CN115216507A - Method for separating highland barley bran polypeptide by using double enzymolysis technology - Google Patents
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- 235000007340 Hordeum vulgare Nutrition 0.000 title claims abstract description 75
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 37
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 37
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 29
- 240000005979 Hordeum vulgare Species 0.000 title 1
- 241000209219 Hordeum Species 0.000 claims abstract description 74
- 108091005658 Basic proteases Proteins 0.000 claims abstract description 43
- 239000000843 powder Substances 0.000 claims abstract description 37
- 108090000145 Bacillolysin Proteins 0.000 claims abstract description 33
- 102000035092 Neutral proteases Human genes 0.000 claims abstract description 33
- 108091005507 Neutral proteases Proteins 0.000 claims abstract description 33
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 29
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 29
- 230000007062 hydrolysis Effects 0.000 claims abstract description 24
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 24
- 239000004365 Protease Substances 0.000 claims abstract description 15
- 102000004142 Trypsin Human genes 0.000 claims abstract description 15
- 108090000631 Trypsin Proteins 0.000 claims abstract description 15
- 239000012588 trypsin Substances 0.000 claims abstract description 15
- 108091005804 Peptidases Proteins 0.000 claims abstract description 8
- 238000001035 drying Methods 0.000 claims abstract description 7
- 239000002994 raw material Substances 0.000 claims abstract description 4
- 102000004190 Enzymes Human genes 0.000 claims description 36
- 108090000790 Enzymes Proteins 0.000 claims description 36
- 239000000243 solution Substances 0.000 claims description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- 230000002779 inactivation Effects 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 12
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 11
- 102000035195 Peptidases Human genes 0.000 claims description 7
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 6
- 238000005238 degreasing Methods 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 5
- 210000000582 semen Anatomy 0.000 claims description 4
- 108010009736 Protein Hydrolysates Proteins 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 230000001376 precipitating effect Effects 0.000 claims description 3
- 239000008213 purified water Substances 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 3
- 238000000354 decomposition reaction Methods 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 abstract description 3
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract 1
- 229940088598 enzyme Drugs 0.000 description 32
- 230000000694 effects Effects 0.000 description 7
- 230000017854 proteolysis Effects 0.000 description 5
- 108090000526 Papain Proteins 0.000 description 3
- 108090000284 Pepsin A Proteins 0.000 description 3
- 102000057297 Pepsin A Human genes 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000007071 enzymatic hydrolysis Effects 0.000 description 3
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 3
- 229940055729 papain Drugs 0.000 description 3
- 235000019834 papain Nutrition 0.000 description 3
- 229940111202 pepsin Drugs 0.000 description 3
- 239000003513 alkali Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 235000019419 proteases Nutrition 0.000 description 2
- 239000012460 protein solution Substances 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 238000007696 Kjeldahl method Methods 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- -1 inc.) Substances 0.000 description 1
- 230000010355 oscillation Effects 0.000 description 1
- 238000005191 phase separation Methods 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- 229960001322 trypsin Drugs 0.000 description 1
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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Abstract
A method for separating highland barley bran polypeptide by using double enzymolysis technology comprises preparing rough highland barley protein powder from highland barley bran, preparing into solution, adding alkaline protease and neutral protease or double protease combination of alkaline protease and trypsin for enzymolysis, and drying to obtain highland barley bran polypeptide powder. The highland barley bran is used as a raw material, alkaline protease and neutral protease are adopted to carry out double-enzyme hydrolysis on the highland barley bran protein, the process means is refined, the resource utilization rate is obviously improved, and the preparation cost is reduced.
Description
Technical Field
The invention relates to a technology in the field of bioengineering, in particular to a method for separating highland barley bran polypeptide by using a double enzymolysis technology.
Background
The existing preparation process of the highland barley bran polypeptide usually carries out enzymolysis on highland barley bran, and then extracts highland barley bran oil by adopting a three-phase separation extraction method, a solvent extraction method or a freezing demulsification extraction method, but the prior art can not separate and further apply protein in highland barley, and loses the possibility of deep processing and utilization of part of highland barley bran.
Disclosure of Invention
The invention provides a method for separating highland barley bran polypeptide by using a double enzymolysis technology aiming at the defects in the prior art, highland barley bran is taken as a raw material, highland barley bran protein is subjected to double-enzyme hydrolysis by using alkaline protease and neutral protease, the process means is refined, the resource utilization rate is obviously improved, and the preparation cost is reduced.
The invention is realized by the following technical scheme:
the invention relates to a method for separating highland barley bran polypeptide by using a double enzymolysis technology, which comprises the steps of preparing rough highland barley protein powder from highland barley bran, preparing into solution, adding alkaline protease and neutral protease or a double protease combination of the alkaline protease and trypsin for enzymolysis, and drying to obtain highland barley bran polypeptide powder.
The crude highland barley protein powder is prepared by taking highland barley bran powder as a raw material, carrying out degreasing water bath, adding hydrochloric acid solution for decomposition, and removing supernatant.
The highland barley bran powder is obtained by completely grinding highland barley bran into powder and collecting the powder through an 80-mesh screen.
The degreasing means that: defatting with n-hexane at a ratio of solvent/highland barley bran powder of 1 (w/v), defatting at 25 deg.C for 6 hr under shaking, drying, and storing at 4 deg.C.
The water bath refers to the following steps: taking the degreased highland barley bran powder, adding purified water, wherein the material-liquid ratio is 1.
The hydrochloric acid solution is added as follows: centrifuging the water-bath highland barley bran solution at 3500r/min and 4 ℃ for 15 minutes; adding 1mol/L HCl solution into the centrifuged supernatant, and adjusting the pH value of the supernatant to 4.5; centrifuging the mixed solution for 20 minutes at 4000r/min and 4 ℃; removing supernatant, precipitating, and lyophilizing to obtain crude semen Avenae Nudae protein powder.
The double-protease combined enzymolysis specifically comprises the following steps:
when the highland barley bran protein powder is subjected to enzymolysis by adopting double enzymes of alkaline protease and trypsin, the enzyme adding amount is 3%, the temperature is 50 ℃, the pH value of the solution is 8, and the time is 2 hours, wherein the alkaline protease is trypsin =1, the preferable enzyme adding amount of the alkaline protease is 12000U/mg, the enzymolysis temperature is 50 ℃, and the pH value of the solution is 9; the optimal enzyme adding amount of the neutral protease is 12000U/mg, the enzymolysis temperature is 45 ℃, and the pH value of the solution is 7;
when the highland barley bran protein powder is subjected to enzymolysis by adopting double enzymes of alkaline protease and neutral protease, the enzyme addition amount is 3-7%, and the alkaline protease: the neutral protease =1, the enzymolysis temperature is 45-55 ℃, the pH value of the solution is 8-9, the time is 2-6 hours, the preferable enzyme adding amount of the trypsin is 12000U/mg, the enzymolysis temperature is 37 ℃, and the pH value of the solution is 7.5.
The double-protease combined enzymolysis is preferably performed by using a combination of alkaline protease and neutral protease, the enzyme adding amount is 3%, the temperature is 50 ℃, the pH of the solution is 8, the time is 2 hours, and the alkaline protease, the neutral protease = 1.
The enzymolysis specifically comprises the following operations: preparing the crude highland barley bran protein powder into a solution with the mass concentration of 5%, adjusting the pH value of the solution to 8-11, carrying out water bath for 10 minutes at a proper temperature, and then adding a double-protease combination for enzymolysis for 2-8 hours; after the enzymolysis is finished, carrying out enzyme inactivation by water bath for 10 minutes at 90 ℃; after inactivation, the hydrolysis degree (%) is determined, and the polypeptide yield (%) is determined; filtering the hydrolysate, ultrafiltering, and drying at low temperature to obtain highland barley bran polypeptide powder.
Technical effects
The highland barley bran polypeptide is obtained by performing combined enzymolysis on highland barley bran protein through alkaline protease and neutral protease, wherein the polypeptide is obtained from the highland barley bran and the used enzymolysis combination does not appear in the related documents of the conventional highland barley bran, and compared with the conventional technical means, the highland barley bran polypeptide has the obviously improved technical details that the yield of the polypeptide subjected to enzymolysis through the double-enzyme combination can reach 12.04 percent, and the technical effect is higher than that of other methods.
Drawings
FIG. 1 is a flow chart of the present invention;
FIG. 2 is a schematic view of the degree of hydrolysis by a single protease;
FIG. 3 is a schematic diagram of the degree of hydrolysis by enzyme hydrolysis in a two-enzyme combination;
FIG. 4 is a graph showing the degree of hydrolysis of alkaline protease and neutral protease in combination at different ratios;
fig. 5 is a schematic diagram of the yield of polypeptide by the combination of two enzymes and single enzyme enzymolysis.
Detailed Description
Example 1
Method for extracting highland barley protein
Completely grinding the highland barley bran into powder, and collecting the powder through a 80-mesh screen; weighing a certain amount of powder, degreasing with n-hexane at a ratio of solvent/highland barley bran powder of 1 (w/v), defatting for 6 hours at 25 deg.C under oscillation, drying, and storing at 4 deg.C. Taking the degreased highland barley bran powder, adding purified water, adjusting the pH value to 11 with 1mol/L NaOH at 40 ℃ in a material-liquid ratio of 1; centrifuging the water-bath highland barley bran solution at 3500r/min and 4 deg.C for 15 min; adding 1mol/L HCl solution into the centrifuged supernatant, and adjusting the pH value of the supernatant to 4.5; centrifuging the mixed solution for 20 minutes at 4000r/min and 4 ℃; removing supernatant, precipitating, and lyophilizing to obtain crude semen Avenae Nudae protein powder. The protein yield is determined by a Kjeldahl method (GB 5009.5-2010).
Under the conditions that the feed-liquid ratio is 1.
Example 2
Effect of different proteases on the degree of proteolysis
Taking the prepared crude highland barley bran protein powder, preparing a solution with the mass concentration of 5%, adjusting the pH of the solution according to the optimal pH range of different enzymes by using a pH detector (Shanghai Yueping scientific instruments, inc.), putting the solution into a water bath pot (Shanghai Jing Macro experimental equipment, inc.), carrying out water bath for 10 minutes at an appropriate temperature, then weighing an appropriate amount of enzyme by using an analytical balance (Aohaus instruments, shanghai, inc.)) and adding the enzyme into the solution for enzymolysis, wherein the enzymolysis time is 2-8 hours; after the enzymolysis is finished, carrying out enzyme inactivation by water bath for 10 minutes at 90 ℃; after inactivation, the degree of hydrolysis (%) was determined and the results of the different enzymatic hydrolysis were compared. The selected single enzymes respectively comprise alkaline protease (Shanghai Maxin Biochemical technology, inc.), neutral protease (Shanghai Yi En chemical technology, inc.), trypsin (Hadamard reagent, inc.), papain (Hadamard reagent, inc.), and pepsin (Hadamard Biochemical technology, inc.).
Degree of Hydrolysis (DH) assay:
according to
h＝V×C b X 1/α x 1/m; h is the amount of peptide bonds hydrolyzed in the highland barley bran protein of unit mass; h is a total of tot The highland barley protein h is the total amount of peptide bonds contained in the protein per unit mass tot Taking 8.3mmg/g; v is the amount of alkali consumed in the enzymolysis process, mL; c b The concentration of the alkali solution is mol/L; m is the mass of protein in the enzymolysis liquid, g; α is the average degree of dissociation of α -NH2, α =10 pH-pk /(1+10 pH-pk )。
As shown in fig. 2, the results showed that the degree of hydrolysis by alkaline protease was 8.26%, that by neutral protease was 7.66%, that by trypsin was 5.89%, that by papain was 5.09%, and that by pepsin was 4.42%. Among them, the use of alkaline protease, neutral protease and trypsin can achieve a higher degree of hydrolysis.
Example 3
Effect of combinations of different proteases on the degree of proteolysis
According to the method of the embodiment 2, respectively adding alkaline protease + neutral protease (1) and alkaline protease + trypsin (1) into a highland barley bran protein solution, adjusting the temperature of the solution, and carrying out enzymolysis on the highland barley bran protein under the condition that the pH value is proper to the enzyme; after the enzymolysis is finished, carrying out enzyme inactivation by water bath for 10 minutes at 90 ℃; after inactivation, the degree of hydrolysis (%) was determined using the pH-stat method. As shown in fig. 3, the results showed that the degree of hydrolysis by alkaline protease + neutral protease was 8.55% and the degree of hydrolysis by alkaline protease + trypsin was 6.42%. The degree of hydrolysis obtained is higher when alkaline protease + neutral protease is used and higher than when any one of alkaline protease, neutral protease, trypsin, papain and pepsin is used.
Example 4
Effect of different ratios of alkaline protease and neutral protease on the degree of proteolysis
According to the method of the example 2, adding alkaline protease and neutral protease into the highland barley bran protein solution according to different ratios (1, 3, 1; after the enzymolysis is finished, carrying out enzyme inactivation by water bath for 10 minutes at 90 ℃; after inactivation, the degree of hydrolysis (%) was determined using the pH-stat method. As shown in fig. 4, the results show that the alkaline protease has a degree of hydrolysis of 5.27%,7.28%,8.55%,6.71%,6.42% when the neutral protease is 1,1. Compared with other ratios, the alkaline protease to neutral protease has the highest hydrolysis degree when the ratio is 1.
Example 5
Effect of combination of enzymes on the degree of proteolysis under different conditions
Selecting four factors of enzymolysis time, enzymolysis temperature, pH and enzyme adding amount, and determining the proteolysis degree of the alkaline protease and the neutral protease by adopting conditions set in a four-factor three-level orthogonal test table. According to the method of the embodiment 2, the temperature and the pH value of the solution are adjusted to the condition that the enzyme is suitable, and alkaline protease and neutral protease with different concentrations are added to carry out enzymolysis on the highland barley bran protein; after the enzymolysis is finished, carrying out enzyme inactivation by water bath for 10 minutes at 90 ℃; after inactivation, the degree of hydrolysis (%) was determined using the pH-stat method. As shown in the following table, when the amount of the added enzyme was 5% (1 in the ratio of alkaline protease to neutral protease), 55 ℃ and pH =9, and the enzymatic hydrolysis was performed for 4 hours, the hydrolysis degree was the highest as compared with other enzymatic hydrolysis conditions.
Example 6
Effect of double and Single enzymes on polypeptide yield
Determination of polypeptide yield (%): taking a certain volume of filtered hydrolysate, adding 15% trichloroacetic acid solution in equal proportion, carrying out water bath reaction at 30 ℃ for 30min, centrifuging at 5000r/min for 15min to remove macromolecular protein, collecting supernatant, diluting by a certain multiple, measuring the polypeptide concentration in the hydrolysate according to a Fulin phenol method,
as shown in fig. 5, the polypeptide yields were 10.32% and 9.39% respectively by enzymolysis with alkaline protease and neutral protease; the polypeptide yield is 12.04% by double-enzyme enzymolysis.
Obviously, the hydrolysis degree and the polypeptide yield obtained by using the double enzymes are obviously higher than the results obtained by using the single enzyme (p is less than 0.005. Although the alkaline protease has strong hydrolysis capacity, the price is higher, the production and use cost is higher, the alkaline protease and the neutral protease with better hydrolysis capacity are combined for use, and the highland barley bran protein is subjected to enzymolysis, so that the polypeptide yield is improved, the production cost of the highland barley bran polypeptide is reduced to a certain extent, and the utilization rate of a highland barley bran processing byproduct is improved.
The foregoing embodiments may be modified in many different ways by those skilled in the art without departing from the spirit and scope of the invention, which is defined by the appended claims and all changes that come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (10)
1. A method for separating highland barley bran polypeptide by using a double enzymolysis technology is characterized in that crude highland barley protein powder is prepared from highland barley bran, prepared into solution, added with alkaline protease and neutral protease or a double protease combination of the alkaline protease and trypsin for enzymolysis, and dried to obtain highland barley bran polypeptide powder.
2. The method for separating highland barley bran polypeptides by using a double enzymolysis technology as claimed in claim 1, wherein the crude highland barley protein powder is obtained by using highland barley bran powder as a raw material, carrying out degreasing water bath, adding hydrochloric acid solution for decomposition, and removing supernatant.
3. The method for separating highland barley bran polypeptides by using the double enzymolysis technology as claimed in claim 1 or 2, wherein the highland barley bran powder is obtained by completely grinding highland barley bran into powder and collecting the powder through an 80-mesh screen.
4. The method for separating highland barley bran polypeptides by using the double enzymolysis technology as claimed in claim 2, wherein the degreasing is: defatting with n-hexane at a ratio of solvent/highland barley bran powder of 1 (w/v), defatting at 25 deg.C for 6 hr under shaking, drying, and storing at 4 deg.C.
5. The method for separating highland barley bran polypeptides by using double enzymolysis technology as claimed in claim 2, wherein the water bath is characterized in that: taking the degreased highland barley bran powder, adding purified water, wherein the material-liquid ratio is 1.
6. The method for separating highland barley bran polypeptides by using the double enzymolysis technology as claimed in claim 2, wherein the adding of hydrochloric acid solution is: centrifuging the water-bath highland barley bran solution at 3500r/min and 4 deg.C for 15 min; adding 1mol/L HCl solution into the centrifuged supernatant, and adjusting the pH value of the supernatant to 4.5; centrifuging the mixed solution for 20 minutes at 4000r/min and 4 ℃; removing supernatant, precipitating, and lyophilizing to obtain crude semen Avenae Nudae protein powder.
7. The method for separating highland barley bran polypeptides by using the double enzymolysis technology as claimed in claim 1, wherein the double protease combination enzymolysis specifically comprises:
when the highland barley bran protein powder is subjected to enzymolysis by adopting double enzymes of alkaline protease and trypsin, the enzyme adding amount is 3%, the temperature is 50 ℃, the pH value of the solution is 8, and the time is 2 hours, wherein the alkaline protease is trypsin = 1;
when the highland barley bran protein powder is subjected to enzymolysis by adopting double enzymes of alkaline protease and neutral protease, the enzyme addition amount is 3-7%, and the alkaline protease: neutral protease =1, the enzymolysis temperature is 45-55 ℃, the pH value of the solution is 8-9, and the time is 2-6 hours.
8. The method for separating highland barley bran polypeptides by using double enzymolysis technology as claimed in claim 7, wherein the enzyme dosage of the alkaline protease is 12000U/mg, the enzymolysis temperature is 50 ℃, and the pH of the solution is 9; the enzyme adding amount of the neutral protease is 12000U/mg, the enzymolysis temperature is 45 ℃, and the pH value of the solution is 7; the adding amount of the trypsin is 12000U/mg, the enzymolysis temperature is 37 ℃, and the pH value of the solution is 7.5.
9. The method for separating highland barley bran polypeptides by using the double enzymolysis technology as claimed in claim 1, wherein the double protease combination is used for enzymolysis, specifically, the combination of alkaline protease and neutral protease, the enzyme adding amount is 3%, the temperature is 50 ℃, the solution pH is 8, the time is 2 hours, and the alkaline protease is neutral protease = 1.
10. The method for separating highland barley bran polypeptides by using double enzymolysis technology as claimed in any one of claims 7 to 9, wherein the enzymolysis specifically comprises the following operations: preparing the crude highland barley bran protein powder into a solution with the mass concentration of 5%, adjusting the pH value of the solution to 8-11, carrying out water bath for 10 minutes at a proper temperature, and then adding a double-protease combination for enzymolysis for 2-8 hours; after the enzymolysis is finished, carrying out enzyme inactivation by water bath for 10 minutes at 90 ℃; after inactivation, the degree of hydrolysis (%) and the polypeptide yield (%) are determined; filtering the hydrolysate, ultrafiltering, and drying at low temperature to obtain semen Avenae Nudae bran polypeptide powder.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101418038A (en) * | 2008-11-14 | 2009-04-29 | 华子昂 | Highland barley peptide and preparation method thereof |
| CN101455241A (en) * | 2008-12-31 | 2009-06-17 | 西藏自治区高原生物研究所 | Highland barley bran oil preparation method from highland barley bran oil using water enzyme method |
| KR20130012688A (en) * | 2011-07-26 | 2013-02-05 | 충남대학교산학협력단 | Manufacture method of mineral binding peptides from barley bran proteins |
| CN109206535A (en) * | 2018-10-08 | 2019-01-15 | 江苏沿江地区农业科学研究所 | A method of preparing high-purity beta glucan from wheat wheat bran |
| CN109548951A (en) * | 2018-11-16 | 2019-04-02 | 潍坊医学院 | The extracting method of quinoa bran protein and its application in terms of food |
| CN113244657A (en) * | 2021-06-18 | 2021-08-13 | 四川省农业科学院农产品加工研究所 | Echelon extraction method of functional components of purple highland barley bran |
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101418038A (en) * | 2008-11-14 | 2009-04-29 | 华子昂 | Highland barley peptide and preparation method thereof |
| CN101455241A (en) * | 2008-12-31 | 2009-06-17 | 西藏自治区高原生物研究所 | Highland barley bran oil preparation method from highland barley bran oil using water enzyme method |
| KR20130012688A (en) * | 2011-07-26 | 2013-02-05 | 충남대학교산학협력단 | Manufacture method of mineral binding peptides from barley bran proteins |
| CN109206535A (en) * | 2018-10-08 | 2019-01-15 | 江苏沿江地区农业科学研究所 | A method of preparing high-purity beta glucan from wheat wheat bran |
| CN109548951A (en) * | 2018-11-16 | 2019-04-02 | 潍坊医学院 | The extracting method of quinoa bran protein and its application in terms of food |
| CN113244657A (en) * | 2021-06-18 | 2021-08-13 | 四川省农业科学院农产品加工研究所 | Echelon extraction method of functional components of purple highland barley bran |
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