CN113249421B - Golden-silk-royal chrysanthemum protein polypeptide and preparation and application thereof - Google Patents
Golden-silk-royal chrysanthemum protein polypeptide and preparation and application thereof Download PDFInfo
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
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Abstract
The invention relates to a golden-silk chrysanthemum polypeptide and application thereof, wherein the preparation method of the golden-silk chrysanthemum polypeptide comprises the following steps: pre-treating; extracting chrysanthemum protein; preparing chrysanthemum polypeptide. The method takes the canary-creeper-wort inulin as a raw material, firstly adopts an alkali-dissolution acid-precipitation method to extract protein in the canary-creeper-wort-creeper inulin, secondly adopts protease to hydrolyze chrysanthemum protein to obtain the chrysanthemum polypeptide with low molecular weight, is green and environment-friendly in the production process, has better alpha-glucosidase inhibitory activity, antioxidation and other functions in the product, can be applied to the fields of functional foods, medicines, cosmetics and the like, and can obviously improve the utilization value of chrysanthemum.
Description
Technical Field
The invention belongs to the technical field of deep processing of drug-derived proteins, and relates to a method for extracting and preparing protein polypeptides, in particular to a method for extracting and preparing protein polypeptides from golden-silk Huang chrysanthemum and application thereof.
Background
The chrysanthemum is used as an important ornamental plant and an edible plant since ancient times, the golden-silk king chrysanthemum is a fine product in the chrysanthemum, the flavone, polyphenol and other substances contained in the golden-silk king chrysanthemum are well known, the protein in the golden-silk king chrysanthemum is about 12 percent and is far higher than other flowers, the protein rich in nutrition in the golden-silk king chrysanthemum is still lack of wide understanding and research, the chrysanthemum protein contains amino acid necessary for a human body, the proportion is balanced, and the utilization rate of an organism is high. However, the protein has the problems of large molecular weight, difficult absorption, easy hydrolysis into amino acid, loss of activity and the like, so the application of the protein in the fields of food, cosmetics, health care products and the like is limited to a great extent, and the research or application of the protein hydrolysis of chrysanthemum into polypeptide is not reported yet. From the industrial and market conditions, the development and utilization of the deep processing products of the golden cypress are relatively lagged, the production added value is low, and no industrial chain is formed. Research shows that the protein content of the golden cypress is as high as 12.9%, the amino acid types are rich, and the golden cypress has high development and utilization values.
The golden emperor chrysanthemum protein polypeptide is a small molecular oligopeptide mixture obtained by hydrolyzing, separating and purifying golden emperor chrysanthemum protein through protease, and is further processed into products such as health products, skin care products and the like, and has important significance for improving the additional value and market competitiveness of golden emperor chrysanthemum.
Through a search, no published patent literature relevant to the present patent application has been found.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a preparation method and application of a golden-silk chrysanthemum protein polypeptide, which is green, simple and convenient and has high hydrolysis degree.
The technical scheme adopted by the invention for solving the technical problem is as follows:
a protein enzymolysis polypeptide of golden cypress, chrysanthemum and the like is prepared by the following steps:
(1) Pretreatment of chrysanthemum: crushing and sieving chrysanthemum raw materials to obtain chrysanthemum crushed materials:
(2) Extracting chrysanthemum protein: extracting chrysanthemum protein in chrysanthemum powder by an alkali-dissolving acid-precipitating method;
(3) Preparing chrysanthemum polypeptide: preparing the chrysanthemum protein powder prepared in the step (2) into chrysanthemum protein aqueous solution, and adding protease for enzymolysis; centrifuging at 8000r/min for 10-20min after enzymolysis, collecting supernatant, decolorizing, and vacuum freeze drying to obtain flos Chrysanthemi polypeptide powder;
the protease comprises papain, flavourzyme and complex enzyme formed by the papain and the flavourzyme.
Further, the alkali-dissolving and acid-precipitating method in the step (2) is to prepare chrysanthemum powder crushed materials into chrysanthemum powder aqueous solution, adjust the pH value to 8.0-9.5, extract at the constant temperature of 50-60 ℃ for 5 hours, and then centrifuge and discard precipitates; regulating the pH value of the supernatant to 3.5-4.5, centrifuging to obtain precipitate, and washing with water to neutrality; and carrying out vacuum freeze drying to obtain the chrysanthemum protein powder.
Further, the mass concentration of the chrysanthemum powder aqueous solution is 1-4%; or, adjusting the pH value to 8.0-9.5 by using 0.5-1 mol/LNaOH solution, reacting at the constant temperature of 50-60 ℃ for 5h, centrifuging at 8000r/min for 10-15 min, and discarding the precipitate; adjusting the pH value of the supernatant to 3.5-4.5 by using 0.5-1 mol/L HCl solution, centrifuging for 10-15 min at 8000r/min, keeping the precipitate, and washing with water to be neutral; vacuum freeze drying, and pulverizing to obtain flos Chrysanthemi protein powder.
Further, the addition amount of the protease in the step (3) is 10000-12000U/g; or when the protease is a compound enzyme consisting of papain and flavourzyme, the mass ratio of the flavourzyme to the papain is 1-1.
Further, the mass concentration of the chrysanthemum protein substrate in the step (3) is 1% -3%.
Further, the pH value is 7.5-8.5 during enzymolysis in the step (3); or, the enzymolysis temperature is 50-55 ℃; or, the enzymolysis time is 4-5h; or taking enzymolysis liquid with the molecular weight less than 3kDa after ultrafiltration and centrifugation;
or adding activated carbon with the final mass concentration of 2-5%, stirring at 55 ℃ for 1-2h, decoloring and concentrating.
The golden cypress protein polypeptide preparation is prepared by utilizing the golden cypress protein enzymolysis polypeptide.
Further, the preparation is granules, tablets, capsules or oral liquid.
The above mentioned protein enzymolysis polypeptide of Hypericum virgatum can be used in nutritional food, health food, athlete food, cosmetics or skin care products.
Further, the nutritional food is a nutritional food for patients or infants; the health food is provided for the elderly or sub-health people; alternatively, the nutritional food is a protein beverage; the cosmetic and skin care product is moisturizer, milk, cream, essence, facial mask, eye cream, foundation liquid or sunscreen cream.
The invention has the advantages and positive effects that:
1. the golden cypress protein polypeptide is prepared by taking dried golden cypress chrysanthemum as a raw material, and performing crushing, sieving, crude protein extraction and freeze drying on the raw material to obtain golden cypress protein; different golden aster polypeptide samples are obtained by carrying out enzymolysis on golden aster protein by using flavourzyme, papain and different compound modes of the flavourzyme and the papain, and a polypeptide sample with high hydrolysis degree is obtained by optimizing conditions, so that the obtained golden aster polypeptide has the effects of reducing blood sugar, resisting oxidation and the like, the production process is green and environment-friendly, the product has good alpha-glucosidase inhibitory activity, resisting oxidation and the like, and the method has wide development and application prospects in the fields of food industry, health care products, cosmetics, medicines and the like. The invention can obviously improve the utilization value of the chrysanthemum.
2. The chrysanthemum protein polypeptide has uniform color, is white to light yellow, has small molecular weight, is easier to be absorbed and utilized by human bodies, has simple preparation method, easy control of enzymolysis degree, good quality and effect of products and high product yield.
3. The method provided by the invention aims to find the potential application value of the golden cypress protein, converts the golden cypress protein into polypeptide by utilizing enzymolysis, obtains the golden cypress protein polypeptide by a preparation method which is environment-friendly, high in product yield and simple and convenient to operate, and applies the golden cypress protein polypeptide to the fields of medicines, health care products, nutriments, foods, cosmetics and the like.
4. The invention aims to further improve the content of effective polypeptide by preparing and purifying chrysanthemum polypeptide, can be used for preparing special diet, health food, food additive, medicament and other fields with the function of regulating blood sugar metabolism and skin care products, cosmetics and other fields with antioxidant function, and is suitable for industrial production.
5. The enzymolysis mode of the protease in the invention is that single papain is used for enzymolysis, single flavourzyme is used for enzymolysis, the complex enzyme composed of flavourzyme and papain is used for synergistic enzymolysis, the complex enzyme composed of flavourzyme and papain is used for stepwise enzymolysis (firstly flavourzyme and then papain), and the complex enzyme composed of flavourzyme and papain is used for stepwise enzymolysis (firstly papain and then flavourzyme).
6. During ultrafiltration and centrifugation, the chrysanthemum polypeptide is subjected to fractionation by an ultrafiltration membrane according to molecular weight to obtain chrysanthemum polypeptide solutions with different molecular weights, and preferably, the cut-off molecular weight is less than 3 KDa.
Drawings
FIG. 1 is a graph showing the results of the antioxidant activity of the polypeptide of the present invention.
Detailed Description
The following detailed description of the embodiments of the present invention is provided for the purpose of illustration and not limitation, and should not be construed as limiting the scope of the invention.
The raw materials used in the invention are conventional commercial products unless otherwise specified; the methods used in the present invention are conventional in the art unless otherwise specified.
A protein enzymolysis polypeptide of golden cypress, chrysanthemum and the like is prepared by the following steps:
(1) Pretreatment of chrysanthemum: crushing and sieving the chrysanthemum raw material to obtain chrysanthemum crushed material:
(2) Extracting chrysanthemum protein: extracting chrysanthemum protein in chrysanthemum powder by an alkali-dissolution acid-precipitation method;
(3) Preparing chrysanthemum polypeptide: preparing the chrysanthemum protein powder prepared in the step (2) into chrysanthemum protein aqueous solution, and adding protease for enzymolysis; centrifuging at 8000r/min for 10-20min after enzymolysis, collecting supernatant, decolorizing, and vacuum freeze drying to obtain flos Chrysanthemi polypeptide powder;
the protease comprises papain, flavourzyme and complex enzyme formed by the papain and the flavourzyme.
Preferably, the alkali-dissolving and acid-precipitating method in the step (2) is to prepare the chrysanthemum powder crushed material into chrysanthemum powder aqueous solution, adjust the pH value to 8.0-9.5, extract at the constant temperature of 50-60 ℃ for 5h, then centrifuge and discard the precipitate; regulating the pH value of the supernatant to 3.5-4.5, centrifuging to obtain precipitate, and washing with water to neutrality; and carrying out vacuum freeze drying to obtain the chrysanthemum protein powder.
Preferably, the mass concentration of the chrysanthemum powder aqueous solution is 1-4%; or, adjusting the pH value to 8.0-9.5 by using 0.5-1 mol/LNaOH solution, reacting at the constant temperature of 50-60 ℃ for 5h, centrifuging at 8000r/min for 10-15 min, and discarding the precipitate; adjusting the pH value of the supernatant to 3.5-4.5 by using 0.5-1 mol/L HCl solution, centrifuging for 10-15 min at 8000r/min, keeping the precipitate, and washing with water to be neutral; and (4) carrying out vacuum freeze drying and crushing to obtain the chrysanthemum protein powder.
Preferably, the addition amount of the protease in the step (3) is 10000-12000U/g; or when the protease is a complex enzyme consisting of papain and flavourzyme, the mass ratio of the flavourzyme to the papain is 1-1.
Preferably, the mass concentration of the chrysanthemum protein substrate in the step (3) is 1-3%.
Preferably, the pH value during enzymolysis in the step (3) is 7.5-8.5; or, the enzymolysis temperature is 50-55 ℃; or, the enzymolysis time is 4-5h; or taking enzymolysis liquid with the molecular weight less than 3kDa after ultrafiltration and centrifugation;
or adding active carbon with the final mass concentration of 2-5%, stirring at 55 ℃ for 1-2h, and concentrating after decoloring.
The golden emperor chrysanthemum protein polypeptide preparation is prepared by utilizing the golden emperor chrysanthemum protein enzymolysis polypeptide.
Preferably, the preparation is a granule, a tablet, a capsule or an oral liquid.
The above mentioned protein enzymolysis polypeptide of Hypericum virgatum can be used in nutritional food, health food, athlete food, cosmetics or skin care products.
Preferably, the nutritional food is a nutritional food for patients or infants; the health food is provided for the elderly or sub-health people; alternatively, the nutritional food is a protein beverage; the cosmetic and skin care product is moisturizer, milk, cream, essence, facial mask, eye cream, foundation liquid or sunscreen cream.
Specifically, the preparation and detection are as follows:
example 1
A golden-silk chrysanthemum proteolysis polypeptide is prepared by the following steps:
pretreatment of chrysanthemum: pulverizing dried golden-silk chrysanthemum by a pulverizer, and sieving by a 60-mesh sieve to obtain chrysanthemum powder for later use.
Extracting chrysanthemum protein: adding water into chrysanthemum powder, adding NaOH for alkali dissolution, centrifuging for 10min at the rotating speed of 8000r/min, regulating the pH of the supernatant to an isoelectric point, centrifuging for 10min at the rotating speed of 8000r/min, centrifuging to obtain a precipitate, washing with water to neutrality, and performing vacuum freeze drying to obtain chrysanthemum protein. Wherein, 5.0g of chrysanthemum powder is taken, 150ml of deionized water is added, the reaction temperature is 55 ℃, the alkali dissolution pH =8, the acid precipitation pH =4, and the reaction time is 5h.
The preparation method of the chrysanthemum polypeptide comprises the following steps: dissolving chrysanthemum protein with deionized water, adjusting pH with 1mol/LNaOH, adding protease, performing enzymolysis at constant temperature, rapidly heating to 90 ℃ to inactivate enzyme for 20min, centrifuging at a rotation speed of 8000r/min for 10min, and taking supernatant to obtain chrysanthemum polypeptide liquid, wherein the pH value and the constant temperature enzymolysis temperature are determined according to the selected protease, and in the embodiment, the protease is selected from flavourzyme (2.5 ten thousand U/g). Wherein, 0.5g of chrysanthemum protein is taken, 20ml of deionized water is added, the adding amount of flavourzyme is 10000U/g, the pH is 8.5, and the enzymolysis temperature is 55 ℃. And roughly separating the molecular weight of the chrysanthemum crude polypeptide subjected to enzymolysis by adopting a membrane filtration technology to remove macromolecular protein, collecting chrysanthemum polypeptide with the molecular weight of less than 3kDa, stirring 2-5% of activated carbon at 55 ℃ for 1-2h to decolor, concentrating, and drying in vacuum to obtain 60.2mg of chrysanthemum polypeptide powder.
Example 2
A golden-silk chrysanthemum proteolysis polypeptide is prepared by the following steps:
pretreatment of chrysanthemum: pulverizing dried flos Chrysanthemi with a pulverizer, and sieving with 60 mesh sieve to obtain flos Chrysanthemi powder.
Extracting chrysanthemum protein: adding water into chrysanthemum powder, adding NaOH for alkali dissolution treatment, centrifuging for 10min at the rotating speed of 8000r/min, adjusting the pH of the supernatant to the isoelectric point, centrifuging for 10min at the rotating speed of 8000r/min, centrifuging to obtain precipitate, washing with water to neutrality, and vacuum freeze-drying to obtain chrysanthemum protein. Wherein, 5.0g of chrysanthemum powder is taken, 150ml of deionized water is added, the reaction temperature is 55 ℃, the alkali dissolution pH =8, the acid precipitation pH =4, and the reaction time is 5h.
The preparation method of the chrysanthemum polypeptide comprises the following steps: dissolving chrysanthemum protein with deionized water, adjusting pH with 1mol/LNaOH, adding protease, performing enzymolysis at constant temperature, rapidly heating to 90 ℃ to inactivate enzyme for 20min, centrifuging at 8000rpm for 10min, and collecting supernatant to obtain chrysanthemum polypeptide solution, wherein the pH value and the constant temperature enzymolysis temperature are determined according to the selected protease, and in the embodiment, the protease is papain (80 ten thousand U/g). Wherein, 0.5g of chrysanthemum protein is taken and added with 20ml of deionized water, the adding amount of papain is 10000U/g, the pH is 7.5, and the enzymolysis temperature is 55 ℃. And coarsely separating the chrysanthemum crude polypeptide subjected to enzymolysis by adopting a membrane filtration technology to remove macromolecular protein, wherein the molecular interception amount of a preset ultrafiltration membrane is 3kDa, collecting chrysanthemum polypeptide with the molecular weight below 3kDa, stirring 2-5% of activated carbon at 55 ℃ for 1-2h, decoloring, concentrating, and vacuum drying to obtain 35.8mg of chrysanthemum polypeptide powder.
Example 3
A golden-silk chrysanthemum proteolysis polypeptide is prepared by the following steps:
pretreatment of chrysanthemum: pulverizing dried flos Chrysanthemi with a pulverizer, and sieving with 60 mesh sieve to obtain flos Chrysanthemi powder.
Extracting chrysanthemum protein: adding water into chrysanthemum powder, adding NaOH for alkali dissolution treatment, centrifuging for 10min at the rotating speed of 8000r/min, adjusting the pH of the supernatant to the isoelectric point, centrifuging for 10min at the rotating speed of 8000r/min, centrifuging to obtain precipitate, washing with water to neutrality, and vacuum freeze-drying to obtain chrysanthemum protein. Wherein, 5.0g of chrysanthemum powder is taken, 150ml of deionized water is added, the reaction temperature is 55 ℃, the alkali dissolution pH =8, the acid precipitation pH =4, and the reaction time is 5h.
The preparation method of the chrysanthemum polypeptide comprises the following steps: dissolving chrysanthemum protein with deionized water, adjusting pH with 1mol/LNaOH, adding protease, performing enzymolysis at constant temperature, inactivating enzyme at 90 ℃ for 20min, centrifuging at 8000r/min for 10min, and collecting supernatant to obtain chrysanthemum polypeptide liquid, wherein the pH value and the constant temperature enzymolysis temperature are determined according to the selected protease. Wherein, 0.5g of chrysanthemum protein is taken and added with 20ml of deionized water, the compound enzyme enzymolysis mode is synergistic enzymolysis, the adding amount ratio of flavourzyme and papain is 1. And coarsely separating the chrysanthemum crude polypeptide subjected to enzymolysis by adopting a membrane filtration technology to remove macromolecular protein, wherein the molecular interception amount of a preset ultrafiltration membrane is 3kDa, collecting chrysanthemum polypeptide with the molecular weight below 3kDa, stirring 2-5% of activated carbon at 55 ℃ for 1-2h, decoloring, concentrating, and vacuum drying to obtain 78.5mg of chrysanthemum polypeptide powder.
Example 4
A golden-silk chrysanthemum proteolysis polypeptide is prepared by the following steps:
pretreatment of chrysanthemum: pulverizing dried flos Chrysanthemi with a pulverizer, and sieving with 60 mesh sieve to obtain flos Chrysanthemi powder.
Chrysanthemum protein extraction: adding water into chrysanthemum powder, adding NaOH for alkali dissolution treatment, centrifuging for 10min at the rotating speed of 8000r/min, adjusting the pH of the supernatant to the isoelectric point, centrifuging for 10min at the rotating speed of 8000r/min, centrifuging to obtain precipitate, washing with water to neutrality, and vacuum freeze-drying to obtain chrysanthemum protein. Wherein, 5.0g of chrysanthemum powder is taken, 150ml of deionized water is added, the reaction temperature is 55 ℃, the alkali dissolution pH =8, the acid precipitation pH =4, and the reaction time is 5h.
The preparation method of the chrysanthemum polypeptide comprises the following steps: dissolving chrysanthemum protein with deionized water, adjusting pH with 1mol/LNaOH, adding one protease to carry out enzymolysis treatment at constant temperature, rapidly heating to 90 ℃ to inactivate the enzyme for 20min, recovering to the original constant temperature, adjusting pH with 1mol/LNaOH, adding another protease to carry out enzymolysis treatment at constant temperature, rapidly heating to 90 ℃ to inactivate the enzyme for 20min, recovering to room temperature, centrifuging at the rotating speed of 8000r/min for 10min, taking supernate to obtain chrysanthemum polypeptide liquid, wherein the pH value and the constant-temperature enzymolysis treatment temperature are determined according to the selected protease. 0.5g of chrysanthemum protein is taken, 20ml of deionized water is added, the compound enzyme enzymolysis mode is stepwise enzymolysis, the adding amount ratio of the flavor protease to the papain is 1, the total enzyme adding amount is 10000U/g, the flavor protease is added firstly, then the papain is added, the pH value of the two stages is 8, and the enzymolysis temperature is 55 ℃. And coarsely separating the chrysanthemum crude polypeptide subjected to enzymolysis by adopting a membrane filtration technology, removing macromolecular protein, collecting chrysanthemum polypeptide with the molecular weight of below 3kDa, stirring 2-5% of activated carbon at 55 ℃ for 1-2h, decoloring, concentrating, and vacuum drying to obtain 94.6mg of chrysanthemum polypeptide powder, wherein the preset molecular cutoff amount of the ultrafiltration membrane is 3 kDa.
Example 5
A protein enzymolysis polypeptide of golden cypress, chrysanthemum and the like is prepared by the following steps:
pretreatment of chrysanthemum: pulverizing dried flos Chrysanthemi with a pulverizer, and sieving with 60 mesh sieve to obtain flos Chrysanthemi powder.
Extracting chrysanthemum protein: adding water into chrysanthemum powder, adding NaOH for alkali dissolution, centrifuging for 10min at the rotating speed of 8000r/min, regulating the pH of the supernatant to an isoelectric point, centrifuging for 10min at the rotating speed of 8000r/min, centrifuging to obtain a precipitate, washing with water to neutrality, and performing vacuum freeze drying to obtain chrysanthemum protein. Wherein, 5.0g of chrysanthemum powder is taken, 150ml of deionized water is added, the reaction temperature is 55 ℃, the alkali dissolution pH =8, the acid precipitation pH =4, and the reaction time is 5h.
The preparation method of the chrysanthemum polypeptide comprises the following steps: dissolving chrysanthemum protein with deionized water, adjusting pH with 1mol/LNaOH, adding one protease, performing enzymolysis at a constant temperature, rapidly heating to 90 ℃ to inactivate the enzyme for 20min, recovering to the original constant temperature, adjusting pH with 1mol/LNaOH, adding another protease, performing enzymolysis at a constant temperature, rapidly heating to inactivate the enzyme for 20min, recovering to room temperature, performing centrifugal treatment at a rotating speed of 8000r/min for 10min, taking supernatant to obtain chrysanthemum polypeptide liquid, and determining the pH value and the constant temperature enzymolysis treatment temperature according to the selected protease. Wherein, 0.5g of chrysanthemum protein is taken and added with 20ml of deionized water, the compound enzyme enzymolysis mode is stepwise enzymolysis, the adding amount ratio of the flavor protease to the papain is 1. And coarsely separating the chrysanthemum crude polypeptide subjected to enzymolysis by adopting a membrane filtration technology, removing macromolecular protein, collecting chrysanthemum polypeptide with the molecular weight of below 3kDa, stirring 2-5% of activated carbon at 55 ℃ for 1-2h, decoloring, concentrating, and vacuum drying to obtain 90.3mg of chrysanthemum polypeptide powder, wherein the molecular cut-off amount of the preset ultrafiltration membrane is 3 kDa.
The chrysanthemum small-molecule polypeptide obtained in the method of the embodiment 1 to 5 is subjected to alpha-glucosidase inhibition activity detection, and the results are shown in table 1:
TABLE 1. Results of alpha-glucosidase inhibitory Activity test
The above table shows that the five polypeptides all have alpha-glucosidase inhibitory activity, wherein the chrysanthemum polypeptide obtained by adding the flavourzyme first and then adding the papain has better alpha-glucosidase inhibitory activity.
The chrysanthemum small-molecule polypeptide obtained by the method in the embodiment 1-2 is subjected to an antioxidant activity test, and the result is shown in fig. 1, so that the survival rate of PC12 cells after oxidative damage by hydrogen peroxide can be remarkably improved by the flavourzyme enzymolysis polypeptide (ZQ-FW-9) and the papain enzymolysis polypeptide (ZQ-MG-5), and the chrysanthemum small-molecule polypeptide has an antioxidant effect, wherein the flavourzyme enzymolysis polypeptide has a stronger antioxidant effect.
Although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, alterations and modifications are possible without departing from the spirit and scope of this disclosure and appended claims, and accordingly, the scope of this disclosure is not limited to the embodiments disclosed.
Claims (5)
1. The protein enzymolysis polypeptide of the golden cypress is characterized in that: the preparation steps are as follows:
(1) Pretreatment of chrysanthemum: crushing and sieving the chrysanthemum raw material to obtain chrysanthemum crushed material:
(2) Extracting chrysanthemum protein: extracting chrysanthemum protein in chrysanthemum powder by an alkali-dissolution acid-precipitation method;
(3) Preparing chrysanthemum polypeptide: preparing the chrysanthemum protein powder prepared in the step (2) into chrysanthemum protein aqueous solution, and adding protease for enzymolysis; centrifuging at 8000r/min for 10-20min after enzymolysis, collecting supernatant, decolorizing, and vacuum freeze drying to obtain flos Chrysanthemi polypeptide powder;
the protease is selected from papain, flavourzyme and complex enzyme consisting of the papain and the flavourzyme;
the alkali-dissolving and acid-precipitating method in the step (2) is to prepare chrysanthemum powder crushed materials into chrysanthemum powder aqueous solution, adjust the pH value to 8.0-9.5, extract at the constant temperature of 50-60 ℃ for 5 hours, and then centrifuge and remove precipitates; regulating the pH value of the supernatant to 3.5-4.5, centrifuging to obtain precipitate, and washing with water to neutrality; carrying out vacuum freeze drying to obtain chrysanthemum protein powder;
the mass concentration of the chrysanthemum powder aqueous solution is 1-4%; adjusting the pH value to 8.0-9.5 by using 0.5-1 mol/L NaOH solution, reacting at the constant temperature of 50-60 ℃ for 5h, centrifuging at 8000r/min for 10-15 min, and discarding the precipitate; adjusting the pH value of the supernatant to 3.5-4.5 by using 0.5-1 mol/L HCl solution, centrifuging for 10-15 min at 8000r/min, keeping the precipitate, and washing with water to be neutral; freeze-drying in vacuum, and pulverizing to obtain flos Chrysanthemi protein powder;
the addition amount of the protease in the step (3) is 10000-12000U/g; when the protease is a compound enzyme consisting of papain and flavourzyme, the adding mass ratio of the flavourzyme to the papain is 1-1;
in the step (3), the mass concentration of the chrysanthemum protein substrate is 1-3%;
the pH value is 7.5-8.5 during enzymolysis in the step (3); the enzymolysis temperature is 50-55 ℃; the enzymolysis time is 4-5h; taking enzymolysis liquid with the molecular weight less than 3kDa after ultrafiltration and centrifugation;
the decolorization is adding active carbon with the final concentration of 2-5% by mass, stirring for 1-2h at 55 ℃, and concentrating after decolorization.
2. The canary chrysanthemum protein polypeptide preparation prepared by utilizing the canary chrysanthemum protein enzymolysis polypeptide as claimed in claim 1.
3. The canary chrysanthemum protein polypeptide formulation as claimed in claim 2, wherein: the preparation is granules, tablets, capsules or oral liquid.
4. Use of the golden emperor protease polypeptide of claim 1 in nutritional food, health food, athlete food or cosmetics.
5. Use according to claim 4, characterized in that: the nutritional food is a nutritional food for patients or infants; the health food is provided for the elderly or sub-health people; the nutritional food is a protein beverage; the cosmetic is moisturizer, milk, cream, essence, facial mask, eye cream, foundation liquid and sunscreen cream.
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| CN104152521A (en) * | 2014-08-11 | 2014-11-19 | 李�杰 | Paeonia suffruticosa pollen protein polypeptide and preparation method and application thereof |
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| CN104152521A (en) * | 2014-08-11 | 2014-11-19 | 李�杰 | Paeonia suffruticosa pollen protein polypeptide and preparation method and application thereof |
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