CN109652488B - Preparation method of sesame polypeptide - Google Patents
Preparation method of sesame polypeptide Download PDFInfo
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- CN109652488B CN109652488B CN201910058562.4A CN201910058562A CN109652488B CN 109652488 B CN109652488 B CN 109652488B CN 201910058562 A CN201910058562 A CN 201910058562A CN 109652488 B CN109652488 B CN 109652488B
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Abstract
The invention discloses a preparation method of sesame polypeptide, which comprises the following steps: mixing sesame protein and distilled water uniformly, and heating, decompressing and preserving heat to obtain sesame protein slurry; adjusting the pH value of the sesame protein slurry to 5-6, adding neutral protease, hemicellulase and pancreatin to perform ultrasonic-assisted enzymolysis, and obtaining primary enzymolysis liquid after the primary enzymolysis reaction is finished; adjusting the pH value of the primary enzymolysis liquid to 8-9, adding saccharifying enzyme and alkaline protease to carry out ultrasonic auxiliary enzymolysis, and obtaining secondary enzymolysis liquid after the secondary enzymolysis reaction is finished; carrying out microwave enzyme deactivation treatment on the secondary enzymolysis liquid to obtain enzyme-deactivated enzymolysis liquid; and sequentially carrying out ultrafiltration, microfiltration and reverse osmosis concentration on the enzyme-inactivated enzymolysis liquid, and freeze-drying to obtain the sesame polypeptide. The preparation method of the sesame polypeptide is safe in process and convenient and fast to prepare.
Description
Technical Field
The invention relates to the technical field of sesame polypeptide, and particularly relates to a preparation method of sesame polypeptide.
Background
It has been shown that proteins are not necessarily absorbed in the form of free amino acids, but in the form of peptides, after the action of digestive enzymes. The research proves that the absorption rate of the peptide is larger than that of the free amino acid, and the peptide is easier and faster to pass through a small intestine drill membrane to be absorbed and utilized by the organism than the amino acid, and the structure of the protein after being enzymolyzed into the polypeptide has diversity and complexity, so that the unique physicochemical property and the biological activity of the protein are determined. A large number of researches prove that some polypeptides have the effects of enhancing immunity, resisting virus, resisting oxidation, reducing blood pressure, reducing blood fat, reducing cholesterol, promoting nutrient absorption, regulating hormone and the like. Therefore, it is necessary to develop a method for preparing sesame polypeptide.
Disclosure of Invention
Aiming at the defects in the technology, the invention provides the preparation method of the sesame polypeptide, which has safe process and convenient and fast preparation.
The technical scheme adopted by the invention for solving the technical problems is as follows: a preparation method of sesame polypeptide comprises the following steps: step one, mixing sesame protein and distilled water uniformly, and heating, decompressing and preserving heat to obtain sesame protein slurry; adjusting the pH value of the sesame protein slurry to 5-6, adding neutral protease, hemicellulase and pancreatin to perform ultrasonic-assisted enzymolysis, and obtaining primary enzymolysis liquid after the primary enzymolysis reaction is finished; regulating the pH value of the primary enzymolysis liquid to 8-9, adding saccharifying enzyme and alkaline protease to carry out ultrasonic-assisted enzymolysis, and obtaining secondary enzymolysis liquid after the secondary enzymolysis reaction is finished; step four, carrying out microwave enzyme deactivation treatment on the secondary enzymatic hydrolysate to obtain enzyme deactivated enzymatic hydrolysate; and fifthly, sequentially carrying out ultrafiltration, microfiltration and reverse osmosis concentration on the enzyme-inactivated enzymolysis liquid, and carrying out freeze drying to obtain the sesame polypeptide.
Preferably, in the step one, the sesame protein and the distilled water are mixed according to a feed-liquid ratio of 1: 16-20, uniformly mixing, heating to 70-80 ℃, and carrying out heat preservation treatment for 14-16 minutes under the pressure of 0.08-0.1 Mpa; and continuously heating to 90-100 ℃, and carrying out heat preservation treatment for 14-16 minutes under the pressure of 0.05-0.07 Mpa to obtain the sesame protein slurry.
Preferably, the conditions of the ultrasound-assisted enzymolysis in the second step are as follows: the enzymolysis reaction temperature is 45-50 ℃, the enzymolysis reaction time is 2-3 hours, and the ultrasonic power is 30-40 kHz.
Preferably, the neutral protease in the second step: hemicellulase (b): the content ratio of pancreatin is 1: 3-4: 2-3, wherein the total enzyme adding amount of the neutral protease, the hemicellulase and the pancreatin is 3-3.5 ug/g of the dosage of the sesame protein slurry.
Preferably, the conditions of the ultrasonic-assisted enzymolysis in the third step are as follows: the enzymolysis reaction temperature is 55-60 ℃, the enzymolysis reaction time is 3-4 hours, and the ultrasonic power is 30-40 kHz.
Preferably, the saccharifying enzyme in step three: the content ratio of the alkaline protease is 1: 2-4, and the total enzyme adding amount of the saccharifying enzyme and the alkaline protease is 2-2.5 ug/g of the dosage of the sesame protein slurry.
Preferably, the microwave enzyme deactivation treatment conditions in the fourth step are as follows: the microwave power is 400-600W, the microwave enzyme deactivation temperature is 90-95 ℃, and the microwave enzyme deactivation time is 3-5 minutes.
Preferably, the aperture of the ultrafiltration membrane used in the ultrafiltration treatment in the fifth step is 20-30 nanometers, the ultrafiltration pressure is 0.3-0.5 MPa, the ultrafiltration temperature is 30-40 ℃, and the ultrafiltration time is 20-30 minutes.
Preferably, in the fifth step, the microfiltration membrane adopted for microfiltration has a pore diameter of 0.4-0.8 micron, a microfiltration pressure of 0.05-0.1 MPa, a microfiltration temperature of 20-30 ℃ and a microfiltration time of 40-50 minutes.
Preferably, the conditions of the reverse osmosis concentration treatment in the step five are as follows: the pressure is 0.3-0.5 MPa, the pH is 7.5-8.5, and the temperature is 15-25 ℃.
Compared with the prior art, the invention has the beneficial effects that: according to the preparation method of the sesame polypeptide, the sesame protein is subjected to heating, pressure reduction and heat preservation pretreatment before enzymolysis, so that a highly compressed structure of the sesame protein can be loosened and fully denatured, the hydrolysis degree is improved, and the reaction speed and the reaction efficiency of later enzymolysis can be remarkably accelerated; the primary compound enzyme consisting of neutral protease, cellulase and diastase in a certain proportion is adopted for enzymolysis during primary enzymolysis, and the secondary compound enzyme consisting of alkaline protease, papain and flavourzyme in a certain proportion is adopted for enzymolysis during secondary enzymolysis, so that some pseudoproteins such as saccharides can be effectively removed, and the purity and hydrolysis rate of the sesame polypeptide are remarkably improved; the sesame polypeptide has good flavor by sequentially adopting ultrafiltration, microfiltration and concentration treatment.
Detailed Description
The present invention is described in further detail below to enable those skilled in the art to practice the invention with reference to the description.
The invention provides a preparation method of sesame polypeptide, which comprises the following steps:
step one, mixing sesame protein with water absorptivity of 2.33-3.12 g/g and oil absorptivity of 2.48-3.06 g/g and distilled water according to a material-liquid ratio of 1: 16-20, uniformly mixing, and heating, decompressing and preserving heat: in the first stage, heating to 70-80 ℃, and carrying out heat preservation treatment for 14-16 minutes under the pressure of 0.08-0.1 Mpa; in the second stage, continuously heating to 90-100 ℃, and carrying out heat preservation treatment for 14-16 minutes under the pressure of 0.05-0.07 Mpa to obtain sesame protein slurry;
adjusting the pH value of the sesame protein slurry to 5-6, adding neutral protease, hemicellulase and pancreatin to perform ultrasonic-assisted enzymolysis, wherein the enzymolysis reaction temperature is 45-50 ℃, the enzymolysis reaction time is 2-3 hours, the ultrasonic power is 30-40 kHz, and a primary enzymolysis liquid is obtained after the primary enzymolysis reaction is finished;
wherein the neutral protease: hemicellulase (b): the content ratio of pancreatin is 1: 3-4: 2-3, wherein the total enzyme adding amount of the neutral protease, the hemicellulase and the pancreatin is 3-3.5 ug/g of the dosage of the sesame protein slurry;
regulating the pH value of the primary enzymolysis liquid to 8-9, adding saccharifying enzyme and alkaline protease to carry out ultrasonic-assisted enzymolysis, wherein the enzymolysis reaction temperature is 55-60 ℃, the enzymolysis reaction time is 3-4 hours, the ultrasonic power is 30-40 kHz, and after the secondary enzymolysis reaction is finished, secondary enzymolysis liquid is obtained;
wherein the saccharifying enzyme: the content ratio of the alkaline protease is 1: 2-4, wherein the total enzyme adding amount of the saccharifying enzyme and the alkaline protease is 2-2.5 ug/g of the dosage of the sesame protein slurry;
step four, carrying out microwave enzyme deactivation treatment on the secondary enzymatic hydrolysate, wherein the microwave power is 400-600W, the microwave enzyme deactivation temperature is 90-95 ℃, and the microwave enzyme deactivation time is 3-5 minutes, so as to obtain enzyme deactivation enzymatic hydrolysate;
fifthly, sequentially carrying out ultrafiltration, microfiltration and reverse osmosis concentration treatment on the enzyme-inactivated enzymolysis liquid, wherein the ultrafiltration treatment adopts an ultrafiltration membrane with the aperture of 20-30 nanometers, the ultrafiltration pressure of 0.3-0.5 MPa, the ultrafiltration temperature of 30-40 ℃ and the ultrafiltration time of 20-30 minutes; the micro-filtration treatment adopts a micro-filtration membrane with the aperture of 0.4-0.8 micron, the micro-filtration pressure of 0.05-0.1 MPa, the micro-filtration temperature of 20-30 ℃ and the micro-filtration time of 40-50 minutes; the conditions of the reverse osmosis concentration treatment are as follows: the pressure is 0.3-0.5 MPa, the pH is 7.5-8.5, and the temperature is 15-25 ℃; and freeze-drying to obtain the sesame polypeptide, wherein the water absorption of the sesame polypeptide is 1.72-2.21 g/g, and the oil absorption of the sesame polypeptide is 2.55-3.02 g/g.
Example 1
Mixing sesame protein and distilled water according to a material-liquid ratio of 1: 16, uniformly mixing, and heating, decompressing and preserving heat: the first stage, heating to 70 deg.C, and maintaining the temperature under 0.08Mpa for 16 min; in the second stage, the sesame seed protein slurry is obtained by continuously heating to 90 ℃ and carrying out heat preservation treatment for 16 minutes under the pressure of 0.05 Mpa; adjusting the pH value of the sesame protein slurry to 5, adding neutral protease, hemicellulase and pancreatin to carry out ultrasonic-assisted enzymolysis, wherein the enzymolysis reaction temperature is 45 ℃, the enzymolysis reaction time is 3 hours, the ultrasonic power is 30kHz, and a primary enzymolysis liquid is obtained after the primary enzymolysis reaction is finished; wherein the neutral protease: hemicellulase (b): the content ratio of pancreatin is 1: 3: 2, the total enzyme adding amount of the neutral protease, the hemicellulase and the pancreatin is 3ug/g of the dosage of the sesame protein slurry; adjusting the pH value of the primary enzymolysis liquid to 8, adding saccharifying enzyme and alkaline protease to carry out ultrasonic-assisted enzymolysis, wherein the enzymolysis reaction temperature is 55 ℃, the enzymolysis reaction time is 4 hours, the ultrasonic power is 30kHz, and after the secondary enzymolysis reaction is finished, secondary enzymolysis liquid is obtained; the saccharifying enzyme: the content ratio of the alkaline protease is 1: 2, the total enzyme adding amount of the saccharifying enzyme and the alkaline protease is 2ug/g of the dosage of the sesame protein slurry; carrying out microwave enzyme deactivation treatment on the secondary enzymolysis liquid, wherein the microwave power is 400W, the microwave enzyme deactivation temperature is 90 ℃, and the microwave enzyme deactivation time is 5 minutes to obtain enzyme-deactivated enzymolysis liquid; sequentially carrying out ultrafiltration, microfiltration and reverse osmosis concentration treatment on the enzyme-inactivated enzymolysis liquid, wherein the ultrafiltration treatment adopts an ultrafiltration membrane with the aperture of 20 nanometers, the ultrafiltration pressure of 0.3MPa, the ultrafiltration temperature of 30 ℃ and the ultrafiltration time of 30 minutes; the micro-filtration treatment adopts a micro-filtration membrane with the aperture of 0.4 micron, the micro-filtration pressure of 0.05MPa, the micro-filtration temperature of 20 ℃ and the micro-filtration time of 50 minutes; the conditions of the reverse osmosis concentration treatment are as follows: the pressure is 0.3MPa, the pH is 7.5, and the temperature is 15 ℃; and freeze-drying to obtain the sesame polypeptide.
Example 2
Mixing sesame protein and distilled water according to a material-liquid ratio of 1: 20, uniformly mixing, and heating, decompressing and preserving heat: the first stage, heating to 80 deg.C, and maintaining the temperature under 0.1Mpa for 14 min; in the second stage, the sesame seed protein slurry is obtained by continuously heating to 100 ℃ and carrying out heat preservation treatment for 14 minutes under the pressure of 0.07 Mpa; adjusting the pH value of the sesame protein slurry to 6, adding neutral protease, hemicellulase and pancreatin to carry out ultrasonic-assisted enzymolysis, wherein the enzymolysis reaction temperature is 50 ℃, the enzymolysis reaction time is 2 hours, the ultrasonic power is 40kHz, and a primary enzymolysis liquid is obtained after the primary enzymolysis reaction is finished; wherein the neutral protease: hemicellulase (b): the content ratio of pancreatin is 1: 4: 3, the total enzyme adding amount of the neutral protease, the hemicellulase and the pancreatin is 3.5ug/g of the dosage of the sesame protein slurry; adjusting the pH value of the primary enzymolysis liquid to 9, adding saccharifying enzyme and alkaline protease to carry out ultrasonic-assisted enzymolysis, wherein the enzymolysis reaction temperature is 60 ℃, the enzymolysis reaction time is 3 hours, the ultrasonic power is 40kHz, and after the secondary enzymolysis reaction is finished, secondary enzymolysis liquid is obtained; the saccharifying enzyme: the content ratio of the alkaline protease is 1: 4, the total enzyme adding amount of the saccharifying enzyme and the alkaline protease is 2.5ug/g of the dosage of the sesame protein slurry; carrying out microwave enzyme deactivation treatment on the secondary enzymolysis liquid, wherein the microwave power is 600W, the microwave enzyme deactivation temperature is 95 ℃, and the microwave enzyme deactivation time is 3 minutes to obtain enzyme-deactivated enzymolysis liquid; sequentially carrying out ultrafiltration, microfiltration and reverse osmosis concentration treatment on the enzyme-inactivated enzymolysis liquid, wherein the ultrafiltration treatment adopts an ultrafiltration membrane with the aperture of 30 nanometers, the ultrafiltration pressure of 0.5MPa, the ultrafiltration temperature of 40 ℃ and the ultrafiltration time of 20 minutes; the micro-filtration treatment adopts a micro-filtration membrane with the aperture of 0.8 micron, the micro-filtration pressure of 0.1MPa, the micro-filtration temperature of 30 ℃ and the micro-filtration time of 40 minutes; the conditions of the reverse osmosis concentration treatment are as follows: the pressure is 0.5MPa, the pH is 8.5, and the temperature is 25 ℃; and freeze-drying to obtain the sesame polypeptide.
Example 3
Mixing sesame protein and distilled water according to a material-liquid ratio of 1: 18, uniformly mixing, and heating, decompressing and preserving heat: in the first stage, heating to 75 ℃ and carrying out heat preservation treatment for 15 minutes under the pressure of 0.09 Mpa; in the second stage, the sesame seed protein slurry is obtained by continuously heating to 95 ℃ and carrying out heat preservation treatment for 15 minutes under the pressure of 0.06 Mpa; adjusting the pH value of the sesame protein slurry to 5.5, adding neutral protease, hemicellulase and pancreatin to carry out ultrasonic-assisted enzymolysis, wherein the enzymolysis reaction temperature is 48 ℃, the enzymolysis reaction time is 2.5 hours, the ultrasonic power is 35kHz, and a primary enzymolysis liquid is obtained after one enzymolysis reaction is finished; wherein the neutral protease: hemicellulase (b): the content ratio of pancreatin is 1: 3.5: 2.5, and the total enzyme adding amount of the neutral protease, the hemicellulase and the pancreatin is 3.2ug/g of the using amount of the sesame protein slurry; adjusting the pH value of the primary enzymolysis liquid to 8.5, adding saccharifying enzyme and alkaline protease to carry out ultrasonic-assisted enzymolysis, wherein the enzymolysis reaction temperature is 58 ℃, the enzymolysis reaction time is 3.5 hours, the ultrasonic power is 35kHz, and after the secondary enzymolysis reaction is finished, secondary enzymolysis liquid is obtained; the saccharifying enzyme: the content ratio of the alkaline protease is 1: 3, the total enzyme adding amount of the saccharifying enzyme and the alkaline protease is 2.2ug/g of the dosage of the sesame protein slurry; carrying out microwave enzyme deactivation treatment on the secondary enzymolysis liquid, wherein the microwave power is 500W, the microwave enzyme deactivation temperature is 92 ℃, and the microwave enzyme deactivation time is 4 minutes, so as to obtain enzyme-deactivated enzymolysis liquid; sequentially carrying out ultrafiltration, microfiltration and reverse osmosis concentration treatment on the enzyme-inactivated enzymolysis liquid, wherein the ultrafiltration treatment adopts an ultrafiltration membrane with the aperture of 25 nanometers, the ultrafiltration pressure of 0.4MPa, the ultrafiltration temperature of 35 ℃ and the ultrafiltration time of 25 minutes; the micro-filtration treatment adopts a micro-filtration membrane with the aperture of 0.6 micron, the micro-filtration pressure of 0.08MPa, the micro-filtration temperature of 25 ℃ and the micro-filtration time of 45 minutes; the conditions of the reverse osmosis concentration treatment are as follows: the pressure is 0.4MPa, the pH is 8 and the temperature is 20 ℃; and freeze-drying to obtain the sesame polypeptide.
While embodiments of the invention have been disclosed above, it is not limited to the applications listed in the description and the embodiments, which are fully applicable in all kinds of fields of application of the invention, and further modifications may readily be effected by those skilled in the art, whereby the invention is not limited to the details given, without departing from the general concept defined by the claims and the scope of equivalents.
Claims (2)
1. The preparation method of the sesame polypeptide is characterized by comprising the following steps:
step one, mixing sesame protein and distilled water according to a material-liquid ratio of 1: 16-20, uniformly mixing, heating to 70-80 ℃, and carrying out heat preservation treatment for 14-16 minutes under the pressure of 0.08-0.1 Mpa; continuously heating to 90-100 ℃, and carrying out heat preservation treatment for 14-16 minutes under the pressure of 0.05-0.07 Mpa to obtain sesame protein slurry;
adjusting the pH value of the sesame protein slurry to 5-6, adding neutral protease, hemicellulase and pancreatin to perform ultrasonic-assisted enzymolysis, wherein the enzymolysis reaction temperature is 45-50 ℃, the enzymolysis reaction time is 2-3 hours, the ultrasonic power is 30-40 kHz, and a primary enzymolysis liquid is obtained after the primary enzymolysis reaction is finished;
the neutral protease is: hemicellulase (b): the content ratio of pancreatin is 1: 3-4: 2-3, wherein the total enzyme adding amount of the neutral protease, the hemicellulase and the pancreatin is 3-3.5 ug/g of the dosage of the sesame protein slurry;
regulating the pH value of the primary enzymolysis liquid to 8-9, adding saccharifying enzyme and alkaline protease to carry out ultrasonic-assisted enzymolysis, wherein the enzymolysis reaction temperature is 55-60 ℃, the enzymolysis reaction time is 3-4 hours, the ultrasonic power is 30-40 kHz, and after the secondary enzymolysis reaction is finished, secondary enzymolysis liquid is obtained;
the saccharifying enzyme: the content ratio of the alkaline protease is 1: 2-4, wherein the total enzyme adding amount of the saccharifying enzyme and the alkaline protease is 2-2.5 ug/g of the dosage of the sesame protein slurry;
step four, carrying out microwave enzyme deactivation treatment on the secondary enzymatic hydrolysate to obtain enzyme deactivated enzymatic hydrolysate;
fifthly, sequentially carrying out ultrafiltration, microfiltration and reverse osmosis concentration on the enzyme-inactivated enzymolysis liquid, and carrying out freeze drying to obtain sesame polypeptide;
the ultrafiltration treatment adopts an ultrafiltration membrane with the aperture of 20-30 nanometers, the ultrafiltration pressure of 0.3-0.5 MPa, the ultrafiltration temperature of 30-40 ℃ and the ultrafiltration time of 20-30 minutes;
the micro-filtration treatment adopts a micro-filtration membrane with the aperture of 0.4-0.8 micron, the micro-filtration pressure of 0.05-0.1 MPa, the micro-filtration temperature of 20-30 ℃ and the micro-filtration time of 40-50 minutes;
the conditions of the reverse osmosis concentration treatment are as follows: the pressure is 0.3-0.5 MPa, the pH is 7.5-8.5, and the temperature is 15-25 ℃.
2. The method for preparing sesame polypeptide according to claim 1, wherein the microwave enzyme deactivation treatment conditions in the fourth step are as follows: the microwave power is 400-600W, the microwave enzyme deactivation temperature is 90-95 ℃, and the microwave enzyme deactivation time is 3-5 minutes.
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| CN105936928A (en) * | 2016-05-30 | 2016-09-14 | 铜陵东晟生态农业科技有限公司 | Technology for preparing sesame polypeptides through enzymatic hydrolysis of sesame seed meal |
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