the supernatant obtained by freezing and crystallization, melt at room temperature into an ice-water mixture, filter with suction, and dry at 70°C for 2 hours to obtain wheat amylose;

S8. Preparation of low GI puffed powder: mix the dietary fiber fine powder prepared by 30 parts by weight of step S6 and the wheat amylose starch prepared by 70 parts by weight of step S7, stir at 300r/min for 15min, and adjust the moisture content to 200g/kg , after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 120°C, the screw speed power is 20Hz, and low GI puffed powder is obtained;

S9. Preparation of a prebiotic sweetener: Mix 5 parts by weight of xylitol and 2 parts by weight of steviol glycoside, and stir at 300 r/min for 10 minutes to obtain a prebiotic sweetener;

S10. Taste adjustment: Mix 30 parts by weight of the sustained-release protein granules prepared in step S5, 45 parts by weight of the low-GI puffed powder prepared in step S8, and 5 parts by weight of the prebiotic sweetener prepared in step S9, 300r/min Stir for 10 minutes to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 3% CO ₂ , 5% O ₂ , the balance is nitrogen, and % is volume percentage.

Example 2

The present embodiment provides a compound cereal composition with high protein and low GI, and its preparation method is as follows:

S1. Primary enzymatic hydrolysis: Mix 10 parts by weight of pea protein powder and 5 parts by weight of white kidney bean extract into 100 parts by weight of water, stir at 500 r/min for 10 minutes, add 2 parts by weight of primary compound protease, and heat to 55°C for enzymolysis 4h, 1500W microwave treatment for 5min to inactivate the enzyme, and obtain the first-level enzymolysis protein peptide product;

Described primary composite protease is the mixture of alkaline protease and papain, and mass ratio is 7:2;

S2. Secondary enzymatic hydrolysis: Mix 20 parts by weight of the primary enzymatic protein peptide product prepared in step S1 into 50 parts by weight of water, stir at 500r/min for 10min, add 2 parts by weight of the secondary compound protease, and heat to 60°C to enzymatically After 3 hours of hydrolysis, 1500W microwave inactivates the enzyme for 5 minutes to obtain the second-level enzymolysis protein peptide product;

The secondary composite protease is a mixture of neutral protease and animal protease, and the mass ratio is 5:1;

S3. Activation of fermented probiotics: inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaulier medium respectively, under micro-anoxic conditions, 38°C, 70r/min, activate and cultivate for 24h, to obtain strain seed liquid, The bacteria content is 10 ⁹ cfu/mL;

S4. Fermentation: Inoculate the seed liquids of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus prepared in step S3 into the secondary enzymatic protein peptide products obtained in step S2 respectively, and the inoculation amounts are 5%, 2%, respectively. %, 3%, under micro-anoxic conditions, 39°C, 70r/min, fermented for 72 hours, freeze-dried to obtain enzymatic fermented protein peptide powder;

S5. Preparation of slow-release protein particles: 7 parts by weight of sodium carboxymethylcellulose, 4 parts by weight of sodium alginate, and 7 parts by weight of water-soluble chitosan are dissolved in 30 parts by weight of water, and 12 parts by weight of step S4 are added to prepare Enzymolysis fermented protein peptide powder, stirred and mixed at 500r/min for 15min, added 2 parts by weight of distilled monoglyceride and 50 parts by weight of sesame oil, quickly emulsified with a pore size of 3000nm, added dropwise 5wt% calcium chloride solution, and solidified at room temperature for 50 minutes , centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain slow-release protein particles;

S6. Colloid mill: Mix 70 parts by weight of oat flour and 50 parts by weight of rice bran powder evenly, grind for 5 hours through a colloid mill, and pass through a 500-mesh sieve to obtain fine dietary fiber powder;

S7. Preparation of wheat amylose: soak the wheat flour in 10wt% acetic acid solution for 4 days, the solid-to-liquid ratio of the wheat flour and the acetic acid solution is 1:5g/mL, after adjusting the pH value to be neutral, filter, filter cake Dry in the sun, gelatinize and centrifuge the supernatant obtained by freezing and crystallization, melt at room temperature into an ice-water mixture, filter with suction, and dry at 70°C for 2 hours to obtain wheat amylose;

S8. Preparation of low GI puffed powder: mix the dietary fiber fine powder prepared by 50 parts by weight of step S6 and the wheat amylose obtained by 120 parts by weight of step S7, stir at 300r/min for 15min, and adjust the moisture content to 300g/kg , after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 130°C, the screw speed power is 40Hz, and low GI puffed powder is obtained;

S9. Preparation of a prebiotic sweetener: Mix 7 parts by weight of xylitol and 4 parts by weight of steviol glycoside, and stir at 300 r/min for 10 minutes to obtain a prebiotic sweetener;

S10. Taste adjustment: mix 50 parts by weight of the sustained-release protein granules prepared in step S5, 70 parts by weight of the low-GI puffed powder prepared in step S8, and 7 parts by weight of the prebiotic sweetener prepared in step S9, 300r/min Stir for 10 minutes to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 5% CO ₂ , 10% O ₂ , the balance is nitrogen, and % is volume percentage.

Example 3

The present embodiment provides a compound cereal composition with high protein and low GI, and its preparation method is as follows:

S1. Primary enzymatic hydrolysis: Mix 7 parts by weight of pea protein powder and 4 parts by weight of white kidney bean extract into 100 parts by weight of water, stir at 500 r/min for 10 minutes, add 1.5 parts by weight of primary compound protease, and heat to 50°C for enzymolysis 3h, 1250W microwave treatment for 4min to inactivate the enzyme, and obtain the first-level enzymolysis protein peptide product;

Described primary composite protease is the mixture of alkaline protease and papain, and mass ratio is 6:2;

S2. Secondary enzymatic hydrolysis: Mix 20 parts by weight of the primary enzymatic protein peptide product prepared in step S1 into 50 parts by weight of water, stir at 500r/min for 10min, add 1.5 parts by weight of the secondary compound protease, and heat to 50°C to enzymatically Decompose for 2 hours, 1250W microwave inactivates the enzyme for 4 minutes, and obtains the second-level enzymatic hydrolysis protein peptide product;

The secondary composite protease is a mixture of neutral protease and animal protease, and the mass ratio is 4:1;

S3. Activation of fermented probiotics: Inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaulier medium respectively, under micro-anoxic conditions, 37°C, 60r/min, activate and cultivate for 22h, to obtain strain seed liquid, The bacteria content is 10 ⁹ cfu/mL;

S4. Fermentation: inoculate the seed liquids of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus prepared in step S3 into the secondary enzymatic protein peptide products obtained in step S2, and the inoculum amounts are 4%, 1.5%, respectively. %, 2.5%, under micro-anoxic conditions, 37°C, 60r/min, fermented for 56 hours, freeze-dried to obtain enzymatic fermented protein peptide powder;

S5. Preparation of slow-release protein particles: 6 parts by weight of sodium carboxymethylcellulose, 3 parts by weight of sodium alginate, and 5 parts by weight of water-soluble chitosan are dissolved in 30 parts by weight of water, and 11 parts by weight of step S4 are added to prepare Enzymolysis fermented protein peptide powder, 500r/min stirring and mixing for 15min, adding 1.5 parts by weight of sucrose fatty ester and 40 parts by weight of corn oil, rapid membrane emulsification with a pore size of 2000nm, adding 4wt% calcium chloride solution dropwise, curing at room temperature for 40 minutes , centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain slow-release protein particles;

S6. Colloid mill: Mix 60 parts by weight of oat flour and 40 parts by weight of rice bran powder evenly, grind for 4 hours through a colloid mill, and pass through a 400-mesh sieve to obtain fine dietary fiber powder;

S7. Preparation of wheat amylose: soak the wheat flour in 7wt% acetic acid solution for 3 days, the solid-to-liquid ratio of the wheat flour and the acetic acid solution is 1:4g/mL, after adjusting the pH value to be neutral, filter, filter cake Dry in the sun, gelatinize and centrifuge the supernatant obtained by freezing and crystallization, melt at room temperature into an ice-water mixture, filter with suction, and dry at 70°C for 2 hours to obtain wheat amylose;

S8. Preparation of low GI puffed powder: mix the dietary fiber fine powder prepared by 40 parts by weight of step S6 and the wheat amylose prepared by 100 parts by weight of step S7, stir at 300r/min for 15min, and adjust the moisture content to 250g/kg , after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 125°C, the screw speed power is 30Hz, and low GI puffed powder is obtained;

S9. Preparation of a prebiotic sweetener: Mix 6 parts by weight of xylitol and 3 parts by weight of steviol glycoside, and stir at 300 r/min for 10 minutes to obtain a prebiotic sweetener;

S10. Taste adjustment: Mix 40 parts by weight of the sustained-release protein granules prepared in step S5, 55 parts by weight of the low-GI puffed powder prepared in step S8, and 6 parts by weight of the prebiotic sweetener prepared in step S9, 300r/min Stir for 10 minutes to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 4% CO ₂ , 7% O ₂ , the balance is nitrogen, and % is volume percentage.

Example 4

Compared with Example 3, the difference is that the primary composite protease is a single alkaline protease.

Example 5

Compared with Example 3, the difference is that the primary composite protease is a single papain.

Example 6

Compared with Example 3, the difference is that the secondary composite protease is a single neutral protease.

Example 7

Compared with Example 3, the difference is that the secondary composite protease is a single animal protease.

Comparative example 1

Compared with Example 3, the difference is that no pea protein powder is added in step S1.

details as follows:

S1. Primary enzymatic hydrolysis: Add 11 parts by weight of white kidney bean extract to 100 parts by weight of water, stir at 500 r/min for 10 minutes, add 1.5 parts by weight of primary compound protease, heat to 50°C for 3 hours, and treat with 1250W microwave for 4 minutes , to obtain a primary enzymolysis protein peptide product;

The primary composite protease is a mixture of alkaline protease and papain, and the mass ratio is 6:2.

Comparative example 2

Compared with Example 3, the difference is that no white kidney bean extract is added in step S1.

details as follows:

S1. Primary enzymatic hydrolysis: Add 11 parts by weight of pea protein powder to 100 parts by weight of water, stir at 500 r/min for 10 minutes, add 1.5 parts by weight of primary compound protease, heat to 50°C for 3 hours, and then 1250W microwave to inactivate the enzyme for 4 minutes. Obtain primary enzymolysis protein peptide products;

The primary composite protease is a mixture of alkaline protease and papain, and the mass ratio is 6:2.

Comparative example 3

Compared with Example 3, the difference is that step S1 is not performed.

details as follows:

S1. Enzymolysis: Mix 7 parts by weight of pea protein powder and 4 parts by weight of white kidney bean extract into 100 parts by weight of water, stir at 500r/min for 10 minutes, add 1.5 parts by weight of compound protease, heat to 50°C for 2 hours, and use 1250W microwave Enzyme inactivation treatment for 4 minutes to obtain the enzymatic protein peptide product;

Described composite protease is the mixture of neutral protease and animal protease, and mass ratio is 4:1;

S2. Activation of fermented probiotics: inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaulier medium respectively, under micro-anoxic conditions, 37°C, 60r/min, activate and cultivate for 22h, and obtain strain seed liquid, The bacteria content is 10 ⁹ cfu/mL;

S3. Fermentation: Inoculate the seed liquids of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus prepared in step S3 into the enzymatic protein peptide products prepared in step S1 respectively, and the inoculum amounts are 4%, 1.5%, 2.5%, under micro-anoxic conditions, 37°C, 60r/min, fermented for 56 hours, freeze-dried to obtain enzymatic fermented protein peptide powder;

S4. Preparation of slow-release protein particles: 6 parts by weight of sodium carboxymethylcellulose, 3 parts by weight of sodium alginate, and 5 parts by weight of water-soluble chitosan are dissolved in 30 parts by weight of water, and 11 parts by weight of step S3 are added to prepare Enzymolysis fermented protein peptide powder, 500r/min stirring and mixing for 15min, adding 1.5 parts by weight of sucrose fatty ester and 40 parts by weight of corn oil, rapid membrane emulsification with a pore size of 2000nm, adding 4wt% calcium chloride solution dropwise, curing at room temperature for 40 minutes , centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain slow-release protein particles;

S5. Colloid mill: Mix 60 parts by weight of oat flour and 40 parts by weight of rice bran powder evenly, grind for 4 hours through a colloid mill, and pass through a 400-mesh sieve to obtain fine dietary fiber powder;

S6. Preparation of wheat amylose: soak the wheat flour in 7wt% acetic acid solution for 3 days, the solid-to-liquid ratio of the wheat flour and the acetic acid solution is 1:4g/mL, after adjusting the pH value to be neutral, filter, filter cake Dry in the sun, gelatinize and centrifuge the supernatant obtained by freezing and crystallization, melt at room temperature into an ice-water mixture, filter with suction, and dry at 70°C for 2 hours to obtain wheat amylose;

S7. Preparation of low GI puffed powder: mix the dietary fiber fine powder prepared by 40 parts by weight of step S5 and the wheat amylose prepared by 100 parts by weight of step S6, stir at 300r/min for 15min, and adjust the moisture content to 250g/kg , after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 125°C, the screw speed power is 30Hz, and low GI puffed powder is obtained;

S8. Preparation of a prebiotic sweetener: Mix 6 parts by weight of xylitol and 3 parts by weight of steviol glycoside, and stir at 300 r/min for 10 minutes to obtain a prebiotic sweetener;

S9. Adjusting mouthfeel: mix 40 parts by weight of slow-release protein granules prepared in step S4, 55 parts by weight of low GI puffed powder prepared in step S7, and 6 parts by weight of prebiotic sweetener prepared in step S8, 300r/min Stir for 10 minutes to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 4% CO ₂ , 7% O ₂ , the balance is nitrogen, and % is volume percentage.

Comparative example 4

Compared with Embodiment 3, the difference is that step S2 is not performed.

details as follows:

S1. Primary enzymatic hydrolysis: Mix 7 parts by weight of pea protein powder and 4 parts by weight of white kidney bean extract into 100 parts by weight of water, stir at 500 r/min for 10 minutes, add 1.5 parts by weight of primary compound protease, and heat to 50°C for enzymolysis 3h, 1250W microwave treatment for 4min to inactivate the enzyme, and obtain the first-level enzymolysis protein peptide product;

Described primary composite protease is the mixture of alkaline protease and papain, and mass ratio is 6:2;

S2. Activation of fermented probiotics: inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaulier medium respectively, under micro-anoxic conditions, 37°C, 60r/min, activate and cultivate for 22h, and obtain strain seed liquid, The bacteria content is 10 ⁹ cfu/mL;

S3. Fermentation: inoculate the seed liquids of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus prepared in step S3 into the first-level enzymatic protein peptide products prepared in step S1, and the inoculum amounts are 4%, 1.5%, respectively. %, 2.5%, under micro-anoxic conditions, 37°C, 60r/min, fermented for 56 hours, freeze-dried to obtain enzymatic fermented protein peptide powder;

S4. Preparation of slow-release protein particles: 6 parts by weight of sodium carboxymethylcellulose, 3 parts by weight of sodium alginate, and 5 parts by weight of water-soluble chitosan are dissolved in 30 parts by weight of water, and 11 parts by weight of step S3 are added to prepare Enzymolysis fermented protein peptide powder, 500r/min stirring and mixing for 15min, adding 1.5 parts by weight of sucrose fatty ester and 40 parts by weight of corn oil, rapid membrane emulsification with a pore size of 2000nm, adding 4wt% calcium chloride solution dropwise, curing at room temperature for 40 minutes , centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain slow-release protein particles;

S5. Colloid mill: Mix 60 parts by weight of oat flour and 40 parts by weight of rice bran powder evenly, grind for 4 hours through a colloid mill, and pass through a 400-mesh sieve to obtain fine dietary fiber powder;

S6. Preparation of wheat amylose: soak the wheat flour in 7wt% acetic acid solution for 3 days, the solid-to-liquid ratio of the wheat flour and the acetic acid solution is 1:4g/mL, after adjusting the pH value to be neutral, filter, filter cake Dry in the sun, gelatinize and centrifuge the supernatant obtained by freezing and crystallization, melt at room temperature into an ice-water mixture, filter with suction, and dry at 70°C for 2 hours to obtain wheat amylose;

S7. Preparation of low GI puffed powder: mix the dietary fiber fine powder prepared by 40 parts by weight of step S5 and the wheat amylose prepared by 100 parts by weight of step S6, stir at 300r/min for 15min, and adjust the moisture content to 250g/kg , after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 125°C, the screw speed power is 30Hz, and low GI puffed powder is obtained;

S8. Preparation of a prebiotic sweetener: Mix 6 parts by weight of xylitol and 3 parts by weight of steviol glycoside, and stir at 300 r/min for 10 minutes to obtain a prebiotic sweetener;

S9. Adjusting mouthfeel: mix 40 parts by weight of slow-release protein granules prepared in step S4, 55 parts by weight of low GI puffed powder prepared in step S7, and 6 parts by weight of prebiotic sweetener prepared in step S8, 300r/min Stir for 10 minutes to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 4% CO ₂ , 7% O ₂ , the balance is nitrogen, and % is volume percentage.

Comparative example 5

Compared with Example 3, the difference is that steps S1 and S2 are not performed.

details as follows:

S1. Activation of fermented probiotics: Inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaulier medium respectively, under micro-anoxic conditions, 37°C, 60r/min, activate and cultivate for 22h, and obtain strain seed liquid, The bacteria content is 10 ⁹ cfu/mL;

S2. Fermentation: Mix 7 parts by weight of pea protein powder and 4 parts by weight of white kidney bean extract into 100 parts by weight of water, inoculate the seed liquid of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus, and the inoculation amounts are 4%, 1.5%, 2.5%, under micro-anoxic conditions, 37°C, 60r/min, fermented for 56 hours, freeze-dried to obtain enzymatic fermented protein peptide powder;

S3. Preparation of slow-release protein particles: 6 parts by weight of sodium carboxymethylcellulose, 3 parts by weight of sodium alginate, and 5 parts by weight of water-soluble chitosan are dissolved in 30 parts by weight of water, and 11 parts by weight of step S2 are added to prepare Enzymolysis fermented protein peptide powder, 500r/min stirring and mixing for 15min, adding 1.5 parts by weight of sucrose fatty ester and 40 parts by weight of corn oil, rapid membrane emulsification with a pore size of 2000nm, adding 4wt% calcium chloride solution dropwise, curing at room temperature for 40 minutes , centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain slow-release protein particles;

S4. Colloid mill: Mix 60 parts by weight of oat flour and 40 parts by weight of rice bran powder evenly, grind for 4 hours through a colloid mill, and pass through a 400-mesh sieve to obtain fine dietary fiber powder;

S5. Preparation of wheat amylose: soak the wheat flour in 7wt% acetic acid solution for 3 days, the solid-to-liquid ratio of the wheat flour and the acetic acid solution is 1:4g/mL, after adjusting the pH value to be neutral, filter, filter cake Dry in the sun, gelatinize and centrifuge the supernatant obtained by freezing and crystallization, melt at room temperature into an ice-water mixture, filter with suction, and dry at 70°C for 2 hours to obtain wheat amylose;

S6. Preparation of low GI puffed powder: mix the dietary fiber fine powder prepared in 40 parts by weight of step S4 and the wheat amylose obtained in step S5 of 100 parts by weight, stir at 300r/min for 15min, and adjust the moisture content to 250g/kg , after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 125°C, the screw speed power is 30Hz, and low GI puffed powder is obtained;

S7. Preparation of a prebiotic sweetener: Mix 6 parts by weight of xylitol and 3 parts by weight of steviol glycoside, and stir at 300 r/min for 10 minutes to obtain a prebiotic sweetener;

S8. Adjusting mouthfeel: mix 40 parts by weight of slow-release protein granules prepared in step S3, 55 parts by weight of low GI puffed powder prepared in step S6, and 6 parts by weight of prebiotic sweetener prepared in step S7, 300r/min Stir for 10 minutes to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 4% CO ₂ , 7% O ₂ , the balance is nitrogen, and % is volume percentage.

Comparative example 6

Compared with Example 3, the difference is that only Lactobacillus plantarum is inoculated in step S4.

details as follows:

S4. Fermentation: Inoculate the seed solution of Lactobacillus plantarum strain prepared in step S3 into the secondary enzymatic protein peptide product obtained in step S2, the inoculation amount is 8%, under micro-anoxic conditions, 37°C, 60r /min, fermented for 56 hours, and freeze-dried to obtain enzymatic fermented protein peptide powder.

Comparative example 7

Compared with Example 3, the difference is that only Saccharomyces cerevisiae is inoculated in step S4.

details as follows:

S4. Fermentation: Inoculate the Saccharomyces cerevisiae strain seed liquid obtained in step S3 into the secondary enzymatic protein peptide product obtained in step S2, the inoculation amount is 8%, under micro-anoxic conditions, 37°C, 60r /min, fermented for 56 hours, and freeze-dried to obtain enzymatic fermented protein peptide powder.

Comparative example 8

Compared with Example 3, the difference is that only Lactobacillus bulgaricus is inoculated in step S4.

details as follows:

S4. Fermentation: Inoculate the seed solution of Lactobacillus bulgaricus prepared in step S3 into the secondary enzymatic protein peptide product obtained in step S2, the inoculum amount is 8%, under micro-anoxic conditions, 37°C, 60r /min, fermented for 56 hours, and freeze-dried to obtain enzymatic fermented protein peptide powder.

Comparative example 9

Compared with Example 3, the difference is that steps S3 and S4 are not performed.

details as follows:

S1. Primary enzymatic hydrolysis: Mix 7 parts by weight of pea protein powder and 4 parts by weight of white kidney bean extract into 100 parts by weight of water, stir at 500 r/min for 10 minutes, add 1.5 parts by weight of primary compound protease, and heat to 50°C for enzymolysis 3h, 1250W microwave treatment for 4min to inactivate the enzyme, and obtain the first-level enzymolysis protein peptide product;

Described primary composite protease is the mixture of alkaline protease and papain, and mass ratio is 6:2;

S2. Secondary enzymatic hydrolysis: Mix 20 parts by weight of the primary enzymatic protein peptide product prepared in step S1 into 50 parts by weight of water, stir at 500r/min for 10min, add 1.5 parts by weight of the secondary compound protease, and heat to 50°C to enzymatically Decompose for 2 hours, treat with 1250W microwave for 4 minutes, and freeze-dry to obtain the second-level enzymolysis protein peptide product;

The secondary composite protease is a mixture of neutral protease and animal protease, and the mass ratio is 4:1;

S3. Preparation of slow-release protein particles: 6 parts by weight of sodium carboxymethylcellulose, 3 parts by weight of sodium alginate, and 5 parts by weight of water-soluble chitosan are dissolved in 30 parts by weight of water, and 11 parts by weight of step S2 are added to prepare The secondary enzymatic hydrolysis protein peptide product, stirred and mixed at 500r/min for 15min, added 1.5 parts by weight of sucrose fatty ester and 40 parts by weight of corn oil, quickly emulsified with a pore size of 2000nm, added dropwise 4wt% calcium chloride solution, and solidified at room temperature 40min, centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain sustained-release protein particles;

S4. Colloid mill: Mix 60 parts by weight of oat flour and 40 parts by weight of rice bran powder evenly, grind for 4 hours through a colloid mill, and pass through a 400-mesh sieve to obtain fine dietary fiber powder;

S5. Preparation of wheat amylose: soak the wheat flour in 7wt% acetic acid solution for 3 days, the solid-to-liquid ratio of the wheat flour and the acetic acid solution is 1:4g/mL, after adjusting the pH value to be neutral, filter, filter cake Dry in the sun, gelatinize and centrifuge the supernatant obtained by freezing and crystallization, melt at room temperature into an ice-water mixture, filter with suction, and dry at 70°C for 2 hours to obtain wheat amylose;

S6. Preparation of low GI puffed powder: mix the dietary fiber fine powder prepared in 40 parts by weight of step S4 and the wheat amylose obtained in step S5 of 100 parts by weight, stir at 300r/min for 15min, and adjust the moisture content to 250g/kg , after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 125°C, the screw speed power is 30Hz, and low GI puffed powder is obtained;

S7. Preparation of a prebiotic sweetener: Mix 6 parts by weight of xylitol and 3 parts by weight of steviol glycoside, and stir at 300 r/min for 10 minutes to obtain a prebiotic sweetener;

S8. Adjusting mouthfeel: mix 40 parts by weight of slow-release protein granules prepared in step S3, 55 parts by weight of low GI puffed powder prepared in step S6, and 6 parts by weight of prebiotic sweetener prepared in step S7, 300r/min Stir for 10 minutes to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 4% CO ₂ , 7% O ₂ , the balance is nitrogen, and % is volume percentage.

Comparative example 10

Compared with Embodiment 3, the difference is that step S5 is not performed.

details as follows:

S1. Primary enzymatic hydrolysis: Mix 7 parts by weight of pea protein powder and 4 parts by weight of white kidney bean extract into 100 parts by weight of water, stir at 500 r/min for 10 minutes, add 1.5 parts by weight of primary compound protease, and heat to 50°C for enzymolysis 3h, 1250W microwave treatment for 4min to inactivate the enzyme, and obtain the first-level enzymolysis protein peptide product;

Described primary composite protease is the mixture of alkaline protease and papain, and mass ratio is 6:2;

S2. Secondary enzymatic hydrolysis: Mix 20 parts by weight of the primary enzymatic protein peptide product prepared in step S1 into 50 parts by weight of water, stir at 500r/min for 10min, add 1.5 parts by weight of the secondary compound protease, and heat to 50°C to enzymatically Decompose for 2 hours, 1250W microwave inactivates the enzyme for 4 minutes, and obtains the second-level enzymatic hydrolysis protein peptide product;

The secondary composite protease is a mixture of neutral protease and animal protease, and the mass ratio is 4:1;

S3. Activation of fermented probiotics: Inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaulier medium respectively, under micro-anoxic conditions, 37°C, 60r/min, activate and cultivate for 22h, to obtain strain seed liquid, The bacteria content is 10 ⁹ cfu/mL;

S4. Fermentation: inoculate the seed liquids of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus prepared in step S3 into the secondary enzymatic protein peptide products obtained in step S2, and the inoculum amounts are 4%, 1.5%, respectively. %, 2.5%, under micro-anoxic conditions, 37°C, 60r/min, fermented for 56 hours, freeze-dried to obtain enzymatic fermented protein peptide powder;

S5. Colloid mill: Mix 60 parts by weight of oat flour and 40 parts by weight of rice bran powder evenly, grind for 4 hours through a colloid mill, and pass through a 400-mesh sieve to obtain fine dietary fiber powder;

S6. Preparation of wheat amylose: soak the wheat flour in 7wt% acetic acid solution for 3 days, the solid-to-liquid ratio of the wheat flour and the acetic acid solution is 1:4g/mL, after adjusting the pH value to be neutral, filter, filter cake Dry in the sun, gelatinize and centrifuge the supernatant obtained by freezing and crystallization, melt at room temperature into an ice-water mixture, filter with suction, and dry at 70°C for 2 hours to obtain wheat amylose;

S7. Preparation of low GI puffed powder: mix the dietary fiber fine powder prepared by 40 parts by weight of step S5 and the wheat amylose prepared by 100 parts by weight of step S6, stir at 300r/min for 15min, and adjust the moisture content to 250g/kg , after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 125°C, the screw speed power is 30Hz, and low GI puffed powder is obtained;

S8. Preparation of a prebiotic sweetener: Mix 6 parts by weight of xylitol and 3 parts by weight of steviol glycoside, and stir at 300 r/min for 10 minutes to obtain a prebiotic sweetener;

S9. Adjusting mouthfeel: mix 40 parts by weight of slow-release protein granules prepared in step S5, 55 parts by weight of low GI puffed powder prepared in step S7, and 6 parts by weight of prebiotic sweetener prepared in step S8, 300r/min Stir for 10 minutes to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 4% CO ₂ , 7% O ₂ , the balance is nitrogen, and % is volume percentage.

Comparative example 11

Compared with Example 3, the difference is that step S5 only adds a single sodium carboxymethylcellulose.

details as follows:

S5. Preparation of slow-release protein granules: Dissolve 14 parts by weight of sodium carboxymethylcellulose in 50 parts by weight of water, add 11 parts by weight of the enzymatic fermented protein peptide powder prepared in step S4, stir and mix at 500r/min for 15min, add 1.5 parts by weight of sucrose fatty ester and 40 parts by weight of corn oil, the pore size is the fast membrane emulsification of 2000nm, the calcium chloride solution of 4wt% is added dropwise, solidified at room temperature for 40min, centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain Slow-release protein granules.

Comparative example 12

Compared with Example 3, the difference is that only a single sodium alginate is added in step S5.

details as follows:

S5. Preparation of sustained-release protein granules: Dissolve 11 parts by weight of sodium alginate in 50 parts by weight of water, add 11 parts by weight of the enzymatic fermented protein peptide powder prepared in step S4, stir and mix at 500 r/min for 15 minutes, add 1.5 parts by weight Sucrose fatty ester and 40 parts by weight of corn oil were emulsified with a fast membrane with a pore size of 2000nm, 4wt% calcium chloride solution was added dropwise, solidified at room temperature for 40min, centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain a slow-release protein particles.

Comparative example 13

Compared with Example 3, the difference is that step S5 only adds a single water-soluble chitosan.

details as follows:

S5. Preparation of slow-release protein granules: 11 parts by weight of water-soluble chitosan was dissolved in 30 parts by weight of water, 11 parts by weight of the enzymolysis fermented protein peptide powder prepared in step S4 was added, stirred and mixed at 500r/min for 15min, and 1.5 parts by weight was added. Parts by weight of sucrose fatty ester and 40 parts by weight of corn oil, the pore diameter is the rapid membrane emulsification of 2000nm, the calcium chloride solution of 4wt% is added dropwise, solidified at room temperature for 40min, centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain slow release protein particles.

Comparative example 14

Compared with Example 3, the difference is that no oat flour is added in step S6.

details as follows:

S6. Colloid mill: Grind 100 parts by weight of rice bran powder through a colloid mill for 4 hours, and pass through a 400-mesh sieve to obtain fine dietary fiber powder.

Comparative example 15

Compared with Example 3, the difference is that no rice bran powder is added in step S6.

details as follows:

S6. Colloid mill: Grind 100 parts by weight of oat flour through a colloid mill for 4 hours, and pass through a 400-mesh sieve to obtain fine dietary fiber powder.

Comparative example 16

Compared with Embodiment 3, the difference is that step S6 is not performed.

details as follows:

S1. Primary enzymatic hydrolysis: Mix 7 parts by weight of pea protein powder and 4 parts by weight of white kidney bean extract into 100 parts by weight of water, stir at 500 r/min for 10 minutes, add 1.5 parts by weight of primary compound protease, and heat to 50°C for enzymolysis 3h, 1250W microwave treatment for 4min to inactivate the enzyme, and obtain the first-level enzymolysis protein peptide product;

Described primary composite protease is the mixture of alkaline protease and papain, and mass ratio is 6:2;

S2. Secondary enzymatic hydrolysis: Mix 20 parts by weight of the primary enzymatic protein peptide product prepared in step S1 into 50 parts by weight of water, stir at 500r/min for 10min, add 1.5 parts by weight of the secondary compound protease, and heat to 50°C to enzymatically Decompose for 2 hours, 1250W microwave inactivates the enzyme for 4 minutes, and obtains the second-level enzymatic hydrolysis protein peptide product;

The secondary composite protease is a mixture of neutral protease and animal protease, and the mass ratio is 4:1;

S3. Activation of fermented probiotics: inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaulier medium respectively, under micro-anoxic conditions, 37°C, 60r/min, activate and cultivate for 22h, to obtain strain seed liquid, The bacteria content is 10 ⁹ cfu/mL;

S4. Fermentation: inoculate the seed liquids of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus prepared in step S3 into the secondary enzymatic protein peptide products obtained in step S2, and the inoculum amounts are 4%, 1.5%, respectively. %, 2.5%, under micro-anoxic conditions, 37°C, 60r/min, fermented for 56 hours, freeze-dried to obtain enzymatic fermented protein peptide powder;

S5. Preparation of slow-release protein particles: 6 parts by weight of sodium carboxymethylcellulose, 3 parts by weight of sodium alginate, and 5 parts by weight of water-soluble chitosan are dissolved in 30 parts by weight of water, and 11 parts by weight of step S4 are added to prepare Enzymolysis fermented protein peptide powder, 500r/min stirring and mixing for 15min, adding 1.5 parts by weight of sucrose fatty ester and 40 parts by weight of corn oil, rapid membrane emulsification with a pore size of 2000nm, adding 4wt% calcium chloride solution dropwise, curing at room temperature for 40 minutes , centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain slow-release protein particles;

S6. Preparation of wheat amylose: soak the wheat flour in 7wt% acetic acid solution for 3 days, the solid-to-liquid ratio of the wheat flour and the acetic acid solution is 1:4g/mL, after adjusting the pH value to be neutral, filter, filter cake Dry in the sun, gelatinize and centrifuge the supernatant obtained by freezing and crystallization, melt at room temperature into an ice-water mixture, filter with suction, and dry at 70°C for 2 hours to obtain wheat amylose;

S7. Preparation of low GI puffed powder: Mix 40 parts by weight of dietary fiber powder (including oat flour and rice bran powder, with a mass ratio of 3:2) and 100 parts by weight of wheat amylose prepared in step S6, and stir at 300r/min 15 minutes, adjusting the moisture content to 250g/kg, after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 125°C, and the screw speed power is 30Hz to obtain low GI puffed powder;

S8. Preparation of a prebiotic sweetener: Mix 6 parts by weight of xylitol and 3 parts by weight of steviol glycoside, and stir at 300 r/min for 10 minutes to obtain a prebiotic sweetener;

S9. Adjusting mouthfeel: mix 40 parts by weight of slow-release protein granules prepared in step S5, 55 parts by weight of low GI puffed powder prepared in step S7, and 6 parts by weight of prebiotic sweetener prepared in step S8, 300r/min Stir for 10 minutes to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 4% CO ₂ , 7% O ₂ , the balance is nitrogen, and % is volume percentage.

Comparative example 17

Compared with Embodiment 3, the difference is that step S7 is not performed.

details as follows:

S1. Primary enzymatic hydrolysis: Mix 7 parts by weight of pea protein powder and 4 parts by weight of white kidney bean extract into 100 parts by weight of water, stir at 500 r/min for 10 minutes, add 1.5 parts by weight of primary compound protease, and heat to 50°C for enzymolysis 3h, 1250W microwave treatment for 4min to inactivate the enzyme, and obtain the first-level enzymolysis protein peptide product;

Described primary composite protease is the mixture of alkaline protease and papain, and mass ratio is 6:2;

S2. Secondary enzymatic hydrolysis: Mix 20 parts by weight of the primary enzymatic protein peptide product prepared in step S1 into 50 parts by weight of water, stir at 500r/min for 10min, add 1.5 parts by weight of the secondary compound protease, and heat to 50°C to enzymatically Decompose for 2 hours, 1250W microwave inactivates the enzyme for 4 minutes, and obtains the second-level enzymatic hydrolysis protein peptide product;

The secondary composite protease is a mixture of neutral protease and animal protease, and the mass ratio is 4:1;

S3. Activation of fermented probiotics: Inoculate Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaulier medium respectively, under micro-anoxic conditions, 37°C, 60r/min, activate and cultivate for 22h, to obtain strain seed liquid, The bacteria content is 10 ⁹ cfu/mL;

S4. Fermentation: inoculate the seed liquids of Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus prepared in step S3 into the secondary enzymatic protein peptide products obtained in step S2, and the inoculum amounts are 4%, 1.5%, respectively. %, 2.5%, under micro-anoxic conditions, 37°C, 60r/min, fermented for 56 hours, freeze-dried to obtain enzymatic fermented protein peptide powder;

S5. Preparation of slow-release protein particles: 6 parts by weight of sodium carboxymethylcellulose, 3 parts by weight of sodium alginate, and 5 parts by weight of water-soluble chitosan are dissolved in 30 parts by weight of water, and 11 parts by weight of step S4 are added to prepare Enzymolysis fermented protein peptide powder, 500r/min stirring and mixing for 15min, adding 1.5 parts by weight of sucrose fatty ester and 40 parts by weight of corn oil, rapid membrane emulsification with a pore size of 2000nm, adding 4wt% calcium chloride solution dropwise, curing at room temperature for 40 minutes , centrifuged at 3000r/min for 15min, washed with deionized water, and freeze-dried to obtain slow-release protein particles;

S6. Colloid mill: Mix 60 parts by weight of oat flour and 40 parts by weight of rice bran powder evenly, grind for 4 hours through a colloid mill, and pass through a 400-mesh sieve to obtain fine dietary fiber powder;

S7. Preparation of low GI puffed powder: 40 parts by weight of the dietary fiber fine powder prepared in step S6 are mixed with 100 parts by weight of wheat flour, stirred at 300r/min for 15min, and the moisture content is adjusted to be 250g/kg. After twin-screw extrusion, The barrel temperature of the twin-screw extrusion is 125° C., and the screw speed power is 30 Hz to obtain low GI puffed powder;

S8. Preparation of a prebiotic sweetener: Mix 6 parts by weight of xylitol and 3 parts by weight of steviol glycoside, and stir at 300 r/min for 10 minutes to obtain a prebiotic sweetener;

S9. Adjusting mouthfeel: mix 40 parts by weight of slow-release protein granules prepared in step S5, 55 parts by weight of low GI puffed powder prepared in step S7, and 6 parts by weight of prebiotic sweetener prepared in step S8, 300r/min Stir for 10 minutes to obtain a compound cereal composition with high protein and low GI;

The micro-anoxic condition is 4% CO ₂ , 7% O ₂ , the balance is nitrogen, and % is volume percentage.

Comparative example 18

Compared with Example 3, the difference is that no dietary fiber fine powder is added in step S8.

details as follows:

S8. Preparation of low GI puffed powder: adjust the moisture content of 140 parts by weight of wheat amylose prepared in step S7 to 250g/kg, and after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 125°C , the screw speed power is 30Hz, and low GI puffed powder is obtained.

Comparative example 19

Compared with Example 3, the difference is that no wheat amylose is added in step S8.

details as follows:

S8. Preparation of low GI puffed powder: adjust the moisture content of 140 parts by weight of the dietary fiber fine powder prepared in step S6 to 250g/kg, after twin-screw extrusion, the barrel temperature of the twin-screw extrusion is 125°C , the screw speed power is 30Hz, and low GI puffed powder is obtained.

Comparative example 20

Compared with Example 3, the difference is that step S8 does not carry out twin-screw extrusion, but only simple mixing.

details as follows:

S8. Preparation of low GI puffed powder: Mix 40 parts by weight of dietary fiber fine powder prepared in step S6 with 100 parts by weight of wheat amylose prepared in step S7, and stir at 300 r/min for 15 minutes to obtain low GI powder.

test case 1

Sustained release effect test

Take 1 g of the slow-release protein granules prepared in step S5 prepared in Examples 1-3 and Comparative Examples 10-13 of the present invention and add them to 10 mL of artificial simulated gastric juice and 10 mL of artificial simulated intestinal juice respectively. Stir and react for 2h and 3h respectively. In addition, take 1g of sustained-release protein particles and add them to 10mL of artificial simulated gastric juice. The simulated intestinal juice was stirred and reacted for 3h. After the reaction, the cells of the probiotic population were counted and weighed.

Among them, artificial gastric juice configuration: dilute hydrochloric acid 16.4mL, pepsin 10g, constant volume to 1L, adjust pH to 1.2; artificial intestinal juice configuration: potassium dihydrogen phosphate 6.8g, add water 500mL, adjust pH to 6 with 0.1mol/L sodium hydroxide .8, add trypsin 10g, dilute to 1L.

The survival rate was calculated according to the following formula:

Survival rate (%)=N _t /N ₀ ×100%

In the formula, N _t is the concentration of probiotics (cfu/g) that survived after being incubated in artificial simulated gastric juice or artificial simulated intestinal juice in vitro for a certain period of time, and N ₀ is the original concentration of probiotics added in artificial simulated gastric juice or artificial simulated intestinal juice (cfu/g). /g).

Calculate the release rate according to the following formula:

Release rate (%)=(W _t -W ₀ )/W ₀ ×100%

In the formula, W _t is the initial weight of the sample; W ₀ is the weight of the sample after incubation in simulated artificial simulated gastric juice and artificial simulated intestinal fluid for a certain period of time.

The results are shown in Figures 1 and 2. The sustained-release protein granules prepared in Examples 1-3 of the present invention can maintain good integrity in artificially simulated gastric juice, and the shell of the particles in artificially simulated intestinal fluid collapses, indicating that the sustained-release protein granules can withstand the effects of gastric acid Erosion, targeted delivery of active active ingredients into the intestinal environment.

Test Example 2 Determination of Anti-nutritional Factors

The high protein and low GI compound cereal compositions prepared in Examples 1-7 and Comparative Examples 1-20 of the present invention were tested for content of anti-nutritional factors.

1. Determination of trypsin inhibitor activity

Accurately weigh 1g of the sample, add 50mL Tris-HCl buffer solution to the Erlenmeyer flask, shake for 3h, centrifuge at 3000r/min for 10min, take the supernatant and dilute to 80 times by 2, 4, 10 times in turn. Take 1mL of the diluted sample extract and add 2.5mL of BAPNA solution to the test tube, place the test tube in a water bath at 37°C to preheat for 10min, add 1mL of bovine trypsin solution to accurately time the reaction, and measure the color at 410nm. Simultaneously measure the blank solution by the same method, except that 30% acetic acid is added first and then bovine trypsin solution is added. The absorbance value of the trypsin standard solution should be 0.39-0.42. The trypsin in the sample solution is 40-60% inhibited, otherwise it should be redone.

A ₁ : absorbance value of standard solution at 410nm; A ₂ : absorbance value of sample solution at 410nm; W: sample mass (g); 0.019: absorbance value of 1µg bovine trypsin at 410nm; D: dilution factor.

2. Determination of phytic acid content

Take 1 g of the sample in a Erlenmeyer flask, add 100 mL each of 5% trichloroacetic acid and 10% sodium sulfate, shake fully, place it overnight at room temperature, centrifuge at 3600 r/min for 10 min, and filter the supernatant with filter paper to obtain the sample to be tested.

Take 0.00, 1.00, 2.00, 3.00, 4.00, 5.00mL of phytic acid standard application solution (200μg phytic acid/mL) into 25mL centrifuge tubes, add 0.05% FeCl _3mL , 0.5% sulfosalicylic acid and buffer solution in sequence Add 5 mL each, add distilled water to make up to 25 mL, mix well and centrifuge for 10 min, take the supernatant, measure its absorbance with a spectrophotometer at a wavelength of 500 nm, and draw a standard curve with a reagent blank as a reference. Put 5.00mL of the sample to be tested in a 25mL centrifuge tube, add 0.05% FeCl3 _3mL , 0.5% sulfosalicylic acid and buffer solution 5mL each, add distilled water to make up to 25mL, mix well and centrifuge for 10min, take For the supernatant, measure its absorbance at a wavelength of 500 nm with a spectrophotometer, using a reagent blank as a reference. Calculate the content of phytic acid in the sample (mg/g).

The results are shown in Table 1.

Table 1

It can be seen from the above table that the high-protein and low-GI compound cereal composition prepared in Examples 1-3 of the present invention hardly contains anti-nutritional factors.

Test example 3 Determination of in vitro GI value

The determination method of GI value in vitro refers to reference literature: CHUNG H J, LIM HS, LIM S T. Effect of partial gelatinization and retrogradation on the enzymatic digestion of waxyrice starch［J］． Journal of Cereal Science, 2006, 43(3): 353-359.

The results are shown in Table 2.

Table 2

It can be seen from the above table that the high-protein and low-GI compound cereal composition prepared in Examples 1-3 of the present invention has a lower GI value.

Test Example 4 Test of Immunity Enhancement

SPF grade mice, male, body weight 16-20g, adaptively fed for 4 days, were divided into 29 groups, 10 in each group, and given the high protein and low GI obtained in Examples 1-7 and Comparative Examples 1-20 by intragastric administration respectively. The compound cereal composition is administered according to 5g/100g mouse body weight, once a day, and administered continuously for 30 days. The positive control group was given 1 mL/100 mice body weight of 10 ^-4 g/mL calcium 6S-5-methyltetrahydrofolate, and the negative control group was given the same amount of distilled water.

1. Mouse body weight and organ/body weight ratio

On the 31st day of the experiment, the body weight was weighed, the thymus and spleen were weighed, and the thymus/body weight ratio and spleen/body weight ratio were calculated. The results are shown in Table 3.

table 3

Note: * is compared with the negative control group, P<0.05, ** is compared with the negative control group, P<0.01.

Spleen and thymus are important immune organs in animals, and the quality of spleen and thymus can reflect the strength of their immune ability to a certain extent.

2. Number of chicken red blood cells phagocytosed by mouse peritoneal macrophages

On the 31st day of the experiment, inject 1ml of 20% (v/v) chicken erythrocyte suspension into the abdominal cavity of the mouse, and kill it after an interval of 30 minutes. 1mL of peritoneal washings was evenly distributed on 2 glass slides, and incubated in a 37°C incubator for 30min. After the incubation, the slides were taken out and rinsed with normal saline to remove cells that were patches. After drying, fix with 1:1 acetone methanol solution for 20 minutes, stain with Giemsa staining solution for 3 minutes, rinse with distilled water and dry, count macrophages under a microscope, and calculate phagocytosis rate and phagocytosis index.

Phagocytosis rate (%) = macrophages that phagocytized chicken red blood cells/counted macrophages × 100%;

Phagocytosis index = total number of phagocytized chicken red blood cells/counted macrophages.

The results are shown in Table 4.

Table 4

Note: * is compared with the negative control group, P<0.05, ** is compared with the negative control group, P<0.01.

It can be seen from the above table that the compound cereal composition with high protein and low GI prepared in Examples 1-3 of the present invention has a better effect of enhancing immunity.

Compared with Example 3, Examples 4 and 6 differ in that the primary composite protease is a single alkaline protease or papain. Compared with Example 3, Comparative Example 3 is different in that step S1 is not performed. The content of anti-nutritional factors increased, and the ability of immune regulation decreased. The primary compound protease of the present invention is a mixture of alkaline protease and papain, which enzymatically hydrolyzes protein substrates under alkaline conditions, and the enzymatic cleavage sites of alkaline protease are all hydrophobic and aromatic compounds on the carboxyl side chain. The amide bond at the carboxyl end of the amino acid catalyzes the hydrolysis of the peptide bond in the protein molecule to produce a peptide with a smaller molecular weight. Papain is a thiol protease. Papain is complementary to the peptide bond catalyzed by alkaline protease, so that it can hydrolyze the protein to the greatest extent.

Compared with Example 3, Examples 6 and 7 differ in that the secondary composite protease is a single neutral protease or animal protease. Compared with Example 3, Comparative Example 4 is different in that step S2 is not performed. The content of anti-nutritional factors increased, and the ability of immune regulation decreased. The secondary composite protease of the present invention is a mixture of neutral protease and animal protease. Neutral protease is a metalloprotease that preferentially cuts the peptide bond between leucine and phenylalanine. Animal protease can hydrolyze animal protein The control of macromolecular protein decomposition can significantly enhance the nutrition of substrate protein.

Compared with Example 3, Comparative Example 5 is different in that the content of anti-nutritional factors is significantly increased without steps S1 and S2, and the immune regulation ability is significantly decreased. After secondary enzymatic hydrolysis, trypsin inhibitors (a type of small molecular protein) can be degraded to the greatest extent, thereby greatly reducing the content of trypsin inhibitors in beans, and at the same time, bean proteins can be better enzymatically hydrolyzed Form into small molecular peptides to improve the absorption rate of protein.

Compared with Example 3, Comparative Examples 1 and 2 are different in that no pea protein powder or white kidney bean extract is added in step S1. The content of anti-nutritional factors increased, and the ability of immune regulation decreased. The pea protein powder and the white kidney bean extract guarantee the protein supply of the composition of the present invention, and can play a good role in regulating immunity.

Compared with Example 3, Comparative Examples 6, 7, and 8 differ in that only Lactobacillus plantarum, Saccharomyces cerevisiae or Lactobacillus bulgaricus are inoculated in step S4. The content of anti-nutritional factors increased slightly, and the ability of immune regulation decreased. Compared with Example 3, Comparative Example 9 is different in that steps S3 and S4 are not performed. The secondary enzymatic protein peptide products are fermented by Lactobacillus plantarum, Saccharomyces cerevisiae, and Lactobacillus bulgaricus. These probiotics can decompose anti-nutritional factors such as tannins, α-galactooligosaccharides, and phytic acid through self-proliferation on the one hand, On the other hand, the growth of the three has a synergistic effect. In the early stage of fermentation, Lactobacillus plantarum can utilize monosaccharides and produce formic acid, propionic acid, butyric acid, folic acid, etc. required for the proliferation of Lactobacillus bulgaricus. In the later stage, Saccharomyces cerevisiae and Lactobacillus bulgaricus produce a large amount of lactic acid and amino acids, which promote the growth of Lactobacillus plantarum, complement each other and promote each other, thereby promoting the fermentation of fermentation bacteria and producing a large amount of beneficial nutrients. At the same time, probiotics proliferate in large quantities. The obtained enzymatically hydrolyzed fermented protein peptide powder is rich in short-chain fatty acids, live probiotic bacteria, amino acids, protein peptides, etc., which consumes almost all anti-nutritional factors and greatly improves the nutritional value of the product.

Compared with Example 3, Comparative Example 10 is different in that step S5 is not performed. Compared with Example 3, Comparative Examples 11, 12, and 13 are different in that step S5 only adds a single sodium carboxymethylcellulose, sodium alginate or water-soluble chitosan. Immunomodulatory ability was significantly decreased. The present invention prepares a shell material with obvious acid resistance through sodium carboxymethyl cellulose, sodium alginate and water-soluble chitosan, and wraps the obtained enzymolysis and fermentation protein peptide in the granules. After entering the human body, it can resist Gastric acid erosion effectively protects live probiotics against acidic environments, protects proteins against pepsin degradation and inactivation, enters the intestinal tract smoothly, absorbs nutrients smoothly by the small intestine, probiotics colonize and colonize the inner wall of the intestinal tract, and plays a very good role in regulating immunity and Blood sugar, enhance the body's anti-sugar, anti-oxidation, and anti-aging functions, and has a certain anti-cancer effect.

Compared with Example 3, Comparative Examples 14 and 15 are different in that no oat flour or rice bran powder is added in step S6. Compared with Example 3, Comparative Example 16 is different in that step S6 is not performed. The GI value is significantly improved. Oat flour and rice bran powder are an excellent dietary fiber. After being ground by a colloid mill, a very fine dietary fiber powder is obtained. By adding the dietary fiber fine powder prepared by the present invention to the wheat amylose, and grinding the colloid to a suitable particle size, the overall GI value can be significantly reduced.

Compared with Example 3, Comparative Example 17 is different in that step S7 is not performed, and the GI value is obviously increased. The invention prepares wheat amylose, on the one hand, after entering the human body, it can form an anti-digestion complex with lipids to reduce the digestibility of starch. On the other hand, amylose can reduce the gelatinization degree of starch, increase crystallinity, and promote starch aging, thereby greatly reducing the GI value of grains.

Compared with Example 3, Comparative Examples 18 and 19 are different in that no dietary fiber fine powder or wheat amylose is added in step S8. Compared with Example 3, Comparative Example 20 is different in that step S8 does not carry out twin-screw extrusion, but only simple mixing. The GI value increased obviously, and the immunomodulatory effect decreased slightly. After mixing and extruding oat flour, rice bran flour, and wheat amylose in an appropriate proportion, the vitamins, trace elements, and amino acids in the grain flour can be well preserved. By adjusting the parameters of the appropriate twin-screw extruder, the obtained lower GI puffed powder has a more delicate taste, better dispersibility, no sedimentation and agglomeration, and better sensory quality.

The above descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included in the scope of the present invention. within the scope of protection.

Claims (10)

1. A preparation method of a high-protein low-GI composite grain composition is characterized in that pea protein powder and a white kidney bean extract are subjected to secondary enzymolysis, fermented by lactobacillus plantarum, saccharomyces cerevisiae and lactobacillus bulgaricus, freeze-dried to obtain enzymolysis fermented protein peptide powder, and embedded by sodium carboxymethylcellulose, sodium alginate and water-soluble chitosan to obtain slow-release protein particles; grinding oat flour and rice bran powder mixed colloid into fine powder, mixing with wheat amylose, adjusting the moisture content, extruding by a double screw, and spray drying to obtain the low-GI puffed powder; uniformly mixing the slow-release protein particles, the low-GI puffed powder and the prebiotics sweetener to obtain a high-protein low-GI composite grain composition;

the second-stage enzymolysis is to carry out first-stage enzymolysis on pea protein powder and a white kidney bean extract by taking a mixture of alkaline protease and papain as first-stage compound protease; then the mixture of neutral protease and animal protease is used as secondary compound protease to carry out secondary enzymolysis.

2. The method of claim 1, comprising the steps of:

s1, primary enzymolysis: mixing and dispersing pea protein powder and white kidney bean extract in water, adding primary compound protease, heating for enzymolysis, and inactivating enzyme to obtain primary enzymolysis protein peptide product;

s2, secondary enzymolysis: dispersing the primary enzymolysis protein peptide product prepared in the step S1 into water, adding secondary compound protease, heating for enzymolysis, and inactivating enzyme to obtain a secondary enzymolysis protein peptide product;

s3, activating fermentation probiotics: respectively inoculating lactobacillus plantarum, saccharomyces cerevisiae and lactobacillus bulgaricus into a high-yield culture medium, and performing activated culture to obtain strain seed liquid;

s4, fermentation: respectively inoculating the seed liquid of the lactobacillus plantarum, saccharomyces cerevisiae and lactobacillus bulgaricus strains prepared in the step S3 into the secondary enzymolysis protein peptide product prepared in the step S2, performing fermentation culture, and performing freeze drying to obtain enzymolysis fermentation protein peptide powder;

s5, preparation of slow-release protein particles: dissolving sodium carboxymethylcellulose, sodium alginate and water-soluble chitosan in water, adding the enzymolysis fermented protein peptide powder prepared in the step S4, stirring and mixing uniformly, adding a food-grade emulsifier and edible oil, performing rapid membrane emulsification, dropwise adding a calcium chloride solution, curing at normal temperature, centrifuging, washing, and freeze-drying to obtain slow-release protein particles;

s6, colloid milling: mixing oat powder and rice bran powder uniformly, and grinding by a colloid mill to obtain dietary fiber fine powder;

s7, preparing wheat amylose: performing acidolysis on wheat flour, adjusting pH value to be neutral, filtering, drying filter cake, gelatinizing, centrifuging to obtain supernatant, freezing for crystallization, melting at room temperature to obtain ice-water mixture, suction filtering, and drying to obtain wheat amylose;

s8, preparing the low GI puffed powder: uniformly mixing the dietary fiber fine powder prepared in the step S6 and the wheat amylose prepared in the step S7, adjusting the moisture content, and performing double-screw extrusion to obtain the low GI puffed powder;

s9, preparation of the prebiotic sweetening agent: uniformly mixing xylitol and stevioside to obtain a prebiotic sweetener;

s10, taste adjustment: and (4) uniformly mixing the slow-release protein particles prepared in the step (S5), the low-GI puffed powder prepared in the step (S8) and the prebiotic sweetener prepared in the step (S9) to obtain the high-protein low-GI composite grain composition.

3. The preparation method according to claim 2, wherein in step S1, the pea protein powder and the white kidney bean extract are mixed at a mass ratio of 5-10; in the step S2, the secondary compound protease is a mixture of neutral protease and animal protease, and the mass ratio is 3-5; the temperature of the enzymolysis is 40-60 ℃, and the time is 1-3h; the enzyme deactivation method comprises 1000-1500W microwave treatment for 3-5min.

4. The method according to claim 2, wherein the activating culture in step S3 is performed under a micro-anoxic condition at 35-38 ℃ and 50-70r/min for 18-24h, and the bacteria content of the strain seed solution is 10 ⁸ -10 ⁹ cfu/mL; in the step S4, the inoculation amounts of the lactobacillus plantarum, the saccharomyces cerevisiae and the lactobacillus bulgaricus are 3-5%, 1-2% and 2-3%, and the fermentation culture conditions are 36-39 ℃ and 50-70r/min under the micro-anoxic condition for 48-72h.

5. The preparation method according to claim 2, wherein in the step S5, the mass ratio of the sodium carboxymethylcellulose, the sodium alginate, the water-soluble chitosan, the zymolytic protein peptide powder, the food-grade emulsifier and the edible oil is 5-7; the food-grade emulsifier is selected from at least one of sodium stearyl lactate, diacetyl tartaric acid monoglyceride, sucrose fatty ester and distilled monoglyceride, and the edible oil is selected from at least one of peanut oil, soybean oil, corn oil, rapeseed oil, sesame oil, linseed oil and olive oil; the aperture of the rapid membrane is 1000-3000nm, the concentration of the calcium chloride solution is 3-5wt%, and the time of normal-temperature curing is 30-50min.

6. The preparation method according to claim 2, wherein the oat flour and the rice bran flour in the step S6 are mixed at a mass ratio of 5-7 and 3-5, and the colloid mill is used for 3-5h; the acidolysis method in the step S7 is to soak wheat flour in 5-10wt% acetic acid solution for 2-4 days, wherein the solid-to-liquid ratio of the wheat flour to the acetic acid solution is 1.

7. The preparation method according to claim 2, wherein the mass ratio of the dietary fiber fine powder to the wheat amylose in the step S8 is 3-5 to 7-12, the regulated moisture content is 200-300g/kg, the barrel temperature of the twin-screw extruder is 120-130 ℃, and the screw rotation speed power is 20-40Hz; in the step S9, the mass ratio of the xylitol to the stevioside is 5-7; the mass ratio of the slow-release protein particles, the low GI puffed powder and the prebiotic sweetener in the step S10 is 30-50.

8. The preparation method according to claim 2, characterized by comprising the following steps:

s1, primary enzymolysis: mixing and dispersing 5-10 parts by weight of pea protein powder and 3-5 parts by weight of white kidney bean extract in 100 parts by weight of water, adding 1-2 parts by weight of primary compound protease, heating to 45-55 ℃ for enzymolysis for 2-4h, and carrying out 1000-1500W microwave enzyme deactivation treatment for 3-5min to obtain a primary enzymolysis protein peptide product;

the primary composite protease is a mixture of alkaline protease and papain, and the mass ratio is 5-7;

s2, secondary enzymolysis: dispersing 20 parts by weight of the first-stage enzymolysis protein peptide product prepared in the step S1 into 50 parts by weight of water, adding 1-2 parts by weight of second-stage compound protease, heating to 40-60 ℃ for enzymolysis for 1-3h, and carrying out microwave enzyme deactivation treatment for 3-5min at 1000-1500W to obtain a second-stage enzymolysis protein peptide product;

the secondary compound protease is a mixture of neutral protease and animal protease, and the mass ratio is 3-5;

s3, activating fermentation probiotics: respectively inoculating Lactobacillus plantarum, saccharomyces cerevisiae, and Lactobacillus bulgaricus into Gaultheria culture medium, activating and culturing at 35-38 deg.C and 50-70r/min under micro-anoxic condition for 18-24 hr to obtain strain seed solution with bacteria content of 10 ⁸ -10 ⁹ cfu/mL；

S4, fermentation: respectively inoculating the seed liquid of the lactobacillus plantarum, saccharomyces cerevisiae and lactobacillus bulgaricus strains prepared in the step S3 into the secondary enzymolysis protein peptide product prepared in the step S2, wherein the inoculation amounts are 3-5%, 1-2% and 2-3%, respectively, fermenting and culturing for 48-72h at 36-39 ℃ at 50-70r/min under the condition of micro-hypoxia, and freeze-drying to obtain enzymolysis and fermentation protein peptide powder;

s5, preparation of slow-release protein particles: dissolving 5-7 parts by weight of sodium carboxymethylcellulose, 2-4 parts by weight of sodium alginate and 3-7 parts by weight of water-soluble chitosan in 30 parts by weight of water, adding 10-12 parts by weight of the enzymolysis fermented protein peptide powder prepared in the step S4, stirring and mixing uniformly, adding 1-2 parts by weight of food-grade emulsifier and 30-50 parts by weight of edible oil, emulsifying by a rapid membrane with the aperture of 1000-3000nm, dropwise adding 3-5wt% of calcium chloride solution, solidifying at normal temperature for 30-50min, centrifuging, washing, and freeze-drying to obtain slow-release protein particles;

s6, colloid milling: uniformly mixing 50-70 parts by weight of oat flour and 30-50 parts by weight of rice bran powder, grinding for 3-5h by a colloid mill, and sieving by a 300-500-mesh sieve to obtain dietary fiber fine powder;

s7, preparing wheat amylose: soaking wheat flour in 5-10wt% acetic acid solution for 2-4 days, wherein the solid-to-liquid ratio of the wheat flour to the acetic acid solution is 1-5 g/mL, adjusting the pH value to be neutral, filtering, drying a filter cake, carrying out gelatinization and centrifugation to obtain supernatant, freezing and crystallizing, melting at room temperature to obtain a water-ice mixture, carrying out suction filtration, and drying to obtain wheat amylose;

s8, preparing the low GI puffed powder: uniformly mixing 30-50 parts by weight of the dietary fiber fine powder prepared in the step S6 and 70-120 parts by weight of the wheat amylose prepared in the step S7, adjusting the water content to 200-300g/kg, and performing twin-screw extrusion, wherein the temperature of a cylinder of the twin-screw extrusion is 120-130 ℃, and the rotating speed power of a screw is 20-40Hz to obtain low GI (glycemic index) puffed powder;

s9, preparation of the prebiotic sweetening agent: uniformly mixing 5-7 parts by weight of xylitol and 2-4 parts by weight of stevioside to obtain a prebiotic sweetener;

s10, taste adjustment: uniformly mixing 30-50 parts by weight of the sustained-release protein granules prepared in the step S5, 45-70 parts by weight of the low GI puffed powder prepared in the step S8 and 5-7 parts by weight of the prebiotic sweetener prepared in the step S9 to obtain a high-protein low-GI composite grain composition;

said micro-anoxic conditions are 3-5% CO ₂ 、5-10%O ₂ The balance is nitrogen,% is volume percent.

9. A high protein low GI composite cereal composition made by the method of any of claims 1-8.

10. Use of the high protein low GI composite cereal composition of claim 9 in the preparation of a low GI, high protein nutritional food.

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