CN109430512B - Preparation method of light-color sunflower seed protein and light-color sunflower seed protein prepared by same - Google Patents
Preparation method of light-color sunflower seed protein and light-color sunflower seed protein prepared by same Download PDFInfo
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- 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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Abstract
The invention provides a preparation method of light-color sunflower seed protein and the light-color sunflower seed protein prepared by the same, wherein the preparation method comprises the following steps: extracting sunflower seed protein S1, mixing sunflower seed meal or sunflower seed cake with NaCl solution, stirring and leaching, centrifuging, removing precipitate, and collecting supernatant to obtain first sunflower seed protein crude extract; a restriction enzyme hydrolysis step S2, adding protease into the first sunflower seed protein crude extract for enzymolysis to obtain a second sunflower seed protein crude extract; an adsorption decoloring step S3, adding adsorption resin into the second sunflower seed protein crude extract, and stirring and adsorbing to obtain a third sunflower seed protein crude extract; and a step S4 of preparing light-color sunflower seed protein, namely standing the third sunflower seed protein crude extract, centrifuging, collecting precipitate, washing the precipitate with water, pulping, and drying to obtain the light-color sunflower seed protein. The sunflower seed protein prepared by the method is light in color and good in performance, and can be applied to the food industry.
Description
Technical Field
The invention relates to the field of agricultural product processing, in particular to the technical field of preparation of light-colored sunflower seed protein.
Background
Sunflower seeds are one of the world's important oil crops, and the global yield reaches 3942 million tons in 2016. The by-product after the sunflower seed oil extraction, namely the sunflower seed meal or the sunflower seed cake, has the protein content of about 40 percent. The sunflower seed protein has less anti-nutrient substance content and no toxic substance, has the protein biological value second to that of the soybean protein, and is a high-quality plant protein resource. The sunflower seed protein contains rich essential amino acids, eight kinds of essential amino acids for human body, only has slightly low content of lysine and tryptophan, the rest is higher than the ideal protein standard provided by the world health organization WHO, and the efficacy ratio of the sunflower seed protein belongs to high-quality protein. The sunflower seed protein is mainly applied to animal feed at present and cannot become a protein resource edible for human beings, because the content of phenolic compounds in sunflower seed meal or sunflower seed cakes is rich, when the sunflower seed protein is extracted under an alkaline condition, phenolic substances are extracted together, the phenolic substances are easily oxidized, the protein is gray, the nutritional value and the functional characteristics are reduced, and the sunflower seed protein cannot be applied to food processing.
Therefore, researchers at home and abroad have made many attempts to produce the light-colored sunflower seed protein without phenol for many years. Common methods include extracting sunflower seed protein with organic solvents or aqueous solutions to reduce the polyphenol content of protein extracted under alkaline conditions; other methods are to exclude oxygen or use antioxidants during alkaline extraction, followed by removal of co-extracted phenolics from the protein extract. However, none of these methods has achieved the desired effect.
Disclosure of Invention
In view of the above problems, the present invention aims to provide a method for preparing light-colored sunflower seed protein and the light-colored sunflower seed protein prepared thereby.
According to the first aspect of the invention, the preparation method of the light-colored sunflower seed protein comprises the following steps: sunflower seed protein extraction step S1: mixing sunflower seed meal or sunflower seed cake with NaCl solution, stirring and leaching, centrifuging, discarding precipitate, and collecting supernatant to obtain a first sunflower seed protein crude extract; restriction step S2: adding protease into the first sunflower seed protein crude extract for enzymolysis, and inactivating enzyme after the enzymolysis is finished to obtain a second sunflower seed protein crude extract; adsorption decoloring step S3: adding adsorption resin into the second sunflower seed protein crude extract, stirring and adsorbing, and then filtering to remove the adsorption resin to obtain a third sunflower seed protein crude extract; preparation of light-colored sunflower seed protein step S4: and standing the third sunflower seed protein crude extract, centrifuging, collecting precipitate, washing and pulping the precipitate, and drying to obtain light-color sunflower seed protein.
Optionally, the sunflower seed meal is defatted sunflower seed meal, and the sunflower seed cake is defatted sunflower seed cake.
Optionally, in the step S1, the concentration of the NaCl solution is 1.3mol/L, the sunflower seed meal or sunflower seed cake is mixed with the NaCl solution according to a mass ratio of 1:10, the pH of the mixed solution is adjusted to 6.0, the mixture is stirred and extracted at room temperature for 60min, after that, centrifugation is performed at a centrifugation rate of 4000r/min for 15min, and after the centrifugation, the first sunflower seed protein crude extract is obtained by discarding the precipitate and collecting the supernatant.
Optionally, the first sunflower seed protein crude extract has a protein concentration of 10% by mass.
Optionally, in step S2, adjusting the pH of the first sunflower seed protein crude extract to 5.0-9.0, adding protease to perform restriction enzyme hydrolysis within 30min at 50 ℃, and inactivating the enzyme for 10min after the completion of the enzymatic hydrolysis to obtain the second sunflower seed protein crude extract.
Optionally, a protease is added for restriction for 10 min.
Optionally, the enzyme deactivation temperature is below 20 ℃.
Optionally, the pH value is 6.0-8.0.
Optionally, the protease is an alkaline protease.
Optionally, in step S2, the addition amount of the protease is 6% (μ L/mg) of the protein content in the first sunflower seed protein crude extract.
Optionally, in step S3, adjusting the pH value of the second sunflower seed protein crude extract to 6.0-8.0, adding macroporous adsorption resin at room temperature, stirring and adsorbing for 90min, and filtering and separating to remove the macroporous adsorption resin after adsorption is finished, so as to obtain the third sunflower seed protein crude extract.
Optionally, the pH is adjusted to 7.0.
Optionally, the macroporous adsorbent resin is an AB-8 type macroporous adsorbent resin.
Optionally, the addition amount of the adsorption resin is 8% to 12% (g/mL) of the second sunflower seed protein crude extract.
Optionally, the addition amount of the adsorption resin is 9% to 11% (g/mL) of the second sunflower seed protein crude extract.
Optionally, in step S4, adjusting the pH of the third sunflower seed protein crude extract to 4.0, standing at room temperature for 20-40 min, then centrifuging for 20min at a centrifugation rate of 4000r/min, collecting the precipitate, adding 10 times of volume of water into the precipitate, washing with water, and performing size mixing to form a uniform dispersion, adjusting the pH of the uniform dispersion to 7.0, and then drying to obtain the light-colored sunflower seed protein.
Optionally, the temperature is 20-25 ℃.
Optionally, the standing time of the third sunflower seed protein crude extract is 30 min.
Optionally, in step S4, the water washing and the slurry mixing are performed by stirring with a homogenizer.
Optionally, in step S4, the drying includes freeze drying or spray drying.
According to a second aspect of the present invention, there is provided a light-colored sunflower seed protein prepared according to the method of any one of the embodiments of the first aspect of the present invention.
Optionally, the protein content of the light-colored sunflower seed protein is 80% by mass or more.
Optionally, the protein content of the light-colored sunflower seed protein is 80-90% by mass.
The technical scheme of the invention has the following advantages:
1) the sunflower seed protein product prepared by the technical scheme of the invention is light in color;
2) the sunflower seed protein product prepared by the technical scheme of the invention has better water absorption, emulsion stability, foamability and the like;
3) the sunflower seed protein product prepared by the technical scheme of the invention has higher protein content, can be directly used as a food additive to be added into common foods such as meat products, flour products, dairy products and the like, can also be used as a human body protein supplement, and has wide application prospect;
4) the technical scheme of the invention has the advantages of low production cost, simple process and the like.
Drawings
Fig. 1 shows a flow chart of a method for preparing light-colored sunflower seed protein according to the invention.
Fig. 2 shows a schematic diagram of the effect of pH of a first sunflower seed protein crude extract according to the invention on product colour.
FIG. 3 shows a schematic representation of the effect of resin addition on product color according to the present invention.
Fig. 4 shows a picture of the product prepared according to the method of the invention and sunflower seed protein powder prepared by a conventional process.
Detailed Description
In the following detailed description of the preferred embodiments of the invention, reference is made to the accompanying drawings that form a part hereof, and in which is shown by way of illustration, specific features of the invention, such that the advantages and features of the invention may be more readily understood and appreciated. The following description is an embodiment of the claimed invention, and other embodiments related to the claims not specifically described also fall within the scope of the claims. Techniques, methods, and apparatus known to those of ordinary skill in the relevant art may not be discussed in detail but are intended to be part of the specification where appropriate.
Fig. 1 shows a flow chart of a method for preparing light-colored sunflower seed protein according to the invention.
As shown in fig. 1, the invention provides a preparation method of light-colored sunflower seed protein, which comprises the following steps: sunflower seed protein extraction step S1: mixing sunflower seed meal or sunflower seed cake with NaCl solution, stirring and leaching, centrifuging, discarding precipitate, and collecting supernatant to obtain a first sunflower seed protein crude extract; restriction step S2: adding protease into the first sunflower seed protein crude extract for enzymolysis, and inactivating enzyme after the enzymolysis is finished to obtain a second sunflower seed protein crude extract; adsorption decoloring step S3: adding adsorption resin into the second sunflower seed protein crude extract, stirring and adsorbing, and then filtering to remove the adsorption resin to obtain a third sunflower seed protein crude extract; preparation of light-colored sunflower seed protein step S4: and standing the third sunflower seed protein crude extract, centrifuging, collecting precipitate, washing and pulping the precipitate, and drying to obtain light-color sunflower seed protein.
The sunflower seed meal or the sunflower seed cake is a byproduct after sunflower seed oil extraction and has the gray or gray black characteristic of sunflower seeds.
The restriction enzyme is to decompose the protein to a low degree of hydrolysis in a short time, not to decompose the protein into peptides and amino acids. The restriction time may be within 30 min.
The sunflower seed protein extracted by the conventional protein extraction method, namely the alkali-soluble acid precipitation method is dark green in color, and the application of the sunflower seed protein in food is seriously influenced. The reason why the sunflower seed protein is dark green may be due to chlorogenic acid. Chlorogenic acid is combined with sunflower seed protein in a form of being oxidized to form monomer quinone, dimer or polymer with higher molecular weight is formed, and is covalently or non-covalently combined with the sunflower seed protein in an enzymatic oxidation or non-enzymatic oxidation mode, so that the conventional method for directly adsorbing and decoloring hardly achieves effective decoloring effect. The restriction enzyme hydrolysis adopted by the invention aims to firstly open or mostly separate the combination of sunflower seed protein and chlorogenic acid, and then adopts adsorption resin to adsorb the chlorogenic acid, namely, the method of using restriction enzyme hydrolysis combined adsorption resin to decolorize.
According to one embodiment of the invention, sunflower seed protein is restricted by alkaline protease for 10min, and then is decolored by AB-8 type macroporous adsorption resin, so that compared with the decoloration effect which is realized by directly using macroporous adsorption resin without restriction enzyme, the decoloration effect is obviously improved. The whiteness value of the sunflower seed protein which is not subjected to the restriction enzyme hydrolysis after being directly decolored is 76.9, and the whiteness value of the sunflower seed protein which is subjected to the decoloration after being subjected to the restriction enzyme hydrolysis is improved to 82.5, which shows that the restriction enzyme hydrolysis plays an important role.
The enzyme deactivation is carried out after the restriction enzyme hydrolysis, so as to reduce or eliminate the activity of protease used for restriction enzyme hydrolysis and ensure the stability of the final light-colored sunflower seed protein product.
The adsorption resin refers to a high molecular polymer, and can be used for removing organic matters in wastewater, decoloring, separating and refining natural products and biochemical products, and the like. The adsorption resin has a plurality of varieties, and the change of the monomer and the change of the functional group on the monomer can endow the resin with various special properties. The adsorption resin is a resin adsorbent which has the characteristic of adsorption and has a porous three-dimensional structure.
The light-color sunflower seed protein obtained according to the technical scheme of the invention can be light-color sunflower seed protein powder.
Optionally, the sunflower seed meal is defatted sunflower seed meal, and the sunflower seed cake is defatted sunflower seed cake. Optionally, the defatted sunflower seed meal can be low-temperature defatted sunflower seed meal, and the defatted sunflower seed cake can be low-temperature defatted sunflower seed cake. The sunflower seed meal degreased at low temperature has low protein denaturation degree and good solubility, and can be effectively separated and extracted. If the defatted protein is defatted at high temperature, the protein is denatured to a high degree and is difficult to extract.
Optionally, in the step S1, the concentration of the NaCl solution is 1.3mol/L, the sunflower seed meal or sunflower seed cake is mixed with the NaCl solution according to a mass ratio of 1:10, the pH of the mixed solution is adjusted to 6.0, the mixture is stirred and extracted at room temperature for 60min, after that, centrifugation is performed at a centrifugation rate of 4000r/min for 15min, and after the centrifugation, the first sunflower seed protein crude extract is obtained by discarding the precipitate and collecting the supernatant.
According to the technical scheme, the concentration of NaCl is increased under a weak acid condition to extract the sunflower seed protein, so that the defects of dark color, low nutritional value and poor functional performance of the sunflower seed protein caused by extraction of phenolic substances together when the sunflower seed protein is extracted under an alkaline condition can be overcome.
Optionally, the first sunflower seed protein crude extract has a protein concentration of 10% by mass.
According to the technical scheme of the invention, the first sunflower seed protein crude extract extracted in the sunflower seed protein extraction step S1 has a protein concentration of 10% by mass through determination. The method for measuring the content of the protein is a micro Kjeldahl method, namely, a sample and concentrated sulfuric acid are heated together, nitrogenous organic matters are decomposed to generate ammonia, and the ammonia reacts with sulfuric acid to become ammonium sulfate. Decomposing the sample by alkalifying with strong alkali to release ammonia, steaming the ammonia into the acid solution with steam, and calculating the nitrogen content of the sample according to the degree of neutralization of the acid solution; kjeldahl determination of nitrogen by total nitrogen content in the sample and the product of the corresponding protein coefficient to determine the protein content, this method is called crude protein content.
Optionally, in step S2, adjusting the pH of the first sunflower seed protein crude extract to 5.0-9.0, adding protease to perform restriction enzyme hydrolysis within 30min at 50 ℃, and inactivating the enzyme for 10min after the completion of the enzymatic hydrolysis to obtain the second sunflower seed protein crude extract.
The adjustment of the pH of the first sunflower seed protein crude extract has a great influence on the whiteness value of the sunflower seed protein of the present invention, and the influence trend is shown in fig. 2, which will be specifically described below with reference to specific embodiments, and will not be described herein again.
The time for adding protease for restriction is controlled within 30min, and the longer the enzymolysis time is, the more the protein is decomposed into peptides and amino acids.
Preferably, the protease is added for 10min of restriction.
According to a preferred embodiment of the present invention, the time for adding protease for restriction is controlled to 10min, so that the restriction can be achieved, the time is saved, and the risk of complete enzymolysis is avoided.
According to one embodiment of the present invention, the temperature at which the enzyme deactivation is carried out after the completion of the restriction enzyme hydrolysis is 20 ℃ or lower. The enzyme deactivation method adopted by the embodiment is low-temperature enzyme deactivation, namely, the temperature of the restriction enzyme solution is reduced to be lower than 20 ℃ for enzyme deactivation, and the enzyme deactivation can keep the solubility of protein in the temperature range and is beneficial to the next step of adsorption decoloration compared with high-temperature enzyme deactivation. The enzyme inactivating temperature is below 20 ℃ and below 0 ℃.
According to an embodiment of the present invention, the pH of the first sunflower seed protein crude extract may be adjusted to 6.0 to 8.0. Under the condition, the prepared product has higher whiteness value.
Alternatively, the protease may be an alkaline protease. The alkaline protease is an enzyme capable of hydrolyzing protein peptide bonds under alkaline conditions, and the optimal pH is within the range of 9-11. The method is widely applied to industries such as detergent, food, medical treatment, brewing, silk and tanning.
According to an embodiment of the invention, sunflower seed protein can be restricted and decolorized by protease such as flavourzyme and neutral protease. Preferably, the alkaline protease is used for decolorization after enzymolysis, and the decolorization effect is optimal, so the alkaline protease can be selected as a tool enzyme in the technical scheme of the invention.
Optionally, in step S2, the addition amount of the protease is 6% (μ L/mg) of the protein content in the first sunflower seed protein crude extract.
The addition amount of the protease is determined according to the mass of the protein in the first sunflower seed protein crude extract, wherein mg is the mass unit of the protein in the first sunflower seed protein crude extract, and μ L is the volume unit of the protease. Namely, 6 mu L of the protease is added for every 100mg of the protein of the first sunflower seed protein crude extract. The protein content of the first sunflower seed protein crude extract is measured by a titration method.
Optionally, in step S3, adjusting the pH value of the second sunflower seed protein crude extract to 6.0-8.0, adding macroporous adsorption resin at room temperature, stirring and adsorbing for 90min, and filtering and separating to remove the macroporous adsorption resin after adsorption is finished, so as to obtain the third sunflower seed protein crude extract.
The pH value of the sunflower seed protein crude extract is reduced due to the restriction enzymolysis process, and the secondary adjustment is to ensure that the pH value is kept in a reasonable range before the macroporous absorption resin is absorbed.
The macroporous adsorption resin is a macromolecular adsorption resin which does not contain exchange groups and has a macroporous structure, has a good macroporous network structure and a larger specific surface area, can selectively adsorb organic matters in an aqueous solution through physics, is a novel organic polymer adsorbent developed in the 60 th century, and has been widely applied to the fields of environmental protection, food, medicine and the like. The macroporous adsorbent resin is generally white spherical particles with the granularity of 20-60 meshes. The macro beads of the macroporous adsorbent resin are composed of a plurality of micro beads with pores among each other.
Optionally, the pH is adjusted to 7.0. When the pH is adjusted to 7.0, the adsorption effect is best, so that the decolorization effect is best, and the whiteness value of the prepared product is highest, namely the color is lightest.
Alternatively, the macroporous adsorbent resin may be an AB-8 type macroporous adsorbent resin.
The AB-8 type macroporous adsorption resin is polystyrene type weak polar adsorption resin, the surface of the resin has a certain ester group, the hydrophilicity is improved, but the adsorption mechanism is still hydrophobic adsorption. The resin has large specific surface area and pore size, is suitable for adsorbing various components with certain hydrophobicity, and has large adsorption capacity, easy elution and good adsorption dynamic performance. Is stable to heat, organic solvents and acid and alkali under common use conditions, so the service life is longer. It can separate general components from protein, saccharide, inorganic acid, alkali, salt and hydrophilic micromolecular organic matter without adsorption.
Optionally, the addition amount of the adsorption resin is 8% to 12% (g/mL) of the second sunflower seed protein crude extract.
And determining the addition amount of the adsorption resin according to the volume of the second sunflower seed protein crude extract. The mL is the volume unit of the second sunflower seed protein crude extract, and the g is the mass unit of the adsorption resin. Namely, 8g to 12g of the adsorption resin is correspondingly added into each 100mL of the second sunflower seed protein crude extract.
The addition amount of the adsorption resin has a great influence on the adsorption effect, so that the whiteness value of the light-colored sunflower seed protein product of the invention is finally influenced, the specific influence trend can be shown in fig. 3, the specific description will be described in detail in the following with reference to fig. 3, and the detailed description is omitted here.
Optionally, the addition amount of the adsorption resin is 9% to 11% (g/mL) of the second sunflower seed protein crude extract.
When the addition amount of the adsorption resin is 9-11% (g/mL) of the second sunflower seed protein crude extract, the obtained product has a good whiteness value, and the addition amount of the adsorption resin can be controlled, so that the cost is saved.
Optionally, in step S4, adjusting the pH of the third sunflower seed protein crude extract to 4.0, standing at room temperature for 20-40 min, then centrifuging for 20min at a centrifugation rate of 4000r/min, collecting the precipitate, adding 10 times of volume of water into the precipitate, washing with water, and performing size mixing to form a uniform dispersion, adjusting the pH of the uniform dispersion to 7.0, and then drying to obtain the light-colored sunflower seed protein.
The room temperature refers to an indoor temperature. Generally, the room temperature may be 15 to 30 ℃. The pH of 4 is the isoelectric point of sunflower seed protein. The isoelectric point of a protein is the isoelectric point of the protein when the protein solution is in a certain pH solution, the protein has equal tendency to dissociate into positive and negative ions, namely, the protein becomes a zwitter ion, the net charge is zero, and the pH of the solution is called the isoelectric point of the protein. And standing the third sunflower seed protein crude extract under an acidic condition for acid precipitation, centrifuging, collecting the precipitate and the like after the precipitate is completely formed. It is right the sediment is washed, washes away the acidic material in the sediment, the size mixing is so that form homodisperse liquid, can obtain likelinely after drying light colour sunflower seed protein to carry out subsequent processing.
Alternatively, the temperature may be 20 to 25 ℃. The third sunflower seed protein crude extract can be kept standing at the temperature of 20-25 ℃.
Optionally, the standing time of the third sunflower seed protein crude extract can be 30 min. When the standing time of the third sunflower seed protein crude extract is 30min, complete precipitation can be ensured under general conditions.
Optionally, in step S4, the water washing and the slurry mixing are performed by stirring with a homogenizer.
The homogenizer can adopt a stainless steel system, can effectively separate the surface of a protective sample from a microorganism uniform sample contained in the sample, the sample is arranged in a disposable sterile homogenizing bag and is not contacted with an instrument, and the homogenizer has the characteristics of soft homogenization, no pollution, no damage, no temperature rise, no need of sterilization treatment and no need of washing and brushing vessels, and meets the requirements of rapidness, accurate result and good repeatability. Is widely applied to the industries of biology, medicine, food and the like.
The homogenizer can homogenize the precipitate to help form a uniform dispersion.
Alternatively, in step S4, the drying means may include freeze drying or spray drying.
The drying mode can comprise freeze drying or spray drying, and other drying modes can also be adopted.
The freeze drying is also called sublimation drying. Drying processes in which an aqueous material is frozen below freezing to convert water to ice and then the ice is removed by converting it to a vapor under a relatively high vacuum. The material can be frozen in a freezing device and then dried. But can also be frozen directly in the drying chamber by rapidly drawing a vacuum. The water vapor formed by sublimation is removed by a condenser. The heat of vaporization required during sublimation is typically supplied by thermal radiation. The material dried by the method can keep the original chemical composition and physical properties, and the heat consumption is less than that of other drying methods.
The spray drying is a method for applying systematic technology to material drying. After the thin material is atomized in the drying chamber, the water content is quickly vaporized in the contact of the thin material and hot air, and then the dried product is obtained. The method can directly dry the solution or emulsion into powder or granular product, and can omit the procedures of evaporation, pulverization, etc. The method has the advantages of rapid drying process, direct drying into powder, certain negative pressure in the drying chamber, guaranteed sanitary conditions in production, no dust flying in the workshop, high product purity, high production capacity and high product quality. The spraying amount per hour can reach hundreds of tons, which is one of the larger treatment capacity of the dryer.
According to a second aspect of the present invention, there is provided a light-colored sunflower seed protein prepared according to the method of any one of the embodiments of the first aspect of the present invention.
Optionally, the protein content of the light-colored sunflower seed protein is 80% by mass or more.
Optionally, the protein content of the light-colored sunflower seed protein is 80-90% by mass.
The protein content of the light-colored sunflower seed protein prepared by the method can reach more than 80% by mass. The protein content is higher, and the application range is wider. The method for measuring the protein content is a micro Kjeldahl method, which has been described in detail above and will not be described herein again.
The following description will be given with reference to specific examples.
Example 1: the low-temperature degreased sunflower seed meal is prepared by the following steps of 1:10, mixing the sunflower seed protein with 1.3mol/L NaCl solution, extracting for 60min at pH 6.0, centrifuging for 15min at 4000r/min, removing the precipitate, and collecting the supernatant to obtain a first sunflower seed protein crude extract. The protein concentration of the first sunflower seed protein crude extract was 10% by mass. Taking the first sunflower seed protein crude extract, adding 6% (v/m) alkaline protease at 50 deg.C and pH of 7.0, and performing restriction hydrolysis for 10 min. Inactivating enzyme at 20 deg.C for 10min after enzymolysis to obtain a second sunflower seed protein crude extract. And (3) adjusting the pH value of the second sunflower seed protein crude extract to 7.0, adding 10% (g/mL) of AB-8 macroporous adsorption resin at room temperature for adsorption, stirring and adsorbing for 90min, and filtering, separating and removing the macroporous adsorption resin to obtain a third sunflower seed protein crude extract. Adjusting the pH value of the third sunflower seed protein crude extract to 4.0, standing for 30min at room temperature for acid precipitation, centrifuging at 4000r/min for 20min, collecting precipitate, adding 10 times of water, stirring with a homogenizer to obtain uniform dispersion liquid, adjusting the pH value to 7.0, and freeze-drying to obtain light-colored sunflower seed protein. The protein content of the light-colored sunflower seed protein is more than 90%.
Example 2: the low-temperature degreased sunflower seed cake is prepared by the following steps of 1:10, mixing the sunflower seed protein with 1.3mol/L NaCl solution, extracting for 60min at pH 6.0, centrifuging for 15min at 4000r/min, removing the precipitate, and collecting the supernatant to obtain a first sunflower seed protein crude extract. Taking the first sunflower seed protein crude extract, adding 6% (v/m) protease at 50 deg.C and pH 6.0, and performing restriction for 30 min. And (3) inactivating enzyme for 10min at 10 ℃ after enzymolysis to obtain a second sunflower seed protein crude extract. Adjusting the pH value of the second sunflower seed protein crude extract to 6.0, adding 9% (g/mL) of AB-8 macroporous adsorption resin for adsorption at room temperature, stirring and adsorbing for 90min, and filtering, separating and removing the macroporous adsorption resin to obtain a third sunflower seed protein crude extract. Adjusting the pH value of the third sunflower seed protein crude extract to 4.0, standing for 20min at 25 ℃ for acid precipitation, centrifuging for 20min at 4000r/min, collecting precipitate, adding 10 times of water, stirring by a homogenizer to obtain a uniform dispersion liquid, adjusting the pH value to 7.0, and spray-drying to obtain light-colored sunflower seed protein. The protein content of the light-colored sunflower seed protein was 85%.
Example 3: mixing sunflower seed meal according to the proportion of 1:10, mixing the sunflower seed protein with 1.3mol/L NaCl solution, extracting for 60min at pH 6.0, centrifuging for 15min at 4000r/min, removing the precipitate, and collecting the supernatant to obtain a first sunflower seed protein crude extract. Taking the first sunflower seed protein crude extract, adding alkaline protease at 50 deg.C and pH of 8.0, and performing restriction hydrolysis for 20 min. And (3) inactivating enzyme for 10min at 0 ℃ after enzymolysis to obtain a second sunflower seed protein crude extract. And (3) adjusting the pH value of the second sunflower seed protein crude extract to 8.0, adding 11% (g/mL) of macroporous adsorption resin at room temperature for adsorption, stirring and adsorbing for 90min, and filtering and separating to remove the macroporous adsorption resin to obtain a third sunflower seed protein crude extract. Adjusting pH of the third sunflower seed protein crude extract to 4.0, standing at 20 deg.C for 40min for acid precipitation, centrifuging at 4000r/min for 20min, collecting precipitate, adding 10 times volume of water, stirring with a homogenizer to obtain uniform dispersion, adjusting pH to 7.0, and freeze drying to obtain light-colored sunflower seed protein. The protein content of the light-colored sunflower seed protein was 90%.
Example 4: mixing sunflower seed cakes according to the weight ratio of 1:10, mixing the sunflower seed protein with 1.3mol/L NaCl solution, extracting for 60min at pH 6.0, centrifuging for 15min at 4000r/min, removing the precipitate, and collecting the supernatant to obtain a first sunflower seed protein crude extract. The protein concentration of the first sunflower seed protein crude extract was 10% by mass. Adding protease into the first sunflower seed protein crude extract at 50 deg.C and pH of 5.0, and performing restriction hydrolysis for 30 min. Inactivating enzyme at-5 deg.C for 10min after enzymolysis to obtain a second sunflower seed protein crude extract. And (3) adjusting the pH value of the second sunflower seed protein crude extract to 7.5, adding 8% (g/mL) of AB-8 macroporous adsorption resin at room temperature for adsorption, stirring and adsorbing for 90min, and filtering, separating and removing the macroporous adsorption resin to obtain a third sunflower seed protein crude extract. Adjusting the pH value of the third sunflower seed protein crude extract to 4.0, standing for 25min at room temperature for acid precipitation, centrifuging at 4000r/min for 20min, collecting precipitate, adding 10 times of water, stirring with a homogenizer to obtain uniform dispersion liquid, adjusting the pH value to 7.0, and spray drying to obtain light-colored sunflower seed protein. The protein content of the light-colored sunflower seed protein was 80%.
Example 5: mixing sunflower seed cakes according to the weight ratio of 1:10, mixing the sunflower seed protein with 1.3mol/L NaCl solution, extracting for 60min at pH 6.0, centrifuging for 15min at 4000r/min, removing the precipitate, and collecting the supernatant to obtain a first sunflower seed protein crude extract. Taking the first sunflower seed protein crude extract, adding alkaline protease at 50 deg.C and pH of 9.0, and performing restriction hydrolysis for 15 min. And (3) inactivating enzyme for 10min at 5 ℃ after enzymolysis to obtain a second sunflower seed protein crude extract. And adjusting the pH value of the second sunflower seed protein crude extract to 6.5, adding 12% (g/mL) of macroporous adsorption resin at room temperature for adsorption, stirring and adsorbing for 90min, and filtering and separating to remove the macroporous adsorption resin to obtain a third sunflower seed protein crude extract. Adjusting the pH value of the third sunflower seed protein crude extract to 4.0, standing at 22 ℃ for 35min for acid precipitation, centrifuging at 4000r/min for 20min, collecting precipitate, adding 10 times of water by volume, stirring to obtain uniform dispersion liquid, adjusting the pH value to 7.0, and spray drying to obtain light-colored sunflower seed protein. The protein content of the light-colored sunflower seed protein was 88%.
Example 6: mixing sunflower seed meal with NaCl solution, stirring and leaching, centrifuging, discarding precipitate, and collecting supernatant to obtain sunflower seed protein crude extract. Adding protease into the crude extract for enzymolysis, inactivating enzyme after the enzymolysis is finished, then adding adsorbent resin, stirring for adsorption, and filtering to remove the adsorbent resin. And standing, centrifuging and collecting the precipitate of the coarse extract subjected to adsorption and decoloration, washing and pulping the precipitate, and drying to obtain the light-color sunflower seed protein.
Fig. 2 shows a schematic diagram of the effect of pH of a first sunflower seed protein crude extract according to the invention on product colour.
The data in fig. 2 are based on the basic experimental conditions of example 1, and the effect of the pH of the first crude sunflower seed protein extract on the final product was determined by changing the pH of the first crude sunflower seed protein extract only in the restriction step to ensure the reliability of the experimental data. As shown in fig. 2, the pH of the first sunflower seed protein crude extract had a significant effect on the product whiteness value. L was 81.9 at pH 5, 84.9 at pH 6, 86.4 at pH 7, 84.6 at pH 8, and 82.5 at pH9. At pH values below 6 or above 8, the L value of the product decreased significantly, whereas at pH 7, L value was maximal. The L value refers to the whiteness value, and the bigger the L value is, the whiter the product is. Therefore, the pH is preferably adjusted to be 6-8, and the optimal scheme is 7.
FIG. 3 shows a schematic representation of the effect of resin addition on product color according to the present invention.
The data in fig. 3 are obtained by changing the addition amount of the macroporous adsorbent resin only under the basic experimental conditions of example 1 to ensure the reliability of the experimental data, thereby determining the influence of the addition amount of the macroporous resin on the final product. As shown in fig. 3, the amount of macroporous resin added has a significant effect on the whiteness value of the product. The value of L is 83.7 when the addition amount of the macroporous resin is 8%, 84.9 when the addition amount of the macroporous resin is 9%, 86.4 when the addition amount of the macroporous resin is 10%, 85.9 when the addition amount of the macroporous resin is 11%, and 85.7 when the addition amount of the macroporous resin is 12%. The L value refers to the whiteness value, and the bigger the L value is, the whiter the product is. When the addition amount of the macroporous resin is 8 percent, the L value is obviously reduced. When the addition amount of the macroporous resin is 12%, the L value is higher, but the L value is not obviously increased, and the addition amount of the macroporous resin is increased, so that unnecessary cost is increased. The influence effect of the addition amount of the macroporous resin on the product and the material cost are comprehensively considered, and the addition amount of the macroporous adsorption resin is preferably 9-11%, and is preferably 10%.
The light-colored sunflower seed protein product prepared by the technical scheme of the example 1 is used as a sample for performance test, and is compared with test data of sunflower seed protein prepared by the traditional process, the test data is shown in the table 1, and a product chromaticity comparison graph is shown in fig. 4. The following is a detailed description with reference to table 1 and fig. 4.
TABLE 1
As can be seen from table 1, the light-colored sunflower seed protein produced by the present invention has a color difference value of L63.1, a 3.9, b 12.3 to L86.4, a 2.6, b 10.7, compared to the sunflower seed protein produced by the conventional process. The L value is a white value, and the larger the L value, the whiter the color, the pure white when the L value is 100, and the pure black when the L value is 0. The a value refers to a red-green value, and when the a value is a positive number, the color is represented as red; when the value of a is negative, the representative color is green; and a represents neutral when the value is 0. The b value refers to a yellow-blue color value, and when the b value is a positive number, the color is yellow; when the b value is negative, the color is blue; b represents neutral when the value is 0. Namely, the light-colored sunflower seed protein prepared by the technical scheme of the invention has larger L value, a value and b value which are all closer to 0, and the light-colored sunflower seed protein is lighter in color. The color difference data in this experiment were tested using the CR-10Plus color difference meter of Konika minolta, Japan.
Fig. 4 shows a picture of the product prepared according to the method of the invention and sunflower seed protein powder prepared by a conventional process. Wherein, a in fig. 4 is sunflower seed protein powder prepared by a traditional process, and b in fig. 4 is light-color sunflower seed protein powder prepared according to the technical scheme in the embodiment 1 of the invention.
As shown in fig. 4, the light color sunflower seed protein powder prepared by the technical scheme of the invention has a color obviously lighter than that of sunflower seed protein powder prepared by the traditional process. The results of the color difference value test are shown in table 1, namely, the L value of the sunflower seed protein prepared by the traditional process is 63.1, and the L value of the light-color sunflower seed protein of the invention is 86.4, and the change is obvious. Therefore, the sunflower seed protein prepared according to the technical scheme of the invention is light in color, whether from visual observation or experimental test data display.
As shown in table 1, the water absorption of the light-colored sunflower seed protein prepared by the technical scheme of the invention is 223%, while the water absorption of the sunflower seed protein extracted by the traditional process is 190%, and the water absorption of the light-colored sunflower seed protein prepared by the technical scheme of the invention is improved by 17%. The water absorption refers to the moisture content of the dried protein at which the moisture equilibrium is reached in a certain humidity. The water absorption and water retention of the vegetable protein are commonly used in meat product processing, so that meat juice can be kept, and good mouthfeel and flavor of the meat product can be kept; when the bread additive is used in baked food, the bread yield can be increased, the shelf life of bread can be prolonged, the shrinkage of cakes can be reduced, and the shelf life of the cakes can be prolonged. The water absorption of the light-color sunflower seed protein prepared by the technical scheme of the invention is obviously higher than that of the traditional process, so that the light-color sunflower seed protein prepared by the technical scheme of the invention has a prospect of being applied to the food industry.
The water absorption was measured by putting a sample on a sand core filter, immersing the sample in an excessive amount of distilled water for 1 hour, filtering out water by natural sedimentation, and weighing the sample so that the water absorption was defined as the mass of protein after immersion/dry weight of the sample × 100%.
As shown in table 1, the emulsibility of sunflower seed protein extracted by the conventional process is 51.12%, and the emulsion stability is 47.9%, whereas the emulsibility of the light-colored sunflower seed protein prepared by the technical scheme of the present invention is 47.9%, and the emulsion stability is 187.0%. Although the emulsifiability is reduced by 6%, the emulsion stability is increased by 290%. The emulsifiability and emulsion stability refer to the characteristic that protein can help oil drops form emulsion in water and keep the emulsion stable. Emulsification of vegetable proteins has important applications in many food processes, such as coffee cream, lemon mayonnaise, salad dressings, and ground meat products.
The emulsifying property was measured by weighing 2.5g of a protein sample, dispersing it in 50ml of water, adding 50ml of salad oil, emulsifying at 200r/min for 1min, centrifuging for 5min, and taking out the emulsified layer height, i.e., the emulsified layer height/total liquid height in the centrifuge tube × 100%.
As shown in table 1, the foamability of sunflower seed protein extracted by the conventional process is 33.3%, and the foam stability is 75%, while the foamability of the light-colored sunflower seed protein prepared by the technical scheme of the present invention is 36.6%, and the foam stability is 82.9%. The foaming property is increased by 10 percent, and the foam stability is increased by 10.5 percent. Foaming is a property of a protein sol that when subjected to rapid mechanical stirring, a large amount of air is mixed into the protein sol to form a large amount of small bubbles surrounded by the protein solution. The vegetable protein with good foamability can replace the traditional foaming agent-egg protein, gelatin and the like, is widely applied to foods such as cakes, candies, western cakes, cold drinks and the like, and endows the foods with loose structures and good mouthfeel.
The foaming property was measured by taking 100ml of a 3% protein solution, stirring the solution at 1000 rpm for 2min in a high-speed homogenizer, and measuring the foam volume (ml), wherein the foaming property was (foam volume-100)/100 × 100%.
In conclusion, the light-color sunflower seed protein prepared according to the technical scheme of the invention has light color, and functional properties such as water absorption, emulsion stability, foamability and the like are greatly improved compared with the traditional process, and the protein content is high, so that the light-color sunflower seed protein can be directly used as a food additive to be added into common foods such as meat products, dairy products, flour products and the like, can also be used as a human protein supplement, and has wide application prospect. Meanwhile, the technical scheme of the invention has the advantages of low production cost, simple process and high utilization rate of sunflower seed meal or sunflower seed cake, and widens the range of deep processing application of sunflower seed protein.
The technical scheme of the invention has the following advantages:
1) the sunflower seed protein product prepared by the technical scheme of the invention is light in color;
2) the sunflower seed protein product prepared by the technical scheme of the invention has better water absorption, emulsion stability, foamability and the like;
3) the sunflower seed protein product prepared by the technical scheme of the invention has higher protein content, can be directly used as a food additive to be added into common foods such as meat products, flour products, dairy products and the like, can also be used as a human body protein supplement, and has wide application prospect;
4) the technical scheme of the invention has the advantages of low production cost, simple process and the like.
It should be noted that the above-mentioned embodiments illustrate rather than limit the invention, and that those skilled in the art will be able to design alternative embodiments without departing from the scope of the appended claims. In the claims, any reference signs placed between parentheses shall not be construed as limiting the claim.
Claims (19)
1. A preparation method of light-colored sunflower seed protein comprises the following steps:
sunflower seed protein extraction step S1:
mixing sunflower seed meal or sunflower seed cake with NaCl solution, stirring and leaching, centrifuging, discarding precipitate, and collecting supernatant to obtain a first sunflower seed protein crude extract;
restriction step S2:
adjusting the pH value of the first sunflower seed protein crude extract to 5.0-9.0, adding protease into the first sunflower seed protein crude extract at the temperature of 50 ℃ for carrying out restriction enzymolysis for less than 30min, and inactivating the enzyme for 10min after the enzymolysis is finished to obtain a second sunflower seed protein crude extract, wherein the protease is alkaline protease;
adsorption decoloring step S3:
adding an adsorption resin into the second sunflower seed protein crude extract, stirring and adsorbing, and then filtering to remove the adsorption resin to obtain a third sunflower seed protein crude extract, wherein the addition amount of the adsorption resin is 8-12% g/mL of the second sunflower seed protein crude extract;
preparation of light-colored sunflower seed protein step S4:
and standing the third sunflower seed protein crude extract, centrifuging, collecting precipitate, washing and pulping the precipitate, and drying to obtain light-color sunflower seed protein.
2. The method of preparing light-colored sunflower seed protein according to claim 1, wherein the sunflower seed meal is defatted sunflower seed meal and the sunflower seed cake is defatted sunflower seed cake.
3. The method for preparing light-colored sunflower seed protein according to claim 1, wherein the concentration of the NaCl solution in step S1 is 1.3mol/L, the sunflower seed meal or sunflower seed cake is mixed with the NaCl solution according to the mass ratio of 1:10, the pH value of the mixed solution is adjusted to 6.0, the mixed solution is stirred and extracted for 60min at room temperature, centrifugation is performed after the completion, the centrifugation speed is 4000r/min, the centrifugation time is 15min, and the first sunflower seed protein crude extract is obtained by discarding the precipitate and collecting the supernatant after the centrifugation.
4. The method for preparing light-colored sunflower seed protein in claim 1, wherein the protein concentration of the first sunflower seed protein crude extract is 10% by mass.
5. The method of claim 1, wherein the protease is added for 10 min.
6. The method for preparing light-colored sunflower seed protein according to claim 1, wherein the pH is 6.0-8.0.
7. The method for preparing light-colored sunflower seed protein in claim 1, wherein in step S2, the protease is added in an amount of 6% μ L/mg of the protein content of the first crude sunflower seed protein extract.
8. The method for preparing light-colored sunflower seed protein according to claim 1, wherein in step S3, the pH of the second crude sunflower seed protein extract is adjusted to 6.0-8.0, macroporous adsorbent resin is added at room temperature, stirring and adsorption are performed for 90min, and after adsorption, the macroporous adsorbent resin is removed by filtration and separation, so as to obtain the third crude sunflower seed protein extract.
9. The process for preparing light-colored sunflower seed protein according to claim 8, wherein the pH is adjusted to 7.0.
10. The method for preparing light-colored sunflower seed protein according to claim 8, wherein the macroporous adsorbent resin is AB-8 type macroporous adsorbent resin.
11. The method for preparing light-colored sunflower seed protein according to claim 1, wherein the addition amount of the adsorption resin is 9-11% g/mL of the second sunflower seed protein crude extract.
12. The method for preparing light-colored sunflower seed protein according to claim 1, wherein in step S4, the pH of the third crude sunflower seed protein extract is adjusted to 4.0, the third crude sunflower seed protein extract is allowed to stand at room temperature for 20-40 min, then centrifuged for 20min at a centrifugation rate of 4000r/min, the precipitate is collected, 10 times the volume of water is added to the precipitate for water washing and size mixing to form a uniform dispersion, the pH of the uniform dispersion is adjusted to 7.0, and then the uniform dispersion is dried to obtain the light-colored sunflower seed protein.
13. The method for preparing light-colored sunflower seed protein according to claim 12, wherein the room temperature is 20-25 ℃.
14. The method for preparing light-colored sunflower seed protein in claim 12, wherein the standing time of the third sunflower seed protein crude extract is 30 min.
15. The method of claim 1, wherein the washing and slurrying are performed by stirring with a homogenizer in step S4.
16. The process of claim 1, wherein the drying step S4 comprises freeze drying or spray drying.
17. A light colored sunflower seed protein produced by the method of any one of claims 1-16.
18. The light-colored sunflower seed protein of claim 17 having a protein content of greater than 80% by mass.
19. The light-colored sunflower seed protein of claim 17 having a protein content of 80% to 90% by mass.
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