CN117568429A - Method for preparing peony seed meal peptide powder by bionic enzymolysis - Google Patents

Method for preparing peony seed meal peptide powder by bionic enzymolysis Download PDF

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CN117568429A
CN117568429A CN202210939852.1A CN202210939852A CN117568429A CN 117568429 A CN117568429 A CN 117568429A CN 202210939852 A CN202210939852 A CN 202210939852A CN 117568429 A CN117568429 A CN 117568429A
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seed meal
peony seed
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孙志强
刘玉军
刘国飞
柏道鸣
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Shandong Yuze Yaokang Industrial Technology Research Institute Co ltd
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Abstract

The invention discloses an industrial production method for preparing peony seed meal peptide powder from peony seed meal protein through bionic enzymolysis. The method is characterized in that peony seed meal peptide powder is obtained by taking peony seed meal protein as a starting material through bionic enzymolysis, high-speed centrifugation, ultrafiltration, electrodialysis, concentration and drying. The peony seed meal peptide powder has the total protein content of more than 90%, and the trichloroacetic acid-soluble small molecular peptide content of more than 85%, has the advantages of meeting the human body absorption selection mechanism, having no toxic or side effect, being low in cost and the like, and can be applied to foods, health-care foods and medicines.

Description

Method for preparing peony seed meal peptide powder by bionic enzymolysis
Technical Field
The invention relates to a preparation method of peony seed meal peptide powder, in particular to a production method for preparing peony seed meal peptide powder from peony seed meal protein by bionic enzymolysis.
Background
Shandong is a great province of peony planting, the planting area of 2015 is 58 ten thousand mu, 58% of the country is occupied, the planting area of the whole province of peony reaches 400 ten thousand mu by 2020, and the peony industry already covers various fields of traditional Chinese medicine, ornamental, oil extraction, food and the like. Through intensive investigation in the lotus region, the weight of the peony seed meal which is the leftover after the peony seed is squeezed accounts for 45% of the weight of the peony seed, the protein content of the peony seed meal is up to 30%, but the peony seed meal has bitter taste and can not meet the requirements of eating (pigs do not eat), so the peony seed meal is discarded as garbage, and the environmental pollution and the resource waste are caused. The peony seed meal is solid residue after oil extraction of the peony seeds, li Jiaxing and the like are used for optimizing and researching the extraction process of the peony seed meal protein alkali extraction and acid precipitation method, and the isoelectric point of the precipitated protein is analyzed, so that the optimal process parameters for extracting the peony seed meal protein are pH11, the extraction time is 100min, the extraction temperature is 55 ℃, the feed liquid ratio is 1:20 (m: V), the protein extraction rate can reach 86.77%, the isoelectric point of the peony seed meal protein is pH4.0, and the precipitation rate can reach 94.55%.
Under normal physiological conditions, reactive Oxygen Species (ROS) generation and scavenging are in dynamic equilibrium, and ROS can be maintained at very low levels that are advantageously harmless. Antioxidant defense systems for eliminating ROS are classified into enzyme systems including superoxide dismutase, glutathione peroxidase, catalase, and the like, and non-enzyme systems including endogenous antioxidants and exogenous antioxidants. The natural antioxidants in organisms are mainly reduced glutathione and carnosine, but the content is very low. Exogenous antioxidants with free radical scavenging and lipid peroxidation inhibiting functions have been the focus of research in biochemical nutrition. At present, most of the used exogenous antioxidants are chemical compositions, such as BHA and BHT, and have good antioxidation effect, but due to toxic and side effects, the government of each country forcedly stipulates ADI values and controls excessive addition of the ADI values. Therefore, peptide substances which gradually turn the eyes to natural antioxidants such as ascorbic acid, alpha-tocopherol, beta-carotene, flavone, catechin and the like and are accumulated in polypeptide chains of proteins are also found to have strong antioxidant activity, the antioxidant activity is often stronger than that of parent proteins and amino acids, and the research of antioxidant peptides as a non-enzymatic free radical scavenger is on the rise. The antioxidant peptide has the characteristics of small molecular weight, easy absorption, strong activity and the like, has wide application prospect in the fields of medicines, cosmetics, health products, food, feed additives and the like, and is hopeful to be developed into a novel functional food
Therefore, if the protein of the peony seed meal is taken as an entry point, the main activity of the symbiotic parts (peony seed oil, cortex moutan extract and the like) of the peony seed meal is taken as a target, the technology of preparing and separating the active peptide by bionic enzymolysis is adopted, the antioxidant activity is taken as an index, the small peptide with high antioxidant activity is found out from the active peptide of the peony seed meal, and the structure-activity relationship between the peptide sequence and the activity is found out by performing test analysis and research on the small peptide. Compared with the traditional peony seed protein, the peony seed meal peptide is easier to digest and absorb, and has unique physiological functions which the protein does not have. The peony seed meal peptide is superior to the peony seed meal protein in physical and chemical properties, nutritional properties and functional properties, so that the peony seed meal peptide has very broad application prospect and huge market potential.
The bionic enzymolysis is to simulate the gastrointestinal tract digestion process of oral protein (the stomach is the digestion part, the absorption is very small, the intestinal tract is the digestion and absorption part), the creative method for decomposing protein into small molecular peptide is to continuously carry out enzymolysis by pepsin and trypsin under proper conditions, and then high-speed centrifugation, ultrafiltration, electrodialysis or nanofiltration is carried out to obtain the small molecular peptide with the relative molecular mass smaller than 3KD as the main component. Compared with the prior art, the effective substance group absorbed into blood is the same as the direct oral protein, the bionic enzymatic hydrolysis effective component is extracted more fully, the bioavailability is high, the curative effect is enhanced, the immunogenic reaction caused by taking the heterologous protein and the like is eliminated, the occurrence rate of adverse reaction is reduced, and the safety is high; compared with other enzymolysis methods, the bionic enzymolysis is more in line with the selection mechanism of the human body and safer.
The preparation of the peony seed meal peptide powder from the peony seed meal protein by the bionic enzymolysis method is an original method of the inventor, and has wide application prospect after amplified industrial production verification.
Disclosure of Invention
The first aim of the invention is to provide an industrial production method for preparing peony seed meal peptide powder from peony seed meal protein by bionic enzymolysis; the second aim is to provide the peony seed meal peptide powder obtained by the industrialized production method; the third purpose is to extract the application of the peony seed meal peptide powder in the fields of food, health-care food and medicines.
The purpose of the invention is realized in the following way:
an industrial production method for preparing peony seed meal peptide powder from peony seed meal protein by bionic enzymolysis comprises the following steps:
(1) Adding 20-40 liters of purified water into a bionic enzymolysis tank, starting stirring slurry, setting the rotating speed to be 30-60 revolutions per minute, taking 2kg of peony seed meal protein powder, slowly adding, continuing stirring and swelling for 30 minutes after the addition is finished, heating an interlayer to 80-85 ℃ by steam, sterilizing for 15-30 minutes, cooling the interlayer by cooling water to about 50-55 ℃ in the tank, starting circulation of the interlayer water and a heater, and heating the heater to 50-55 ℃ for heat preservation;
(2) Starting a pH detection terminal, adding dilute hydrochloric acid into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 1.8-2.2, adding pepsin accounting for 1-3% of the protein content of peony seed meal, preserving heat and carrying out enzymolysis for 1-3 hours, monitoring the pH value at any time, and supplementing the dilute hydrochloric acid to keep the pH value within the range of 1.8-2.2;
(3) Adding 40% sodium hydroxide into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 7.8-8.2, adding 1-3% trypsin, preserving heat and carrying out enzymolysis for 3-6 hours, monitoring the pH value at any time, and supplementing 40% sodium hydroxide to keep the pH value within the range of 7.8-8.2;
(4) Adding dilute hydrochloric acid into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 6.8-7.2, discharging interlayer water, boiling the interlayer by steam, heating to 80-85 ℃, inactivating enzyme for 15-30 minutes, and discharging liquid;
(5) Centrifuging in a tubular high-speed centrifuge at 8000-12000 rpm, ultrafiltering supernatant with 1812 type ultrafilter, desalting the supernatant with ultrafiltration membrane of 3KD, concentrating the desalted solution under reduced pressure at 30-90 v to obtain fluid extract with relative density of 1.10-1.15 at 60deg.C, spray drying at 180-200deg.C, and air-out at 90-100deg.C to obtain peony seed meal peptide powder, and packaging with composite membrane bag.
The protein content in the peony seed meal protein in the step (1) is more than 90%, and the moisture is less than 10%;
the pepsin activity in the step (2) is 3000U/g;
the trypsin enzyme activity in the step (3) is 100000U/g;
the total protein content in the peony seed meal peptide powder in the step (5) is more than 90%, the content of trichloroacetic acid-soluble small molecular peptide is more than 85%, the content of free amino acid is less than 5%, and the moisture is less than 7%.
A peony seed meal peptide powder is obtained by the industrial production method. The peony seed meal peptide powder obtained by the method has the main relative molecular mass below 3KD, extremely strong water solubility, total protein content above 90%, trichloroacetic acid soluble small molecular peptide content above 85%, free amino acid content less than 5% and water content less than 7%.
The bionic enzymolysis tank and the enzymolysis system refer to equipment involved in CN205874433U, CN205590709U, and the consumption of various materials in the method can be increased along with the increase of the volume of the equipment. The method is also suitable for other devices with similar functions.
The raw material peony seed meal protein powder can be a finished product raw material purchased from the market, or can be a liquid product obtained by hydrolyzing peony seed meal to obtain the peony seed meal protein powder or the peony seed meal protein. In a word, no matter what raw material is used for the production (peony seed meal, peony seed meal protein powder, peony seed meal protein liquid and the like), the technical content of the invention is considered to be adopted as long as the protection content of the invention is related in the production process.
The application of the peony seed meal peptide powder obtained by the industrial production method in foods, health foods and medicines comprises but is not limited to the following applications: according to the nutrition, function and processing characteristics of the produced peony seed meal peptide powder, a plurality of high-quality, nutritional and healthy foods can be developed by taking the peony seed meal peptide powder as a basic material. The peony seed meal polypeptide has quite high water retention, oil absorption and good gel forming property, and can be used as an excellent additive of meat products; the peony seed meal polypeptide has certain foamability and can be added into cake products to replace eggs partially; the peony seed meal polypeptide has very good emulsifying property and can be used as a food emulsifying agent; the peony seed meal peptide can be used for enhancing fragrance and strengthening protein, and can be developed into health-care foods with different functions; the peony seed meal peptide can also be used for flour products, so that the nutritional value, strength and strength are improved, and the appearance and taste of flour foods can be improved; the peony seed meal peptide is made into wine, which not only can be directly absorbed by human body, but also has the function of enhancing human body immunity; can be used for cosmetic to inhibit elastase, protect elastin, improve skin elasticity, and increase skin firmness.
The beneficial effects of the invention are as follows: the industrial production method for preparing the peony seed meal peptide powder from the peony seed meal protein through bionic enzymolysis accords with a human body selection mechanism, and has the advantages of simple and convenient preparation process, feasibility, low cost and environment-friendly production process; the total protein content of the obtained peony seed meal peptide powder is more than 90%, the content of small molecular peptides is more than 85%, the relative molecular mass is mainly less than 3KD, no toxic or side effect is caused, the pure natural requirement of green food is met, and good economic benefit can be obtained.
Detailed Description
The invention is further illustrated by the following examples, which are given solely for the purpose of illustration and are not intended to be limiting of the invention:
example 1 this example includes the following steps:
(1) Adding purified water 20L into a bionic enzymolysis tank, starting stirring slurry, setting the rotating speed to be 60 r/min, taking 2kg of peony seed meal protein powder (the protein content of the peony seed meal is more than 90% and the water content is less than 10%), slowly adding, continuing stirring and swelling for 30 minutes after the adding is finished, heating an interlayer to 80-85 ℃ for sterilization for 15 minutes, cooling the interlayer to the temperature of about 55 ℃ in the tank, starting circulation of the interlayer water and a heater, and heating the heater to 55 ℃ for heat preservation;
(2) Starting a pH detection terminal, adding dilute hydrochloric acid into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 2.0, adding pepsin (the enzyme activity is 3000U/g) accounting for 1% of the protein amount of the peony seed meal, preserving heat and carrying out enzymolysis for 1 hour, monitoring the pH value at any time, and supplementing the dilute hydrochloric acid to keep the pH value within a range of 1.8-2.2;
(3) Adding 40% sodium hydroxide into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 8.0, adding 1% trypsin (the enzyme activity is 100000U/g), preserving the heat and carrying out enzymolysis for 3 hours, monitoring the pH value at any time, and supplementing 40% sodium hydroxide to keep the pH value within the range of 7.8-8.2;
(4) Adding dilute hydrochloric acid into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 6.8, discharging interlayer water, boiling the interlayer by steam, heating to 80-85 ℃, inactivating enzyme for 30 minutes, and discharging liquid;
(5) Centrifuging in a tubular high-speed centrifuge at 8000-12000 rpm, ultrafiltering supernatant with 1812 type ultrafilter, desalting the supernatant with ultrafiltration membrane of 3KD, concentrating the desalted solution under reduced pressure at 30 v to obtain fluid extract with relative density of 1.10-1.15 at 60deg.C, spray drying at 180-200deg.C and air outlet temperature of 90-100deg.C to obtain peony seed meal peptide powder, and packaging with composite membrane bag.
Example 2 this example includes the following steps:
(1) Adding purified water 40L into a bionic enzymolysis tank, starting stirring slurry, setting the rotating speed to be 30 r/min, taking 2kg of peony seed meal protein powder, slowly adding, continuing stirring and swelling for 30 min after the addition is finished, heating an interlayer to 80-85 ℃ by steam, sterilizing for 15 min, cooling the interlayer by cooling water to about 50 ℃ in the tank, starting circulation of the interlayer water and a heater, and heating the heater to 50 ℃ for heat preservation;
(2) Starting a pH detection terminal, adding dilute hydrochloric acid into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 1.8, adding pepsin accounting for 3% of the protein content of peony seed meal, preserving heat and carrying out enzymolysis for 3 hours, monitoring the pH value at any time, and supplementing the dilute hydrochloric acid to keep the pH value within the range of 1.8-2.2;
(3) Adding 40% sodium hydroxide into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 8.2, adding 3% trypsin, preserving heat and carrying out enzymolysis for 6 hours, monitoring the pH value at any time, and supplementing 40% sodium hydroxide to keep the pH value within the range of 7.8-8.2;
(4) Adding dilute hydrochloric acid into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 7.2, discharging interlayer water, boiling the interlayer by steam, heating to 80-85 ℃, inactivating enzyme for 15 minutes, and discharging liquid;
(5) Centrifuging in a tubular high-speed centrifuge at 8000-12000 rpm, ultrafiltering supernatant with 1812 type ultrafilter, desalting the supernatant with ultrafiltration membrane of 3KD, concentrating the desalted solution under reduced pressure at 90 v to obtain fluid extract with relative density of 1.10-1.15 at 60deg.C, spray drying at 180-200deg.C and air outlet temperature of 90-100deg.C to obtain peony seed meal peptide powder, and packaging with composite membrane bag.
Example 3 peony seed meal peptide powder:
the peony seed meal peptide powder obtained by the industrial production method described in the embodiment 1 is taken and detected: the total protein content is 92%, the trichloroacetic acid soluble small molecular peptide content is 86%, the free amino acid content is 4.2%, the water content is 6.1%, and the relative molecular weight distribution is:
range of relative molecular weight Percent peak area
>3000D 7.8%
3000-2000D 21.1%
2000-1000D 32.3%
1000-300D 34.7%
<300D 4.1%
Example 4 peony seed meal peptide powder prepared product:
400g of the peony seed meal peptide powder obtained in the method of example 1 is taken, the mixture of the peony seed meal peptide powder, corn oil, beeswax and lecithin in the proportion of 10:36:3:1 is ground for 30 minutes by a colloid mill, and gelatin, glycerol, purified water and iron oxide black are mixed according to the proportion of 100:40:100:1, preparing soft capsule shell glue solution in proportion, controlling the temperature of a glue box to be 55+/-5 ℃, keeping the temperature of a heat preservation tank to be 40+/-5 ℃, spraying the glue solution to be 36+/-5 ℃, pressing the soft capsule by a pill pressing machine, drying the soft capsule with cold air at the temperature of 18-26 ℃ for 4 hours (the relative humidity is 30-40%) with the weight of 0.50g for each content, shaping, washing pills, drying, selecting pills, and packaging to prepare the soft capsule finished product.
Example 5 application of peony seed meal peptide powder:
according to the nutrition, function and processing characteristics of the produced peony seed meal peptide powder, a plurality of high-quality, nutritional and healthy foods can be developed by taking the peony seed meal peptide powder as a basic material. The peony seed meal polypeptide has quite high water retention, oil absorption and good gel forming property, and can be used as an excellent additive of meat products; the peony seed meal polypeptide has certain foamability and can be added into cake products to replace eggs partially; the peony seed meal polypeptide has very good emulsifying property and can be used as a food emulsifying agent; the peony seed meal peptide can be used for enhancing fragrance and strengthening protein, and can be developed into health-care foods with different functions; the peony seed meal peptide can also be used for flour products, so that the nutritional value, strength and strength are improved, and the appearance and taste of flour foods can be improved; the peony seed meal peptide is made into wine, which not only can be directly absorbed by human body, but also has the function of enhancing human body immunity; can be used for cosmetic to inhibit elastase, protect elastin, improve skin elasticity, and increase skin firmness.

Claims (5)

1. A method for preparing peony seed meal peptide powder by bionic enzymolysis is characterized by comprising the following steps:
(1) Adding 20-40 liters of purified water into a bionic enzymolysis tank, starting stirring slurry, setting the rotating speed to be 30-60 revolutions per minute, taking 2kg of peony seed meal protein powder, slowly adding, continuing stirring and swelling for 30 minutes after the addition is finished, heating an interlayer to 80-85 ℃ by steam, sterilizing for 15-30 minutes, cooling the interlayer by cooling water to about 50-55 ℃ in the tank, starting circulation of the interlayer water and a heater, and heating the heater to 50-55 ℃ for heat preservation;
(2) Starting a pH detection terminal, adding dilute hydrochloric acid into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 1.8-2.2, adding pepsin accounting for 1-3% of the protein content of peony seed meal, preserving heat and carrying out enzymolysis for 1-3 hours, monitoring the pH value at any time, and supplementing the dilute hydrochloric acid to keep the pH value within the range of 1.8-2.2;
(3) Adding 40% sodium hydroxide into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 7.8-8.2, adding 1-3% trypsin, preserving heat and carrying out enzymolysis for 3-6 hours, monitoring the pH value at any time, and supplementing 40% sodium hydroxide to keep the pH value within the range of 7.8-8.2;
(4) Adding dilute hydrochloric acid into a bionic enzymolysis tank by a peristaltic pump to adjust the pH value to 6.8-7.2, discharging interlayer water, boiling the interlayer by steam, heating to 80-85 ℃, inactivating enzyme for 15-30 minutes, and discharging liquid;
(5) Centrifuging in a tube high-speed centrifuge at 8000-12000 rpm, ultrafiltering supernatant with 1812 type ultrafiltration device, ultrafiltering with ultrafiltration membrane with relative molecular weight of 3KD, desalting the permeate with electrodialysis at 30-90 v, concentrating the desalted liquid under reduced pressure to obtain fluid extract with relative density of 1.10-1.15 at 60deg.C, spray drying at 180-200deg.C, and air-out at 90-100deg.C to obtain peony seed meal peptide powder, and packaging with composite membrane bag.
2. The method for preparing peony seed meal peptide powder by bionic enzymatic hydrolysis according to claim 1, which is characterized in that the pepsin activity is 3000U/g; trypsin activity is 100000U/g; the content of protein in the peony seed meal protein is more than 30%, and the moisture is less than 10%.
3. The method for preparing peony seed meal peptide powder by bionic enzymatic hydrolysis according to claim 1, wherein the total protein content in the obtained peony seed meal peptide powder is more than 90%, the trichloroacetic acid-soluble small molecular peptide content is more than 85%, the free amino acid content is less than 5%, and the water content is less than 7%.
4. A peony seed meal peptide powder obtained by the industrial production method of claims 1-3.
5. The use of the peony seed meal peptide powder of claim 4 in the fields of food, health food and medicines.
CN202210939852.1A 2022-08-05 2022-08-05 Method for preparing peony seed meal peptide powder by bionic enzymolysis Pending CN117568429A (en)

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