CN110771890A - Small-molecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity and preparation method thereof - Google Patents

Abstract

The invention discloses a micromolecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity, a preparation method and a preparation method thereof, wherein the micromolecule polypeptide compound nutrient comprises the following components in parts by weight: 15-20 parts of soybean peptide powder, 10-15 parts of pumpkin seed extract, 5-8 parts of poria cocos, 3-5 parts of selenium-rich cordyceps militaris powder, 2-3 parts of yeast extract, 3-5 parts of spina date seed, 5-8 parts of gardenia fruit oil, 1-2 parts of trehalose, 5-8 parts of lily, 2-3 parts of radix ophiopogonis, 8-10 parts of walnut kernel, 3-5 parts of black bean powder and 5-8 parts of mannitol. The pumpkin seed extract, the yeast extract and the gardenia oil extract are prepared by a special extraction process, so that the active ingredients of the pumpkin seed extract, the yeast extract and the gardenia oil extract are fully released, and the pumpkin seed extract, the yeast extract and the gardenia oil extract are reasonably matched with other components to achieve good effects of enhancing immunity, soothing nerves and helping sleep.

Description

Small-molecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity and preparation method thereof

Technical Field

The invention relates to the field of health-care food. More specifically, the invention relates to a micromolecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity and a preparation method thereof.

Background

Insomnia refers to the difficulty in ensuring sleep quality during night sleep, difficulty in falling asleep or short sleep time, light dreaminess and easy wakening. Modern medicine considers that the main reason for insomnia is that nerves in the brain are in an overstrained state or a mood fluctuation state for a long time, such as long-time work, high psychological stress, frequent shift, overfatigue and sadness. The long-term insomnia causes patients to have cachexia, nervous disturbance, immunity reduction, hypomnesis and the like, and is more likely to cause the attack of various diseases such as cerebral hemorrhage, hypertension, hemiplegia, diabetes and the like, thereby increasing the death probability of the patients.

To alleviate the imbalance of excitation and inhibition of the cerebral cortex, relaxation therapy and drug therapy are commonly used. The relaxation therapy mainly aims at guiding patients, relaxing mood and enabling the patients to know that insomnia is caused by normal anxiety or emotion and can cure and relieve the anxiety, and the method has good effect on some patients, but has large limitation and varies from person to person. The drug therapy is often combined with relaxation therapy, and tranquilizers are adopted to inhibit the abnormal activity of cerebral nerves, such as fluoroamine, but drug resistance is generated after long-term administration, insomnia is aggravated after drug withdrawal, and the drugs increase the burden of liver and kidney, and the liver and kidney functions need to be checked regularly.

In addition, the low immunity can be seen in people of various ages, such as children, the old, people taking immunosuppressants for a long time after organ transplantation, malignant tumor patients receiving radiotherapy and chemotherapy, people with reduced immunity caused by lack of nutrition, medical workers with high probability of contacting infectious diseases for a long time, middle-aged people under great deep life pressure, and the like, and the low immunity is widely distributed among people. Among them, for middle-aged people with great deep life and workplace pressure, the thinking and brain activity of middle-aged people are frequent after the middle-aged people, so the pressure is increased, the sleep is less and less, the sleep quality is poorer and poorer, the immunity of the organism is too low, the bacteria and virus infection is easily caused, and the middle-aged people easily feel fatigue, the gastrointestinal function is reduced, the self-protection capability is poor, the middle-aged people are easily attacked by infectious diseases, the spirit is low, the mind and the body are pallid, the serious infection is easily caused, and the like.

Disclosure of Invention

In order to solve the technical problems, the invention provides a micromolecule polypeptide compound nutrient for soothing nerves and helping sleep and improving immunity and a preparation method thereof.

To achieve these objects and other advantages in accordance with the present invention, there is provided a small molecule polypeptide complex nutrient for soothing nerves, aiding sleep and improving immunity, comprising: 15-20 parts of oat flour, 10-15 parts of pumpkin seed extract, 3-8 parts of cored lotus seed, 5-8 parts of poria cocos, 3-5 parts of selenium-rich cordyceps militaris powder, 2-3 parts of yeast extract, 8-10 parts of tomato powder, 3-5 parts of honey, 10-15 parts of denucleated dried longan, 5-8 parts of gardenia fruit oil, 5-8 parts of lily, 2-3 parts of radix ophiopogonis, 8-10 parts of walnut kernel, 3-5 parts of black bean powder and 5-8 parts of mannitol.

Preferably, the preparation method of the pumpkin seed extract comprises the following steps:

s11, taking fresh pumpkin seeds, soaking at 25 ℃ for 10-12h, taking out, washing with deionized water for 2-3 times, drying, grinding, and sieving with a 100-mesh sieve to obtain pumpkin seed powder;

s12, adding deionized water with the weight 10-12 times that of the pumpkin seed powder into the pumpkin seed powder to obtain an enzymolysis raw material, and carrying out enzymolysis on the enzymolysis raw material to obtain an enzymolysis system; wherein, the enzymolysis process comprises the following steps:

carrying out first enzymolysis: adjusting the pH value of the enzymolysis raw material to 8.0-9.0, adding trypsin which accounts for 3-4% of the weight of the enzymolysis raw material, fully stirring, heating to 40-45 ℃ while stirring, and preserving heat for 35-40min 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 3.5-4.5, adding pectinase according to 3.5-4.5% of the weight of the first enzymolysis system, fully stirring, heating to 45-50 ℃ while stirring, and preserving heat 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, adding cellulase according to 2-4% of the weight of the second enzymolysis system, fully stirring, heating to 50-65 ℃ while stirring, and preserving heat for 25-35min to obtain a third enzymolysis system;

s13, 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 a pumpkin seed enzymolysis system;

s14, adding phospholipase accounting for 0.01% of the weight of the pumpkin seed enzymolysis system and pancreatin accounting for 0.02% of the weight of the pumpkin seed enzymolysis system, and recording the total volume value of the pumpkin seed enzymolysis system; then carrying out temperature treatment on the pumpkin seed enzymolysis system to obtain a pumpkin seed extraction system;

the temperature treatment process comprises the following steps:

heating to 35-45 ℃, preserving heat for 60-90min, cooling to 30-40 ℃, preserving heat for 25-35min, and recording a first volume value of the reaction system at the moment; adding a first mixed solution containing deionized water, phospholipase and pancreatin according to (total volume-first volume) 65%, wherein the first mixed solution contains 97.5% of deionized water, 1% of phospholipase and 1.5% of pancreatin according to weight ratio; after the first mixed solution is added, heating to 45-50 ℃, preserving heat for 75-85min, then cooling to 40-45 ℃, preserving heat for 30-35min, and recording a second volume value of the reaction system at the moment; adding a second mixed solution containing deionized water, phospholipase and pancreatin according to (total volume-second volume) 45%, wherein the second mixed solution contains 98% of deionized water, 1% of phospholipase and 1% of pancreatin according to weight ratio; adding the second mixed solution, heating to 55-65 deg.C, maintaining the temperature for 80-90min, cooling to 50-60 deg.C, and maintaining the temperature for 40-45 min;

s15, adding activated carbon in the pumpkin seed extraction system according to 4% of the weight of the pumpkin seed extraction system, uniformly stirring, keeping the temperature at 65 ℃ for 65-85min, centrifuging, and removing sediments to obtain a pumpkin seed crude extract; filtering the crude extract of pumpkin seeds by diatomite to obtain a first-grade pumpkin seed clear liquid, wherein the filtering pressure is 0.25-0.35 MPa; adding 3% of active carbon into the first-stage pumpkin seed clear liquid according to the weight, standing for 45-50min, centrifuging, and removing sediments to obtain a second-stage pumpkin seed clear liquid;

s16, filtering the second-level pumpkin seed clear liquid by a microfiltration ceramic membrane with the filtering aperture of 0.2-1.0 mu m, wherein the filtering temperature is 50-60 ℃ to obtain microfiltration membrane permeate;

filtering the microfiltration membrane permeate through a 220kDa roll-type ultrafiltration membrane with the molecular weight cutoff of 120-;

and concentrating the ultrafiltration membrane retentate through a roll-type high-pressure reverse osmosis membrane with the molecular weight cutoff of 200-.

Preferably, in step S14, the temperature treatment is performed on the mixed solution and the ultrasonic treatment is performed at the same time, wherein the ultrasonic power is 200-400W, and the ultrasonic treatment time is 10-15 min.

Preferably, the preparation method of the yeast extract comprises the following steps:

s21, putting active dry yeast into a fermentation tank, wherein the diameter of the fermentation tank is 30-40cm, adding water which is 3-5 times of the weight of the active dry yeast into the fermentation tank, and uniformly stirring to obtain a yeast stock solution;

s22, irradiating the yeast stock solution by adopting a plurality of pulse laser light sources for 20-30min, stirring the yeast stock solution under the conditions of 200-300 r/min while irradiating, and carrying out ultrasonic treatment on the yeast stock solution at the same time, wherein the ultrasonic power is 100KW and the ultrasonic treatment temperature is 30-35 ℃; the diameter of a light spot formed on the liquid surface of the yeast stock solution by each pulse laser is 0.5-1cm, and the laser energy of each pulse laser acting on the yeast stock solution is 50 mJ;

s23, carrying out enzymolysis treatment on the yeast stock solution treated in the step S22; and the enzymolysis treatment process comprises the following steps:

(1) and 1, enzymolysis: raising the temperature of the yeast stock solution treated in the step S22 to 20-25 ℃, then lowering the temperature to-45 ℃ at the speed of 15 ℃/min, maintaining the temperature for 1min after lowering the temperature to-45 ℃, and then raising the temperature of the yeast stock solution to 20-25 ℃ at the speed of 10 ℃/min; heating to 20-25 deg.C, adding 1 wt% of alkaline protease and 1.5 wt% of papain into the yeast stock solution, adjusting pH to 7.0-8.0 and temperature to 50-60 deg.C, and performing hydrolysis reaction for 10-12 hr;

adjusting the temperature of yeast stock solution treated by alkaline protease and papain to 40-55 ℃, adding snailase accounting for 1 percent of the weight of the yeast stock solution treated by the alkaline protease and the papain and pancreatin accounting for 2.0 percent of the weight of the yeast stock solution treated by the alkaline protease and the papain, adjusting the pH to 6.5-7.5, filling ozone, stirring the yeast stock solution under the condition of 200 plus 300 revolutions per minute, and carrying out hydrolysis reaction for 3-4h to obtain first enzymatic hydrolysate;

(2) and (3) enzymolysis: adjusting the temperature of the first enzymolysis liquid to 20-25 ℃, reducing the temperature to-45 ℃ at the speed of 10 ℃/min, maintaining the temperature for 0.5min after reducing the temperature to-45 ℃, and increasing the temperature of the first enzymolysis liquid to 20-25 ℃ at the speed of 10 ℃/min; after the temperature is raised to 20-25 ℃, adding 0.5 percent of acid protease and 1.5 percent of nuclease by weight into the first enzymolysis liquid, adjusting the pH to 4.5-6.0 and the temperature to 45-50 ℃, and reacting for 8-10 h;

adjusting the temperature of the first enzymolysis liquid treated by acid protease and nuclease to 40-55 ℃, adding helicase accounting for 0.5 percent of the weight of the yeast stock solution treated by the acid protease and the papain and pancreatin accounting for 1.0 percent of the weight of the yeast stock solution treated by the acid protease and the papain, adjusting the pH to 6.0-7.5, filling ozone, and stirring the first enzymolysis liquid under the condition of 200-;

(3) and (3) enzymolysis: adjusting the temperature of the second enzymolysis liquid to 20-25 ℃, reducing the temperature to-45 ℃ at the speed of 5 ℃/min, maintaining the temperature for 0.5min after reducing the temperature to-45 ℃, and increasing the temperature of the second enzymolysis liquid to 20-25 ℃ at the speed of 12 ℃/min; after the temperature is raised to 20-25 ℃, adding glutaminase and deaminase which are 0.5 percent and 1.0 percent of the weight of the second enzymolysis liquid into the second enzymolysis liquid, adjusting the pH to 5.5-6.0 and the temperature to 45-50 ℃, and reacting for 3-4 hours;

regulating the temperature of the second enzymatic hydrolysate treated by the glutaminase and the deaminase to 40-55 ℃, adding the snailase accounting for 1 percent of the weight of the second enzymatic hydrolysate treated by the glutaminase and the deaminase and the pancreatin accounting for 1 percent of the weight of the second enzymatic hydrolysate treated by the glutaminase and the deaminase, regulating the pH to 6.5-7.5, filling ozone, and stirring the second enzymatic hydrolysate under the condition of 200-300 revolutions per minute for 2-3 hours to obtain a third enzymatic hydrolysate;

s24, after the enzymolysis is finished, heating the obtained third enzymolysis liquid to 90 ℃, and maintaining for 10min to finish the enzyme deactivation process to obtain a yeast extract stock solution;

s25, adding activated carbon 3% of the yeast extract stock solution by weight, stirring uniformly, keeping the temperature at 55 ℃ for 60-90min, centrifuging, removing sediments to obtain yeast extract clear solution, and filtering under the pressure of 0.25-0.35 MPa;

s26, filtering the clear yeast extract liquid by a microfiltration ceramic membrane, and controlling the operation temperature to be 55-65 ℃ to obtain microfiltration membrane permeate;

filtering the microfiltration membrane permeate through a roll-type ultrafiltration membrane, and controlling the operation temperature to be 55-65 ℃ to obtain ultrafiltration membrane permeate;

concentrating the ultrafiltration membrane retentate by a roll-type high-pressure reverse osmosis membrane to remove water and partial residual inorganic salt and small molecular impurities, and controlling the operation temperature to be below 40 ℃ to obtain a yeast extract concentrated solution;

s27, drying the yeast extract concentrated solution by a vacuum freeze drying method to obtain the yeast extract.

Preferably, the active dry yeast cell has a protein content of 40-45%, a carbohydrate content of 8-10%, and a water content of 4-5%.

Preferably, the extraction method of the gardenia fruit oil comprises the following steps:

s31, putting fresh gardenia fruits into water, soaking at 25 ℃ for 24-36h, taking out, washing with water for 2-3 times, drying, grinding, and sieving with a 100-mesh sieve to obtain gardenia fruit powder;

s32, taking gardenia fruit powder, adding deionized water with the weight 5-10 times of that of the gardenia fruit powder to obtain an enzymolysis raw material, and carrying out enzymolysis on the enzymolysis raw material; wherein, the enzymolysis process comprises the following steps:

carrying out first enzymolysis: adding trypsin in an amount of 5% of the weight of the gardenia fruit powder and adding a permeability regulating solution for regulating the permeability of cell membranes and/or cell walls in an amount of 45-55% of the weight of the gardenia fruit powder into the enzymolysis raw materials, regulating the pH value to 6.5-7.5, fully stirring, heating to 42-45 ℃ while stirring, and preserving heat for 30-45min to obtain a first enzymolysis system; the permeability regulating liquid consists of acid solution, glycerol, sodium chloride and lysozyme, and the acid solution comprises the following components in percentage by weight: glycerol: sodium chloride: lysozyme is 1: (0.7-1.0): (0.02-0.05): (0.03-0.06);

and (3) carrying out second enzymolysis: after the temperature of the first enzymolysis system is reduced to 20-25 ℃, adjusting the pH value to 3.5-4.5, adding pectinase according to 4% of the weight of the first system, fully stirring, heating to 50-60 ℃ while stirring, and preserving heat 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, adding cellulase according to 3.5% of the weight of the second enzymolysis system, fully stirring, heating to 50-65 ℃ while stirring, and preserving heat for 25-35min to obtain a third enzymolysis system;

s33, after the enzymolysis process in the step S22 is finished, heating the obtained third enzymolysis system to 85 ℃, and maintaining for 10min to finish the enzyme deactivation process to obtain a gardenia fruit enzymolysis system;

s34, adding activated carbon in an enzymolysis system of the gardenia fruit according to 3% of the weight of the gardenia fruit, uniformly stirring, keeping the temperature at 65 ℃ for 65-85min, centrifuging, and removing sediments to obtain a crude extract of gardenia fruit oil; filtering the crude gardenia fruit oil extract by diatomite to obtain a gardenia fruit oil extract, wherein the filtering pressure is 0.3-0.4 MPa; adding 3% of active carbon into the gardenia fruit oil extracting solution according to the weight, standing for 45-50min, centrifuging, and removing sediments; standing for 2-3h, and collecting the upper oil layer to obtain the fructus Gardeniae fruit oil.

Also provides a preparation method of the micromolecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity, which comprises 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 75-85 ℃, and the diameter of a die hole of the granulator is 1.5-3 mm;

s300, sieving the crude product by using an 8-mesh sieve to finally obtain the micromolecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity and the preparation method thereof.

The invention at least comprises the following beneficial effects:

according to the invention, through the extraction process of staged heating, repeated enzymolysis combined with ultrasonic treatment and multiple filtration, the yield and purity of the pumpkin seed extract (containing pumpkin seed protein and the like) and the gardenia oil extract are greatly improved, meanwhile, the yeast autolysis is promoted by adopting a mode of pulse laser, ultrasonic treatment and multiple enzymolysis, so that the yeast cell content is fully released, and further, after the multiple active ingredients are reasonably compounded and used, the formed small molecular polypeptide compound nutrient can remarkably improve the organism immunity of metabolic diseases such as diabetes and the like, crowds with various symptoms such as palpitation, insomnia, uneasiness and the like, and middle-aged people crowds, improve the sleep quality and delay aging.

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 micromolecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity in the embodiment comprises the following components in parts by weight: 15 parts of oat flour, 10 parts of pumpkin seed extract, 3 parts of lotus seed without core, 5 parts of poria cocos, 3 parts of selenium-rich cordyceps militaris powder, 2 parts of yeast extract, 8 parts of tomato powder, 3 parts of honey, 10 parts of dried denucleated longan, 5 parts of gardenia fruit oil, 5 parts of lily, 2 parts of radix ophiopogonis, 8 parts of walnut kernel, 3 parts of black bean powder and 5 parts of mannitol.

Further, the preparation method of the pumpkin seed extract comprises the following steps:

s11, taking fresh pumpkin seeds, soaking at 25 ℃ for 10-12h, taking out, washing with deionized water for 2-3 times, drying, grinding, and sieving with a 100-mesh sieve to obtain pumpkin seed powder;

s12, adding deionized water which is 10-12 times of the weight of the pumpkin seed powder (preferably the pumpkin seed powder) into the pumpkin seed powder to obtain an enzymolysis raw material, and carrying out enzymolysis on the enzymolysis raw material to obtain an enzymolysis system; wherein, the enzymolysis process comprises the following steps:

carrying out first enzymolysis: adjusting the pH value of the enzymolysis raw material to 8.0-9.0 (preferably 8.5), adding trypsin 3-4% (preferably 3.5%) of the enzymolysis raw material, stirring, heating to 40-45 deg.C (preferably 42 deg.C) while stirring, and keeping the temperature for 35-40min 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 3.5-4.5 (preferably 4.0), adding pectinase according to 3.5-4.5% (preferably 4.0%) of the weight of the first enzymolysis system, fully stirring, heating to 45-50 ℃ while stirring, and preserving heat 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 2-4% (preferably 3.0%) of the weight of the second enzymolysis system, fully stirring, heating to 50-65 ℃ while stirring, and preserving heat for 25-35min to obtain a third enzymolysis system;

s13, 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 a pumpkin seed enzymolysis system;

s14, adding 0.01% phospholipase and 0.02% pancreatin into the pumpkin seed enzymolysis system, and recording

The total volume value of the pumpkin seed enzymolysis system; then carrying out temperature treatment on the pumpkin seed enzymolysis system at the moment, and simultaneously carrying out ultrasonic treatment on the pumpkin seed enzymolysis system, wherein the ultrasonic power is 200-400W (preferably 300W), and the ultrasonic treatment time is 10-15min (preferably 12min), so as to obtain a pumpkin seed extraction system;

the temperature treatment process comprises the following steps:

heating to 35-45 deg.C (preferably 40 deg.C), maintaining for 60-90min (preferably 70min), cooling to 30-40 deg.C (preferably 35 deg.C), maintaining for 25-35min (preferably 35min), and recording the first volume value of the reaction system; adding a first mixed solution containing deionized water, phospholipase and pancreatin according to (total volume-first volume) 65%, wherein the first mixed solution contains 97.5% of deionized water, 1% of phospholipase and 1.5% of pancreatin according to weight ratio; adding the first mixed solution, heating to 45-50 deg.C, maintaining the temperature for 75-85min (preferably 80min), cooling to 40-45 deg.C (preferably 42 deg.C), maintaining the temperature for 30-35min, and recording the second volume value of the reaction system; adding a second mixed solution containing deionized water, phospholipase and pancreatin according to (total volume-second volume) 45%, wherein the second mixed solution contains 98% of deionized water, 1% of phospholipase and 1% of pancreatin according to weight ratio; adding the second mixed solution, heating to 55-65 deg.C, maintaining the temperature for 80-90min (preferably 85min), cooling to 50-60 deg.C (preferably 55 deg.C), and maintaining the temperature for 40-45 min; in the step, water, phospholipase and pancreatin in corresponding proportion are supplemented after each temperature rise and heat preservation stage is finished, so that a reaction system after water evaporation and partial enzyme inactivation is compensated, and the reaction system is always in a better enzymolysis environment to achieve the best enzymolysis effect;

s15, adding activated carbon in the pumpkin seed extraction system according to 4% of the weight of the pumpkin seed extraction system, uniformly stirring, keeping the temperature at 65 ℃ for 65-85min (preferably 75min), centrifuging for 5-8min under the conditions of 8000-; filtering the crude extract of pumpkin seeds by diatomite to obtain a first-grade pumpkin seed clear liquid, wherein the filtering pressure is 0.25-0.35MPa (preferably 0.3 MPa); adding 3% of active carbon into the first-stage pumpkin seed clear liquid according to the weight, standing for 45-50min, centrifuging, and removing sediments to obtain a second-stage pumpkin seed clear liquid;

s16, filtering the second-level pumpkin seed clear liquid by a microfiltration ceramic membrane with the filtering aperture of 0.2-1.0 μm, wherein the filtering temperature is 50-60 ℃ (preferably 55 ℃) to obtain microfiltration membrane permeate;

filtering the microfiltration membrane permeate through a 220kDa roll-type ultrafiltration membrane with the molecular weight cut-off of 120-;

concentrating the ultrafiltration membrane retentate through a roll-type high-pressure reverse osmosis membrane with the molecular weight cutoff of 200-;

the cell wall tissues of the pumpkin seed cells are rich, the pectinase and the cellulase can destroy the cell wall structures, the phospholipase can destroy the cell membrane structures, and the trypsin and the pancreatin can decompose proteins, so that the cell wall and the cell membrane structures of the pumpkin seed cells can be fully destroyed by the enzymes in combination with independent enzymolysis and ultrasonic treatment conditions, the content of the pumpkin seed cells is released, macromolecular proteins are further decomposed into micromolecular polypeptides which can be easily absorbed by a human body, and the synthesis of immunoglobulin is promoted through the micromolecular pumpkin seed proteins, so that the humoral immunity is effectively enhanced; the T lymphocyte proliferation can be improved, the cellular immune response can be enhanced, the release of various endocrine hormones can be promoted, the T lymphocyte proliferation comprises insulin, growth hormone, prolactin, antidiuretic hormone, catecholamine and the like, the effect of regulating immunity is achieved, meanwhile, the cell structure (such as cell membranes and the like) composition of the components can be repeatedly impacted and destroyed in different temperature change environments through staged temperature rise and heat preservation, meanwhile, water, pancreatin and phospholipase in corresponding proportions are supplemented after each temperature rise and heat preservation stage is finished, so that the reaction system after water evaporation and enzyme inactivation is compensated, and the reaction system is always in a better enzymolysis environment, and the optimal enzymolysis effect can be achieved.

Further, yeast extract (yeast extract) is a substance which is rich in natural active ingredients such as small molecular amino acids, peptides, nucleotides, vitamins and the like and is formed by extracting proteins, nucleic acids, vitamins and the like in yeast after wall breaking and carrying out biological enzymolysis. Wherein, the amino acid is the main component of the natural moisturizing factor in the skin cuticle, is easy to be absorbed by the skin, restores the elasticity of the aged epidermis and delays the skin aging; nucleic acid and nucleotide are main genetic materials of human body, and have the functions of promoting metabolism, increasing protein synthesis speed, etc., so as to enhance immune function and SOD activity and improve anti-aging capability. To this end, the present embodiment also provides a method for preparing a yeast extract, which comprises:

s21, putting active dry yeast into a fermentation tank, wherein the diameter of the fermentation tank is 30-40cm (preferably 35cm), adding water which is 3-5 times (preferably 4 times) the weight of the active dry yeast into the fermentation tank, and uniformly stirring to obtain a yeast stock solution; the active dry yeast cell has the protein content of 40-45%, the carbohydrate content of 8-10% and the water content of 4-5%;

s22, irradiating the yeast stock solution by adopting a plurality of (preferably 8-10) pulse laser light sources for 20-30min (preferably 25cm), stirring the yeast stock solution under the conditions of 200-300 r/min while irradiating, and simultaneously carrying out ultrasonic treatment on the yeast stock solution, wherein the ultrasonic power is 100KW, and the ultrasonic treatment temperature is 30-35 ℃ (preferably 32 ℃); the diameter of a light spot formed on the liquid surface of the yeast stock solution by each pulse laser is 0.5-1cm, and the laser energy of each pulse laser acting on the yeast stock solution is 50 mJ;

the pulse laser and the ultrasonic wave can generate vacuoles in liquid, near cell membranes/cell walls and in cell sap, high temperature and high pressure (5000K, 50Mpa) are generated at the moment of collapsing the vacuoles, so that the cell walls and the plasma membranes of the surrounding yeast cells are broken down or the permeability of the plasma membranes is changed, and the contents of the yeast cells are separated out;

s23, carrying out enzymolysis treatment on the yeast stock solution treated in the step S22; and the enzymolysis treatment process comprises the following steps:

(1) and 1, enzymolysis: raising the temperature of the yeast stock solution treated in the step S22 to 20-25 ℃, then lowering the temperature to-45 ℃ at the speed of 15 ℃/min, maintaining the temperature for 1min after lowering the temperature to-45 ℃, and then raising the temperature of the yeast stock solution to 20-25 ℃ at the speed of 10 ℃/min; heating to 20-25 deg.C, adding alkaline protease and papain 1.5% of the yeast stock solution, adjusting pH to 7.0-8.0 (preferably 7.5) and temperature to 50-60 deg.C (preferably 55 deg.C), and performing hydrolysis reaction for 10-12h (preferably 10.5 h);

adjusting the temperature of yeast stock solution treated by alkaline protease and papain to 40-55 ℃ (preferably 45 ℃), adding helicase 1 wt% and pancreatin 2.0 wt% of the yeast stock solution treated by alkaline protease and papain, adjusting the pH to 6.5-7.5 (preferably 7.0), introducing ozone, stirring the yeast stock solution under the conditions of 200-300 r/min, and performing hydrolysis reaction for 3-4h (preferably 3.5h) to obtain a first enzymolysis solution;

(4) and (3) enzymolysis: adjusting the temperature of the first enzymolysis liquid to 20-25 ℃, reducing the temperature to-45 ℃ at the speed of 10 ℃/min, maintaining the temperature for 0.5min after reducing the temperature to-45 ℃, and increasing the temperature of the first enzymolysis liquid to 20-25 ℃ at the speed of 10 ℃/min; after the temperature is raised to 20-25 ℃, adding 0.5 percent of acid protease and 1.5 percent of nuclease by weight into the first enzymolysis liquid, adjusting the pH to 4.5-6.0 (preferably 5.5) and the temperature to 45-50 ℃ (preferably 48 ℃), and reacting for 8-10h (preferably 9 h);

adjusting the temperature of the first enzymolysis solution treated by acid protease and nuclease to 40-55 ℃ (preferably 42 ℃), adding helicase accounting for 0.5% of the weight of the yeast stock solution treated by the acid protease and the papain and pancreatin accounting for 1.0%, adjusting the pH to 6.0-7.5 (preferably 7.0), filling ozone, and stirring the first enzymolysis solution under the conditions of 200-;

(5) and (3) enzymolysis: adjusting the temperature of the second enzymolysis liquid to 20-25 ℃, reducing the temperature to-45 ℃ at the speed of 5 ℃/min, maintaining the temperature for 0.5min after reducing the temperature to-45 ℃, and increasing the temperature of the second enzymolysis liquid to 20-25 ℃ at the speed of 12 ℃/min; after the temperature is raised to 20-25 ℃, adding glutaminase and deaminase which are 0.5 percent and 1.0 percent of the weight of the glutaminase and deaminase into the second enzymolysis liquid, adjusting the pH to 5.5-6.0 (preferably 5.8) and the temperature to 45-50 ℃ (preferably 42 ℃), and reacting for 3-4h (preferably 3.5 h);

regulating the temperature of the second enzymolysis solution treated by the glutaminase and the deaminase to 40-55 ℃ (preferably 42 ℃), adding the snailase accounting for 1 percent of the weight of the second enzymolysis solution treated by the glutaminase and the deaminase and the pancreatin accounting for 1 percent of the weight of the second enzymolysis solution treated by the glutaminase and the deaminase, regulating the pH to 6.5-7.5, filling ozone, and stirring the second enzymolysis solution according to the condition of 200-300 r/min for hydrolysis for 2-3h to obtain a third enzymolysis solution;

in the step, through the stepwise sudden temperature rise and the sudden temperature drop, ice crystals are generated in the yeast cells, so that the cell structures of the yeast cells are damaged through the expansion and piercing of the ice crystals, and meanwhile, the alkaline protease, the papain, the acidic protease, the helicase, the pancreatin, the glutaminase and the deaminase can all decompose protein, so that the cell membrane structure can be further damaged, macromolecular protein released in the yeast cells is hydrolyzed into micromolecular amino acid, the activity of the micromolecular amino acid is enhanced, and the effects of resisting aging, improving the immunity and the like are fully exerted; in addition, ozone can destroy the cell membrane structure through oxidation, further promote the destruction of the yeast cell structure and promote the release of the content thereof;

s24, after the enzymolysis is finished, heating the obtained third enzymolysis liquid to 90 ℃, and maintaining for 10min to finish the enzyme deactivation process to obtain a yeast extract stock solution;

s25, adding activated carbon 3% of the yeast extract stock solution by weight, stirring uniformly, keeping the temperature at 55 ℃ for 60-90min, centrifuging, removing sediments to obtain yeast extract clear solution, and filtering under the pressure of 0.25-0.35 MPa;

s26, filtering the clear yeast extract liquid by a microfiltration ceramic membrane, and controlling the operation temperature to be 55-65 ℃ to obtain microfiltration membrane permeate;

filtering the microfiltration membrane permeate through a roll-type ultrafiltration membrane, and controlling the operation temperature to be 55-65 ℃ to obtain ultrafiltration membrane permeate;

concentrating the ultrafiltration membrane retentate by a roll-type high-pressure reverse osmosis membrane to remove water and partial residual inorganic salt and small molecular impurities, and controlling the operation temperature to be below 40 ℃ to obtain a yeast extract concentrated solution;

s27, drying the yeast extract concentrated solution by a vacuum freeze drying method to obtain the yeast extract.

In addition, the gardenia fruits are rich in resources, low in price and easy to obtain, have multiple effects and play more and more important roles in modern food industry, and particularly mainly contain compounds such as flavonoids, iridoid mushrooms and cycloenoximones, and components such as crocin, pectin, tannin, polysaccharide, crocin acid, volatile oil, wherein the flavonoids have an auxiliary treatment effect on diseases such as hypertension, and can realize the effects of reducing blood pressure, blood sugar and the like. Therefore, the present embodiment also provides a method for extracting gardenia fruit oil, which comprises:

s31, putting fresh gardenia fruits into water, soaking at 25 ℃ for 24-36h, taking out, washing with water for 2-3 times, drying, grinding, and sieving with a 100-mesh sieve to obtain gardenia fruit powder;

s32, taking gardenia fruit powder, adding deionized water with the weight 5-10 times (preferably 8 times) of that of the gardenia fruit powder to obtain an enzymolysis raw material, and carrying out enzymolysis on the enzymolysis raw material; wherein, the enzymolysis process comprises the following steps:

carrying out first enzymolysis: adding trypsin in an amount of 5% of the weight of the gardenia fruit powder and adding a permeability regulating solution for regulating the permeability of cell membranes and/or cell walls in an amount of 45-55% (preferably 50%) of the weight of the gardenia fruit powder into the enzymolysis raw material, regulating the pH value to 6.5-7.5 (preferably 7.0), fully stirring, raising the temperature to 42-45 ℃ (preferably 43.5 ℃) while stirring, and preserving the temperature for 30-45min (preferably 35min) to obtain a first enzymolysis system; the permeability regulating liquid consists of acid solution, glycerol, sodium chloride and lysozyme, and the acid solution comprises the following components in percentage by weight: glycerol: sodium chloride: lysozyme is 1: (0.7-1.0): (0.02-0.05): (0.03-0.06) (preferably an acid solution: glycerin: sodium chloride: lysozyme: 1: 0.8: 0.03: 0.04, and the acid solution is a citric acid solution);

and (3) carrying out second enzymolysis: after the temperature of the first enzymolysis system is reduced to 20-25 ℃, adjusting the pH value to 3.5-4.5 (preferably 4.0), adding pectinase according to 4% of the weight of the first system, fully stirring, heating to 50-60 ℃ (preferably 55 ℃) while stirring, and keeping the temperature for 30-35min (preferably 32min) 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 3.5% of the weight of the second enzymolysis system, fully stirring, heating to 50-65 ℃ (preferably 60 ℃) while stirring, and preserving heat for 25-35min (preferably 30min) to obtain a third enzymolysis system;

s33, after the enzymolysis process in the step S22 is finished, heating the obtained third enzymolysis system to 85 ℃, and maintaining for 10min to finish the enzyme deactivation process to obtain a gardenia fruit enzymolysis system;

s34, adding activated carbon in an enzymolysis system of the gardenia fruit according to 3% of the weight of the gardenia fruit, uniformly stirring, carrying out heat preservation at 65 ℃ for 65-85min (preferably 75min), centrifuging, and removing sediments to obtain a crude extract of gardenia fruit oil; filtering the crude extract with diatomaceous earth to obtain fructus Gardeniae extract at 0.3-0.4MPa (preferably 0.35 MPa); adding 3% of active carbon into the gardenia fruit oil extracting solution according to the weight, standing for 45-50min, centrifuging, and removing sediments; standing for 2-3h (preferably 2.5h), and collecting the upper oil layer to obtain the fructus Gardeniae fruit oil.

< example 2>

The difference between the embodiment and the embodiment 1 is that the small molecular polypeptide compound nutrient for soothing the nerves, helping sleep and improving immunity consists of the following components in parts by weight: 20 parts of oat flour, 15 parts of pumpkin seed extract, 8 parts of lotus seed without core, 8 parts of poria cocos, 5 parts of selenium-rich cordyceps militaris powder, 3 parts of yeast extract, 10 parts of tomato powder, 5 parts of honey, 15 parts of dried denucleated longan, 8 parts of gardenia fruit oil, 8 parts of lily, 3 parts of radix ophiopogonis, 10 parts of walnut kernel, 5 parts of black bean powder and 8 parts of mannitol.

< example 3>

The difference between the embodiment and the embodiment 1 is that the small molecular polypeptide compound nutrient for soothing the nerves, helping sleep and improving immunity consists of the following components in parts by weight: 18 parts of oat flour, 12 parts of pumpkin seed extract, 6 parts of lotus seed without core, 7 parts of poria cocos, 4 parts of selenium-rich cordyceps militaris powder, 2.5 parts of yeast extract, 9 parts of tomato powder, 4 parts of honey, 12 parts of dried longan without core, 7 parts of gardenia fruit oil, 6 parts of lily, 2.5 parts of radix ophiopogonis, 9 parts of walnut kernel, 4 parts of black bean powder and 7 parts of mannitol.

< detection result of pumpkin seed extract >

Pumpkin seed protein was extracted by the method described in example 1 of the patent application No. 201810995144.3 ("a method for preparing pumpkin seed protein") as comparative example 1, and it was subjected to determination of free amino acid content by formaldehyde titration method and determination of total nitrogen content according to the micro kjeldahl method with the pumpkin seed extract obtained by the method for preparing pumpkin seed extract in examples 1-3 of the present invention, and the degree of hydrolysis was calculated from the Degree of Hydrolysis (DH) ═ free amino acid/total nitrogen × 100%, the results of which are shown in table 1; and detecting the molecular weight and the distribution range of the pumpkin seed protein by high performance gel filtration chromatography, wherein the results are shown in table 2.

TABLE 1 determination of the degree of hydrolysis of pumpkin seed extracts

	Comparative example 1	Example 1	Example 2	Example 3
					Degree of Hydrolysis (DH) (%)	19.74	10.47	9.45	9.58

TABLE 2 pumpkin seed protein molecular weight and distribution

As can be seen from tables 1-2, in the preparation method of the pumpkin seed extract, macromolecular proteins can be decomposed into small molecular polypeptides by trypsin, pancreatin and the like, wherein the content of free amino acids in the extract prepared by the invention is low (only half of that in comparative example 1), but meanwhile, in the pumpkin seed extract prepared by the invention, the average molecular weight of protein peptides is 800Da, 90% of polypeptides below 1000Da and 75% of polypeptides below 500Da, so that the pumpkin seed extract can be rapidly and efficiently absorbed by human bodies, and the effects of the pumpkin seed extract can be fully exerted.

< results of detection of Yeast extract >

The yeast extract prepared by the method described in example 1 of the patent application No. 201510416294.0 ("a yeast extract and a method for preparing the same") was used as comparative example 2, and the wall-breaking rate, and the protein content, the amino acid content, the solid content, the reducing sugar content, and the nucleotide content of the contents were measured with the yeast extract prepared by the method for preparing a yeast extract according to examples 1 to 3 of the present invention, and the above-mentioned measurements were performed in the prior art, and are not repeated herein, and the measurement results are shown in table 3.

TABLE 3 Yeast extract index determination

Protein content (%)

Amino acid content (%)

Solid content (%)

Reducing sugar content (%)

Nucleotide content (%)

Wall breaking ratio (%)

Comparative example 2

7.58±1.21

20.12±1.21

42.43±0.72

10.15±0.68

5.79±0.21

36.31±1.21

Example 1

10.21±0.84

31.33±1.32

67.52±0.58

15.23±0.58

7.11±0.31

67.24±1.20

Example 2

11.44±0.34

29.41±0.98

68.71±0.74

16.14±0.79

6.78±0.78

71.34±1.58

Example 3

10.54±0.21

32.14±1.24

70.02±1.31

15.98±1.06

7.34±0.11

69.55±2.12

As can be seen from table 3, in the yeast extract method of the present invention, through reasonable use of cavitation effect generated by pulsed laser and ultrasonic wave, mechanical force applied by ice crystal generated by temperature shock, ozone, various enzymes, and enzymolysis conditions, cell wall/cell membrane structure of yeast cells can be sufficiently destroyed, wall breaking rate can be substantially increased, yeast cells can be sufficiently autolyzed, and content substances, such as protein content, amino acid content, solid content, reducing sugar content, and nucleotide content, can be sufficiently released, specifically, compared with comparative example 2, wall breaking rate in the present invention is increased by about 67%, protein content, amino acid content, solid content, reducing sugar content, and nucleotide content are increased by about 30%, 50%, 62%, 50%, and 22%, and further, respective effects thereof are sufficiently volatilized.

< detection result of Gardenia fruit oil >

The gardenia fruit oil extracted by the method described in example 1 of the patent application with application number 201110321487.X ("method for extracting gardenia oil by aqueous enzymatic method") was used as a comparative example 4, and the content of several main blood sugar and blood lipid lowering active ingredients, i.e., crocin, chlorogenic acid, flavone and gardenoside, was obtained by detecting the gardenia fruit oil obtained by the method for extracting gardenia fruit oil in examples 1 to 3 of the present invention, and the results are shown in table 4.

TABLE 4 Gardenia oil crocin, chlorogenic acid, flavone and geniposide content

	Crocin (mg/g)	Flavone (mg/g)	Chlorogenic acid (mg/g)	Geniposide (mg/g)
					Comparative example 2	1.24±0.31	3.51±0.25	3.14±0.25	3.42±0.37
Example 1	2.24±0.41	6.58±0.44	4.15±0.58	3.92±0.21
					Example 2	2.39±0.29	7.25±0.38	4.21±0.13	3.82±0.35
Example 3	2.18±0.11	6.95±0.57	4.23±0.34	4.12±0.50

Similarly, the cell walls of the gardenia fruits 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 meanwhile, the acid solution, the glycerol, the sodium chloride and the lysozyme can change the permeability of the cell walls or the cell membrane by changing the cell walls or the cell membrane structure, so that the cell walls and/or the cell wall permeability are regulated by adopting a permeability regulating solution for regulating the permeability of the cell membranes and/or the cell wall, the cell walls and/or the cell wall structure can be destroyed, effective components (such as crocin, flavonoids and the like) in the content of the gardenia fruits are fully released, the blood sugar reducing effect of the gardenia fruits is further exerted, and the small molecular polypeptide compound nutrient is suitable for people with diabetes.

< example 4>

This example also provides a method for preparing the small molecule polypeptide compound nutrient for soothing nerves, aiding sleep and improving immunity of any one of examples 1 to 3, comprising:

s100, weighing the raw material components according to the component dosage in one of the embodiments 1-3, and mixing for 4-6 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 0.6Mpa), the granulation temperature is 75-85 ℃ (preferably 80 ℃), and the diameter of a die hole of the granulator is 1.5-3mm (preferably 2.0 mm);

s300, sieving the crude product by using an 8-mesh sieve to finally obtain the micromolecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity.

< evaluation test of Immunity-improving Effect >

Female mice of 1 month old were selected for the experiment and divided into 4 groups by weight. The human body recommended dose of the small molecular polypeptide compound nutrient (hereinafter referred to as polypeptide compound nutrient) for soothing the nerves, helping sleep and improving immunity is a shaped product with 0.4 kg/d.kg of body weight, and the equivalent dose of a mouse is 10 times of the human body recommended dose. The human body recommended dose is 5 times, 10 times and 30 times respectively as low, medium and high dose groups. The gavage method is adopted, the gavage is performed once a day, and the control group is filled with distilled water. After each group of mice was continuously administered with the polypeptide complex nutrient for 30 days, the proliferation ability of lymphocytes, the difference in toe swelling, the phagocytic rate of phagocytic cells, and the phagocytic index were measured, and the results are shown in table 5.

TABLE 5 Effect of polypeptide micronutrient complexes on mouse lymphocyte proliferation potency, toe swelling differential, phagocytic rate and phagocytic index

As shown in table 5, after the mice are orally administered with different dosages of the polypeptide complex nutrient for 30 days, compared with the blank control group, the low dose group, the medium dose group and the high dose group have obvious differences in lymphocyte proliferation capacity, toe swelling difference, phagocytic rate and phagocytic index of phagocytic cells, especially the high dose group, the differences are obvious (p is less than 0.05), which indicates that the polypeptide complex nutrient of the present invention can enhance the body immunity by improving the lymph transformation capacity and phagocytic function of phagocytic cells of the mice.

< evaluation test of sleep improvement efficacy >

Male mice of 1 month of age are selected, the weight is 18-22g, the mice are randomly divided into 4 groups according to the weight, and the weights between the groups have no significant difference through t test (p is more than 0.05). According to the polypeptide compound nutrient, the human recommended dose is 0.4 kg/d.kg, and the equivalent dose of the mouse is 10 times of the human recommended dose. 2 times, 5 times and 10 times of the recommended dose of the human body are respectively used as low, medium and high dose. The following tests were carried out by gavage and distilled water injection in the control group, respectively:

(1) test for prolonging sleep time of mice induced by pentobarbital sodium: after the animals continuously give the polypeptide compound nutrient for 26 days, 30mg/kg of sodium pentobarbital is injected into the abdominal cavity of each group of animals, and the injection amount is 0.1ml/10 g. And (3) observing whether the polypeptide compound nutrient can prolong the sleep time induced by the pentobarbital sodium by taking the disappearance of the righting reflex of the mouse as a sleep index. The test was performed at night.

(2) Subthreshold dose sodium pentobarbital sleep incidence test: after the animals continuously give the polypeptide compound nutrient for 26 days, 26mg/kg of sodium pentobarbital is injected into the abdominal cavity of each group of animals, and the injection amount is 0.1ml/10 g. And (3) observing whether the polypeptide compound nutrient can prolong the sleep time induced by the pentobarbital sodium by taking the disappearance of the righting reflex of the mouse as a sleep index. The test was performed at night.

(3) Barbiturate sodium sleep latency test: after the animals continuously give the polypeptide compound nutrient for 26 days, 240mg/kg of barbital sodium is injected into the abdominal cavity of each animal group, and the injection amount is 0.1ml/10 g. And (3) observing whether the polypeptide compound nutrient can prolong the sleep time induced by the pentobarbital sodium by taking the disappearance of the righting reflex of the mouse as a sleep index.

After the test, the indexes were measured, and specific results are shown in table 6.

TABLE 6 Effect of polypeptide Complex Nutrients on sleep in Normal mice

As can be seen from Table 6, after the mice are orally administered with different doses of the polypeptide compound nutrient for 26 days, compared with the control group, the sleep time induced by the low, medium and high dose of the pentobarbital sodium is remarkably improved (p is less than 0.01), which indicates that the polypeptide compound nutrient and the threshold dose of the pentobarbital sodium have synergistic sleep effect; the incidence rate of sleep induced by the low, medium and high dose of the mice in the sub-threshold dose of the pentobarbital sodium is remarkably improved, which shows that the incidence rate of sleep induced by the sub-threshold dose of the pentobarbital sodium in the polypeptide compound nutrient can be increased; meanwhile, the sleep latency of the mice induced by the barbital sodium in the low, medium and high dose groups is obviously shortened, which shows that the polypeptide compound nutrient can prolong the sleep latency of the barbital sodium induced by the barbital sodium.

< test for evaluating anti-aging Effect >

Healthy weaning male and female rats with the weight of 80 +/-20 g are selected, the rats are randomly divided into 4 groups according to the weight, 6 rats in each group of male and female rats are fed with the polypeptide compound nutrients in the embodiments 1-3 of the invention according to 0.5g/kg respectively in the morning and evening after the blank control group feeds the normal diet. After 28 days of feeding, abdominal blood was collected (fasting without water prohibition the day before blood sampling), blood was centrifuged, and serum was taken for measurement of Malondialdehyde (MDA) and vitamin E, and the results are shown in table 7.

Determination of MDA and vitamin E: the MDA was measured for 535nm wavelength by spectrophotometry. Vitamin E was measured by fluorescence spectrophotometry at an excitation wavelength of 295nm and an emission wavelength of 325nm, and the whole operation was carried out in a dark room.

TABLE 7 serum MDA and vitamin E contents of rats raised for 4 weeks

	Number of rats (only)	MDA content (mol/ml)	Vitamin E content (mg/dl)
				Blank control group	12	5.13±1.05	1.28±0.16
Example 1	12	2.63±1.28	1.74±0.32
				Example 2	12	2.47±1.14	1.82±0.48
Example 3	12	2.33±0.96	1.93±0.12

Vitamin E is localized in cells, especially on cell membranes, and can directly act with free radicals generated in cells, thereby blocking the lipid peroxidation process initiated by the free radicals. MDA is one of the lipid peroxidation products, and its level can reflect the lipid peroxidation condition. As can be seen from table 7, after the polypeptide complex nutrient of the present invention is administered, the MDA content of the mice is significantly reduced and the vitamin E content is significantly increased compared to the blank control group, thereby indicating that the polypeptide complex nutrient of the present invention contributes to the peroxidation of less cell membranes, and further has the effect of delaying aging.

It should be noted that the technical features of the above embodiments 1 to 3 can be arbitrarily combined, and the technical solutions obtained by combining the technical features belong to the scope of the present invention.

In conclusion, the invention greatly improves the yield and purity of the pumpkin seed extract (containing pumpkin seed protein and the like) and the gardenia oil extract by the extraction process of staged heating, repeated enzymolysis combined with ultrasonic treatment and multiple filtration, simultaneously promotes yeast autolysis by adopting a mode of pulse laser, ultrasonic treatment and multiple enzymolysis, fully releases yeast cell contents, and further, after multiple active ingredients are reasonably compounded and used, the formed small molecular polypeptide compound nutrient can remarkably enhance the body immunity of metabolic disease people such as diabetes and the like, people with various symptoms such as palpitation, insomnia, uneasiness and the like and middle-aged people, improves the sleep quality of the people, calms the heart and tranquilizes the mind and simultaneously delays aging.

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 (7)

1. The micromolecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity is characterized by comprising the following components in parts by weight: 15-20 parts of oat flour, 10-15 parts of pumpkin seed extract, 3-8 parts of cored lotus seed, 5-8 parts of poria cocos, 3-5 parts of selenium-rich cordyceps militaris powder, 2-3 parts of yeast extract, 8-10 parts of tomato powder, 3-5 parts of honey, 10-15 parts of denucleated dried longan, 5-8 parts of gardenia fruit oil, 5-8 parts of lily, 2-3 parts of radix ophiopogonis, 8-10 parts of walnut kernel, 3-5 parts of black bean powder and 5-8 parts of mannitol.

2. The small molecule polypeptide complex nutrient of claim 1, wherein the preparation method of the pumpkin seed extract comprises the following steps:

s11, taking fresh pumpkin seeds, soaking at 25 ℃ for 10-12h, taking out, washing with deionized water for 2-3 times, drying, grinding, and sieving with a 100-mesh sieve to obtain pumpkin seed powder;

s12, adding deionized water with the weight 10-12 times that of the pumpkin seed powder into the pumpkin seed powder to obtain an enzymolysis raw material, and carrying out enzymolysis on the enzymolysis raw material to obtain an enzymolysis system; wherein, the enzymolysis process comprises the following steps:

carrying out first enzymolysis: adjusting the pH value of the enzymolysis raw material to 8.0-9.0, adding trypsin which accounts for 3-4% of the weight of the enzymolysis raw material, fully stirring, heating to 40-45 ℃ while stirring, and preserving heat for 35-40min 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 3.5-4.5, adding pectinase according to 3.5-4.5% of the weight of the first enzymolysis system, fully stirring, heating to 45-50 ℃ while stirring, and preserving heat 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, adding cellulase according to 2-4% of the weight of the second enzymolysis system, fully stirring, heating to 50-65 ℃ while stirring, and preserving heat for 25-35min to obtain a third enzymolysis system;

s13, 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 a pumpkin seed enzymolysis system;

s14, adding phospholipase accounting for 0.01% of the weight of the pumpkin seed enzymolysis system and pancreatin accounting for 0.02% of the weight of the pumpkin seed enzymolysis system, and recording the total volume value of the pumpkin seed enzymolysis system; then carrying out temperature treatment on the pumpkin seed enzymolysis system to obtain a pumpkin seed extraction system;

the temperature treatment process comprises the following steps:

heating to 35-45 ℃, preserving heat for 60-90min, cooling to 30-40 ℃, preserving heat for 25-35min, and recording a first volume value of the reaction system at the moment; adding a first mixed solution containing deionized water, phospholipase and pancreatin according to (total volume-first volume) 65%, wherein the first mixed solution contains 97.5% of deionized water, 1% of phospholipase and 1.5% of pancreatin according to weight ratio; after the first mixed solution is added, heating to 45-50 ℃, preserving heat for 75-85min, then cooling to 40-45 ℃, preserving heat for 30-35min, and recording a second volume value of the reaction system at the moment; adding a second mixed solution containing deionized water, phospholipase and pancreatin according to (total volume-second volume) 45%, wherein the second mixed solution contains 98% of deionized water, 1% of phospholipase and 1% of pancreatin according to weight ratio; adding the second mixed solution, heating to 55-65 deg.C, maintaining the temperature for 80-90min, cooling to 50-60 deg.C, and maintaining the temperature for 40-45 min;

s15, adding activated carbon in the pumpkin seed extraction system according to 4% of the weight of the pumpkin seed extraction system, uniformly stirring, keeping the temperature at 65 ℃ for 65-85min, centrifuging, and removing sediments to obtain a pumpkin seed crude extract; filtering the crude extract of pumpkin seeds by diatomite to obtain a first-grade pumpkin seed clear liquid, wherein the filtering pressure is 0.25-0.35 MPa; adding 3% of active carbon into the first-stage pumpkin seed clear liquid according to the weight, standing for 45-50min, centrifuging, and removing sediments to obtain a second-stage pumpkin seed clear liquid;

s16, filtering the second-level pumpkin seed clear liquid by a microfiltration ceramic membrane with the filtering aperture of 0.2-1.0 mu m, wherein the filtering temperature is 50-60 ℃ to obtain microfiltration membrane permeate;

filtering the microfiltration membrane permeate through a 220kDa roll-type ultrafiltration membrane with the molecular weight cutoff of 120-;

and concentrating the ultrafiltration membrane retentate through a roll-type high-pressure reverse osmosis membrane with the molecular weight cutoff of 200-.

3. The small molecule polypeptide composite nutrient of claim 2, wherein in step S14, the temperature treatment and the ultrasonic treatment are performed on the mixed solution, the ultrasonic power is 200-400W, and the ultrasonic treatment time is 10-15 min.

4. The small molecule polypeptide compound nutrient as claimed in claim 1, wherein the preparation method of the yeast extract comprises:

s21, putting active dry yeast into a fermentation tank, wherein the diameter of the fermentation tank is 30-40cm, adding water which is 3-5 times of the weight of the active dry yeast into the fermentation tank, and uniformly stirring to obtain a yeast stock solution;

s22, irradiating the yeast stock solution by adopting a plurality of pulse laser light sources for 20-30min, stirring the yeast stock solution under the conditions of 200-300 r/min while irradiating, and carrying out ultrasonic treatment on the yeast stock solution at the same time, wherein the ultrasonic power is 100KW and the ultrasonic treatment temperature is 30-35 ℃; the diameter of a light spot formed on the liquid surface of the yeast stock solution by each pulse laser is 0.5-1cm, and the laser energy of each pulse laser acting on the yeast stock solution is 50 mJ;

s23, carrying out enzymolysis treatment on the yeast stock solution treated in the step S22; and the enzymolysis treatment process comprises the following steps:

(1) and 1, enzymolysis: raising the temperature of the yeast stock solution treated in the step S22 to 20-25 ℃, then lowering the temperature to-45 ℃ at the speed of 15 ℃/min, maintaining the temperature for 1min after lowering the temperature to-45 ℃, and then raising the temperature of the yeast stock solution to 20-25 ℃ at the speed of 10 ℃/min; heating to 20-25 deg.C, adding 1 wt% of alkaline protease and 1.5 wt% of papain into the yeast stock solution, adjusting pH to 7.0-8.0 and temperature to 50-60 deg.C, and performing hydrolysis reaction for 10-12 hr;

adjusting the temperature of yeast stock solution treated by alkaline protease and papain to 40-55 ℃, adding snailase accounting for 1 percent of the weight of the yeast stock solution treated by the alkaline protease and the papain and pancreatin accounting for 2.0 percent of the weight of the yeast stock solution treated by the alkaline protease and the papain, adjusting the pH to 6.5-7.5, filling ozone, stirring the yeast stock solution under the condition of 200 plus 300 revolutions per minute, and carrying out hydrolysis reaction for 3-4h to obtain first enzymatic hydrolysate;

(2) and (3) enzymolysis: adjusting the temperature of the first enzymolysis liquid to 20-25 ℃, reducing the temperature to-45 ℃ at the speed of 10 ℃/min, maintaining the temperature for 0.5min after reducing the temperature to-45 ℃, and increasing the temperature of the first enzymolysis liquid to 20-25 ℃ at the speed of 10 ℃/min; after the temperature is raised to 20-25 ℃, adding 0.5 percent of acid protease and 1.5 percent of nuclease by weight into the first enzymolysis liquid, adjusting the pH to 4.5-6.0 and the temperature to 45-50 ℃, and reacting for 8-10 h;

adjusting the temperature of the first enzymolysis liquid treated by acid protease and nuclease to 40-55 ℃, adding helicase accounting for 0.5 percent of the weight of the yeast stock solution treated by the acid protease and the papain and pancreatin accounting for 1.0 percent of the weight of the yeast stock solution treated by the acid protease and the papain, adjusting the pH to 6.0-7.5, filling ozone, and stirring the first enzymolysis liquid under the condition of 200-;

(3) and (3) enzymolysis: adjusting the temperature of the second enzymolysis liquid to 20-25 ℃, reducing the temperature to-45 ℃ at the speed of 5 ℃/min, maintaining the temperature for 0.5min after reducing the temperature to-45 ℃, and increasing the temperature of the second enzymolysis liquid to 20-25 ℃ at the speed of 12 ℃/min; after the temperature is raised to 20-25 ℃, adding glutaminase and deaminase which are 0.5 percent and 1.0 percent of the weight of the second enzymolysis liquid into the second enzymolysis liquid, adjusting the pH to 5.5-6.0 and the temperature to 45-50 ℃, and reacting for 3-4 hours;

regulating the temperature of the second enzymatic hydrolysate treated by the glutaminase and the deaminase to 40-55 ℃, adding the snailase accounting for 1 percent of the weight of the second enzymatic hydrolysate treated by the glutaminase and the deaminase and the pancreatin accounting for 1 percent of the weight of the second enzymatic hydrolysate treated by the glutaminase and the deaminase, regulating the pH to 6.5-7.5, filling ozone, and stirring the second enzymatic hydrolysate under the condition of 200-300 revolutions per minute for 2-3 hours to obtain a third enzymatic hydrolysate;

s24, after the enzymolysis is finished, heating the obtained third enzymolysis liquid to 90 ℃, and maintaining for 10min to finish the enzyme deactivation process to obtain a yeast extract stock solution;

s25, adding activated carbon 3% of the yeast extract stock solution by weight, stirring uniformly, keeping the temperature at 55 ℃ for 60-90min, centrifuging, removing sediments to obtain yeast extract clear solution, and filtering under the pressure of 0.25-0.35 MPa;

s26, filtering the clear yeast extract liquid by a microfiltration ceramic membrane, and controlling the operation temperature to be 55-65 ℃ to obtain microfiltration membrane permeate;

filtering the microfiltration membrane permeate through a roll-type ultrafiltration membrane, and controlling the operation temperature to be 55-65 ℃ to obtain ultrafiltration membrane permeate;

concentrating the ultrafiltration membrane retentate by a roll-type high-pressure reverse osmosis membrane to remove water and partial residual inorganic salt and small molecular impurities, and controlling the operation temperature to be below 40 ℃ to obtain a yeast extract concentrated solution;

s27, drying the yeast extract concentrated solution by a vacuum freeze drying method to obtain the yeast extract.

5. The small molecule polypeptide complex nutrient of claim 4, wherein the active dry yeast cell has a protein content of 40-45%, a carbohydrate content of 8-10%, and a moisture content of 4-5%.

6. The small molecule polypeptide complex nutrient of claim 1, wherein the extraction method of gardenia fruit oil comprises:

s31, putting fresh gardenia fruits into water, soaking at 25 ℃ for 24-36h, taking out, washing with water for 2-3 times, drying, grinding, and sieving with a 100-mesh sieve to obtain gardenia fruit powder;

s32, taking gardenia fruit powder, adding deionized water with the weight 5-10 times of that of the gardenia fruit powder to obtain an enzymolysis raw material, and carrying out enzymolysis on the enzymolysis raw material; wherein, the enzymolysis process comprises the following steps:

carrying out first enzymolysis: adding trypsin in an amount of 5% of the weight of the gardenia fruit powder and adding a permeability regulating solution for regulating the permeability of cell membranes and/or cell walls in an amount of 45-55% of the weight of the gardenia fruit powder into the enzymolysis raw materials, regulating the pH value to 6.5-7.5, fully stirring, heating to 42-45 ℃ while stirring, and preserving heat for 30-45min to obtain a first enzymolysis system; the permeability regulating liquid consists of acid solution, glycerol, sodium chloride and lysozyme, and the acid solution comprises the following components in percentage by weight: glycerol: sodium chloride: lysozyme is 1: (0.7-1.0): (0.02-0.05): (0.03-0.06);

and (3) carrying out second enzymolysis: after the temperature of the first enzymolysis system is reduced to 20-25 ℃, adjusting the pH value to 3.5-4.5, adding pectinase according to 4% of the weight of the first system, fully stirring, heating to 50-60 ℃ while stirring, and preserving heat 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, adding cellulase according to 3.5% of the weight of the second enzymolysis system, fully stirring, heating to 50-65 ℃ while stirring, and preserving heat for 25-35min to obtain a third enzymolysis system;

s33, after the enzymolysis process in the step S22 is finished, heating the obtained third enzymolysis system to 85 ℃, and maintaining for 10min to finish the enzyme deactivation process to obtain a gardenia fruit enzymolysis system;

s34, adding activated carbon in an enzymolysis system of the gardenia fruit according to 3% of the weight of the gardenia fruit, uniformly stirring, keeping the temperature at 65 ℃ for 65-85min, centrifuging, and removing sediments to obtain a crude extract of gardenia fruit oil; filtering the crude gardenia fruit oil extract by diatomite to obtain a gardenia fruit oil extract, wherein the filtering pressure is 0.3-0.4 MPa; adding 3% of active carbon into the gardenia fruit oil extracting solution according to the weight, standing for 45-50min, centrifuging, and removing sediments; standing for 2-3h, and collecting the upper oil layer to obtain the fructus Gardeniae fruit oil.

7. A preparation method of a micromolecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity 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 75-85 ℃, and the diameter of a die hole of the granulator is 1.5-3 mm;

s300, sieving the crude product by using an 8-mesh sieve to finally obtain the micromolecule polypeptide compound nutrient for soothing nerves, helping sleep and improving immunity and the preparation method thereof.