CN110583778A - Preparation method of normal-temperature health-care hemp milk - Google Patents
Preparation method of normal-temperature health-care hemp milk Download PDFInfo
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- CN110583778A CN110583778A CN201910960576.5A CN201910960576A CN110583778A CN 110583778 A CN110583778 A CN 110583778A CN 201910960576 A CN201910960576 A CN 201910960576A CN 110583778 A CN110583778 A CN 110583778A
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- PXQLVRUNWNTZOS-UHFFFAOYSA-N sulfanyl Chemical compound [SH] PXQLVRUNWNTZOS-UHFFFAOYSA-N 0.000 description 1
- 235000019605 sweet taste sensations Nutrition 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 230000035922 thirst Effects 0.000 description 1
- 229930003799 tocopherol Natural products 0.000 description 1
- 235000010384 tocopherol Nutrition 0.000 description 1
- 239000011732 tocopherol Substances 0.000 description 1
- 229960001295 tocopherol Drugs 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000002936 tranquilizing effect Effects 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
- 238000009941 weaving Methods 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 description 1
- 239000002478 γ-tocopherol Substances 0.000 description 1
- QUEDXNHFTDJVIY-DQCZWYHMSA-N γ-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1 QUEDXNHFTDJVIY-DQCZWYHMSA-N 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
- A23C9/1526—Amino acids; Peptides; Protein hydrolysates; Nucleic acids; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/006—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from vegetable materials
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/63—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from plants
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/22—Cysteine endopeptidases (3.4.22)
- C12Y304/22002—Papain (3.4.22.2)
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- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Molecular Biology (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
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Abstract
The invention belongs to the field of food, and particularly relates to a preparation method of hemp-containing milk. The invention comprises the following steps: (1) preparing hemp protein polypeptide powder; (2) preparing skim raw milk; (3) blending the hemp protein polypeptide powder; (4) emulsifying, homogenizing and sterilizing; (5) and filling to obtain the normal-temperature health-care hemp milk. The small molecular peptide of the hydrolyzed hemp protein in the milk can increase the protein content in the milk by 20 percent, and the hydrolyzed small molecular peptide is easier to be absorbed by human bodies and has no anaphylactic reaction. The small hemp protein peptide can increase the content of polyunsaturated fatty acids such as omega-3, omega-6 and the like and dietary fiber, can delay senility, prevent cardiovascular diseases, promote brain development, resist inflammation and cancer, keep the vitality of the heart and the thinking agility of the brain and keep the digestive tract smooth and healthy.
Description
Technical Field
The invention belongs to the field of food, and particularly relates to a preparation method of yoghurt containing cannabis.
Background
The cannabis sativa is also called hemp, hemp and the like, annual herbaceous plants are originally produced with tingold, India and Zhongyaya, and various places in China are also cultivated. The hemp stalk skin fiber is firm and long, and can be used for making paper, weaving hemp cloth, making ropes and the like; the oil content of the hemp seeds is 30 percent, and the hemp seeds can be eaten and also can be used as paint, coating and the like; fructus cannabis can be used as a medicine, has mild nature and sweet taste, and has the efficacy of moistening intestines and relaxing bowels. With the continuous development of biochemical technology, various bioactive components have been extracted and separated from cannabis sativa resources, and the substances have prevention effects on hypertension, diabetes, constipation and the like due to good antioxidation and health care effects. In recent years, various cannabis products such as cannabis oil, cannabis protein powder, cannabis beverage, cannabis tortoise jelly and the like are developed, and the green health food is gradually favored by people. The hemp is planted on a large scale in the global scope, and compared with China, the hemp resources are more abundant, and a large amount of raw materials can be provided for scientific research, development and application of hemp products.
The hemp seeds are rich in oil and fat, which are slightly different with the difference of planting climatic conditions, years and regions of the hemp plants. The hemp seed oil is a yellow green semitransparent oily liquid and has a special faint scent of hemp seeds. In the fatty acid composition of cannabis oil, the saturated fatty acid content is about 10%, the unsaturated fatty acid content is as high as approximately 90%, with the polyunsaturated fatty acid content ranging from 70-80%. The content of two polyunsaturated essential fatty acids, linoleic acid (LA, 18:2n-6) and alpha-linolenic acid (ALA, 18:3n-3) is typically 50-70% and 15-25%. The two polyunsaturated fatty acids have anti-inflammatory, antithrombotic, antiarrhythmic, and hypolipidemic effects. The ratio of n-6 to n-3 fatty acids in hemp oil is about 3:1, and this balance is unique among common vegetable oils and is most beneficial to human health. The cannabis oil is rich in tocopherol and chlorophyll, wherein the content of gamma-tocopherol is highest, and the cannabis oil has a strong antioxidation effect in grease. Hemp oil is widely used in baking and cooking and is an important raw material for cheese, salad dressing, margarine, cake, dessert, and the like.
The hemp seeds are rich in nutrient components, contain 20-25% of protein, the protein mass of the soluble hemp seeds is about 30%, and the protein content of the shelled seeds is more than 40%. The hemp seed protein contains 21 amino acids, including 8 amino acids essential for human body. Nearly 2/3 of the hemp seed proteins are edestin which can promote the digestion and absorption of human body; further, 2/3 is albumin. The hemp seed protein has arginine of about 18.8mg/g and glycine of about 9.7mg/g, and has high content. Arginine can maintain the normal growth and development of infants. Most importantly, the hemp seed protein is easy to be absorbed by human body, and does not cause the advantages of gastrectasia, regurgitation, anaphylactic reaction and the like, so the hemp seed protein is very helpful for the nutrition supplement of infants, old people, sick and cured groups and weak and lean people.
A large number of research results show that the protein is not easy to digest and absorb after being taken into a human body due to large molecular weight and complex structure, thereby influencing the effective exertion of the physiological function and the nutritional value of the protein. The hemp protein is hydrolyzed into protein peptide, and the protein peptide has physicochemical properties such as good solubility, computational or thermal stability and low viscosity which are not possessed by the hemp seed protein; and has various physiological functions of easy absorption, blood pressure reduction, blood sugar reduction, antioxidation and the like, and can be used for nutritious food, functional health food, athlete food and the like.
The milk is rich in protein, fat, carbohydrate, vitamin A, lactose, lecithin, pigment, etc. The milk has effects of invigorating lung and stomach, promoting fluid production and moisturizing intestine, tranquilizing and allaying excitement, and is beneficial to patients with diabetes, thirst, constipation, asthenia, deficiency of qi and blood, and incoordination between spleen and stomach; milk drinking can promote sleep stability; iodine, zinc and lecithin in the milk can greatly improve the working efficiency of the brain; magnesium in milk can promote fatigue resistance of heart and nervous system; the milk can moisten skin, and can make the skin white and smooth and increase elasticity after being drunk frequently; based on the function of the enzyme, the milk also has the effects of diminishing inflammation, reducing swelling and relieving skin tension; children often drink fresh milk to help the development of the body, because calcium can promote the development of bones; the old people can supplement calcium demand by drinking milk, reduce skeleton atrophy, reduce the occurrence probability of osteoporosis and increase body flexibility. Through the search of the inventor, 4 prior art documents are detected in total, which are respectively as follows: an treatise on the safety and application research of the hemp seed oil in yoghourt in 2015, an article published in the Chinese dairy industry 2009 by Lishoubbo and the like, "development of hemp milk", a hemp seed extract, hemp seed yoghourt and a preparation method thereof in application No. CN201710303268.6, a jet cooking combined ultrafiltration prepared hemp protein and functional characteristic characterization thereof published in Junfeng in 2017 by Wufeng in China fat and 2017, a response surface method optimized hemp seed protein extracted in the first stage published in 2017 by Liushuxian Xixia in China hemp science 2017, and the like. In addition, a great number of patents and documents about the extraction of the hemp nutrient substances exist, and are not repeated any more because the technical relevance is not high.
Disclosure of Invention
The invention aims to provide a preparation method of normal-temperature health-care hemp milk.
The purpose of the invention is realized as follows:
a preparation method of normal-temperature health-care hemp milk comprises the following steps:
(1) preparing hemp protein polypeptide powder;
the preparation of the hemp protein polypeptide powder comprises the following steps:
(1.1) weighing 5-10g of fructus cannabis meal, dissolving in 50-100ml of distilled water, adding alkaline protease, adjusting the pH value to 8-11 at 55-65 ℃, adding 3-5% of enzyme, and carrying out first enzymolysis for 2-4h at a material-liquid ratio of 1: 5;
(1.2) after the time of the first enzymolysis is reached, inactivating enzymes at high temperature, cooling, adjusting the pH value to 5, adding papain, carrying out second enzymolysis at the temperature of 70-80 ℃ with the enzyme addition amount of 0.4-0.6%, after 1-2h, inactivating enzymes at high temperature, cooling, centrifuging, extracting supernatant, carrying out microwave detoxification, and carrying out spray drying to prepare the hemp protein peptide powder;
(2) preparing skim raw milk;
(3) blending the hemp protein polypeptide powder;
(4) emulsifying, homogenizing and sterilizing;
(5) and filling to obtain the normal-temperature health-care hemp milk.
The preparation of the skim raw milk comprises the following steps: taking 1000g of raw milk, preheating the raw milk to 35-40 ℃ after flash evaporation treatment, and degreasing in a disc separator to control the fat content of the degreased raw milk to be below 0.5%. The hemp protein polypeptide blended powder comprises: pouring the skim raw milk obtained in the step (2) into a blending tank, heating to 55-60 ℃, adding the hemp protein polypeptide powder obtained in the step (1) and 4g of honey, and stirring and dissolving for 10-15min to obtain the base material. The emulsification, homogenization and sterilization comprises the following steps: heating the obtained base material to 65-70 deg.C, homogenizing, and pasteurizing at 95 + -1 deg.C for 10-15 s. The filling comprises the following steps: and cooling the sterilized product to 2-6 ℃ within 3-15min, and filling to obtain the normal-temperature health-care cannabis sativa milk.
The alkaline protease is solid alkaline protease, and the preparation steps of the alkaline protease are as follows: firstly, preparing 15ml of agarose-phosphoric acid solution with the mass fraction of 3% by using phosphoric acid solution with the pH value of 7-8; boiling for more than 5min under microwave condition, cooling to room temperature, adding 60g/L alkaline protease solution 2ml, cooling to 13-16 deg.C to obtain gel, cutting into 1cm x 1cm pieces, and storing at 3-7 deg.C as solid alkaline protease. Screening out the agarose block after the first enzymolysis, wherein the papain is solid papain during the second enzymolysis, and the preparation steps of the papain are as follows: firstly, preparing 15ml of agarose-phosphoric acid solution with the mass fraction of 3% by using phosphoric acid solution with the pH value of 7.5; boiling for more than 5min under microwave condition, cooling to room temperature, adding 65g/L alkaline protease solution 2ml, cooling to 10-14 deg.C to form gel, cutting into 1cm x 1cm pieces, and storing at 2-5 deg.C as solid alkaline protease. After cutting into 1cm x 1cm small pieces, adding 0.15% by weight of 1,2,3,4, 5-pentanol solution as protectant into alkaline protease or papain before storing at low temperature, and soaking for more than 5 min.
The flash evaporation uses a flash evaporation cavity, the microwave power in the flash evaporation cavity is 1k W-1.25kW, the raw milk flow is 15L/h-30L/h, the inlet temperature of the flash evaporation cavity is 50-55 ℃, and the vacuum degree of the system is 0.095 MPa. The microwave detoxification is carried out under the conditions that the microwave intensity is 4.375w/g, the time is 6min, and the water vapor flow at 50 ℃ is 2 m/s.
The invention has the beneficial effects that:
1. the small molecular peptide after the hemp protein hydrolysis can increase the protein content in the milk by 20 percent, and the small molecular peptide after the hydrolysis is easier to be absorbed by human bodies and has no anaphylactic reaction.
2. The small hemp protein peptide can increase the content of polyunsaturated fatty acids such as omega-3, omega-6 and the like and dietary fiber, can delay senility, prevent cardiovascular diseases, promote brain development, resist inflammation and cancer, keep the vitality of the heart and the thinking agility of the brain and keep the digestive tract smooth and healthy.
3. The hemp protein can increase the content of linoleic acid in the milk, thereby reducing cholesterol and triglyceride in blood, reducing blood viscosity and improving blood microcirculation.
4. The physiological activity, transportation physiological activity, carrier physiological activity, dynamic physiological activity, transmitter physiological activity and the like of the decomposed small molecular peptide can promote the absorption of all functional elements in the milk.
Drawings
FIG. 1 is a flow chart of the method of the present invention;
FIG. 2 is a comparison of activity of solid alkaline protease with 1,2,3,4, 5-pentanol protectant added at different temperatures;
FIG. 3 is a comparison of activity of solid alkaline protease at different temperatures;
FIG. 4 is a comparison of activity of liquid alkaline protease at different temperatures;
FIG. 5 is a comparison of activity of solid alkaline protease with 1,2,3,4, 5-pentanol protectant added at different temperatures;
FIG. 6 is a graph showing the decolorization effect of the hemp clear solution.
Detailed Description
The invention is further described below with reference to the accompanying drawings. The invention discloses a preparation method of hemp protein peptide health milk, which comprises the following steps:
(1) preparing the hemp protein peptide; (2) preparing skim raw milk; (3) blending the hemp protein peptide milk; (4) emulsifying, homogenizing and sterilizing; (5) and (4) quickly cooling the sterilized product to 2-6 ℃ for filling.
These 5 steps are described in detail below:
(1) preparing hemp protein polypeptide powder;
the preparation of the hemp protein polypeptide powder comprises the following steps:
(1.1) weighing 5-10g of fructus cannabis meal, dissolving in 50-100ml of distilled water, adding alkaline protease, adjusting the pH value to 8-11 at 55-65 ℃, adding 3-5% of enzyme, and carrying out first enzymolysis for 2-4h at a material-liquid ratio of 1: 5; the alkaline protease is solid alkaline protease, and the preparation steps of the alkaline protease are as follows: firstly, preparing 15ml of agarose-phosphoric acid solution with the mass fraction of 3% by using phosphoric acid solution with the pH value of 6.5; boiling for more than 5min under microwave condition, cooling to room temperature, adding 60g/L alkaline protease solution 2ml, cooling to 13-16 deg.C to obtain gel, cutting into 1cm x 1cm pieces, and storing at 3-7 deg.C as solid alkaline protease.
(1.2) after the time of the first enzymolysis is reached, inactivating enzymes at high temperature, cooling, adjusting the pH value to 5, adding papain, carrying out second enzymolysis at the temperature of 70-80 ℃ with the enzyme addition amount of 0.4-0.6%, after 1-2h, inactivating enzymes at high temperature, cooling, centrifuging, extracting supernatant, carrying out microwave detoxification, and carrying out spray drying to prepare the hemp protein peptide powder; screening out the agarose block after the first enzymolysis, wherein the papain is solid papain during the second enzymolysis, and the preparation steps of the papain are as follows: firstly, preparing 15ml of agarose-phosphoric acid solution with the mass fraction of 3% by using phosphoric acid solution with the pH value of 7.5; boiling for more than 5min under microwave condition, cooling to room temperature, adding 65g/L alkaline protease solution 2ml, cooling to 10-14 deg.C to obtain gel, cutting into 1cm x 1cm pieces, and storing at 2-5 deg.C to obtain solid papain.
After the solid material is cut into small blocks of 1cm x 1cm, 1,2,3,4, 5-pentadiol solution with the mass fraction of 0.15% is added into alkaline protease or papain to be used as a protective agent before low-temperature storage, and the mixture is soaked for more than 5 min. The microwave detoxification is carried out under the conditions that the microwave intensity is 4.375w/g, the time is 6min, and the water vapor flow at 50 ℃ is 2 m/s.
(2) Preparing skim raw milk; the preparation of the skim raw milk comprises the following steps: taking 1000g of raw milk, preheating the raw milk to 35-40 ℃ after flash evaporation treatment, and degreasing in a disc separator to control the fat content of the degreased raw milk to be below 0.5%. The flash evaporation uses a flash evaporation cavity, the microwave power in the flash evaporation cavity is 1k W-1.25kW, the raw milk flow is 15L/h-30L/h, the inlet temperature of the flash evaporation cavity is 50-55 ℃, and the vacuum degree of the system is 0.095 MPa.
(3) Blending the hemp protein polypeptide powder; the hemp protein polypeptide blended powder comprises: pouring the skim raw milk obtained in the step (2) into a blending tank, heating to 55-60 ℃, adding the hemp protein polypeptide powder obtained in the step (1) and 4g of honey, and stirring and dissolving for 10-15min to obtain the base material.
(4) Emulsifying, homogenizing and sterilizing; the emulsification, homogenization and sterilization comprises the following steps: heating the obtained base material to 65-70 deg.C, homogenizing, and pasteurizing at 95 + -1 deg.C for 10-15 s.
(5) And filling to obtain the normal-temperature health-care hemp milk. The filling comprises the following steps: and cooling the sterilized product to 2-6 ℃ within 3-15min, and filling to obtain the normal-temperature health-care cannabis sativa milk.
The hemp milk of the invention has the following effects: 1. the small molecular peptide after the hemp protein hydrolysis can increase the protein content in the milk by 20 percent, and the small molecular peptide after the hydrolysis is easier to be absorbed by human bodies and has no anaphylactic reaction. 2. The small hemp protein peptide can increase the content of polyunsaturated fatty acids such as omega-3, omega-6 and the like and dietary fiber, can delay senility, prevent cardiovascular diseases, promote brain development, resist inflammation and cancer, keep the vitality of the heart and the thinking agility of the brain and keep the digestive tract smooth and healthy. 3. The hemp protein can increase the content of linoleic acid in the milk, thereby reducing cholesterol and triglyceride in blood, reducing blood viscosity and improving blood microcirculation. 4. The physiological activity, transportation physiological activity, carrier physiological activity, dynamic physiological activity, transmitter physiological activity and the like of the decomposed small molecular peptide can promote the absorption of all functional elements in the milk.
The invention provides a secondary protease hydrolysis method, which hydrolyzes the hemp protein into small molecular peptides which are easier to be absorbed by human bodies and avoid anaphylactic reaction. Meanwhile, the alkaline protease and the papain provided by the invention carry out enzymolysis at the temperature of about 55-65 ℃, the traditional enzymolysis means mostly directly adopts a liquid enzyme preparation for enzymolysis, namely a protease solution is prepared and mixed into protein for liquid enzymolysis, the enzymolysis speed of the protein has obvious speed reduction fluctuation from 40 ℃ to 70 ℃, because the activity of the alkaline protease is obviously reduced after the temperature exceeds 60 ℃, the enzymolysis process is in a lower appropriate temperature such as 40-50 ℃ in order to ensure the enzyme activity, but once the temperature is too low, the performance of the hemp protein tends to be stable and even solidified, and the enzymolysis reaction speed is reduced along with the reduction of the temperature, so that the enzymolysis is not facilitated; if the temperature is too high, the protein has allergic reaction and also has solid property, thereby influencing the effective storage of the protein, reducing the content of the protein and reducing the health care function. Therefore, for liquid enzyme preparations, the reaction conditions are harsh, and the inactivation failure is easily caused by the influence of factors such as high temperature, acid and alkali, and the like, and the free enzyme is difficult to recycle in the reaction system, so the application in large-scale industrial production is limited. In addition, in the catalysis process, the protease possibly has a cross-linking effect with the enzyme under the action of carbodiimide, so that the enzyme activity of the protease is limited, and the catalytic hydrolysis of a substrate is influenced. Therefore, the invention provides that the protease is solidified, the molecule of the protease is locked in the solidified structure, so that the active center of the protease is protected by the heat-insulating layer, the stability to temperature change is improved, and the protease molecule can be positioned to avoid the crosslinking effect.
Therefore, the invention provides a solid stating step for controlling protease: firstly, preparing 15ml of agarose-phosphoric acid solution with the mass fraction of 3% by using phosphoric acid solution with the pH value of 6; boiling under microwave condition for more than 5min, cooling to room temperature, adding 60g/L alkaline protease solution 2ml, cooling to 13-16 deg.C to obtain gel, cutting into 1cm x 1cm pieces, and storing at 3-7 deg.C. The alkaline protease of the present invention is immobilized in agarose gel blocks.
After the first enzymolysis is finished, the agarose block needs to be screened out, and then the second enzymolysis is carried out. The papain in the secondary enzymolysis process is solidified by adopting the method, which comprises the following steps: firstly, preparing 15ml of agarose-phosphoric acid solution with the mass fraction of 3% by using phosphoric acid solution with the pH value of 7.5; boiling under microwave condition for more than 5min, cooling to room temperature, adding 65g/L alkaline protease solution 2ml, cooling to 10-14 deg.C to obtain gel, cutting into 1cm x 1cm pieces, and storing at 2-5 deg.C. The papain is solidified into the agarose gel block.
Papain is called papain for short, and is also called papain. Is a pure natural biological enzyme product which is refined by using milk in immature papaya fruits and adopting the modern bioengineering technology. It is a sulfhydryl (-SH) containing endopeptidase, has the activity of proteinase and esterase, has extensive specificity, and has strong hydrolytic power for animal and plant protein, polypeptide, ester and amide, at the same time, it also has synthesis function, and can synthesize protein hydrolysate into protein-like product.
Agarose agar, abbreviated as AG, added for the purposes of the present invention is an uncharged neutral constituent of agar and is also known as agarosine or Agarose. The chemical structure of the agar sugar is formed by long chains formed by alternately connecting 1, 3-linked beta-D-galactose and 1, 4-linked 3, 6-lacto-L-galactose. Can be used in the food industry for the production of: soft candy, shaped soft candy, aquatic product, canned meat, fruit juice beverage, fruit pulp beverage, rice wine beverage, dairy beverage, essence, and dairy cake. The addition of the agarose can not only solidify the protease, but also preserve the residues in the milk to improve the liquid viscosity and improve the taste.
The invention designs an experiment by combining liquid enzymolysis reaction, and liquid and solid alkaline proteases with the same mass (50mg) are respectively used for carrying out enzymolysis on the hemp protein feed liquid with the same mass (15mg) to reflect the effects of two methods, as shown in a table 1:
meanwhile, the activity measurement is also carried out on the protease with different performances, and because the data amount is excessive and the data trends of the alkaline protease and the papain are similar, only the full data of the alkaline protease and the typical data of the papain are given here, as shown in tables 2-5 and figures 2-5.
Analysis shows that the activity of the solid protease can be stabilized to be more than 70% in a certain time, and the activity is reduced very quickly without the enzymolysis process of the method, so the method has obvious effect. In addition, after the protease is solidified, the protease is easy to dehydrate, the self space structure of the protease collapses, and the enzyme activity is further lost, so that a 0.15 mass percent solution of 1,2,3,4, 5-pentanol is added before freezing and storing as a protective agent, and the influence of a dehydration process on the activity of the alkaline protease is reduced. And then the agarose gel block is put into 1,2,3,4, 5-pentadiol solution with the mass fraction of 0.15% to be soaked for more than 5min, and is taken out to be refrigerated into the agarose gel block, and the agarose gel block is analyzed. The taste of the invention can be effectively improved by 1,2,3,4, 5-pentanol and agarose, so that the invention can improve the performance and combine the taste, and the inventor does not find that other technical systems can solve the problem at present.
It should be noted that, the prior art proposed in the background art by using agarose as a solid protease carrier and using a 1,2,3,4, 5-pentadiol solution as a protective agent in the present invention is not described, but the technical personnel in the art do not suggest a scheme of secondary solid enzymolysis and using a protective agent to improve the enzymolysis efficiency.
In addition, the flash evaporation of the invention uses a flash evaporation cavity, the microwave power in the flash evaporation cavity is 1k W-1.25kW, the raw milk flow is 15L/h-30L/h, the inlet temperature of the flash evaporation cavity is 50-55 ℃, and the vacuum degree of the system is 0.095 MPa. The microwave detoxification is carried out under the conditions that the microwave intensity is 4.375w/g, the time is 6min, and the water vapor flow at 50 ℃ is 2 m/s. Through the experimental comparison of microwave working condition and conventional working condition, the microwave has the effect of promotion in the flash evaporation process, so that the evaporation effect of the single-stage system is greatly improved.
The linseed contains rich linolenic acid, and is a special oil material with extremely high nutritional value. However, the flax seeds contain cyanogenic glucoside which is harmful to human body, and almost all the cyanogenic glucoside is remained in the pressed cake after the flax seeds are cold-pressed to prepare grease. Cyanogenic glycosides are formed by the condensation of the hydroxyl groups of cyanohydrin derivatives and D-glucose, and include beta-gentiobiose acetone cyanohydrin, beta-gentiobiose methyl ethyl ketone cyanohydrin, linkurin and picroside. Cyanogenic glycoside has strong toxicity because it can generate hydrocyanic acid in the presence of hydrolase, and hydrocyanic acid can cause toxic hypoxia, heart rate disorder, muscle paralysis and other symptoms in cells. The principle of microwave detoxification is that the property of selective absorption of microwaves is utilized, according to the difference of loss factors of different substances, during microwave irradiation, the rapid rising of the temperature of water molecules in flaxseeds excites the activity of beta-glycosidase, so that cyanogenic glycoside is subjected to enzymolysis reaction to form cyanohydrin, and the cyanohydrin is decomposed to compounds such as hydrocyanic acid and the like. These compounds are released to the outside of the flaxseed along with the moisture emitted during the microwave process. The general microwave detoxification can lead the detoxification rate of cyanogenic glucoside to reach more than 75 percent. The microwave detoxification time is short and the efficiency is high. But the detoxification rate of the flaxseeds is still difficult to meet the edible requirement at present, and the cyanogenic glucoside generation after detoxification is still more than 5.7 mg/g. The microwave detoxification method disclosed by the invention has the advantages that the microwave detoxification is carried out under the conditions that the microwave intensity is 4.375w/g, the time is 6min, and the water vapor flow at 50 ℃ is 2m/s, so that the microwave detoxification process can be met, and meanwhile, the water vapor can ensure the content of water in the hemp seeds and the stability of other substances. Meanwhile, the steam can rapidly blow away the evaporated hydrocyanic acid to reduce the concentration of the surface of the hemp seeds, and accelerate the enzymolysis reaction of the cyanogenic glycosidase.
The above technical solution according to the present invention provides the following two embodiments.
Example 1
Preparing the cannabis polypeptide:
weighing 5g of fructus cannabis meal, dissolving in 50ml of distilled water, adding alkaline protease, adjusting the pH value to 11 at 55 ℃, adding 3% of enzyme, and carrying out first enzymolysis for 2 hours at a material-liquid ratio of 1: 10. And (3) reaching enzymolysis time, inactivating enzyme at high temperature, cooling and adjusting the pH value to 5. Adding papain, performing enzymolysis at 70 deg.C with 0.4% enzyme, inactivating enzyme at high temperature after 1 hr, cooling, centrifuging, extracting supernatant, and spray drying to obtain fructus Cannabis protein peptide powder.
Preparing skim raw milk: taking 10kg of raw milk, preheating the raw milk to 35-40 ℃ after flash evaporation treatment, and degreasing in a disc separator (the fat content of the raw milk after degreasing is controlled below 0.5%).
Blending the hemp milk: pouring the defatted raw milk into a blending tank, heating to 55-60 deg.C, adding hemp protein peptide powder, adding 2% Mel, stirring and dissolving for 10-15 min.
Emulsification and homogenization: heating the mixed base material to 65-70 ℃ for homogenizing treatment.
Pasteurization: pasteurizing the homogenized base material at 95 + -1 deg.C for 10-15 s.
Filling: and (4) quickly cooling the sterilized product to 2-6 ℃ for filling.
The product has the following sense: is light yellow, has pure and rich flavor of cow milk and honey, has palatable sweetness, delicate and smooth mouthfeel, and is health-care and nutritious.
Example 2
Preparing the cannabis polypeptide:
weighing 10g of fructus cannabis meal, dissolving in 100ml of distilled water, adding alkaline protease, adjusting the pH value to 11 at 55 ℃, adding 5% of enzyme, and carrying out first enzymolysis for 1.5h at a material-liquid ratio of 1: 10. And (3) reaching enzymolysis time, inactivating enzyme at high temperature, cooling and adjusting the pH value to 5. Adding papain, performing enzymolysis at 75 deg.C with enzyme amount of 0.6% for the second time, inactivating enzyme at high temperature after 1.5 hr, cooling, centrifuging, extracting supernatant, and spray drying to obtain fructus Cannabis protein peptide powder;
preparing skim raw milk: taking 50kg of raw milk, preheating the raw milk to 35-40 ℃ after flash evaporation treatment, and degreasing in a disc separator (the fat content of the raw milk after degreasing is controlled below 0.5%).
Blending the hemp milk: pouring the defatted raw milk into a blending tank, heating to 55-60 deg.C, adding hemp protein peptide powder, adding 2% Mel, stirring and dissolving for 10-15 min.
Emulsification and homogenization: heating the mixed base material to 65-70 ℃ for homogenizing treatment.
Pasteurization: pasteurizing the homogenized base material at 95 + -1 deg.C for 10-15 s.
Filling: and (4) quickly cooling the sterilized product to 2-6 ℃ for filling.
The product has the following sense: is light yellow, has pure and rich flavor of cow milk and honey, has palatable sweetness, delicate and smooth mouthfeel, and is health-care and nutritious.
Removing color before extracting supernatant: putting the container containing the supernatant into a reaction kettle, putting a rotor, and adding activated clay. The reaction kettle is sealed and vacuumized to 0.1 MPa. Placing on a magnetic stirrer for fully stirring, heating at a constant temperature of 50 ℃ for 2 hours, stopping heating and stirring, opening the reaction kettle after the temperature of the supernatant is reduced, centrifuging the oil-soil mixture for 20 minutes at 6000r/min, separating the decolored supernatant, cooling, filling nitrogen, placing in a refrigerator at the temperature of minus 20 ℃ for 20 minutes, and then carrying out reduced pressure concentration.
The decolorization rates of the supernatant liquid were measured at the mass fractions of 7%, 9%, 11%, 13%, and 15% of the added amount of the white soil, respectively, as shown in fig. 6. We considered that the effect of decoloring at 11% was the best. The decolorization helps to reduce the removal of the color of the supernatant of the hemp and helps to improve the product phase.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A preparation method of normal-temperature health-care hemp milk is characterized by comprising the following steps:
(1) preparing hemp protein polypeptide powder;
the preparation of the hemp protein polypeptide powder comprises the following steps:
(1.1) weighing 5-10g of fructus cannabis meal, dissolving in 50-100ml of distilled water, adding alkaline protease, adjusting the pH value to 8-11 at 55-65 ℃, adding 3-5% of enzyme, and carrying out first enzymolysis for 2-4h at a material-liquid ratio of 1: 5;
(1.2) after the time of the first enzymolysis is reached, inactivating enzymes at high temperature, cooling, adjusting the pH value to 5, adding papain, carrying out second enzymolysis at the temperature of 70-80 ℃ with the enzyme addition amount of 0.4-0.6%, after 1-2h, inactivating enzymes at high temperature, cooling, centrifuging, extracting supernatant, carrying out microwave detoxification, and carrying out spray drying to prepare the hemp protein peptide powder;
(2) preparing skim raw milk;
(3) blending the hemp protein polypeptide powder;
(4) emulsifying, homogenizing and sterilizing;
(5) and filling to obtain the normal-temperature health-care hemp milk.
2. The preparation method of normal-temperature health-care hemp milk according to claim 1, which is characterized by comprising the following steps: the preparation of the skim raw milk comprises the following steps: taking 1000g of raw milk, preheating the raw milk to 35-40 ℃ after flash evaporation treatment, and degreasing in a disc separator to control the fat content of the degreased raw milk to be below 0.5%.
3. The preparation method of normal-temperature health-care hemp milk according to claim 1, which is characterized by comprising the following steps: the hemp protein polypeptide blended powder comprises: pouring the skim raw milk obtained in the step (2) into a blending tank, heating to 55-60 ℃, adding the hemp protein polypeptide powder obtained in the step (1) and 4g of honey, and stirring and dissolving for 10-15min to obtain the base material.
4. The preparation method of normal-temperature health-care hemp milk according to claim 1, which is characterized by comprising the following steps: the emulsification, homogenization and sterilization comprises the following steps: heating the obtained base material to 65-70 deg.C, homogenizing, and pasteurizing at 95 + -1 deg.C for 10-15 s.
5. The preparation method of normal-temperature health-care hemp milk according to claim 1, which is characterized by comprising the following steps: the filling comprises the following steps: and cooling the sterilized product to 2-6 ℃ within 3-15min, and filling to obtain the normal-temperature health-care cannabis sativa milk.
6. The preparation method of normal-temperature health-care hemp milk according to claim 1, wherein the alkaline protease is solid alkaline protease, and the preparation steps of the alkaline protease are as follows: firstly, preparing 15ml of agarose-phosphoric acid solution with the mass fraction of 3% by using phosphoric acid solution with the pH value of 7-8; boiling for more than 5min under microwave condition, cooling to room temperature, adding 60g/L alkaline protease solution 2ml, cooling to 13-16 deg.C to obtain gel, cutting into 1cm x 1cm pieces, and storing at 3-7 deg.C as solid alkaline protease.
7. The preparation method of normal-temperature health-care hemp milk according to claim 1, which is characterized by comprising the following steps: screening out the agarose block after the first enzymolysis, wherein the papain is solid papain during the second enzymolysis, and the preparation steps of the papain are as follows: firstly, preparing 15ml of agarose-phosphoric acid solution with the mass fraction of 3% by using phosphoric acid solution with the pH value of 7.5; boiling for more than 5min under microwave condition, cooling to room temperature, adding 65g/L alkaline protease solution 2ml, cooling to 10-14 deg.C to form gel, cutting into 1cm x 1cm pieces, and storing at 2-5 deg.C as solid alkaline protease.
8. The preparation method of normal-temperature health-care hemp milk according to claim 6 or 7, which is characterized in that: after cutting into 1cm by 1cm pieces, adding glucose or sucrose solution as protectant to the alkaline protease of claim 6 or papain of claim 7, and soaking for more than 5min before low-temperature storage.
9. The preparation method of normal-temperature health-care hemp milk according to claim 2, which is characterized by comprising the following steps: the flash evaporation uses a flash evaporation cavity, the microwave power in the flash evaporation cavity is 1k W-1.25kW, the raw milk flow is 15L/h-30L/h, the inlet temperature of the flash evaporation cavity is 50-55 ℃, and the vacuum degree of the system is 0.095 MPa.
10. The preparation method of normal-temperature health-care hemp milk according to claim 1, which is characterized by comprising the following steps: the microwave detoxification is carried out under the conditions that the microwave intensity is 4.375w/g, the time is 6min, and the water vapor flow at 50 ℃ is 2 m/s.
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