CN114395606A - Method for preparing sesame oligopeptide from sesame oil by-product by water extraction method - Google Patents
Method for preparing sesame oligopeptide from sesame oil by-product by water extraction method Download PDFInfo
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
Abstract
The invention discloses a method for preparing sesame oligopeptide from a sesame oil byproduct by an aqueous extraction method, belonging to the technical field of biology. The invention takes the byproduct sesame seed meal of the sesame oil preparation by the water generation method as the raw material, adopts double-enzyme synchronous enzymolysis, and is separated and purified by activated carbon decoloration, microfiltration and ultrafiltration, and spray drying to obtain the sesame oligopeptide product, wherein the hydrolysis degree is 33.9 percent, the peptide segment of the oligopeptide product is less than 1000 Da, the percentage of the peptide segment is 87.5 percent, and the purity is 91.4 percent. The preparation method of the sesame oligopeptide disclosed by the invention is high in hydrolysis degree, short in time consumption, high in product purity, light yellow in color and has good application value, and is suitable for large-scale production.
Description
Technical Field
The invention relates to a method for preparing sesame oligopeptide from a sesame oil byproduct by an aqueous extraction method, belonging to the technical field of biology.
Background
Sesame is one of four oil crops in China, and contains 45% -63% of grease, 20% -27% of protein and 50% of sesame seeds for preparing oil. The water substitution method is a traditional method for preparing sesame oil in China, accounts for more than 40% of the annual processing amount of the sesame oil, and the by-product generated in the production process is sesame meal. The sesame seed meal is produced by 1 ton per 1 ton of sesame oil, and is about 16 ten thousand tons per year in China. The sesame seed meal contains about 40% of protein and 2.3-11.0% of crude fat (based on the dry weight of the sesame seed meal), and is rich in protein resources. The sesame protein has the highest content of glutamic acid and aspartic acid, the content of cysteine and methionine is higher than that of soybean protein and corn protein, hydroxyl free radicals and superoxide anion free radicals can be eliminated, and the sesame protein has strong oxidation resistance. The molecular weight of the oligopeptide is below 1000 Da, and the oligopeptide can be directly absorbed and utilized by a human body and has biological functions. Therefore, the preparation of oligopeptide with antioxidant activity by using sesame seed meal as a raw material becomes a current research hotspot and draws extensive attention of academia.
At present, experts and scholars research the technology for preparing sesame peptide by using sesame meal generated by different oil making processes as raw materials.
The condition optimization of the preparation of short-peptide chelated zinc by utilizing cold-pressed sesame protein zymolyte [ J ] grain and oil food science and technology, 2014, 22(6): 83-87.) uses cold-pressed sesame seed meal as a raw material, obtains sesame protein by alkali dissolution and acid precipitation and freeze drying, and then carries out enzymolysis on the sesame protein by using papain, wherein the hydrolysis degree is 14.98%. Wang Shoturn, Liuming Chuan, Yangjie, a sesame peptide powder and its preparation method and use [ P ]. Zhejiang province: CN110669099B, 2021-12-03.) the sesame peptide powder with purity of 91.6% is prepared by using cold-pressed sesame seed dregs as raw material, alkali extracting protein for 4 h, neutral proteinase enzymolysis for 3 h, microfiltration of 0.1 μm, and spray drying, and the yield of the polypeptide is 22%. The method selects cold-pressed sesame seed meal as a raw material, but the oil prepared by a cold pressing method in the existing sesame oil production technology only accounts for 11.1 percent, the source of the raw material is less, the protein is firstly extracted and then the neutral protease or papain is used for enzymolysis, the hydrolysis degree is low, and the process is complex.
Taking hot-pressed sesame cake meal as a raw material, degreasing by normal hexane, carrying out ultrasonic-assisted alkali extraction, freezing and drying to obtain sesame protein, carrying out enzymolysis by alkaline protease, centrifuging and freeze-drying to prepare the sesame polypeptide, wherein the hydrolysis degree is 20.3%. The method uses hot-pressed sesame seed meal as a raw material, protein is seriously denatured through high-temperature extrusion in the hot-pressing process, pigment is combined with the protein, the method does not relate to the decoloring problem of active peptide, the product is dark in color, the method still uses the technology of preparing polypeptide by extracting the protein firstly and then performing enzymolysis, the hydrolysis degree is still low, the process is still complex, and the environmental pollution is serious because organic solvent is used for removing oil in the process.
Research on extraction of proteins from subcritical sesame seed meal and preparation of ACE inhibitory peptides [ D ]. Henan university, 2015.) from Yuandong (Yuandong. subcritical sesame seed meal), carrying out alkali extraction and freeze drying on subcritical sesame seed meal to obtain sesame protein, carrying out enzymolysis by using alkaline protease and membrane separation to prepare sesame peptide with the function of reducing blood pressure, wherein the hydrolysis degree is 19.51%, and three peptide segments with molecular weights of 1416.6 Da, 1746.5 Da and 2017.6 Da are obtained. Research on separation, purification and structure identification of sesame antioxidant peptide [ J ] of China grain and oil institute, 2019, 34(3): 45-60.) on subcritical sesame meal as a raw material, preparing sesame protein by an alkali-soluble acid precipitation method, and preparing the sesame antioxidant peptide with molecular weights of 1328.7 Da, 1585.7 Da, 1738.8 Da, 1679.8 Da and 1774.8 Da through trypsin enzymolysis and membrane separation, wherein the hydrolysis degree is 17.54%. The method selects subcritical sesame seed meal as a raw material, the technology of subcritical sesame oil extraction is still in a research stage, large-scale production is not realized, the source of the raw material is less, the preparation process still adopts the steps of firstly extracting protein and then preparing sesame peptide by trypsin or alkaline protease single enzyme, the hydrolysis degree is still lower, and the process is still complex.
Lijia (Lijia, research on protein extraction and protein peptide preparation from sesame oil processing by-products [ D ]. 2016, university of Shandong agriculture) uses water-substituted sesame seed meal as raw material, prepares sesame protein by alkali-soluble acid precipitation, prepares sesame peptide with antioxidant activity by alkaline protease enzymolysis and centrifugal freeze-drying, and has hydrolysis degree of 20.35%, and the obtained peptide segment has molecular weight ranges of 92-. Although the raw material of the method is sesame meal by a water-substitution method and has rich sources, the process is not improved, the method is to extract protein and perform single enzyme enzymolysis, the hydrolysis degree is low, the molecular weight of the obtained sesame peptide is high, and the problem of deep color of the sesame peptide is not solved.
Xue Sa Jiang (Xue Jiang. development of sesame protein peptide and its process design [ D ]. Henan university of Industrial, Sanguan.) uses cold pressed sesame seed meal as raw material, and is leached by 70% ethanol to obtain sesame protein, then the sesame peptide is prepared by stepwise enzymolysis and centrifugation with alkaline protease and flavourzyme, the enzymolysis is 6 h, the hydrolysis degree is 31.78%, and the content of soluble peptide is 85.14%. Although the hydrolysis degree is improved, the method still extracts protein and then carries out enzymolysis, the time consumption is long, and the sesame peptide is purified by centrifugation, so that the product purity is low.
The method firstly prepares the sesame protein and then carries out enzymolysis, and has the disadvantages of complex production process, long preparation period and low production efficiency.
Shaojiawei (Shaojiawei, sesame seed meal protein peptide preparation separation and activity research [ D ]. Jinan university, 2021.) uses alkaline protease to directly carry out enzymolysis on subcritical sesame seed meal to prepare the sesame peptide with the antioxidant function, wherein the yield is 12.38%. The method has the advantages of less raw material sources and lower product yield.
Disclosure of Invention
Aiming at the defects of the technology, the invention provides the preparation method of the sesame oligopeptide, which has the advantages of simple process, convenient preparation and environmental protection, simplifies the preparation process of the sesame peptide, shortens the preparation time, improves the hydrolysis degree, reduces the pigment content and improves the product purity.
The invention relates to a method for preparing sesame oligopeptide from a sesame oil byproduct by an aqueous method, which comprises the following steps:
first, double enzyme synchronous enzymolysis
Uniformly mixing sesame seed meal and distilled water according to a material-liquid ratio of 1:20-1:30(g: mL), adjusting the pH value to 9.0-10.0 by 1 mol/L sodium hydroxide solution, adding alkaline protease and papain (food grade) according to the enzyme activity unit of the alkaline protease and the papain of 1:1-2:1(U: U), adding 6000-8000U per gram of sesame seed meal, synchronously performing enzymolysis at 45-55 ℃ for 2-4 h by using double enzymes, and performing enzyme deactivation at 95 ℃ for 15 min to obtain an enzymolysis liquid;
the sesame seed meal is a byproduct of sesame oil preparation by a water generation method, the water content is 67.3%, and the protein content is 12.6%;
the double-enzyme synchronous enzymolysis is that alkaline protease and papain directly hydrolyze sesame meal at the same time;
the enzyme deactivation is to deactivate alkaline protease and papain at 95 deg.C for 15 min;
② separation and purification of enzymolysis liquid
Regulating the pH value of the enzymolysis liquid obtained in the step (I) to 5.0-6.0, adding 1-2% (mass fraction) of activated carbon powder, and adsorbing at a constant temperature of 50-60 ℃ for 30-60 min; centrifuging at 7000 rpm for 15 min, microfiltering the supernatant (0.45 μm), ultrafiltering (molecular weight cut-off of 3000 Da), and spray drying the filtrate (inlet temperature of 170 deg.C and outlet temperature of 90 deg.C) to obtain semen Sesami oligopeptide product;
the constant-temperature adsorption is to add activated carbon powder to adsorb macromolecular protein, pigment and grease in the enzymolysis liquid, so that the purity of the oligopeptide is improved;
the microfiltration adopts a 0.45 mu m microporous filter membrane;
the ultrafiltration is an ultrafiltration membrane of Beijing Asahi Bangkong membrane equipment, Limited liability company, and is made of polysulfone material with the molecular weight cutoff of 3000 Da;
the hydrolysis degree of the present invention is 32.9-33.9%, the sesame oligopeptide yield is 30.8-33.2% (sesame oligopeptide mass/sesame meal protein content), the peptide segment of oligopeptide product < 1000 Da accounts for 85.9-87.5%, and the purity is 91.0-91.4% (mass fraction).
Compared with the prior art, the method for preparing the sesame oligopeptide has the following remarkable characteristics:
(1) the invention takes the sesame seed meal which is the byproduct of the sesame oil by the water extraction method as the raw material, has wide source, low price and easy obtainment and is suitable for large-scale production;
(2) the sesame seed meal is directly and synchronously enzymolyzed by double enzymes, so that the loss of intermediate extracted protein is reduced, the production process is simplified, the enzymolysis time is shortened, the enzymolysis efficiency is improved, and the oligopeptide preparation time is shortened to 4 hours;
(3) adsorbing macromolecular protein, pigment and grease by using active carbon, performing microfiltration, and performing ultrafiltration (3000 Da) to obtain high-purity sesame oligopeptide, so that the production cost is saved, the energy consumption is reduced, the sesame oligopeptide is light yellow, and the product has good color and high purity;
(4) the highest hydrolysis degree of the obtained sesame oligopeptide can reach 33.9%, the yield is 33.2% (the quality of active sesame peptide/the content of sesame meal protein), the peptide segment of the oligopeptide product is less than 1000 Da and accounts for 87.5%, the purity is 91.4% (mass fraction), and the hydroxyl radical clearance rate of each microgram g of the oligopeptide product is 28.50%, which is 2 times of the hydroxyl radical clearance rate of the same Vc.
Detailed Description
The invention is further described below with reference to examples, but the scope of the invention is not limited to the examples.
Example 1:
first, double enzyme synchronous enzymolysis
Uniformly mixing sesame seed meal and distilled water according to a feed-liquid ratio of 1:25(g: mL), adjusting the pH value to 9.5 by using 1 mol/L sodium hydroxide solution, adding alkaline protease and papain (food grade) according to an enzyme activity unit of 1.5:1(U: U) of the alkaline protease and the papain, adding 7000U of the alkaline protease and the papain into each gram of the sesame seed meal, synchronously performing enzymolysis for 3 hours at 50 ℃, and performing enzyme deactivation for 15 minutes at 95 ℃ to obtain an enzymolysis solution;
the sesame seed meal is a byproduct of sesame oil preparation by a water generation method, the water content is 67.3%, and the protein content is 12.6%;
the double-enzyme synchronous enzymolysis is that alkaline protease and papain directly hydrolyze sesame meal at the same time;
the enzyme deactivation is to deactivate alkaline protease and papain at 95 deg.C for 15 min;
② separation and purification of enzymolysis liquid
Regulating the pH value of the enzymolysis liquid obtained in the step I to 5.5, adding 1.5 percent (mass fraction) of activated carbon powder, and adsorbing at the constant temperature of 55 ℃ for 40 min; centrifuging at 7000 rpm for 15 min, microfiltering the supernatant (0.45 μm), ultrafiltering (molecular weight cut-off of 3000 Da), and spray drying the filtrate (inlet temperature of 170 deg.C and outlet temperature of 90 deg.C) to obtain semen Sesami oligopeptide product;
the constant-temperature adsorption is to add activated carbon powder to adsorb macromolecular protein, pigment and grease in the enzymolysis liquid, so that the purity of the oligopeptide is improved;
the microfiltration adopts a 0.45 mu m microporous filter membrane;
the ultrafiltration is an ultrafiltration membrane of Beijing Asahi Bangkong membrane equipment, Limited liability company, and is made of polysulfone material with the molecular weight cutoff of 3000 Da;
the hydrolysis degree of the present invention is 33.9%, the yield of the sesame oligopeptide is 33.2%, the peptide segment of the oligopeptide product is less than 1000 Da is 87.5%, and the purity is 91.4%.
Example 2:
first, double enzyme synchronous enzymolysis
Uniformly mixing sesame seed meal and distilled water according to a feed-liquid ratio of 1:30(g: mL), adjusting the pH value to 9.0 by 1 mol/L sodium hydroxide solution, adding alkaline protease and papain (food grade) according to an enzyme activity unit of 1:1(U: U) of the alkaline protease and the papain, adding 6000U into each gram of sesame seed meal, synchronously performing enzymolysis for 2 hours at 45 ℃, and performing enzyme deactivation for 15 minutes at 95 ℃ to obtain an enzymolysis liquid;
the sesame seed meal is a byproduct of sesame oil preparation by a water generation method, the water content is 67.3%, and the protein content is 12.6%;
the double-enzyme synchronous enzymolysis is that alkaline protease and papain directly hydrolyze sesame meal at the same time;
the enzyme deactivation is to deactivate alkaline protease and papain at 95 deg.C for 15 min;
② separation and purification of enzymolysis liquid
Regulating the pH value of the enzymolysis liquid obtained in the step (I) to 5.0, adding 1 percent (mass fraction) of activated carbon powder, and adsorbing for 30 min at a constant temperature of 50 ℃; centrifuging at 7000 rpm for 15 min, microfiltering the supernatant (0.45 μm), ultrafiltering (molecular weight cut-off of 3000 Da), and spray drying the filtrate (inlet temperature of 170 deg.C and outlet temperature of 90 deg.C) to obtain semen Sesami oligopeptide product;
the constant-temperature adsorption is to add activated carbon powder to adsorb macromolecular protein, pigment and grease in the enzymolysis liquid, so that the purity of the oligopeptide is improved;
the microfiltration adopts a 0.45 mu m microporous filter membrane;
the ultrafiltration is an ultrafiltration membrane of Beijing Asahi Bangkong membrane equipment, Limited liability company, and is made of polysulfone material with the molecular weight cutoff of 3000 Da;
the hydrolysis degree of the present invention is 32.9%, the yield of the sesame oligopeptide is 30.8%, the peptide segment of the oligopeptide product is less than 1000 Da accounts for 86.8%, and the purity is 91.0%.
Example 3:
first, double enzyme synchronous enzymolysis
Uniformly mixing sesame seed meal and distilled water according to a material-liquid ratio of 1:20(g: mL), adjusting the pH value to 10.0 by 1 mol/L sodium hydroxide solution, adding alkaline protease and papain (food grade) according to an enzyme activity unit of 2:1(U: U) of the alkaline protease and the papain, adding 8000U of the sesame seed meal per gram, synchronously performing enzymolysis at 55 ℃ for 4 hours by using double enzymes, and inactivating the enzymes at 95 ℃ for 15 min to obtain an enzymolysis solution;
the sesame seed meal is a byproduct of sesame oil preparation by a water generation method, the water content is 67.3%, and the protein content is 12.6%;
the double-enzyme synchronous enzymolysis is that alkaline protease and papain directly hydrolyze sesame meal at the same time;
the enzyme deactivation is to deactivate alkaline protease and papain at 95 deg.C for 15 min;
② separation and purification of enzymolysis liquid
Regulating the pH value of the enzymolysis liquid obtained in the step (I) to 6.0, adding 2 percent (mass fraction) of activated carbon powder, and adsorbing at the constant temperature of 60 ℃ for 60 min; centrifuging at 7000 rpm for 15 min, microfiltering the supernatant (0.45 μm), ultrafiltering (molecular weight cut-off of 3000 Da), and spray drying the filtrate (inlet temperature of 170 deg.C and outlet temperature of 90 deg.C) to obtain semen Sesami oligopeptide product;
the constant-temperature adsorption is to add activated carbon powder to adsorb macromolecular protein, pigment and grease in the enzymolysis liquid, so that the purity of the oligopeptide is improved;
the microfiltration adopts a 0.45 mu m microporous filter membrane;
the ultrafiltration is an ultrafiltration membrane of Beijing Asahi Bangkong membrane equipment, Limited liability company, and is made of polysulfone material with the molecular weight cutoff of 3000 Da;
the hydrolysis degree of the present invention is 33.5%, the yield of the sesame oligopeptide is 31.6%, the peptide segment of the oligopeptide product is less than 1000 Da is 85.9%, and the purity is 91.2%.
Claims (6)
1. A method for preparing sesame oligopeptide from a sesame oil byproduct by an aqueous extraction method comprises the following steps:
first, double enzyme synchronous enzymolysis
Uniformly mixing sesame seed meal and distilled water according to a material-liquid ratio of 1:20-1:30(g: mL), adjusting the pH value to 9.0-10.0 by 1 mol/L sodium hydroxide solution, adding alkaline protease and papain (food grade) according to the enzyme activity unit of the alkaline protease and the papain of 1:1-2:1(U: U), adding 6000-8000U per gram of sesame seed meal, synchronously performing enzymolysis at 45-55 ℃ for 2-4 h by using double enzymes, and performing enzyme deactivation at 95 ℃ for 15 min to obtain an enzymolysis liquid;
② separation and purification of enzymolysis liquid
Regulating the pH value of the enzymolysis liquid obtained in the step (I) to 5.0-6.0, adding 1-2% (mass fraction) of activated carbon powder, and adsorbing at a constant temperature of 50-60 ℃ for 30-60 min; centrifuging at 7000 rpm for 15 min, microfiltering the supernatant (0.45 μm), ultrafiltering (molecular weight cut-off of 3000 Da), and spray drying the filtrate (inlet temperature of 170 deg.C and outlet temperature of 90 deg.C) to obtain semen Sesami oligopeptide product.
2. The method for preparing sesame oligopeptide from by-products of sesame oil by an aquation method according to claim 1, wherein the sesame seed meal is a by-product of sesame oil by an aquation method, has a water content of 67.3% and a protein content of 12.6%.
3. The method for preparing sesame oligopeptide from the by-product of sesame oil by aqueous extraction according to claim 1, wherein the simultaneous enzymolysis of the two enzymes is alkaline protease and papain which simultaneously hydrolyze sesame meal directly.
4. The method for preparing sesame oligopeptide from the by-product of sesame oil by aqueous extraction according to claim 1, wherein the enzyme deactivation is carried out under the conditions of 95 ℃ and 15 min to inactivate alkaline protease and papain.
5. The method for preparing sesame oligopeptide from sesame oil by-product of aqueous extraction method according to claim 1, wherein the constant temperature adsorption is adding activated carbon powder to adsorb macromolecular protein, pigment and grease in the enzymatic hydrolysate, thereby improving the purity of oligopeptide.
6. The method for preparing sesame oligopeptide from sesame oil by-product of aqueous extraction method according to claim 1, wherein the hydrolysis degree of the sesame oligopeptide product is 32.9-33.9%, the yield is 30.8-33.2% (mass of sesame oligopeptide/content of sesame seed meal protein), the peptide segment of the oligopeptide product is less than 1000 Da is 85.9-87.5%, and the purity is 91.0-91.4% (mass fraction).
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103250863A (en) * | 2013-03-11 | 2013-08-21 | 武汉工业学院 | Preparation method of sesame polypeptide |
| CN104388513A (en) * | 2014-12-02 | 2015-03-04 | 东莞市荷花食品有限公司 | Method of preparing salty sesame peptide from sesame cake meal by compound enzyme |
| CN108239666A (en) * | 2016-12-23 | 2018-07-03 | 江苏久思乡食品科技有限公司 | A kind of composite zymoprotein peptide extracting method for sesame |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103250863A (en) * | 2013-03-11 | 2013-08-21 | 武汉工业学院 | Preparation method of sesame polypeptide |
| CN104388513A (en) * | 2014-12-02 | 2015-03-04 | 东莞市荷花食品有限公司 | Method of preparing salty sesame peptide from sesame cake meal by compound enzyme |
| CN108239666A (en) * | 2016-12-23 | 2018-07-03 | 江苏久思乡食品科技有限公司 | A kind of composite zymoprotein peptide extracting method for sesame |
Non-Patent Citations (4)
| Title |
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| ROSHNI CHATTERJEE等: "Enzymatic modification of sesame seed protein, sourced from waste resource for nutraceutical application" * |
| XIN LU等: "Extraction, identification and structure-activity relationship of antioxidant peptides from sesame (Sesamum indicum L.) protein hydrolysate" * |
| 康媛媛等: "酶法制备芝麻肽及其抑菌活性的研究" * |
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