CN110801020A - Prebiotics polypeptide composite probiotics for relieving gastrointestinal discomfort and abdominal distension and diarrhea and preparation method thereof - Google Patents

Abstract

The invention discloses a prebiotics polypeptide composite probiotic for relieving gastrointestinal discomfort and abdominal distension and diarrhea and a preparation method thereof, wherein the prebiotics polypeptide composite probiotic comprises the following components in parts by weight: 3-5 parts of maltodextrin, 5-8 parts of corn starch, 1-2 parts of fructo-oligosaccharide, 20-25 parts of arrowhead powder, 10-15 parts of apple powder, 5-10 parts of soybean polypeptide powder, 10-15 parts of bitter gourd peptide powder, 0.1-0.2 part of lactobacillus plantarum, 0.2-0.4 part of lactobacillus salivarius, 0.1-0.2 part of lactobacillus acidophilus and 1-2 parts of anhydrous citric acid. The content of small molecular balsam pear peptide and soybean peptide is improved by a special extraction process of balsam pear peptide powder and soybean peptide powder, and better effects of reducing blood sugar, increasing intestinal motility, promoting digestion and absorption of intestines and stomach, regulating functions of intestines and stomach and the like are achieved by matching with other natural plant components.

Description

Prebiotics polypeptide composite probiotics for relieving gastrointestinal discomfort and abdominal distension and diarrhea and preparation method thereof

Technical Field

The invention relates to the field of nutritional functional food and biological fermentation. More specifically, the invention relates to a prebiotics polypeptide composite probiotic for relieving gastrointestinal discomfort and abdominal distension and diarrhea and a preparation method thereof.

Background

The spleen and the stomach are the basis of health, and if the spleen and the stomach are weak, the functions of other internal organs such as heart, liver, lung and kidney can be further affected, so that the five internal organs are not regulated; and the weakness of the spleen and the stomach is easy to invade by exogenous pathogenic factors, the risk of infecting bacteria such as helicobacter pylori and the like is increased, and then diseases such as gastritis, gastric ulcer and the like are caused, even gastric cancer is caused. The intestinal tract is the largest digestive organ of the human body, 99% of the nutrition of the human body needs to be absorbed by the intestinal tract, and the intestinal tract is also the organ most susceptible to the influence of external pressure. Under the pressure of antibiotic treatment, poor diet and fast-paced lifestyle, the proportion of probiotic flora in the intestinal tract is gradually reduced, and abdominal pain, abdominal distension, diarrhea and constipation caused by poor micro-ecological environment in the intestinal tract occur.

Diarrhea is a common frequently occurring disease, and the greatest damage to health is the resulting malnutrition. In diarrhea, not only water and electrolytes, but also a large amount of proteins and other nutrients are lost, and severe diarrhea causes death due to electrolyte disturbance. The current methods for correcting electrolyte disturbance mainly comprise intravenous electrolyte supplement and oral electrolyte supplement. The vein supplement requires special conditions and techniques, sometimes causes transfusion reaction and brings pain to patients. The oral supplement mainly utilizes the infusion salt recommended by WHO, which is only suitable for treating patients with slight illness state and is ineffective for patients with severe dehydration, vomiting, abdominal distension and shock, although the infusion salt makes up the defect of intravenous infusion and brings convenience to the patients. In addition, it has high sodium content, and is easy to cause hypernatremia and natriuresis.

Therefore, it is very important to provide a food or health care product which can effectively relieve gastrointestinal dyspepsia and dysfunction and improve the conditions of flatulence and diarrhea.

Disclosure of Invention

In order to solve the technical problems, the invention provides prebiotics polypeptide composite probiotics for relieving gastrointestinal discomfort and abdominal distension and diarrhea and a preparation method thereof, wherein the contents of small molecular bitter gourd peptide and soybean peptide are improved through a special bitter gourd peptide powder and soybean peptide powder extraction process, and the prebiotics polypeptide composite probiotics is matched with other natural plant components to achieve better effects of reducing blood sugar, increasing intestinal motility, promoting gastrointestinal digestion and absorption, regulating gastrointestinal functions and the like.

To achieve these objects and other advantages in accordance with the present invention, there is provided a prebiotic polypeptide complex probiotic that relieves gastrointestinal discomfort and abdominal distension and diarrhea, comprising, in parts by weight: 3-5 parts of corn dextrin, 5-8 parts of eggplant powder, 10-15 parts of pumpkin powder, 1-2 parts of oligomeric maltose, 20-25 parts of arrowhead powder, 10-15 parts of apple powder, 5-10 parts of soybean polypeptide powder, 10-15 parts of bitter gourd peptide powder, 0.1-0.2 part of lactobacillus plantarum, 0.2-0.4 part of lactobacillus salivarius, 0.1-0.2 part of lactobacillus acidophilus, 0.2-0.3 part of bifidobacterium and 1-2 parts of potassium sorbate.

Preferably, the preparation method of the momordica charantia peptide powder comprises the following steps:

s11, taking one or more of fresh bitter gourds, dried bitter gourds and bitter gourds as bitter gourds, adding deionized water with the weight 5 times of that of the bitter gourds, soaking for 8-12 hours at the water temperature of 25 ℃, taking out and washing for 2-3 times by using the deionized water;

s12, drying the cleaned bitter gourd raw material, smashing and grinding the bitter gourd raw material into pulp to obtain bitter gourd pulp;

s13, taking the balsam pear pulp and the buffer solution to mix so as to obtain a mixed system, wherein the weight ratio of the balsam pear pulp is as follows: buffer solution is 1 (4.5-5.5); recording the total volume value of the mixed system, adjusting the pH value to 7, and then carrying out temperature treatment on the mixed system to obtain an extract;

the temperature treatment process comprises the following steps:

heating to 45-55 ℃, preserving heat for 45-60min, cooling to 20-25 ℃, preserving heat for 25-30min, and recording the first volume value of the whole mixed system; a first mixed solution containing deionized water and a buffer was supplemented at 60% (total volume-first volume), and the deionized water: buffer 4: 1; after the first mixed solution is added, heating to 60-75 ℃, preserving heat for 60-75min, then cooling to 45-55 ℃, preserving heat for 30-35min, and recording a second volume value of the whole mixed system at the moment; a second mixed solution containing deionized water and a buffer was supplemented by 75% (total volume-second volume), and the deionized water: buffer 3: 1; adding the second mixed solution, heating to 80-90 deg.C, maintaining the temperature for 75-85min, cooling to 60-75 deg.C, and maintaining the temperature for 35-45 min;

s14, reducing the temperature of the extract to 20-25 ℃, and then carrying out enzymolysis on the extract to obtain a momordica charantia peptidase hydrolysis system; wherein, the enzymolysis process comprises the following steps:

carrying out first enzymolysis: adjusting the pH value of the extract to 8.0, adding trypsin according to 4.5% of the weight of the extract, stirring at 80-100 rpm, heating to 35-40 ℃ while stirring, and keeping the temperature for 45-60min to obtain a first enzymolysis system;

and (3) carrying out second enzymolysis: after the temperature of the first enzymolysis system is reduced to 20-25 ℃, adjusting the pH value to 3.5, adding pectinase according to 3 percent of the weight of the first enzymolysis system, stirring at 80-100 r/min, heating to 45-55 ℃ while stirring, and preserving heat for 40-60min to obtain a second enzymolysis system;

and (3) carrying out third enzymolysis: after the temperature of the second enzymolysis system is reduced to 20-25 ℃, adjusting the pH value to 4.8, adding cellulase according to 2% of the weight of the second enzymolysis system, stirring at 80-100 r/min, heating to 55-60 ℃ while stirring, and preserving heat for 30-45min to obtain a third enzymolysis system;

s15, after the enzymolysis process in the step S14 is finished, heating the obtained third enzymolysis system to 90 ℃, and maintaining for 10min to finish the enzyme deactivation process to obtain a bitter gourd peptide crude extraction system; adding activated carbon in the crude extract system of the bitter gourd peptide according to 4-5% of the weight of the bitter gourd peptide, uniformly stirring, keeping the temperature at 65 ℃ for 60-90min, centrifuging, and removing sediments to obtain a crude extract of the bitter gourd peptide;

filtering the crude extract of the bitter gourd peptide by diatomite to obtain a bitter gourd peptide clear solution, wherein the filtering pressure is 0.2-0.3 MPa; adding 4.5% of active carbon into the bitter gourd peptide clear liquid by weight, standing for 45-50min, centrifuging, and removing sediments;

s16, filtering the bitter gourd peptide clear liquid after removing the sediment by a microfiltration ceramic membrane with the filtering aperture of 0.5-0.8 mu m, wherein the filtering temperature is 60 ℃ to obtain microfiltration membrane permeate;

filtering the microfiltration membrane permeate through a 200kDa roll-type ultrafiltration membrane with the molecular weight cutoff of 100-;

concentrating the ultrafiltration membrane retentate through a roll-type high-pressure reverse osmosis membrane with the molecular weight cutoff of 150-;

s17, drying the bitter gourd peptide concentrated solution by a vacuum freeze drying method to obtain bitter gourd peptide powder with the bitter gourd polypeptide protein content not less than 30%.

Preferably, the buffer is a phosphate buffer.

Preferably, the preparation method of the soybean polypeptide powder comprises the following steps:

s21, inoculating the aspergillus niger strains and the aspergillus oryzae strains on independent potato glucose agar culture media respectively, then putting the culture media into an incubator respectively, and activating the culture media for 48 to 72 hours at the temperature of between 28 and 35 ℃ respectively to obtain activated aspergillus niger strains and activated aspergillus oryzae strains;

culturing the activated aspergillus niger species with the prepared first liquid culture medium to obtain a liquid aspergillus niger seed solution, and culturing the activated aspergillus oryzae species with the prepared second liquid culture medium to obtain a liquid aspergillus oryzae seed solution;

inoculating the liquid Aspergillus niger seed liquid into a first fermentation culture medium in a fermentation tank for amplification culture at 30-35 ℃, the amplification culture temperature is 30.5-7.0, the rotation speed is 500-; and inoculating the liquid Aspergillus oryzae seed liquid into a second fermentation culture medium in a fermentation tank for amplification culture at 30-40 deg.C under stirring at 400-600rpm at pH6.5-7.0 for 1-2d to obtain liquid Aspergillus oryzae spore suspension;

s22, adding 30-45 parts of soybean protein powder, 250-350 parts of deionized water, 20-25 parts of Aspergillus niger spore suspension, 15-20 parts of Aspergillus oryzae spore suspension and La (NO) into a reaction kettle in parts by weight₃)₃.6H₂0.5-0.6 part of O and NH₄Cl 1.5-2.5 parts, KNO₃1-2 parts to obtain a fermentation system, and adjusting the pH of the fermentation system to 6.8-7.0;

sequentially carrying out three-stage fermentation treatment on the fermentation system:

in the first fermentation stage treatment, the fermentation temperature is 30-35 ℃, the fermentation system is stirred under the condition of the rotation speed of 300-400rpm, and simultaneously the first illumination treatment and the first magnetic field treatment are carried out on the fermentation system for 1-2 d; the first illumination treatment is as follows: the light intensity is 25-30 mu mol.m^-2.s^-1The red light and the light intensity are 30-35 mu mol^-2.s^-1Irradiating the fermentation system with blue light for 45-60 min; the first magnetic field treatment is as follows: performing magnetic field treatment on the fermentation system with 50Hz alternating magnetic field with magnetic field strength of 0.45-0.55mT for 45-60 min;

in the second fermentation stage treatment, the fermentation temperature is 25-35 ℃, the fermentation system is stirred under the condition of the rotation speed of 500-600rpm, and simultaneously the second illumination treatment and the second magnetic field treatment are carried out on the fermentation system for 1-2 d; the second illumination treatment is as follows: the light intensity is 20-24 mu mol.m^-2.s^-1The red light and the light intensity are 24-28 mu mol^-2.s^-1Irradiating the fermentation system with blue light for 35-45 min; the second magnetic field treatment is; fermenting with 50Hz alternating magnetic field with magnetic field strength of 0.35-0.45mTTreating the system with a magnetic field for 35-45 min;

in the third fermentation stage treatment, the fermentation temperature is 25-35 ℃, the fermentation system is stirred under the condition of the rotation speed of 300-400rpm, and simultaneously the third illumination treatment and the third magnetic field treatment are carried out on the fermentation system for 1-2 d; the third illumination treatment is as follows: the light intensity is 16-20 mu mol.m^-2.s^-1The red light and the light intensity are 20-22 mu mol^-2.s^-1Irradiating the fermentation system with blue light for 25-35 min; the third magnetic field treatment is as follows: treating the fermentation system with 50Hz alternating magnetic field with magnetic field strength of 0.2-0.3mT for 25-35 min;

s23, heating the fermentation system subjected to the fermentation treatment in the step S22 to 85 ℃, and maintaining for 10min to complete the enzyme deactivation process to obtain the soybean peptidase hydrolysate;

s24, dispersing the soybean peptidase hydrolyzed solution into an ethanol solution with volume fraction of 95% and mass of 5-6 times of the soybean peptidase hydrolyzed solution, and simultaneously performing ultrasonic treatment and stirring, wherein the ultrasonic power is 600-800W, the ultrasonic treatment time is 10-15min, and the stirring speed is 100-120 r/min;

s25, filtering the reaction system processed in the step S24, controlling the filtering pressure at 0.3-0.4MPa and the filtering temperature at 50-60 ℃, and removing filter residues to obtain soybean peptide clear liquid;

s26, carrying out freeze drying on the soybean peptide clear liquid to obtain the soybean peptide powder.

Preferably, the first liquid medium comprises, by weight: 1% of tea polyphenol, 1.5% of cane sugar, 2.5% of glucose, 5% of malt extract and CoCl.6H₂O 0.1％、CuSO₄.5H₂O 0.05％、FeNaEDTA 2％、H₃BO₃0.25％、Na₂MoO₄.2H₂O 0.15％、ZnSO₄.7H₂O 0.1％、MgSO₄0.1％、KNO₃0.1％、KNO₃0.1％、MnSO₄.H₂0.2 percent of O and 86.85 percent of deionized water.

Preferably, the first fermentation medium bag is measured by weightComprises the following steps: 10% of tomato juice, 2% of soluble starch, 2% of sucrose, 2% of glucose, 2.5% of corn flour, 0.5% of yeast powder and FeSO₄.7H₂O 0.1％、MgSO₄0.05％、KNO₃0.05％、CuCl₂.2H₂O 0.1％、(NH4)₆Mo₇O₂₄.7H₂O 0.15％、MnSO₄.H₂0.1 percent of O and 80.45 percent of deionized water.

Preferably, the second liquid culture medium comprises, by weight: 8% of bean sprout juice, 1.5% of cane sugar, 2.5% of glucose, 2% of peptone, 2% of FeNaEDTA, and H₃BO₃0.25％、Na₂MoO₄.2H₂O 0.15％、MgSO₄0.1％、KNO₃0.1％、MnSO₄.H₂0.2 percent of O and 83.2 percent of deionized water.

Preferably, the second fermentation medium comprises, by weight: 5% of maltodextrin, 8% of corn flour, 2% of xylose, 1.5% of yeast extract and MgSO₄0.05％、KNO₃0.1％、NH₄NO₃0.15％、MnSO₄.H₂0.1 percent of O and 83.1 percent of deionized water.

Preferably, in step S21, when the liquid aspergillus niger seed solution and/or the liquid aspergillus oryzae seed solution is subjected to scale-up culture, the ultrasonic treatment is performed on the corresponding fermentation medium inoculated with the aspergillus niger seed solution or the liquid aspergillus oryzae seed solution every day, the ultrasonic treatment frequency is 20-40Khz, and the treatment time is 30 min.

Also provides a preparation method of the prebiotics polypeptide composite probiotics for relieving gastrointestinal discomfort and abdominal distension and diarrhea, which comprises the following steps:

s100, weighing the components according to the using amounts of the components, and mixing for 4-6 minutes to obtain a mixture;

s200, feeding the mixture into a granulator for granulation to obtain a coarse finished product, wherein the steam pressure in the granulator is 0.5-0.8Mpa, the granulation temperature is 40-50 ℃, and the diameter of a die hole of the granulator is 1.5-3 mm;

s300, sieving the crude product by a 300-mesh sieve to finally obtain the prebiotics polypeptide composite probiotics for relieving gastrointestinal discomfort and abdominal distension and diarrhea.

The invention at least comprises the following beneficial effects:

the invention enables the content of the balsam pear polypeptide protein in the balsam pear peptide to be not less than 20% through the extraction process of staged heating, repeated enzymolysis and multiple filtration, simultaneously greatly improves the content of the micromolecular balsam pear peptide and the micromolecular soybean peptide, enables the micromolecular balsam pear peptide and the micromolecular soybean peptide to be easier to digest and absorb, improves the gastrointestinal function, and simultaneously, the balsam pear peptide and the soybean peptide are compounded with other natural plant components rich in dietary fibers and probiotics for use, thereby effectively promoting the gastrointestinal absorption, preventing constipation, regulating the intestinal immunity function and relieving various uncomfortable symptoms such as flatulence and the like.

Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.

Detailed Description

The present invention is further described in detail below with reference to examples to enable those skilled in the art to practice the invention with reference to the description.

It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.

It is to be noted that the test methods described in the following embodiments are conventional methods unless otherwise specified, and the reagents and materials are commercially available without otherwise specified.

< example 1>

The prebiotics polypeptide composite probiotics for relieving gastrointestinal discomfort and abdominal distension and diarrhea in the embodiment comprise the following components in parts by weight: 3 parts of corn dextrin, 5 parts of eggplant powder, 10 parts of pumpkin powder, 1 part of oligomeric maltose, 20 parts of arrowhead powder, 10 parts of apple powder, 5 parts of soybean polypeptide powder, 10 parts of bitter gourd peptide powder, 0.1 part of lactobacillus plantarum, 0.2 part of lactobacillus salivarius, 0.1 part of lactobacillus acidophilus, 0.2 part of bifidobacterium and 1 part of potassium sorbate.

Further, the preparation method of the bitter gourd peptide powder comprises the following steps:

s11, taking one or more of fresh bitter gourds, dried bitter gourds and bitter gourds as bitter gourds, adding deionized water with the weight 5 times of that of the bitter gourds, soaking for 8-10 hours (preferably 8.5 hours) at the temperature of 25 ℃, taking out, and washing for 2-3 times by using the deionized water to remove pesticide residues and impurities;

s12, drying the washed bitter gourd raw materials in air, taking out, smashing and grinding to obtain bitter gourd pulp;

s13, mixing the balsam pear pulp with a buffer solution (the buffer solution is a buffer system containing reagents such as acid, alkali, salt and the like, such as a phosphate buffer solution) to obtain a mixed system, wherein the weight ratio of the balsam pear pulp is as follows: buffer 1 (4.5-5.5) (preferably 1: 5); recording the total volume value of the mixed system, adjusting the pH value to 7, and then carrying out temperature treatment on the mixed system to obtain an extract;

wherein the temperature treatment process comprises:

heating to 45-55 ℃ (preferably 50 ℃), keeping the temperature for 45-60min (preferably 55min), cooling to 20-25 ℃ (preferably 22 ℃), keeping the temperature for 25-30min (preferably 28min), and recording the first volume value of the whole mixed system at the moment; since water, acid, etc. in the reaction system may be evaporated during the aforementioned temperature rising and holding process, which may cause the solubility of acid, alkali, and inorganic ions to change, thereby affecting the leaching effect, after the aforementioned temperature rising and holding process, a first mixed solution containing deionized water and the buffer solution is added according to (total volume — first volume) × 60%, and the deionized water is calculated according to the weight ratio: buffer 4: 1, compensating the reaction system after evaporation of water, acid and the like, so that the reaction system is always in a better leaching environment; adding the first mixed solution, heating to 60-75 ℃ (preferably 65 ℃), keeping the temperature for 60-75min (preferably 65min), cooling to 45-55 ℃ (preferably 50 ℃), keeping the temperature for 30-35min (preferably 32 ℃), and recording a second volume value of the whole mixed system at the moment; a second mixed solution containing deionized water and a buffer was supplemented by 75% (total volume-second volume), and the deionized water: buffer 3: 1, the temperature of the temperature rise and the heat preservation is increased compared with the first time, so that the evaporation effect of water, acid and the like in the reaction system is more obvious, the proportion of the second mixed solution supplemented at this time is increased (to 75%), and the proportion of the buffer solution in the second mixed solution is increased; adding the second mixture, heating to 80-90 deg.C (preferably 85 deg.C), maintaining the temperature for 75-85min (preferably 80min), cooling to 60-75 deg.C (preferably 70 deg.C), and maintaining the temperature for 35-45min (preferably 40 min);

in the step, cell structure (such as cell walls and the like) compositions of the components can be repeatedly impacted and destroyed in different temperature change environments through staged temperature rise and heat preservation, and meanwhile, water and buffer solution in corresponding proportion are supplemented after each temperature rise and heat preservation stage is finished, so that a reaction system after water, acid and the like are evaporated is compensated, the reaction system is always in a better leaching environment, and the best leaching effect is achieved;

s14, reducing the temperature of the extract to 20-25 ℃, and then carrying out enzymolysis on the extract to obtain a momordica charantia peptidase hydrolysis system; wherein, the enzymolysis process comprises the following steps:

carrying out first enzymolysis: adjusting pH of the extract to 8.0, adding trypsin 4.5 wt% of the extract, stirring at 80-100 rpm, heating to 35-40 deg.C (preferably 37 deg.C) while stirring, and maintaining for 45-60min (preferably 55min) to obtain a first enzymolysis system;

and (3) carrying out second enzymolysis: after the temperature of the first enzymolysis system is reduced to 20-25 ℃, adjusting the pH value to 3.5, adding pectinase according to 3 percent of the weight of the first enzymolysis system, stirring at 80-100 r/min, heating to 45-55 ℃ (preferably 50 ℃) while stirring, and preserving heat for 40-60min (preferably 50min) to obtain a second enzymolysis system;

and (3) carrying out third enzymolysis: after the temperature of the second enzymolysis system is reduced to 20-25 ℃, adjusting the pH value to 4.8, adding cellulase according to 2% of the weight of the second enzymolysis system, stirring at 80-100 r/min, heating to 55-60 ℃ (preferably 58 ℃) while stirring, and preserving heat for 30-45min (preferably 35min) to obtain a third enzymolysis system;

in the invention, the components are plant components, and the cell structure of the plant components contains cell walls, so that in the step, the cell walls are subjected to full enzymolysis by adopting different enzymes and enzymolysis conditions at different stages, so that cellulose, pectin and other components in the cell walls are completely destroyed, and effective components (such as balsam pear polypeptide protein) in the cell walls can be fully released, thereby improving the extraction efficiency;

s15, after the enzymolysis process in the step S14 is finished, heating the obtained third enzymolysis system to 90 ℃, and maintaining for 10min to finish the enzyme deactivation process to obtain a bitter gourd peptide crude extraction system; adding activated carbon in the crude extract system according to 4-5% of the weight of the crude extract system, stirring uniformly, keeping the temperature at 65 ℃ for 60-90min (preferably 75min), centrifuging, and removing residues to obtain a crude extract of the bitter gourd peptide;

filtering the crude extract with diatomaceous earth to obtain fructus Momordicae Charantiae peptide clear solution, with filtering pressure of 0.2-0.3MPa (preferably 0.25 MPa); adding 4-5% of active carbon into the bitter gourd peptide clear liquid by weight, standing for 45-50min, centrifuging, and removing sediments;

through the adsorption treatment of the active carbon and the diatomite, the impurities such as pigment, suspended particles, colloid and the like in the momordica charantia peptidase hydrolyzed liquid ensure that the finally obtained finished product has higher purity;

s16, filtering the bitter gourd peptide clear liquid after removing the sediment by a microfiltration ceramic membrane with the filtering aperture of 0.5-0.8 μm, wherein the filtering temperature is 55-65 ℃ (preferably 60 ℃) to obtain microfiltration membrane permeate; furthermore, the microfiltration ceramic membrane adopts three membranes which are used in parallel;

filtering the microfiltration membrane permeate through a 200kDa roll-type ultrafiltration membrane with the molecular weight cutoff of 100-; wherein the roll-type ultrafiltration membrane is a roll-type ultrafiltration membrane with the molecular weight cutoff of 100-200kDa, and the roll-type ultrafiltration membrane adopts two membranes which are used in parallel;

concentrating the ultrafiltration membrane retentate through a roll-type high-pressure reverse osmosis membrane with the molecular weight cutoff of 150-; the roll-type high-pressure reverse osmosis membrane system is a high-pressure concentration membrane, is specifically made of composite material membranes such as (PS) polysulfone or (PFS) polyethersulfone materials and is used by connecting four membranes in series;

in the step, the bitter gourd polypeptide protein is separated and purified by adopting a multi-layer membrane separation and purification technology, the concentration temperature is low, and the natural activity and high content of the bitter gourd polypeptide are effectively ensured;

s17, drying the bitter gourd peptide concentrated solution by a vacuum freeze drying method to obtain bitter gourd peptide powder with the bitter gourd polypeptide protein content not less than 30%.

Meanwhile, the soybean polypeptide is a small molecular protein, is easy to be absorbed by a human body, is suitable for people with poor digestion and absorption of the protein, and has the effects of improving immunity, enhancing physical strength, relieving fatigue, reducing high blood pressure, high blood sugar and high blood fat and the like. Therefore, the present embodiment also provides a method for preparing soybean polypeptide powder, which comprises the following steps:

s21, inoculating the aspergillus niger strains and the aspergillus oryzae strains on independent potato glucose agar culture mediums respectively, then putting the culture mediums into an incubator respectively, and activating the culture mediums for 48 to 72 hours (preferably 60 hours) under the condition of 28 to 35 ℃ (preferably 32 ℃) so as to obtain activated aspergillus niger strains and aspergillus oryzae strains;

culturing the activated aspergillus niger species with the prepared first liquid culture medium to obtain a liquid aspergillus niger seed solution, and culturing the activated aspergillus oryzae species with the prepared second liquid culture medium to obtain a liquid aspergillus oryzae seed solution; wherein the first liquid culture medium comprises, by weight: 1% of tea polyphenol, 1.5% of cane sugar, 2.5% of glucose, 5% of malt extract and CoCl.6H₂O 0.1％、CuSO₄.5H₂O 0.05％、FeNaEDTA 2％、H₃BO0.25％、Na₂MoO₄.2H₂O0.15％、ZnSO₄.7H₂O 0.1％、MgSO₄0.1％、KNO₃0.1％、KNO₃0.1％、MnSO₄.H₂0.2% of O, 86.85% of deionized water, and the second liquid culture medium comprises: 8% of bean sprout juice, 1.5% of cane sugar, 2.5% of glucose, 2% of peptone, FeNaEDTA 2%, and H₃BO₃0.25％、Na₂MoO₄.2H₂O 0.15％、MgSO₄0.1％、KNO₃0.1％、MnSO₄.H₂0.2 percent of O and 83.2 percent of deionized water;

then inoculating the liquid Aspergillus niger seed liquid into a first fermentation culture medium in a fermentation tank for amplification culture at the temperature of 30-35 ℃ (preferably 32 ℃), the pH value of the amplification culture is 6.5-7.0, the rotation speed is 500-800rpm (preferably 650rpm), and the culture time is 1-2d, so as to obtain Aspergillus niger spore suspension; and inoculating the liquid Aspergillus oryzae seed liquid into a second fermentation culture medium in a fermentation tank for amplification culture at 30-40 deg.C (preferably 35 deg.C), pH6.5-7.0, and stirring at 400-600rpm (preferably 500rpm) for 1-2d to obtain liquid Aspergillus oryzae spore suspension; wherein the first fermentation medium comprises, by weight: 10% of tomato juice, 2% of soluble starch, 2% of sucrose, 2% of glucose, 2.5% of corn flour, 0.5% of yeast powder and FeSO₄.7H₂O 0.1％、MgSO₄0.05％、KNO₃0.05％、CuCl₂.2H₂O 0.1％、(NH4)₆Mo₇O₂₄.7H₂O 0.15％、MnSO₄.H₂0.1% of O and 80.45% of deionized water, wherein the second fermentation medium comprises: 5% of maltodextrin, 8% of corn flour, 2% of xylose, 1.5% of yeast extract and MgSO₄0.05％、KNO₃0.1％、NH₄NO₃0.15％、MnSO₄.H₂0.1% of O and 83.1% of deionized water;

preferably, in order to promote the growth of aspergillus niger and/or aspergillus oryzae and improve the enzyme production efficiency, when the liquid aspergillus niger seed liquid and/or liquid aspergillus oryzae seed liquid are subjected to amplification culture, the ultrasonic treatment is carried out on the corresponding fermentation culture medium inoculated with the aspergillus niger seed liquid or the liquid aspergillus oryzae seed liquid every day, the ultrasonic treatment frequency is 20-40Khz (preferably 30Khz), and the treatment time is 30 min;

s22, adding 30-45 parts (preferably 35 parts) of soybean protein powder, 250 parts (preferably 280 parts) of deionized water and 350 parts (preferably 280 parts) of Aspergillus niger spore suspension into a reaction kettle by weight part, and adding 20-25 parts (preferably 22.5 parts) of Aspergillus niger spore suspension) 15-20 parts (preferably 10 parts) of Aspergillus oryzae spore suspension, and La (NO)₃)₃.6H₂O0.5-0.6 part (preferably 0.55 part), NH₄Cl 1.5-2.5 parts (preferably 2.0 parts), KNO₃1-2 parts (preferably 1.5 parts) to obtain a fermentation system and adjusting the pH of the fermentation system to 6.8-7.0; in the step, the soybean protein powder is prepared from non-transgenic soybeans, and the content of polypeptides with molecular weight more than or equal to 8000Da is more than or equal to 45 percent;

sequentially carrying out three-stage fermentation treatment on the fermentation system:

in the first fermentation stage treatment, the fermentation temperature is 30-35 ℃ (preferably 32 ℃), the fermentation system is stirred under the condition of the rotation speed of 300-; the first illumination treatment is as follows: the light intensity is 25-30 mu mol.m^-2.s^-1(preferably 28. mu. mol. m)^-2.s^-1) The red light and the light intensity are 30-35 mu mol^-2.s^-1(preferably 32. mu. mol. m)^-2.s^-1) The blue light irradiates the fermentation system, and the irradiation treatment time is 45-60min (preferably 50 min); the first magnetic field treatment is as follows: treating the fermentation system with 50Hz alternating magnetic field with magnetic field strength of 0.45-0.55mT (preferably 0.5mT) for 45-60min (preferably 50 min);

in the second fermentation stage treatment, the fermentation temperature is 25-35 ℃ (preferably 30 ℃), the fermentation system is stirred under the condition of the rotation speed of 500-600rpm (preferably 550rpm), and simultaneously the fermentation system is subjected to second illumination treatment and second magnetic field treatment for 1-2 d; the second illumination treatment is as follows: the light intensity is 20-24 mu mol.m^-2.s^-1(preferably 22. mu. mol. m)^-2.s^-1) The red light and the light intensity are 24-28 mu mol^-2.s^-1(preferably 25. mu. mol. m)^-2.s^-1) The blue light irradiates the fermentation system, and the irradiation treatment time is 35-45min (preferably 40 min); the second magnetic field treatment is; performing magnetic field treatment on the fermentation system with 50Hz alternating magnetic field with magnetic field intensity of 0.35-0.45mT (preferably 0.4mT) for 35-45min (preferably 40 min);

in the third fermentation stage treatment, the fermentation temperature is 25-35 ℃ (preferably 30 ℃), the fermentation system is stirred under the condition of the rotation speed of 300-; the third illumination treatment is as follows: the light intensity is 16-20 mu mol.m^-2.s^-1(preferably 18. mu. mol. m)^-2.s^-1) The red light and the light intensity are 20-22 mu mol^-2.s^-1(preferably 21. mu. mol. m)^-2.s^-1) The blue light irradiates the fermentation system, and the irradiation treatment time is 25-35min (preferably 30 min); the third magnetic field treatment is as follows: treating the fermentation system with 50Hz alternating magnetic field with magnetic field strength of 0.2-0.3mT (preferably 0.25mT) for 25-35min (preferably 30 min);

s23, heating the fermentation system subjected to the fermentation treatment in the step S22 to 85 ℃, and maintaining for 10min to complete the enzyme deactivation process to obtain the soybean peptidase hydrolysate;

s24, dispersing the soybean peptidase hydrolyzed solution into an ethanol solution with volume fraction of 95% and mass of 5-6 times of the soybean peptidase hydrolyzed solution, and simultaneously performing ultrasonic treatment and stirring, wherein the ultrasonic power is 600-800W (preferably 750W), the ultrasonic treatment time is 10-15min, and the stirring speed is 100-120 r/min;

s25, filtering the reaction system processed in the step S24, controlling the filtering pressure at 0.3-0.4MPa (preferably 0.35MPa), the filtering temperature at 50-60 ℃ (preferably 55 ℃), and discarding the filter residue to obtain the soybean peptide clear solution;

s26, carrying out freeze drying on the soybean peptide clear liquid to obtain the soybean peptide powder.

The Aspergillus niger and Aspergillus oryzae can produce β -glucosidase, amylase, neutral protease, acid protease, alkali protease, saccharifying enzyme, cellulase and other enzymes, and may be further used in decomposing cellulose, lactose, macromolecular protein, starch and other material in soybean protein powder into small molecular nutritious components easy to be absorbed by stomach and intestine to improve the immunity of stomach and intestineAfter transformation and expansion culture, La (NO) is added₃)₃.6H₂Carrying out enzymolysis fermentation on the soybean protein powder containing macromolecular protein in a culture medium of O, wherein La³⁺The growth and metabolism of aspergillus niger and aspergillus oryzae are promoted, the growth efficiency of aspergillus niger and aspergillus oryzae is improved, the yield and the enzyme activity of various enzymes of aspergillus niger and aspergillus oryzae are improved, and therefore macromolecular substances such as cellulose, starch, protein and the like in soybean protein powder can be effectively decomposed. Furthermore, in the three-stage fermentation treatment, the red light and the blue light can promote assimilation of cytoplasmic membranes by adjusting permeability of the cytoplasmic membranes and improve enzyme activity, and the alternating magnetic field promotes growth of the biomass of aspergillus niger and aspergillus oryzae by releasing calcium ions on the cellular membranes so as to further improve the enzyme yield. Meanwhile, in order to avoid adverse effects on the growth of aspergillus niger and aspergillus oryzae caused by the fact that red light, blue light and an alternating magnetic field are constant values for a long time, three-stage fermentation treatment is adopted in the step, and the light intensity of the red light and the blue light and the intensity of the alternating magnetic field are gradually decreased in each stage, so that the aspergillus niger and aspergillus oryzae are always in strong growth vigor, various high-vigor enzymes are continuously and efficiently generated, macromolecules such as cellulose, starch, protein and the like are further decomposed, and the release of active ingredients (such as micromolecular polypeptide and the like) in the soybean protein powder is promoted.

In addition, in order to fully exert the synergistic effect of the comprehensive arrowhead powder and the probiotics so as to improve the gastrointestinal digestion promoting and immunity enhancing effects, the embodiment further provides a preparation method of the comprehensive arrowhead powder, which specifically comprises the following steps:

s31, putting arrowhead raw materials (including leaf diameters and bulbs) into water with the weight 5-8 times (preferably 6.5 times) of that of the arrowhead raw materials, soaking for 10-12h (preferably 10.5h), taking out, draining, and then carrying out ultrasonic cleaning, wherein the ultrasonic power is 180-200W, the ultrasonic frequency is 40-50KHz, and the ultrasonic cleaning time is 10-15 min; ultrasonically cleaning, draining, drying in an oven at 100 deg.C for 8-10 hr, pulverizing at 35-40 deg.C (preferably 37 deg.C), and sieving with 100 mesh sieve to obtain arrowhead raw material powder;

s32, inoculating the Corning Trichoderma strain on a potato glucose agar culture medium, putting into an incubator, and activating for 48-72h under the condition of 28-35 ℃ (preferably 32 ℃) to obtain an activated Corning Trichoderma strain; wherein,

culturing the activated kangning trichoderma strain by adopting a prepared liquid culture medium to obtain liquid kangning trichoderma strain seed liquid; wherein the liquid culture medium comprises the following components in percentage by weight: 1% of tea polyphenol, 1.5% of cane sugar, 2.5% of glucose, 5% of malt extract and CoCl.6H₂O 0.1％、CuSO₄.5H₂O 0.05％、FeNaEDTA 2％、H₃BO₃0.15％、Na₂MoO₄.2H₂O 0.15％、ZnSO₄.7H₂O 0.1％、MgSO₄0.1％、KNO₃0.1％、MnSO₄.H₂0.25% of O and 87% of deionized water;

then inoculating the liquid Trichoderma koningii seed liquid to a fermentation medium in a fermentation tank for amplification culture, adjusting the pH to 6.5-7.5 (preferably 7.0), and stirring at the amplification culture temperature of 25-35 ℃ (preferably 30 ℃) and the rotation speed of 500-800rpm for 1-2d to obtain Trichoderma koningii spore suspension; wherein the fermentation medium comprises the following components in percentage by weight: 10% of bran, 2% of soluble starch, 2% of sucrose, 2% of lactose, 2.5% of corn flour and MgSO₄0.05％、KH₂PO₄0.05％、(NH4)₂SO₄0.15 percent and 81.25 percent of deionized water;

s33, taking the arrowhead raw material powder, adding Trichoderma koningii spore suspension with the weight of 5-10% (preferably 8%) of the arrowhead raw material powder and water with the weight of 4-5 times of the arrowhead raw material powder, and performing enzymolysis to obtain an enzymolysis system; wherein, the enzymolysis process comprises the following steps:

carrying out first enzymolysis: adding lipase according to 0.03-0.04% (preferably 0.035%) of raw material powder of rhizoma Sagittariae Sagittifoliae, adjusting pH to 6.5-7.5 (preferably 7.0), stirring, heating to 30-45 deg.C (preferably 37 deg.C) while stirring, and maintaining the temperature for 35-40min to obtain a first enzymolysis system;

performing second enzymolysis, namely cooling the first enzymolysis system to 20-25 ℃, adjusting the pH value to 4.0-4.5 (preferably 4.2), adding β -glucanase according to 0.02-0.025% (preferably 0.022%) of the weight of the first enzymolysis system, fully stirring, heating to 40-50 ℃ (preferably 45 ℃) while stirring, and preserving the temperature for 30-35min to obtain a second enzymolysis system;

and (3) carrying out third enzymolysis: after the temperature of the second enzymolysis system is reduced to 20-25 ℃, adjusting the pH value to 4.0-5.5 (preferably 5.0), adding cellulase according to 0.02-0.03% (preferably 0.025%) of the weight of the second enzymolysis liquid, fully stirring, heating to 50-65 ℃ (preferably 60 ℃) while stirring, and preserving the temperature for 25-35min to obtain a third enzymolysis system;

s34, after the enzymolysis is finished, heating the obtained third enzymolysis system to 85 ℃, and maintaining for 10min to finish the enzyme deactivation process to obtain arrowhead enzymolysis liquid;

s235, adding activated carbon which accounts for 4% of the weight of the arrowhead enzymatic hydrolysate, uniformly stirring, carrying out heat preservation at 65 ℃ for 65-85min (preferably 75min), centrifuging, and removing sediments to obtain a crude arrowhead extracting solution;

and S36, carrying out vacuum concentration on the crude arrowhead extracting solution, stopping concentration when the volume of the crude arrowhead extracting solution is reduced to 12-15% of the original volume to obtain an arrowhead concentrated solution, carrying out vacuum drying on the arrowhead concentrated solution, and sieving to obtain the comprehensive arrowhead powder.

< example 2>

The difference between this example and example 1 is that the prebiotic polypeptide composite probiotic for relieving gastrointestinal discomfort and abdominal distension and diarrhea in this example is composed of the following components: 3 parts of corn dextrin, 5 parts of eggplant powder, 10 parts of pumpkin powder, 1 part of oligomeric maltose, 20 parts of arrowhead powder, 10 parts of apple powder, 5 parts of soybean polypeptide powder, 10 parts of bitter gourd peptide powder, 0.1 part of lactobacillus plantarum, 0.2 part of lactobacillus salivarius, 0.1 part of lactobacillus acidophilus, 0.2 part of bifidobacterium and 1 part of potassium sorbate.

< example 3>

The difference between this example and example 1 is that the prebiotic polypeptide composite probiotic for relieving gastrointestinal discomfort and abdominal distension and diarrhea in this example is composed of the following components: 4 parts of corn dextrin, 7 parts of eggplant powder, 12 parts of pumpkin powder, 1.5 parts of oligomeric maltose, 22 parts of arrowhead powder, 12 parts of apple powder, 7 parts of soybean polypeptide powder, 12 parts of bitter gourd peptide powder, 0.15 part of lactobacillus plantarum, 0.3 part of lactobacillus salivarius, 0.15 part of lactobacillus acidophilus, 0.25 part of bifidobacterium and 1.5 parts of potassium sorbate.

< measurement of molecular weight of Momordica charantia peptide >

The method of example 1 of the application No. 201710832199.8 ("a new method for producing momordica charantia polypeptide protein extract at low temperature throughout, momordica charantia polypeptide protein extract and its use") was used to extract momordica charantia peptide as comparative example 1, which was subjected to high performance gel filtration chromatography with momordica charantia peptide powder prepared by the method of preparing momordica charantia peptide powder of examples 1-3 of the present invention to obtain the molecular weight and distribution range of momordica charantia peptide, and the results are shown in table 1.

TABLE 1 molecular weight and distribution of bitter gourd peptides

Therefore, in the preparation method of the bitter gourd peptide powder, firstly, the cell structure (such as cell walls and the like) composition of the components can be repeatedly impacted and destroyed under different temperature change environments through staged heating and heat preservation, meanwhile, water and buffer solution in corresponding proportions are supplemented after each heating and heat preservation stage is finished, so that a reaction system after water, acid and the like are evaporated is compensated, the reaction system is always in a better extraction environment, and further, the bitter gourd polypeptide protein is prepared by performing staged heating, repeated enzymolysis and multi-level membrane separation and purification technology extraction processes, so that the content of the bitter gourd polypeptide protein in the obtained bitter gourd polypeptide extract is higher than 30%, and the extract product does not contain polypeptide proteins which are not derived from bitter gourd, such as soybean protein polypeptide and the like. As can be seen from Table 1, the proportion of the momordica charantia polypeptide fragments in the range of 1000-plus 5000Da prepared by the invention is close to 35%, and the momordica charantia polypeptide fragments in the range of 1000-plus 5000Da can promote intestinal tract peristalsis, increase gastrointestinal motility, clear up intestinal tract garbage, and further play roles in relaxing bowel and relieving constipation.

< results of measuring molecular weight of Soybean peptide >

Soybean peptide powder prepared according to the protocol described in example 1 of the invention patent application No. 201310478523.2 ("a method for improving the yield of soybean peptide having low molecular weight") was first used as comparative example 2. The molecular weight and distribution range of the soybean peptide were measured by high performance gel filtration chromatography with the soybean peptide powder prepared by the preparation method of the present invention in examples 1 to 3, and the results are shown in table 2.

TABLE 2 molecular weight and distribution Range of Soybean peptides

As can be seen from Table 2, in the method for preparing soybean peptide powder of the present invention, various enzymes such as β -glucosidase, amylase, neutral protease, acid protease, alkaline protease, glucoamylase, cellulase, etc. can be generated by inoculating Aspergillus niger strains and Aspergillus oryzae strains, and further can be used for decomposing raw materials such as cellulose, lactose, macromolecular protein, starch, etc. in soybean protein powder into small molecular nutrients (such as small molecular polypeptides, etc.) which are more easily absorbed by the gastrointestinal tract, thereby improving the gastrointestinal immune function and relieving various gastrointestinal discomfort symptoms, wherein in the soybean peptide powder prepared by the present invention, the average molecular weight of protein peptide is about 500Da, wherein the polypeptide below 1000Da accounts for 85%, and the polypeptide below 500Da accounts for 37%, thereby facilitating the rapid and efficient absorption of the gastrointestinal tract of human body and fully exerting the efficacy thereof.

< detection of comprehensive arrowhead powder >

Firstly, adding a proper amount of water into the mixed arrowhead raw materials, pulping the materials by a conventional crushing and pulping machine, pumping the materials into a microchannel crusher for superfine crushing, pumping the materials under the pressure of 80Mpa and the flow rate of 120m/s, drying and crushing the obtained superfine crushed arrowhead slurry to obtain the comprehensive arrowhead powder serving as a comparative example 3, and carrying out sensory evaluation and detection on cellulose content, pectin content, soluble β -glucose content, soluble xylan content and dissolution rate on the comprehensive arrowhead powder obtained by the preparation method in the embodiments 1 to 3 of the invention, wherein the sensory evaluation values are 20 minutes in appearance, 25 minutes in texture, 30 minutes in flavor and 25 minutes in taste, and the results are shown in Table 3.

TABLE 3 comprehensive detection of arrowhead powder

Performance index	Example 1	Example 2	Example 3	Comparative example 1
					Cellulose content (%)	34.31±1.25	35.12±0.58	34.33±1.41	72.61±0.88
Pectin content (%)	14.21±0.54	16.25±1.43	15.75±0.74	7.82.25±0.33
					Soluble β -glucose content (%)	7.51±0.22	8.35±0.56	7.35±0.76	4.26±0.28
Soluble xylan content (%)	8.65±1.01	9.45±0.12	8.35±0.99	5.61±0.91
					Appearance of the product	19	20	20	18
Texture of	24	24	25	22
					Flavor (I) and flavor (II)	28	29	30	24
Taste of the product	23	24	24	15
					Dissolution rate (%)	97.56±1.57	98.33±0.78	97.26±1.16	34.27±1.46

As can be seen from the above Table 3, the content of insoluble cellulose in the comprehensive arrowhead powder prepared by the method of the present invention is reduced by more than 50% compared with that in the comparative example 3, but the content of three soluble dietary fibers, namely pectin, soluble β -glucose and soluble xylan, is significantly increased, so that the gastric emptying time is prolonged, the gastrointestinal motility is increased, the absorption of glucose is delayed, the blood cholesterol level is reduced, the retention time of excrement in the intestinal tract is reduced, the volume is increased, the bowel relaxing is achieved, and various uncomfortable symptoms such as flatulence and the like are relieved through the synergistic effect of the insoluble cellulose.

< example 4>

Meanwhile, the present embodiment also provides a method for preparing the prebiotic polypeptide complex probiotic bacteria according to any one of embodiments 1 to 3, which comprises the following steps:

s100, weighing the components according to the component usage amount in one of the embodiments 1-3, and mixing for 4-6 minutes (preferably 5 minutes) to obtain a mixture;

s200, feeding the mixture into a granulator for granulation to obtain a crude finished product, wherein the steam pressure in the granulator is 0.5-0.8Mpa (preferably 0.65Mpa), the granulation temperature is 40-50 ℃ (preferably 45 ℃), and the diameter of a die hole of the granulator is 1.5-3 mm;

s300, sieving the crude product by a 300-mesh sieve to finally obtain the prebiotics polypeptide composite probiotics for relieving gastrointestinal discomfort and abdominal distension and diarrhea.

< digestion promoting efficacy evaluation test >

200 male SD rats with the weight of 120-150 g are selected as cleaning male SD rats and randomly divided into 10 groups, namely a blank control group, a low dose group, a medium dose group and a high dose group. Wherein, blank control group: feeding purified water; low dose group: 1.67g/(kg · bw) of the prebiotic polypeptide composite probiotic (hereinafter, "prebiotic polypeptide composite probiotic") of the invention and purified water with the same volume are prepared into suspension, and the suspension is fed to rats; the medium dose group: 3.33 g/(kg. bw) of the prebiotics polypeptide composite probiotics and purified water with the same volume are prepared into suspension to feed rats; high dose group: 6.67g/(kg · bw) of the prebiotics polypeptide composite probiotics and purified water with the same volume are prepared into suspension to feed rats. 1 time per day for 30 days. During the experiment, rats measured 2 times per week the body weight and food intake, and adjusted the dosage according to the body weight change. Before the experiment, the animals in each group were fasted for 24 hours, gastric juice discharged within 3 hours was collected by a rat pyloric ligation method, and the amount of gastric juice and the pepsin activity per unit time were measured, and the results are shown in table 4.

Wherein, the pepsin determination comprises: sucking egg white (filtering with gauze) with capillary glass tube with inner diameter of 2mm and length of 10cm, and coagulating protein in hot water at 70 deg.C to obtain protein tube. Accurately taking 1mL of gastric juice, putting into 15mL0.05mol/L hydrochloric acid solution, shaking up, putting into two protein tubes, keeping the temperature for 48 hours in a 37 ℃ thermostat, measuring the lengths (mm) of transparent parts at two ends of the protein tubes, and calculating the activity of the pepsin. Pepsin Activity (U/mL) ═ four terminal protein tube clear section Length average²×16。

TABLE 4 Effect of prebiotic polypeptide Complex probiotics on gastric juice amount and pepsin Activity in rats

As can be seen from the results in table 4, compared with the blank control group, in the rats gavaged with the prebiotic polypeptide complex probiotic bacteria, the gastric juice amount and the pepsin activity in unit time are both significantly increased (up to 107% and 68.9% respectively), and the significant increase of the gastric juice amount and the pepsin activity can greatly promote the digestive ability of the intestines and the stomach, and have the functions of promoting digestion and relieving gastrointestinal discomfort.

< evaluation test for improving efficacy of gastrointestinal System >

240 healthy mice with the weight of 18-22g and males are selected for the experiment and randomly divided into a blank control group, a model control group, a positive control group, a low-dose group, a medium-dose group and a high-dose group, and 20 mice are selected for each group. Adaptive feeding for 7d, and starting molding. Except for the blank group, 60mg/kg of acetylphenylhydrazine is injected subcutaneously on the 1 st, 4 th, 7 th and 10 th days of mice in other groups, 40mg/kg of cyclophosphamide is injected intraperitoneally on the 10 th, 11 th, 12 th, 13 th and 14 th days, and 25mg/kg of compound diphenoxylate tablet suspension (5d is 1 administration period, 2d is stopped during the period, and 4 administration periods are used for total) begins to be infused in stomach.

After the molding is successful, from day 14, the high, medium and low dose groups are respectively prepared into suspension by using equal volume of physiological saline according to the dosage of 4.38g/kg, 2.92g/kg and 1.46g/kg of the prebiotic polypeptide composite probiotics of the invention, and then the suspension is administrated, the normal control group and the model control group are filled with equal volume of physiological saline, the positive control group is prepared into 0.25mg/ml suspension by using domperidone tablet after being ground into powder and the equal volume of physiological saline, and the suspension is continuously filled with stomach for 28 days, once a day.

After 28 days, the feed is fed according to the following method: blank control group: 1.5% normal saline of the weight of the gavage mouse; positive control group: grinding domperidone tablets, and preparing the powder and isometric physiological saline into 0.25mg/ml suspension for intragastric administration; high dose group: preparing a probiotic polypeptide composite probiotic sample of the invention and isometric physiological saline into suspension according to 4.38g/kg, and performing intragastric administration; the medium dose group: preparing a probiotic polypeptide composite probiotic sample of the invention and physiological saline with the same volume as the sample into suspension according to the ratio of 2.92g/kg, and performing intragastric administration; low dose group: the probiotic polypeptide composite probiotic sample of the invention and physiological saline with the same volume are prepared into suspension according to 1.46g/kg, and the suspension is administrated by gastric lavage.

After 0.5h, 2ml of ink (containing 5% of activated carbon and 10% of arabic resin) was administered to each mouse, and 30min later, the mouse was immediately sacrificed by removing the cervical vertebrae, opening the abdominal cavity to separate mesentery, cutting off the intestinal canal from the pylorus at the upper end and from the lower end to the ileocecal portion, measuring the length of the intestinal canal as the total length of the small intestine, and the ink advancing length from the pylorus to the ink front edge, and the results of observation are shown in table 5.

TABLE 5 Effect of prebiotic polypeptide Complex Probiotics on Small bowel movement

As can be seen from Table 5, the ink propulsion rate of the model control group was reduced relative to the blank control group, indicating that the model was successfully constructed. The ink propulsion rates of the low, medium and high dose groups are all obviously higher than those of the model control group, and have no obvious difference with that of the blank control group, which indicates that the probiotic polypeptide composite probiotic bacteria can obviously improve gastrointestinal motility and gradually recover the damaged gastrointestinal functions. Wherein, the ink propelling rate of the high-dose group is obviously higher than that of the positive control group, and the effect is optimal. Therefore, the probiotic polypeptide composite probiotics can greatly enhance the gastric motility and regulate the flatulence by relieving the gastrointestinal discomfort and abdominal distension.

It should be noted that the technical solutions in the above embodiments 1 to 4 can be arbitrarily combined, and the technical solutions obtained after the combination all belong to the protection scope of the present invention.

In conclusion, the momordica charantia peptide has the advantages that the content of momordica charantia polypeptide protein in the momordica charantia peptide is not lower than 20% through the extraction process of staged heating, repeated enzymolysis and multiple filtration, meanwhile, the content of small-molecule momordica charantia peptide and small-molecule soybean peptide is greatly increased, the momordica charantia peptide and the small-molecule soybean peptide are easier to digest and absorb, the gastrointestinal function is improved, and meanwhile, the momordica charantia peptide and the soybean peptide are compounded with other natural plant components rich in dietary fibers and probiotics for use, so that the gastrointestinal absorption is effectively promoted, constipation is prevented, the intestinal immunity function is adjusted, and meanwhile, various uncomfortable.

The number of apparatuses and the scale of the process described herein are intended to simplify the description of the present invention. Applications, modifications and variations of the present invention will be apparent to those skilled in the art.

While embodiments of the invention have been described above, it is not limited to the applications set forth in the description and the embodiments, which are fully applicable to various fields of endeavor for which the invention may be embodied with additional modifications as would be readily apparent to those skilled in the art, and the invention is therefore not limited to the details given herein and to the examples shown and described without departing from the generic concept as defined by the claims and their equivalents.

Claims (10)

1. The prebiotics polypeptide composite probiotics for relieving gastrointestinal discomfort and abdominal distension and diarrhea is characterized by comprising the following components in parts by weight: 3-5 parts of corn dextrin, 5-8 parts of eggplant powder, 10-15 parts of pumpkin powder, 1-2 parts of oligomeric maltose, 20-25 parts of arrowhead powder, 10-15 parts of apple powder, 5-10 parts of soybean polypeptide powder, 10-15 parts of bitter gourd peptide powder, 0.1-0.2 part of lactobacillus plantarum, 0.2-0.4 part of lactobacillus salivarius, 0.1-0.2 part of lactobacillus acidophilus, 0.2-0.3 part of bifidobacterium and 1-2 parts of potassium sorbate.

2. The prebiotic polypeptide complex probiotic as claimed in claim 1, wherein the preparation method of the momordica charantia peptide powder comprises the following steps:

s11, taking one or more of fresh bitter gourds, dried bitter gourds and bitter gourds as bitter gourds, adding deionized water with the weight 5 times of that of the bitter gourds, soaking for 8-12 hours at the water temperature of 25 ℃, taking out and washing for 2-3 times by using the deionized water;

s12, drying the cleaned bitter gourd raw material, smashing and grinding the bitter gourd raw material into pulp to obtain bitter gourd pulp;

s13, taking the balsam pear pulp and the buffer solution to mix so as to obtain a mixed system, wherein the weight ratio of the balsam pear pulp is as follows: buffer solution is 1 (4.5-5.5); recording the total volume value of the mixed system, adjusting the pH value to 7, and then carrying out temperature treatment on the mixed system to obtain an extract;

the temperature treatment process comprises the following steps:

heating to 45-55 ℃, preserving heat for 45-60min, cooling to 20-25 ℃, preserving heat for 25-30min, and recording the first volume value of the whole mixed system; a first mixed solution containing deionized water and a buffer was supplemented at 60% (total volume-first volume), and the deionized water: buffer 4: 1; after the first mixed solution is added, heating to 60-75 ℃, preserving heat for 60-75min, then cooling to 45-55 ℃, preserving heat for 30-35min, and recording a second volume value of the whole mixed system at the moment; a second mixed solution containing deionized water and a buffer was supplemented by 75% (total volume-second volume), and the deionized water: buffer 3: 1; adding the second mixed solution, heating to 80-90 deg.C, maintaining the temperature for 75-85min, cooling to 60-75 deg.C, and maintaining the temperature for 35-45 min;

s14, reducing the temperature of the extract to 20-25 ℃, and then carrying out enzymolysis on the extract to obtain a momordica charantia peptidase hydrolysis system; wherein, the enzymolysis process comprises the following steps:

carrying out first enzymolysis: adjusting the pH value of the extract to 8.0, adding trypsin according to 4.5% of the weight of the extract, stirring at 80-100 rpm, heating to 35-40 ℃ while stirring, and keeping the temperature for 45-60min to obtain a first enzymolysis system;

and (3) carrying out second enzymolysis: after the temperature of the first enzymolysis system is reduced to 20-25 ℃, adjusting the pH value to 3.5, adding pectinase according to 3 percent of the weight of the first enzymolysis system, stirring at 80-100 r/min, heating to 45-55 ℃ while stirring, and preserving heat for 40-60min to obtain a second enzymolysis system;

and (3) carrying out third enzymolysis: after the temperature of the second enzymolysis system is reduced to 20-25 ℃, adjusting the pH value to 4.8, adding cellulase according to 2% of the weight of the second enzymolysis system, stirring at 80-100 r/min, heating to 55-60 ℃ while stirring, and preserving heat for 30-45min to obtain a third enzymolysis system;

s15, after the enzymolysis process in the step S14 is finished, heating the obtained third enzymolysis system to 90 ℃, and maintaining for 10min to finish the enzyme deactivation process to obtain a bitter gourd peptide crude extraction system; adding activated carbon in the crude extract system of the bitter gourd peptide according to 4-5% of the weight of the bitter gourd peptide, uniformly stirring, keeping the temperature at 65 ℃ for 60-90min, centrifuging, and removing sediments to obtain a crude extract of the bitter gourd peptide;

filtering the crude extract of the bitter gourd peptide by diatomite to obtain a bitter gourd peptide clear solution, wherein the filtering pressure is 0.2-0.3 MPa; adding 4.5% of active carbon into the bitter gourd peptide clear liquid by weight, standing for 45-50min, centrifuging, and removing sediments;

s16, filtering the bitter gourd peptide clear liquid after removing the sediment by a microfiltration ceramic membrane with the filtering aperture of 0.5-0.8 mu m, wherein the filtering temperature is 60 ℃ to obtain microfiltration membrane permeate;

filtering the microfiltration membrane permeate through a 200kDa roll-type ultrafiltration membrane with the molecular weight cutoff of 100-;

concentrating the ultrafiltration membrane retentate through a roll-type high-pressure reverse osmosis membrane with the molecular weight cutoff of 150-;

s17, drying the bitter gourd peptide concentrated solution by a vacuum freeze drying method to obtain bitter gourd peptide powder with the bitter gourd polypeptide protein content not less than 30%.

3. The prebiotic polypeptide complex probiotic of claim 2, wherein the buffer is a phosphate buffer.

4. The prebiotic polypeptide complex probiotic as claimed in claim 1, wherein the preparation method of the soybean polypeptide powder comprises the following steps:

s21, inoculating the aspergillus niger strains and the aspergillus oryzae strains on independent potato glucose agar culture media respectively, then putting the culture media into an incubator respectively, and activating the culture media for 48 to 72 hours at the temperature of between 28 and 35 ℃ respectively to obtain activated aspergillus niger strains and activated aspergillus oryzae strains;

culturing the activated aspergillus niger species with the prepared first liquid culture medium to obtain a liquid aspergillus niger seed solution, and culturing the activated aspergillus oryzae species with the prepared second liquid culture medium to obtain a liquid aspergillus oryzae seed solution;

inoculating the liquid Aspergillus niger seed liquid into a first fermentation culture medium in a fermentation tank for amplification culture at 30-35 ℃, the amplification culture temperature is 30.5-7.0, the rotation speed is 500-; and inoculating the liquid Aspergillus oryzae seed liquid into a second fermentation culture medium in a fermentation tank for amplification culture at 30-40 deg.C under stirring at 400-600rpm at pH6.5-7.0 for 1-2d to obtain liquid Aspergillus oryzae spore suspension;

s22, calculated by weight portion,30-45 parts of soybean protein powder, 350 parts of deionized water 250-₃)₃.6H₂0.5-0.6 part of O and NH₄Cl 1.5-2.5 parts, KNO₃1-2 parts to obtain a fermentation system, and adjusting the pH of the fermentation system to 6.8-7.0;

sequentially carrying out three-stage fermentation treatment on the fermentation system:

in the first fermentation stage treatment, the fermentation temperature is 30-35 ℃, the fermentation system is stirred under the condition of the rotation speed of 300-400rpm, and simultaneously the first illumination treatment and the first magnetic field treatment are carried out on the fermentation system for 1-2 d; the first illumination treatment is as follows: the light intensity is 25-30 mu mol.m^-2.s^-1The red light and the light intensity are 30-35 mu mol^-2.s^-1Irradiating the fermentation system with blue light for 45-60 min; the first magnetic field treatment is as follows: performing magnetic field treatment on the fermentation system with 50Hz alternating magnetic field with magnetic field strength of 0.45-0.55mT for 45-60 min;

in the second fermentation stage treatment, the fermentation temperature is 25-35 ℃, the fermentation system is stirred under the condition of the rotation speed of 500-600rpm, and simultaneously the second illumination treatment and the second magnetic field treatment are carried out on the fermentation system for 1-2 d; the second illumination treatment is as follows: the light intensity is 20-24 mu mol.m^-2.s^-1The red light and the light intensity are 24-28 mu mol^-2.s^-1Irradiating the fermentation system with blue light for 35-45 min; the second magnetic field treatment is; treating the fermentation system with 50Hz alternating magnetic field with magnetic field strength of 0.35-0.45mT for 35-45 min;

in the third fermentation stage treatment, the fermentation temperature is 25-35 ℃, the fermentation system is stirred under the condition of the rotation speed of 300-400rpm, and simultaneously the third illumination treatment and the third magnetic field treatment are carried out on the fermentation system for 1-2 d; the third illumination treatment is as follows: the light intensity is 16-20 mu mol.m^-2.s^-1The red light and the light intensity are 20-22 mu mol^-2.s^-1Blue light irradiation of fermentation bodyThe irradiation treatment time is 25-35 min; the third magnetic field treatment is as follows: treating the fermentation system with 50Hz alternating magnetic field with magnetic field strength of 0.2-0.3mT for 25-35 min;

s23, heating the fermentation system subjected to the fermentation treatment in the step S22 to 85 ℃, and maintaining for 10min to complete the enzyme deactivation process to obtain the soybean peptidase hydrolysate;

s24, dispersing the soybean peptidase hydrolyzed solution into an ethanol solution with volume fraction of 95% and mass of 5-6 times of the soybean peptidase hydrolyzed solution, and simultaneously performing ultrasonic treatment and stirring, wherein the ultrasonic power is 600-800W, the ultrasonic treatment time is 10-15min, and the stirring speed is 100-120 r/min;

s25, filtering the reaction system processed in the step S24, controlling the filtering pressure at 0.3-0.4MPa and the filtering temperature at 50-60 ℃, and removing filter residues to obtain soybean peptide clear liquid;

s26, carrying out freeze drying on the soybean peptide clear liquid to obtain the soybean peptide powder.

5. The prebiotic polypeptide complex probiotic of claim 4, wherein the first liquid culture medium comprises, by weight: 1% of tea polyphenol, 1.5% of cane sugar, 2.5% of glucose, 5% of malt extract and CoCl.6H₂O 0.1％、CuSO₄.5H₂O 0.05％、FeNaEDTA 2％、H₃BO₃0.25％、Na₂MoO₄.2H₂O 0.15％、ZnSO₄.7H₂O 0.1％、MgSO₄0.1％、KNO₃0.1％、KNO₃0.1％、MnSO₄.H₂0.2 percent of O and 86.85 percent of deionized water.

6. The prebiotic polypeptide complex probiotic of claim 4, wherein the first fermentation medium comprises, by weight: 10% of tomato juice, 2% of soluble starch, 2% of sucrose, 2% of glucose, 2.5% of corn flour, 0.5% of yeast powder and FeSO₄.7H₂O 0.1％、MgSO₄0.05％、KNO₃0.05％、CuCl₂.2H₂O 0.1％、(NH4)₆Mo₇O₂₄.7H₂O 0.15％、MnSO₄.H₂0.1 percent of O and 80.45 percent of deionized water.

7. The prebiotic polypeptide complex probiotic of claim 4, wherein the second liquid culture medium comprises, by weight: 8% of bean sprout juice, 1.5% of cane sugar, 2.5% of glucose, 2% of peptone, 2% of FeNaEDTA, and H₃BO₃0.25％、Na₂MoO₄.2H₂O 0.15％、MgSO₄0.1％、KNO₃0.1％、MnSO₄.H₂0.2 percent of O and 83.2 percent of deionized water.

8. The prebiotic polypeptide complex probiotic of claim 4, wherein the second fermentation medium comprises, by weight: 5% of maltodextrin, 8% of corn flour, 2% of xylose, 1.5% of yeast extract and MgSO₄0.05％、KNO₃0.1％、NH₄NO₃0.15％、MnSO₄.H₂0.1 percent of O and 83.1 percent of deionized water.

9. The prebiotic polypeptide complex probiotic as claimed in claim 4, wherein in step S21, when the liquid Aspergillus niger seed solution and/or the liquid Aspergillus oryzae seed solution is subjected to scale-up culture, the corresponding fermentation medium inoculated with the Aspergillus niger seed solution or the liquid Aspergillus oryzae seed solution is subjected to ultrasonic treatment every day, wherein the ultrasonic treatment frequency is 20-40Khz, and the treatment time is 30 min.

10. A preparation method of prebiotics polypeptide composite probiotics for relieving gastrointestinal discomfort and abdominal distension and diarrhea is characterized by comprising the following steps:

s100, weighing the components according to the component dosage in the claim 1, and mixing for 4-6 minutes to obtain a mixture;

s200, feeding the mixture into a granulator for granulation to obtain a coarse finished product, wherein the steam pressure in the granulator is 0.5-0.8Mpa, the granulation temperature is 40-50 ℃, and the diameter of a die hole of the granulator is 1.5-3 mm;

s300, sieving the crude product by a 300-mesh sieve to finally obtain the prebiotics polypeptide composite probiotics for relieving gastrointestinal discomfort and abdominal distension and diarrhea.