CN110810619A - Extraction process for extracting selenoprotein from soybeans - Google Patents
Extraction process for extracting selenoprotein from soybeans Download PDFInfo
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- CN110810619A CN110810619A CN201911092364.6A CN201911092364A CN110810619A CN 110810619 A CN110810619 A CN 110810619A CN 201911092364 A CN201911092364 A CN 201911092364A CN 110810619 A CN110810619 A CN 110810619A
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- selenoprotein
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
- A23J1/148—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds by treatment involving enzymes or microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
- A23J1/146—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds by using wave energy or electric current
Abstract
The invention discloses an extraction process for extracting selenoprotein from soybeans, which comprises the steps of 1) material selection, 2) deoiling, 3) crushing, 4) alkaline leaching, 5) acid precipitation, 6) filtration, neutralization and bleaching, 7) enzymatic modification, and 8) filtration and drying. The invention carries out twice leaching and twice enzymolysis on the selenoprotein emulsion, the extraction is more thorough, the purity of the selenoprotein is higher, no peculiar smell is generated, and the technical problems of various extraction procedures, inconvenient use and low efficiency in the prior art are effectively solved.
Description
Technical Field
The invention belongs to the technical field of biological fermentation extraction, and particularly relates to an extraction process for extracting selenoprotein from soybeans.
Background
According to the investigation of the dietary structure of urban residents in China, the daily intake of adult selenium is in the range of 20-30 mu g, which is seriously lower than the average intake of 50 mu g specified by the national academy of nutrition and national standards, and the daily intake of selenium of people in sub-health state or in sick state is recommended to be 60 mu g/d. Therefore, scientific selenium supplement is imperative. The selenoprotein can be used as additive for food, medicine and health product, and has antioxidant, antiaging, antitumor, radioprotective, and cardiovascular and cerebrovascular diseases preventing effects. The soybeans planted in selenium-rich areas such as the northwest of Hu and the south of Shanxi in China contain a large amount of selenium-rich protein.
When the selenoprotein is extracted, the steps mainly comprise crushing, sieving, alkaline extraction, acidic precipitation and atomization drying. In the extraction process, the acid consumption and alkali consumption are large, the product yield is low, and the use is very inconvenient.
The soybean selenoprotein primary extract powder which is not technically improved has a strange taste and heavy fishy smell, and the taste of people is influenced in the eating process. The selenium protein powder can destroy the structure of soybean protein during processing if solvent with too high temperature is used, thereby reducing the nutritional value. Meanwhile, the substances such as soybean isoflavone and the like contained in the selenoprotein have certain peculiar smell in the eating process.
Disclosure of Invention
The invention aims to solve the technical problems of low efficiency and inconvenience in extracting selenoprotein from soybeans in the prior art, and provides an extraction process for extracting selenoprotein from soybeans so as to solve the defects in the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme:
an extraction process for extracting selenoprotein from soybeans comprises the steps of 1) material selection, 2) deoiling, 3) crushing, 4) alkaline leaching, 5) acid precipitation, 6) filtration, neutralization and bleaching, 7) enzymatic modification and 8) filtration and drying.
Further, the process comprises the following steps:
1) selecting mature and healthy soybeans, removing sand and stones by using a vibrating screen, drying at the temperature of 45-50 ℃ for 4-6 hours, and then removing skins;
2) deoiling, namely deoiling soybeans by adopting a cold rolling method, wherein the oil content of squeezed soybean meal is less than 4%.
3) Crushing, namely crushing the soybean meal into 40-mesh powder, and filtering and screening the powder by using a sieve body with 20-40 meshes to prepare soybean meal;
4) alkaline leaching to prepare a sodium hydroxide solution with the Ph value of 9;
and (3) primary leaching, namely placing the soybean meal and a sodium hydroxide solution into a leaching tank for primary leaching, wherein the solid-liquid weight ratio of the soybean meal to the sodium hydroxide solution is 1: 10-15, the leaching temperature is 60 ℃, the stirring speed is 80-120 r/min, and the leaching time is 2-3 hours; after leaching, carrying out solid-liquid filtration separation to obtain a solution a and a solid material;
and (3) secondary leaching, wherein the solid material is subjected to secondary leaching, and the solid-liquid weight ratio of the solid material to the sodium hydroxide solution is 1: 5-8, wherein the leaching temperature is 60 ℃, the stirring speed is 80-120 r/min, and the leaching time is 1-1.5 hours, so as to obtain a solution b and material residues;
5) acid precipitation, namely mixing the solution a and the solution b at 50 ℃, adding hydrochloric acid into the mixed solution, slightly stirring to ensure that the pH value reaches 4.5, and then precipitating for 2 hours;
6) filtering, neutralizing and bleaching, centrifuging the precipitated solution at 6000 rpm for 5-10 min to obtain selenoprotein emulsion, adding lime water and sodium hydroxide solution into the selenoprotein emulsion to adjust the pH value to 6.5-7, wherein the lime water is saturated lime water, and the weight ratio of the lime water to the sodium hydroxide solution is 1: 2.
7) Carrying out enzymatic modification, carrying out enzymolysis on the bleached selenoprotein emulsion, adding composite protease into the solution, wherein the weight ratio of the addition amount of the composite protease to the selenoprotein emulsion is 1: 400-;
adding flavourzyme into the cooled selenoprotein emulsion, wherein the weight ratio of the addition amount of the flavourzyme to the selenoprotein emulsion is 1:200-350, the pH value of the selenoprotein emulsion is 6.5-7, the temperature is 40 ℃, continuously stirring, and carrying out enzymolysis for 8 hours;
8) filtering and drying, performing centrifugal filtration on the selenium protein emulsion after enzymolysis, wherein the centrifugal rotation speed is 6000 r/min, performing centrifugation for 10 min, performing atomization drying on the filtered selenium protein emulsion, and drying at the temperature of 65-80 ℃ to obtain powder of selenium protein.
Further, in the material selection process, after sand and stones are removed, hydrogen peroxide or chlorine is added for sterilization and cleaning, then the material is washed by clean water, and then the material is dried and peeled.
Further, in the alkaline leaching, ultrasonic wave is adopted for assisting the leaching, the frequency of the ultrasonic wave is adjusted to be 50-55kHz, and the power is 500-600 w.
Further, in the enzymatic modification, the compound protease and the flavourzyme can be added simultaneously, the ratio of the compound protease to the flavourzyme is 1:2, the pH value of the selenoprotein emulsion is 6.5-7, the temperature is 40-50 ℃, the mixture is continuously stirred, the enzymolysis time is 8 hours, the temperature of the selenoprotein emulsion after enzymolysis is raised to 75 ℃, and the temperature is kept for 3 minutes for enzyme deactivation.
Further, in the filtration, neutralization, bleaching and filtration drying, the centrifugal filtration can also adopt membrane filtration, and a microporous filtration membrane is adopted, and the membrane pores are 15-200 μm.
Further, in the filtering and drying, the selenoprotein emulsion is subjected to freeze drying at the temperature of-55 ℃, the freezing time is 20 hours, the vacuum pressure is 20 Pa, the water content is less than 3 percent, and the dried powder is selenoprotein.
Further, the selenoprotein powder still contains 1-3% of impurities, the selenoprotein powder and 30% ethanol solution are mixed according to the solid-to-liquid ratio of 1:20, the temperature is 40 ℃, centrifugal stirring is carried out for 5 minutes, the stirring speed is 5000-6000 r/min, then standing is carried out for 35 minutes, non-protein components are separated out, then filtration and separation are carried out through a filter membrane with the aperture of 25-50 mu m, after separation, the solution is evaporated and concentrated to 1/3 of the original quantity, and then the pus is atomized and dried to obtain the purified selenoprotein.
The invention carries out twice leaching and twice enzymolysis on the selenoprotein emulsion, the extraction is more thorough, the purity of the selenoprotein is higher, no peculiar smell is generated, and the technical problems of various extraction procedures, inconvenient use and low efficiency in the prior art are effectively solved.
Detailed Description
The technical solutions of the present invention will be described clearly and completely below, and it is obvious that the embodiments described below are some, but not all embodiments of the present invention in the description of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1: an extraction process for extracting selenoprotein from soybeans comprises the steps of 1) material selection, 2) deoiling, 3) crushing, 4) alkaline leaching, 5) acid precipitation, 6) filtration, neutralization and bleaching, 7) enzymatic modification and 8) filtration and drying.
Further, the process comprises the following steps:
1) selecting mature and healthy soybeans, removing sand and stones by using a vibrating screen, drying at the temperature of 45-50 ℃ for 4-6 hours, and then removing skins;
2) deoiling, namely deoiling soybeans by adopting a cold rolling method, wherein the oil content of squeezed soybean meal is less than 4%.
3) Crushing, namely crushing the soybean meal into 40-mesh powder, and filtering and screening the powder by using a sieve body with 20-40 meshes to prepare soybean meal;
4) alkaline leaching to prepare a sodium hydroxide solution with the Ph value of 9;
and (3) primary leaching, namely placing the soybean meal and a sodium hydroxide solution into a leaching tank for primary leaching, wherein the solid-liquid weight ratio of the soybean meal to the sodium hydroxide solution is 1: 10-15, the leaching temperature is 60 ℃, the stirring speed is 80-120 r/min, and the leaching time is 2-3 hours; after leaching, carrying out solid-liquid filtration separation to obtain a solution a and a solid material;
and (3) secondary leaching, wherein the solid material is subjected to secondary leaching, and the solid-liquid weight ratio of the solid material to the sodium hydroxide solution is 1: 5-8, wherein the leaching temperature is 60 ℃, the stirring speed is 80-120 r/min, and the leaching time is 1-1.5 hours, so as to obtain a solution b and material residues;
5) acid precipitation, namely mixing the solution a and the solution b at 50 ℃, adding hydrochloric acid into the mixed solution, slightly stirring to ensure that the pH value reaches 4.5, and then precipitating for 2 hours;
6) filtering, neutralizing and bleaching, centrifuging the precipitated solution at 6000 rpm for 5-10 min to obtain selenoprotein emulsion, adding lime water and sodium hydroxide solution into the selenoprotein emulsion to adjust the pH value to 6.5-7, wherein the lime water is saturated lime water, and the weight ratio of the lime water to the sodium hydroxide solution is 1: 2.
7) Carrying out enzymatic modification, carrying out enzymolysis on the bleached selenoprotein emulsion, adding composite protease into the solution, wherein the weight ratio of the addition amount of the composite protease to the selenoprotein emulsion is 1: 400-;
adding flavourzyme into the cooled selenoprotein emulsion, wherein the weight ratio of the addition amount of the flavourzyme to the selenoprotein emulsion is 1:200-350, the pH value of the selenoprotein emulsion is 6.5-7, the temperature is 40 ℃, continuously stirring, and carrying out enzymolysis for 8 hours;
8) filtering and drying, performing centrifugal filtration on the selenium protein emulsion after enzymolysis, wherein the centrifugal rotation speed is 6000 r/min, performing centrifugation for 10 min, performing atomization drying on the filtered selenium protein emulsion, and drying at the temperature of 65-80 ℃ to obtain powder of selenium protein.
In the material selecting process, after sand and stones are removed, hydrogen peroxide or chlorine is added for sterilization and cleaning, then the materials are washed by clean water, and then the materials are dried and peeled after being cleaned.
In the alkaline leaching, ultrasonic wave is adopted for assisting in leaching, the frequency of the ultrasonic wave is adjusted to be 50-55kHz, and the power is 500-600 w.
In the enzymatic modification, the compound protease and the flavourzyme can be added simultaneously, the ratio of the compound protease to the flavourzyme is 1:2, the pH value of the selenoprotein emulsion is 6.5-7, the temperature is 40-50 ℃, the mixture is continuously stirred, the enzymolysis time is 8 hours, the temperature of the selenoprotein emulsion after enzymolysis is raised to 75 ℃, and the temperature is kept for 3 minutes for enzyme deactivation.
In the filtration, neutralization, bleaching and filtration drying, the centrifugal filtration can also adopt membrane filtration, and a microporous filtration membrane is adopted, and the membrane pores are 15-200 mu m.
In the filtering and drying, the selenoprotein emulsion is subjected to freeze drying at the temperature of-55 ℃, the freezing time is 20 hours, the vacuum pressure is 20 Pa, the water content is less than 3 percent, and the dried powder is selenoprotein.
Example 2, the selenoprotein powder still contains 1-3% of impurities, the selenoprotein powder and 30% ethanol solution are mixed according to the solid-to-liquid ratio of 1:20, the temperature is 40 ℃, centrifugal stirring is carried out for 5 minutes, the stirring speed is 5000-6000 r/min, then standing is carried out for 35 minutes, non-protein components are separated out, then filtration and separation are carried out through a filter membrane with the pore diameter of 25-50 mu m, after separation, the solution is evaporated and concentrated to 1/3 of the original quantity, and then the pus is atomized and dried to obtain the purified selenoprotein.
Example 3, selenoprotein was prepared using the following steps:
1) selecting mature and healthy soybeans, removing sand and stones by using a vibrating screen, sterilizing and cleaning by using 1-3% chlorine water, then washing by using clear water, drying at the temperature of 45-50 ℃ for 4-6 hours, and then removing skins;
2) deoiling, namely deoiling soybeans by a cold rolling method, wherein the squeezed soybean meal can be leached for the second time to deoil, and the oil content of the filtered soybean meal is 1-2%.
3) Crushing, namely crushing the soybean meal into 40-mesh powder particles, filtering and screening the powder particles by using a 40-mesh screen, crushing the screened large particles again, and dropping the soybean meal by using a screen for later use;
4) alkaline leaching to prepare a sodium hydroxide solution with the Ph value of 9;
the first leaching, namely placing the soybean meal and a sodium hydroxide solution into a leaching tank for the first leaching, weighing 200 kg of soybean meal and 3000L of a sodium hydroxide solution into the leaching tank, heating to 60 ℃, continuously stirring in the leaching process, wherein the stirring speed is 120 revolutions per minute, and the leaching time is 3 hours; after leaching, solid-liquid filtration separation is carried out to obtain 3050L of solution a and 150 kg of solid materials;
performing secondary leaching, namely performing secondary leaching on the solid material, putting 150 kg of the solid material and 1000L of sodium hydroxide solution into a leaching tank for continuous leaching, wherein the leaching temperature is 60 ℃, the stirring speed is 80 rpm, and the leaching time is 1.5 hours to obtain 1020L of solution b and 130 kg of material residues;
5) acid precipitation, namely mixing the solution a and the solution b, wherein the total amount is 3070 liters, the temperature is 50 ℃, adding dilute hydrochloric acid into the mixed solution, slightly stirring to ensure that the pH value reaches 4.5, and then precipitating for 2 hours;
6) filtering, neutralizing and bleaching, performing centrifugal filtration on the precipitated solution at the rotating speed of 6000 rpm for 10 minutes to obtain 120 liters of selenoprotein emulsion, adding lime water and sodium hydroxide solution into the selenoprotein emulsion to adjust the pH value to 6.5-7, wherein the lime water is saturated lime water, and the weight ratio of the lime water to the sodium hydroxide solution is 1: 2.
7) Carrying out enzymatic modification, carrying out enzymolysis on the bleached selenoprotein emulsion, adding composite protease into the solution, wherein the addition amount of the composite protease is 0.1 liter, the pH value of the selenoprotein emulsion is 7, the temperature is 50 ℃, continuously stirring, carrying out enzymolysis for 4 hours, heating the selenoprotein emulsion after the enzymolysis to 75 ℃, keeping for 3 minutes for enzyme deactivation, and then cooling to 40 ℃;
adding flavourzyme into the cooled selenoprotein emulsion, wherein the addition amount of the flavourzyme is 0.3 liter, the pH value of the selenoprotein emulsion is 7, the temperature is 40 ℃, continuously stirring, and carrying out enzymolysis for 8 hours;
8) and (3) filtering and drying, performing centrifugal filtration on the selenoprotein emulsion subjected to enzymolysis, wherein the centrifugal rotation speed is 6000 rpm, performing centrifugation for 10 minutes, performing atomization drying on the filtered selenoprotein emulsion, and drying at the drying temperature of 65-80 ℃ to obtain 65 kg of selenoprotein powder.
9) And (2) purifying, namely mixing 65 kg of selenoprotein powder with 1300 liters of 30% ethanol solution, putting the mixture into a reaction kettle, carrying out centrifugal stirring at the temperature of 40 ℃ for 5 minutes at the stirring speed of 5000 r/min, standing for 35 minutes, separating out non-protein components, filtering and separating through a filter membrane with the aperture of 50 mu m, filtering out 1.5 kg of slag, evaporating and concentrating the residual solution to 450 liters, and then carrying out atomization drying on the pus to obtain 63 kg of purified selenoprotein.
The purified selenoprotein has a purity of over 99 percent through detection, and can be used as an additive product of various foods or health care products.
While the invention has been described with reference to specific embodiments, it will be apparent to those skilled in the art that the invention is not limited thereto, and that various changes and modifications can be made without departing from the spirit and scope of the invention.
Claims (8)
1. An extraction process for extracting selenoprotein from soybeans is characterized by comprising the steps of 1) material selection, 2) oil removal, 3) crushing, 4) alkaline leaching, 5) acid precipitation, 6) filtration, neutralization and bleaching, 7) enzymatic modification and 8) filtration and drying.
2. The process of claim 1, wherein the process comprises the steps of:
1) selecting mature and healthy soybeans, removing sand and stones by using a vibrating screen, drying at the temperature of 45-50 ℃ for 4-6 hours, and then removing skins;
2) deoiling, namely deoiling soybeans by adopting a cold rolling method, wherein the oil content of squeezed soybean meal is less than 4%;
3) crushing, namely crushing the soybean meal into 40-mesh powder, and filtering and screening the powder by using a sieve body with 20-40 meshes to prepare soybean meal;
4) alkaline leaching to prepare a sodium hydroxide solution with the Ph value of 9;
and (3) primary leaching, namely placing the soybean meal and a sodium hydroxide solution into a leaching tank for primary leaching, wherein the solid-liquid weight ratio of the soybean meal to the sodium hydroxide solution is 1: 10-15, the leaching temperature is 60 ℃, the stirring speed is 80-120 r/min, and the leaching time is 2-3 hours; after leaching, carrying out solid-liquid filtration separation to obtain a solution a and a solid material;
and (3) secondary leaching, wherein the solid material is subjected to secondary leaching, and the solid-liquid weight ratio of the solid material to the sodium hydroxide solution is 1: 5-8, wherein the leaching temperature is 60 ℃, the stirring speed is 80-120 r/min, and the leaching time is 1-1.5 hours, so as to obtain a solution b and material residues;
5) acid precipitation, namely mixing the solution a and the solution b at 50 ℃, adding hydrochloric acid into the mixed solution, slightly stirring to ensure that the pH value reaches 4.5, and then precipitating for 2 hours;
6) filtering, neutralizing and bleaching, performing centrifugal filtration on the precipitated solution at the rotating speed of 6000 revolutions per minute for 5-10 minutes to obtain a selenoprotein emulsion, adding lime water and a sodium hydroxide solution into the selenoprotein emulsion to adjust the pH value to 6.5-7, wherein the lime water is saturated lime water, and the weight ratio of the lime water to the sodium hydroxide solution is 1: 2;
7) carrying out enzymatic modification, carrying out enzymolysis on the bleached selenoprotein emulsion, adding composite protease into the solution, wherein the weight ratio of the addition amount of the composite protease to the selenoprotein emulsion is 1: 400-;
adding flavourzyme into the cooled selenoprotein emulsion, wherein the weight ratio of the addition amount of the flavourzyme to the selenoprotein emulsion is 1:200-350, the pH value of the selenoprotein emulsion is 6.5-7, the temperature is 40 ℃, continuously stirring, and carrying out enzymolysis for 8 hours;
8) filtering and drying, performing centrifugal filtration on the selenium protein emulsion after enzymolysis, wherein the centrifugal rotation speed is 6000 r/min, performing centrifugation for 10 min, performing atomization drying on the filtered selenium protein emulsion, and drying at the temperature of 65-80 ℃ to obtain powder of selenium protein.
3. The process of claim 2, wherein in the step of selecting, after removing sand, hydrogen peroxide or chlorine is added for sterilization and cleaning, and then the materials are washed with clean water, and then dried and peeled.
4. The extraction process of selenoprotein from soybean as claimed in claim 2, wherein in the enzymatic modification, composite protease and flavourzyme can be added simultaneously, the ratio of the composite protease to the flavourzyme is 1:2, the pH value of the selenoprotein emulsion is 6.5-7, the temperature is 40-50 ℃, the mixture is continuously stirred, the enzymolysis time is 8 hours, the temperature of the selenoprotein emulsion after enzymolysis is raised to 75 ℃, and the temperature is kept for 3 minutes for enzyme deactivation.
5. The process for extracting selenium protein from soybean as claimed in claim 2, wherein the filtration, neutralization, bleaching and drying are performed by membrane filtration with a pore size of 15-200 μm.
6. The process of claim 2, wherein the selenoprotein emulsion is freeze-dried at-55 deg.C under 20 hr, vacuum pressure of 20 Pa, and water content of less than 3%, and the dried powder is selenoprotein.
7. The extraction process of selenoprotein from soybean as claimed in claim 2, wherein in said alkaline leaching, ultrasonic wave is used for assisting leaching, the frequency of ultrasonic wave is adjusted to 50-55kHz, and the power is 500-600 w.
8. The extraction process of selenoprotein from soybean as claimed in claim 1 or 2, wherein the filtration and drying process further comprises step 9) of purification, mixing selenoprotein powder with 30% ethanol solution according to a solid-to-liquid ratio of 1:20, at 40 ℃, carrying out centrifugal stirring for 5 minutes, wherein the stirring speed is 5000-6000 r/min, then standing for 35 minutes, separating out non-protein components, then carrying out filtration and separation through a filter membrane with a pore size of 25-50 μm, evaporating and concentrating the solution to 1/3 of the original amount, and then carrying out atomization and drying on pus to obtain purified selenoprotein.
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