CN115813798A - Preparation process of oat kernel extract - Google Patents
Preparation process of oat kernel extract Download PDFInfo
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- CN115813798A CN115813798A CN202211583331.3A CN202211583331A CN115813798A CN 115813798 A CN115813798 A CN 115813798A CN 202211583331 A CN202211583331 A CN 202211583331A CN 115813798 A CN115813798 A CN 115813798A
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- 239000000284 extract Substances 0.000 title claims abstract description 54
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
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- 239000004365 Protease Substances 0.000 claims abstract description 30
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 30
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- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 12
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 7
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 6
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 6
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 6
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- 229920002774 Maltodextrin Polymers 0.000 claims description 6
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- 229910001628 calcium chloride Inorganic materials 0.000 claims description 6
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- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims description 6
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Abstract
The invention relates to the technical field of oat processing, in particular to a preparation process of an oat kernel extract; the research of the inventor finds that the protease prepared by the strain can enable the prepared oat kernel extract to have higher content of soluble oat polypeptide when the protease prepared by the strain is applied to the enzymolysis reaction of the oat kernel extract.
Description
Technical Field
The invention relates to the technical field of oat processing, in particular to a preparation process of an oat kernel extract.
Background
Oat kernel extract is widely used in cosmetic formulations, the risk of which is registered as grade 1, belongs to one of the safest cosmetic formulations, and mainly plays the roles of an abrasive agent, a softening agent, a skin conditioner and the like in cosmetics.
The oat kernel extract is a natural product extracted from oat kernels, has the advantages of whitening, moisturizing and caring skin, promoting the generation of natural collagen, improving the elasticity of the skin, having the oxidation resistance, effectively delaying the skin aging, and being used as an antioxidant and a humectant.
The active ingredients of the oat kernel extract are water-soluble oat polypeptides, namely the preparation process of the oat kernel extract is an important process for preparing the oat kernel extract through proteolysis, the content of the water-soluble oat polypeptides is determined, finished alkaline protease is generally selected in the process of forming the oat polypeptides through proteolysis in the oat kernel extract, and the existing alkaline protease products are various, but reports on preparation and screening of the alkaline protease in the preparation process of the oat kernel extract in the prior art are few, but the selection of the protease is an important factor influencing the preparation process of the oat kernel extract and is a key influencing the content of the water-soluble oat polypeptides in the preparation process of the oat kernel extract.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: how to improve the content of water-soluble oat polypeptide in oat kernel extract by improving the preparation process of the oat kernel extract.
In order to solve the technical problems, the invention adopts the technical scheme that:
a preparation process of oat kernel extract comprises the following steps:
preparing oat kernel;
soaking oat kernel in water solution with pH of 9-10 at 30-50 deg.C for 4-6 hr, pulverizing to obtain oat kernel slurry, and filtering with 80-100 mesh sieve to obtain filtrate;
adding protease pure enzyme powder into the filtrate, wherein the addition amount of the protease pure enzyme powder is 65-80g/L, and reacting for 1.5-2h under the conditions that the temperature is 40-50 ℃ and the pH is 9-10 to obtain oat kernel enzymatic hydrolysate;
freeze-drying the enzymolysis product to obtain oat kernel extract;
the protease is prepared by the following process:
selecting Bacillus clausii (Bacillus clausii) L-7 with a preservation number of CGMCC No.12953;
activating and culturing the L-7 strain, and fermenting and culturing to obtain protease; the fermentation culture conditions are as follows:
transferred into a fermenter in accordance with an inoculum amount of 5% v/v, the amount of the fermenter charged was 70% (fermentation medium), temperature: 34 ℃; rotating speed: 100-600 r/min; ventilation quantity: 1: 0.5-1: 2.5; the dissolved oxygen is maintained at 20-30%; automatically feeding ammonia water and 20% (v/v) hydrochloric acid in the fermentation process to maintain the pH value of the fermentation liquor at 7.0; the fermentation time is 50-60h; the fermentation medium is as follows: 17g/L of yeast extract powder, 30g/L of cottonseed cake powder, 100g/L of maltodextrin, 3g/L of sodium citrate, 2.6g/L of calcium chloride and K 2 HPO 4 18g/L；
The fermentation broth was centrifuged at 100ml and 8000r/min for 10min, the supernatant was collected and salted out with solid sulfuric acid having a saturation of 70%, the precipitate was collected by centrifugation at 8000r/min for 10min, and the precipitate was dissolved in Tris-HCI (pH 8.8) buffer and dialyzed in the same buffer. Centrifuging to remove a small amount of insoluble substances, collecting supernatant, purifying with DEAE-Sepharose CL-6B ion exchange column, eluting with 20mM Tris-HCl (pH 8.8, containing 10mM NaCl) buffer solution, eluting without adsorbing alkaline protease, collecting enzyme solution, and vacuum freeze drying to obtain pure enzyme powder.
Further, in the preparation process of the oat kernel extract, the activation culture specifically comprises the following steps: inoculating into seed culture medium, culturing at 34-40 deg.C for 11-13h at 200 r/min.
Further, in the preparation process of the oat kernel extract, the activation medium is: 5g/L yeast extract powder, 5g/L tryptone, 10g/L glucose, K 2 HPO 4 18g/L。
Further, in the preparation process of the oat kernel extract, the freeze drying process specifically comprises the following steps: freezing the oat kernel enzymolysis product at-5 to-10 ℃ for 4-5h, carrying out ultrasonic treatment on the coffee extract by an ultrasonic generator in the freezing process, opening a vacuum pump, and vacuumizing a freeze dryer; starting the heating function of the freeze dryer, adjusting the temperature of the shelf plate to-20 ℃, heating for 12-18h, continuously adjusting the temperature to 20 ℃, heating for 3-4h, continuously adjusting the temperature to 35 ℃, and heating for 1-2h; in the heating process, far infrared radiation treatment and magnetic treatment are assisted on the laying plate to obtain the oat kernel extract.
Further, in the preparation process of the oat kernel extract, the steps of preparing the oat kernel are as follows: the oat kernel is prepared by milling the oat kernel into the hull, and the hull milling rate is 15-16%.
The invention has the beneficial effects that: the research of the inventor finds that the protease prepared by the strain can enable the prepared oat kernel extract to have higher content of soluble oat polypeptide when the protease prepared by the strain is applied to the enzymolysis reaction of the oat kernel extract under the specific enzymolysis condition.
Detailed Description
In order to explain the technical content, the objects and the effects of the present invention in detail, the following description will be given with reference to the embodiments.
The specific embodiment of the invention relates to a preparation process of oat kernel extract, which comprises the following steps:
preparing oat kernel;
soaking oat kernel in water solution with pH of 9-10 at 30-50 deg.C for 4-6 hr, pulverizing to obtain oat kernel slurry, and filtering with 80-100 mesh sieve to obtain filtrate;
adding protease pure enzyme powder into the filtrate, wherein the addition amount of the protease pure enzyme powder is 65-80g/L, and reacting for 1.5-2h under the conditions that the temperature is 40-50 ℃ and the pH is 9-10 to obtain oat kernel enzymatic hydrolysate;
freeze-drying the enzymolysis product to obtain oat kernel extract;
the protease is prepared by the following process:
selecting Bacillus clausii (Bacillus clausii) L-7 with a preservation number of CGMCC No.12953;
activating and culturing the L-7 strain, and fermenting and culturing to obtain protease; the fermentation culture conditions are as follows:
transferred into a fermenter in accordance with an inoculum amount of 5% v/v, the amount of the fermenter charged was 70% (fermentation medium), temperature: 34 ℃; rotating speed: 100-600 r/min; ventilation volume: 1: 0.5-1: 2.5; the dissolved oxygen is maintained at 20-30%;automatically feeding ammonia water and 20% (v/v) hydrochloric acid in the fermentation process to maintain the pH value of the fermentation liquor at 7.0; the fermentation time is 50-60h; the seed culture medium is as follows: 5g/L yeast extract powder, 5g/L tryptone, 10g/L glucose, K 2 HPO 4 18g/L; the fermentation medium comprises: 17g/L of yeast extract powder, 30g/L of cottonseed cake powder, 100g/L of maltodextrin, 3g/L of sodium citrate, 2.6g/L of calcium chloride and K 2 HPO 4 18g/L；
The fermentation broth was centrifuged at 100ml and 8000r/min for 10min, the supernatant was collected and salted out with solid sulfuric acid having a saturation of 70%, the precipitate was collected by centrifugation at 8000r/min for 10min, and the precipitate was dissolved in Tris-HCI (pH 8.8) buffer and dialyzed in the same buffer. Centrifuging to remove a small amount of insoluble substances, collecting supernatant, purifying with DEAE-Sepharose CL-6B ion exchange column, eluting with 20mM Tris-HCl (pH 8.8, containing 10mM NaCl) buffer solution, eluting without adsorbing alkaline protease, collecting enzyme solution, and vacuum freeze drying to obtain pure enzyme powder.
In the above embodiment, bacillus clausii (bacillus clausii) L-7 is an existing strain, and the invention is characterized in that the existing strain is selected as an enzyme-producing bacterium, and the prepared protease is applied to an enzymolysis reaction of an oat kernel extract under a specific fermentation process, so that the yield of oat polypeptide in the oat kernel extract is improved.
In the above embodiment, bacillus clausii (Bacillus clausii) L-7 was deposited in the general microbiological culture Collection center of China Committee for culture Collection of microorganisms at 2016, 9, 12, addresses: the microbial research institute of China academy of sciences, no. 3 of West Lu No.1 of Beijing, chaozhou, chao code 100101, with the collection number of CGMCC No.12953. The strain is referred to by the applicant, tianjin science and technology university, in the patent document having application number 201610886191.5.
As an alternative embodiment, the activation culture is specifically: inoculating into seed culture medium, culturing at 34-40 deg.C for 11-13h at 200 r/min.
As an alternative embodiment, the seed medium is: 5g/L yeast extract powder, 5g/L tryptone, 10g/L glucose, K 2 HPO 4 18g/L。
As an optional embodiment, the freeze-drying process specifically comprises: freezing the oat kernel enzymolysis product at-5 to-10 ℃ for 4-5h, carrying out ultrasonic treatment on the coffee extract by an ultrasonic generator in the freezing process, opening a vacuum pump, and vacuumizing a freeze dryer; starting the heating function of the freeze dryer, adjusting the temperature of the shelf plate to-20 ℃, heating for 12-18h, continuously adjusting the temperature to 20 ℃, heating for 3-4h, continuously adjusting the temperature to 35 ℃, and heating for 1-2h; in the heating process, far infrared radiation treatment and magnetic treatment are assisted on the shelf board to obtain the oat kernel extract.
As an alternative embodiment, the step of preparing oat kernel is: the oat kernel is prepared by milling the oat kernel peel, and the milling rate is 15-16%.
In the above embodiment, the oat kernel is obtained by milling the hull, which can further increase the content of soluble oat polypeptide in the oat kernel extract.
In the above embodiment, the detection of the content of the water-soluble oat polypeptide in the oat kernel extract:
a2 g sample of the prepared oat kernel extract was added to 100mL of water, the pH was adjusted with 0.1mol/L hydrochloric acid and 0.1mol/L sodium hydroxide solution, stirred at 1000rpm/min at room temperature for 1h, centrifuged at 3000rpm/min for 20min, and the polypeptide content in the supernatant was measured.
The method for detecting the content of the polypeptide in the supernatant can specifically adopt a biuret method, the detection method is the prior art, and the detailed process is not repeated here.
Content of soluble oat polypeptide (%) = polypeptide content in supernatant (g)/oat kernel extract sample (g) × 100.
Example 1
The oat kernel is prepared by grinding the oat kernel into the skin, and the skin grinding rate is 15 percent;
soaking oat kernel in water solution of pH9.8 at 40 deg.C for 5 hr, pulverizing to obtain oat kernel slurry, and filtering with 100 mesh sieve to obtain filtrate;
adding protease pure enzyme powder into the filtrate, wherein the addition amount of the protease pure enzyme powder is 70%g/L, reacting for 2 hours at the temperature of 45 ℃ and the pH value of 9.5 to obtain an oat kernel enzymolysis product; the activation medium is as follows: 5g/L yeast extract powder, 5g/L tryptone, 10g/L glucose, K 2 HPO 4 18g/L；
Freeze-drying the enzymolysis product to obtain oat kernel extract;
the freeze drying process specifically comprises the following steps: freezing the oat kernel enzymolysis product at-10 deg.C for 5h, performing ultrasonic treatment on coffee extract with an ultrasonic generator during freezing process, turning on a vacuum pump, and vacuumizing a freeze dryer; starting the heating function of the freeze dryer, adjusting the temperature of the shelving plate to-20 ℃, heating for 16h, continuously adjusting the temperature to 20 ℃, heating for 3.5h, continuously adjusting the temperature to 35 ℃, and heating for 1.5h; in the heating process, the laying plate is assisted with far infrared radiation treatment and magnetic treatment to obtain oat kernel extract;
the protease is prepared by the following process:
selecting Bacillus clausii (Bacillus clausii) L-7 with a preservation number of CGMCC No.12953;
activating and culturing the L-7 strain, and fermenting and culturing to obtain protease;
the activation culture specifically comprises the following steps: inoculating into seed culture medium, culturing at 37 deg.C for 12 hr at 200 r/min;
the fermentation culture conditions are as follows:
transferred into a fermenter in accordance with an inoculum amount of 5% v/v, the amount of the fermenter charged was 70% (fermentation medium), temperature: 34 ℃; rotating speed: 400r/min; ventilation quantity: 1: 0.5-1: 2.5; the dissolved oxygen is maintained at 20-30%; automatically feeding ammonia water and 20% (v/v) hydrochloric acid in the fermentation process to maintain the pH value of the fermentation liquor at 7.0; the fermentation time is 50-60h; the fermentation medium comprises: 17g/L of yeast extract powder, 30g/L of cottonseed cake powder, 100g/L of maltodextrin, 3g/L of sodium citrate, 2.6g/L of calcium chloride and K 2 HPO 4 18g/L；
The fermentation broth was centrifuged at 100ml and 8000r/min for 10min, the supernatant was collected and salted out with solid sulfuric acid having a saturation of 70%, the precipitate was collected by centrifugation at 8000r/min for 10min, and the precipitate was dissolved in Tris-HCI (pH 8.8) buffer and dialyzed in the same buffer. Centrifuging to remove a small amount of insoluble substances, taking supernatant, passing through a DEAE-Sepharose CL-6B ion exchange column for purification, eluting with 20mM Tris-HCl (pH 8.8, containing 10mM NaCl) buffer solution, eluting alkaline protease without adsorption to obtain colorless transparent liquid, collecting enzyme solution, and vacuum freeze-drying to obtain pure enzyme powder, wherein the content of soluble oat polypeptide in the pure enzyme powder is 19.8%.
Example 2
The oat kernel is prepared by grinding the oat kernel into the hull, and the hull grinding rate is 16%;
soaking oat kernel in water solution with pH of 9 at 30 deg.C for 4 hr, pulverizing to obtain oat kernel slurry, and filtering with 80 mesh sieve to obtain filtrate;
adding protease pure enzyme powder into the filtrate, wherein the addition amount of the protease pure enzyme powder is 65g/L, and reacting for 1.5h under the conditions that the temperature is 40 ℃ and the pH value is 9 to obtain oat kernel enzymolysis products; the activation medium is as follows: 5g/L yeast extract powder, 5g/L tryptone, 10g/L glucose, K 2 HPO 4 18g/L；
Freeze-drying the enzymolysis product to obtain oat kernel extract;
the freeze drying process specifically comprises the following steps: freezing the oat kernel enzymolysis product at-5 deg.C for 4h, performing ultrasonic treatment on coffee extract with an ultrasonic generator during freezing process, turning on a vacuum pump, and vacuumizing a freeze dryer; starting the heating function of the freeze dryer, adjusting the temperature of the placing plate to-20 ℃, heating for 12h, continuously adjusting the temperature to 20 ℃, heating for 3h, continuously adjusting the temperature to 35 ℃, and heating for 1h; in the heating process, the laying plate is assisted with far infrared radiation treatment and magnetic treatment to obtain oat kernel extract;
the protease is prepared by the following process:
selecting Bacillus clausii (Bacillus clausii) L-7 with a preservation number of CGMCC No.12953;
activating and culturing the L-7 strain, and fermenting and culturing to obtain protease;
the activation culture specifically comprises the following steps: inoculating into seed culture medium, culturing at 34 deg.C for 11-13 hr at 200 r/min;
the fermentation culture conditions are as follows:
transferred into a fermenter in accordance with an inoculum amount of 5% v/v, the amount of the fermenter charged was 70% (fermentation medium), temperature: 34 ℃; rotating speed: 100r/min; ventilation volume: 1: 0.5-1: 2.5; the dissolved oxygen is maintained at 20-30%; automatically feeding ammonia water and 20% (v/v) hydrochloric acid in the fermentation process to maintain the pH value of the fermentation liquor at 7.0; the fermentation time is 50h; the fermentation medium is as follows: 17g/L of yeast extract powder, 30g/L of cottonseed cake powder, 100g/L of maltodextrin, 3g/L of sodium citrate, 2.6g/L of calcium chloride and K 2 HPO 4 18g/L；
The fermentation broth was centrifuged at 100ml and 8000r/min for 10min, the supernatant was collected and salted out with solid sulfuric acid having a saturation of 70%, the precipitate was collected by centrifugation at 8000r/min for 10min, and the precipitate was dissolved in Tris-HCI (pH 8.8) buffer and dialyzed in the same buffer. Centrifuging to remove a small amount of insoluble substances, taking supernatant, purifying by using a DEAE-Sepharose CL-6B ion exchange column, eluting by using 20mM Tris-HCl (pH 8.8, containing 10mM NaCl) buffer solution, not adsorbing alkaline protease, eluting firstly to obtain colorless transparent liquid, collecting enzyme solution, and carrying out vacuum freeze drying to obtain pure enzyme powder, wherein the content of soluble oat polypeptide in the pure enzyme powder is 17.6%.
Example 3
The oat kernel is prepared by grinding the oat kernel into the skin, and the skin grinding rate is 15-16%;
soaking oat kernel in water solution with pH of 10 at 50 deg.C for 6 hr, pulverizing to obtain oat kernel slurry, and filtering with 100 mesh sieve to obtain filtrate;
adding protease pure enzyme powder into the filtrate, wherein the addition amount of the protease pure enzyme powder is 80g/L, and reacting for 2h under the conditions that the temperature is 40-50 ℃ and the pH value is 10 to obtain oat kernel enzymatic hydrolysis products; the activation medium is as follows: 5g/L yeast extract powder, 5g/L tryptone, 10g/L glucose, K 2 HPO 4 18g/L；
Freeze-drying the enzymolysis product to obtain oat kernel extract;
the freeze drying process specifically comprises the following steps: freezing the oat kernel enzymolysis product at-10 deg.C for 5h, performing ultrasonic treatment on coffee extract with an ultrasonic generator during freezing process, turning on a vacuum pump, and vacuumizing a freeze dryer; starting the heating function of the freeze dryer, adjusting the temperature of the shelf plate to-20 ℃, heating for 18h, continuously adjusting the temperature to 20 ℃, heating for 4h, continuously adjusting the temperature to 35 ℃, and heating for 2h; in the heating process, the laying plate is assisted with far infrared radiation treatment and magnetic treatment to obtain oat kernel extract;
the protease is prepared by the following process:
selecting Bacillus clausii (Bacillus clausii) L-7 with a preservation number of CGMCC No.12953;
activating and culturing the L-7 strain, and fermenting and culturing to obtain protease;
the activation culture specifically comprises the following steps: inoculating into seed culture medium, culturing at 40 deg.C for 13 hr at 200 r/min;
the fermentation culture conditions are as follows:
transferred into a fermenter in accordance with an inoculum amount of 5% v/v, the amount of the fermenter charged was 70% (fermentation medium), temperature: 34 ℃; rotating speed: 600r/min; ventilation quantity: 1: 0.5-1: 2.5; the dissolved oxygen is maintained at 20-30%; automatically feeding ammonia water and 20% (v/v) hydrochloric acid in the fermentation process to maintain the pH value of the fermentation liquor at 7.0; the fermentation time is 60h; the fermentation medium is as follows: 17g/L of yeast extract powder, 30g/L of cottonseed cake powder, 100g/L of maltodextrin, 3g/L of sodium citrate, 2.6g/L of calcium chloride and K 2 HPO 4 18g/L；
The fermentation broth was centrifuged at 100ml and 8000r/min for 10min, the supernatant was collected and salted out with solid sulfuric acid having a saturation of 70%, the precipitate was collected by centrifugation at 8000r/min for 10min, and the precipitate was dissolved in Tris-HCI (pH 8.8) buffer and dialyzed in the same buffer. Centrifuging to remove a small amount of insoluble substances, taking supernatant, purifying by using a DEAE-Sepharose CL-6B ion exchange column, eluting by using 20mM Tris-HCl (pH 8.8, containing 10mM NaCl) buffer solution, not adsorbing alkaline protease, eluting firstly to obtain colorless transparent liquid, collecting enzyme solution, and carrying out vacuum freeze drying to obtain pure enzyme powder, wherein the content of soluble oat polypeptide in the pure enzyme powder is 18.9%.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all equivalent modifications made by the present invention in the specification or directly or indirectly applied to the related technical field are included in the scope of the present invention.
Claims (5)
1. The preparation process of the oat kernel extract is characterized by comprising the following steps:
preparing oat kernel;
soaking oat kernel in water solution with pH of 9-10 at 30-50 deg.C for 4-6 hr, pulverizing to obtain oat kernel slurry, and filtering with 80-100 mesh sieve to obtain filtrate;
adding protease pure enzyme powder into the filtrate, wherein the addition amount of the protease pure enzyme powder is 65-80g/L, and reacting for 1.5-2h under the conditions that the temperature is 40-50 ℃ and the pH is 9-10 to obtain oat kernel enzymatic hydrolysate;
freeze-drying the enzymolysis product to obtain oat kernel extract;
the protease is prepared by the following process:
selecting Bacillus clausii (Bacillus clausii) L-7 with a preservation number of CGMCC No.12953;
activating and culturing the L-7 strain, and fermenting and culturing to obtain protease; the fermentation culture conditions are as follows:
transferred into a fermenter in accordance with an inoculum amount of 5% v/v, the amount of the fermenter charged was 70% (fermentation medium), temperature: 34 ℃; rotating speed: 100-600 r/min; ventilation quantity: 1: 0.5-1: 2.5; the dissolved oxygen is maintained at 20-30%; automatically feeding ammonia water and 20% (v/v) hydrochloric acid in the fermentation process to maintain the pH value of the fermentation liquor at 7.0; the fermentation time is 50-60h; the fermentation medium is as follows: 17g/L of yeast extract powder, 30g/L of cottonseed cake powder, 100g/L of maltodextrin, 3g/L of sodium citrate, 2.6g/L of calcium chloride and K 2 HPO 4 18g/L；
Centrifuging the fermentation broth 100ml at 8000r/min for 10min, collecting supernatant, salting out with solid sulfuric acid with saturation of 70%, centrifuging at 8000r/min for 10min, collecting precipitate, dissolving the precipitate in Tris-HCI (pH8.8) buffer, and dialyzing in the same buffer to remove salt. Centrifuging to remove a small amount of insoluble substances, collecting supernatant, purifying with DEAE-Sepharose CL-6B ion exchange column, eluting with 20mM Tris-HCl (pH 8.8, containing 10mM NaCl) buffer solution, eluting without adsorbing alkaline protease, collecting enzyme solution, and vacuum freeze drying to obtain pure enzyme powder.
2. The process for preparing oat kernel extract according to claim 1, wherein the activation culture is specifically: inoculating into seed culture medium, culturing at 34-40 deg.C for 11-13h at 200 r/min.
3. The process for preparing oat kernel extract according to claim 2, wherein the activation medium is: 5g/L yeast extract powder, 5g/L tryptone, 10g/L glucose, K 2 HPO 4 18g/L。
4. The process for preparing an oat kernel extract according to claim 1, wherein the freeze-drying process is specifically: freezing the oat kernel enzymolysis product at-5 to-10 ℃ for 4-5h, carrying out ultrasonic treatment on the coffee extract by an ultrasonic generator in the freezing process, opening a vacuum pump, and vacuumizing a freeze dryer; starting the heating function of the freeze dryer, adjusting the temperature of the shelf plate to-20 ℃, heating for 12-18h, continuously adjusting the temperature to 20 ℃, heating for 3-4h, continuously adjusting the temperature to 35 ℃, and heating for 1-2h; in the heating process, far infrared radiation treatment and magnetic treatment are assisted on the shelf board to obtain the oat kernel extract.
5. The process for preparing oat kernel extract as claimed in claim 1, wherein the step of preparing oat kernel comprises: the oat kernel is prepared by milling the oat kernel peel, and the milling rate is 15-16%.
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