CN116287077A - Selenium polypeptide extraction method - Google Patents
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- CN116287077A CN116287077A CN202310235052.6A CN202310235052A CN116287077A CN 116287077 A CN116287077 A CN 116287077A CN 202310235052 A CN202310235052 A CN 202310235052A CN 116287077 A CN116287077 A CN 116287077A
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- BUGBHKTXTAQXES-UHFFFAOYSA-N Selenium Chemical compound [Se] BUGBHKTXTAQXES-UHFFFAOYSA-N 0.000 title claims abstract description 62
- 239000011669 selenium Substances 0.000 title claims abstract description 62
- 229910052711 selenium Inorganic materials 0.000 title claims abstract description 62
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 45
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 45
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 45
- 238000000605 extraction Methods 0.000 title claims abstract description 25
- 239000011347 resin Substances 0.000 claims abstract description 58
- 229920005989 resin Polymers 0.000 claims abstract description 58
- 238000001179 sorption measurement Methods 0.000 claims abstract description 47
- MYRTYDVEIRVNKP-UHFFFAOYSA-N 1,2-Divinylbenzene Chemical compound C=CC1=CC=CC=C1C=C MYRTYDVEIRVNKP-UHFFFAOYSA-N 0.000 claims abstract description 28
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Chemical compound C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000002904 solvent Substances 0.000 claims abstract description 21
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 claims abstract description 15
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims abstract description 15
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 15
- 235000009566 rice Nutrition 0.000 claims abstract description 15
- 239000013067 intermediate product Substances 0.000 claims abstract description 14
- -1 amino acrylic ester Chemical class 0.000 claims abstract description 12
- 238000002360 preparation method Methods 0.000 claims abstract description 11
- 238000002791 soaking Methods 0.000 claims abstract description 11
- 238000005238 degreasing Methods 0.000 claims abstract description 8
- WSLDOOZREJYCGB-UHFFFAOYSA-N 1,2-Dichloroethane Chemical compound ClCCCl WSLDOOZREJYCGB-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000003054 catalyst Substances 0.000 claims abstract description 7
- 238000006243 chemical reaction Methods 0.000 claims abstract description 7
- NKDDWNXOKDWJAK-UHFFFAOYSA-N dimethoxymethane Chemical compound COCOC NKDDWNXOKDWJAK-UHFFFAOYSA-N 0.000 claims abstract description 7
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims abstract description 7
- 238000010438 heat treatment Methods 0.000 claims abstract description 6
- 150000003254 radicals Chemical class 0.000 claims abstract description 5
- 230000000379 polymerizing effect Effects 0.000 claims abstract description 3
- 240000007594 Oryza sativa Species 0.000 claims abstract 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 34
- 239000000243 solution Substances 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 16
- 239000011550 stock solution Substances 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 235000019750 Crude protein Nutrition 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 12
- 238000005406 washing Methods 0.000 claims description 10
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 8
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical group CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 8
- 238000007873 sieving Methods 0.000 claims description 7
- 239000012043 crude product Substances 0.000 claims description 6
- 239000000706 filtrate Substances 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 4
- 108090000790 Enzymes Proteins 0.000 claims description 4
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 4
- 239000004365 Protease Substances 0.000 claims description 4
- 229940088598 enzyme Drugs 0.000 claims description 4
- ZSIAUFGUXNUGDI-UHFFFAOYSA-N hexan-1-ol Chemical compound CCCCCCO ZSIAUFGUXNUGDI-UHFFFAOYSA-N 0.000 claims description 4
- 239000003208 petroleum Substances 0.000 claims description 4
- YTLYLLTVENPWFT-UPHRSURJSA-N (Z)-3-aminoacrylic acid Chemical compound N\C=C/C(O)=O YTLYLLTVENPWFT-UPHRSURJSA-N 0.000 claims description 3
- 108091005804 Peptidases Proteins 0.000 claims description 3
- 108010059892 Cellulase Proteins 0.000 claims description 2
- 108090001060 Lipase Proteins 0.000 claims description 2
- 239000004367 Lipase Substances 0.000 claims description 2
- 102000004882 Lipase Human genes 0.000 claims description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 2
- 229940106157 cellulase Drugs 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 235000019421 lipase Nutrition 0.000 claims description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 2
- 230000001476 alcoholic effect Effects 0.000 claims 1
- 108010074686 Selenoproteins Proteins 0.000 abstract description 12
- 102000008114 Selenoproteins Human genes 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 6
- 238000003795 desorption Methods 0.000 abstract description 4
- 241000209094 Oryza Species 0.000 description 13
- 230000000052 comparative effect Effects 0.000 description 10
- 239000000047 product Substances 0.000 description 10
- 101100313763 Arabidopsis thaliana TIM22-2 gene Proteins 0.000 description 6
- 239000000126 substance Substances 0.000 description 5
- 108091005658 Basic proteases Proteins 0.000 description 4
- 230000003197 catalytic effect Effects 0.000 description 4
- 238000004108 freeze drying Methods 0.000 description 4
- 239000000178 monomer Substances 0.000 description 4
- 235000013312 flour Nutrition 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 229920000642 polymer Polymers 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 238000005292 vacuum distillation Methods 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- CHRJZRDFSQHIFI-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;styrene Chemical compound C=CC1=CC=CC=C1.C=CC1=CC=CC=C1C=C CHRJZRDFSQHIFI-UHFFFAOYSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 238000005727 Friedel-Crafts reaction Methods 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 239000003531 protein hydrolysate Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
- Peptides Or Proteins (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The invention discloses a selenium polypeptide extraction method, which comprises a modified porous adsorption resin preparation method, and comprises the following steps: l1, polymerizing styrene, divinylbenzene, acrylic acid and amino acrylic ester by free radicals to obtain an intermediate product; and L2, soaking the intermediate product in 1, 2-dichloroethane, then adding dimethoxymethane and a catalyst ferric trichloride, heating for reaction, and removing the solvent to obtain the modified porous adsorption resin. According to the selenium polypeptide extraction method disclosed by the invention, the selenium polypeptide is obtained through degreasing of selenium-enriched broken rice, resin adsorption and desorption and enzymolysis, and the extraction efficiency of the obtained selenium polypeptide is high; and the purity is high, the selenium content of the selenium polypeptide is high, and the extraction effect is good. This is mainly because the adsorption resin of the present invention has a similar structure to selenoprotein and has a good specific surface area and porosity, thereby efficiently adsorbing selenoprotein.
Description
Technical Field
The invention belongs to the technical field of polypeptide extraction, and particularly relates to a selenium polypeptide extraction method.
Background
Selenium is an essential trace element in humans and animals, and has the effects of preventing tumors and strengthening body constitution. Selenium in selenium-enriched rice exists mostly in the form of selenoprotein, and if the selenium is further decomposed into selenium polypeptides, specific enzymes are needed for catalysis. For example Cao Hui et al hydrolyzed selenium rice with papain to obtain selenium polypeptide with strong reducing ability, and confirmed that rice protein hydrolysate is a natural antioxidant polypeptide; shen Yanhao et al used five different proteases to hydrolyze rice proteins, obtained the best conditions, and successfully obtained the product with scavenging effect on human free radicals. Therefore, the efficient extraction and the obtaining of the selenium polypeptide have important roles in scientific research and life.
The method for collecting selenium polypeptide precursor selenium protein mainly comprises porous adsorption resin adsorption and desorption method. The resin commonly used at present is HP20 resin. Typically, the resin is packed, then the selenoprotein sample is allowed to slowly permeate through the resin, and finally the desorbed solution is collected by alcohol solution desorption, and the desired product is obtained by post-treatment. Then, most of the HP20 resin products are held by foreign enterprises, and the adsorption effect also has a certain bottleneck. Therefore, how to extract selenium polypeptide precursor by high-efficiency extraction method, and then obtain selenium polypeptide product with high yield and less impurity is always the target pursued by domestic scientific researchers.
Therefore, there is a need to find a technical method to solve the above-mentioned problems.
Disclosure of Invention
Aiming at the prior art, most of HP20 resin products are adopted as a technical means for purifying selenoprotein, the invention replaces the mature resin with the novel phenyl polar resin disclosed by the invention as equipment for adsorbing selenoprotein. The molecular structure of the resin is a three-dimensional network structure microscopically, wherein the resin contains a benzene ring group which is a common structure, and in addition, the resin contains a small amount of two polar groups of carboxyl and amino, so that the resin has excellent affinity to related polar groups in selenoprotein, and the adsorption efficiency of the selenoprotein is greatly improved. It is noted that the adsorption effect of the resin containing both carboxyl and amino groups is superior to that of the adsorption resin containing only one polar group, probably because the simultaneous presence of both groups has a synergistic effect, promoting the progress of adsorption.
The invention discloses a selenium polypeptide extraction method, which comprises the following steps:
s1, degreasing selenium-enriched crushed rice with a solvent, removing the solvent, and adding water to obtain a mixed stock solution;
s2, homogenizing the mixed stock solution, sieving, passing the obtained filtrate through a modified porous adsorption resin, washing the adsorbed modified porous adsorption resin with water, desorbing with an alcohol solution, and removing the alcohol solution to obtain crude protein;
s3, injecting the crude protein into an immobilized enzymolysis tank for enzymolysis, centrifuging the obtained crude product, taking supernatant, and drying the supernatant to obtain the selenium polypeptide;
the preparation method of the modified porous adsorption resin comprises the following steps:
l1, polymerizing styrene, divinylbenzene, acrylic acid and amino acrylic ester by free radicals to obtain an intermediate product;
and L2, soaking the intermediate product in 1, 2-dichloroethane, then adding dimethoxymethane and a catalyst ferric trichloride, heating for reaction, and removing the solvent to obtain the modified porous adsorption resin.
The resin synthesized by the invention is a random polymer with a main chain synthesized in a free radical mode, and then a reticular porous polymer is formed after grafting an alkyl chain through Friedel-crafts reaction, and the obtained polymer is ball-milled until the particle size is generally between 2 and 3 mm; the aperture is about 25-30 nm; the porosity is about 45-55%.
Further, in step S1, the solvent is selected from n-hexane or petroleum ether.
Further, in the step S3, the enzyme used for the enzymolysis is one or more selected from protease, cellulase or lipase.
Further, in the step S2, the alcohol solution is selected from an ethanol solution, a propanol solution, a butanol solution, or a hexanol solution.
Further, in the step L1, the molar ratio of the styrene to the divinylbenzene to the acrylic acid to the amino acrylic acid ester is 1:1:1:1 to 1:5:0.2:0.2.
Further, the enzymolysis conditions in the immobilized enzymolysis tank are as follows: the pH is 8-10, and the temperature is 40-50 ℃.
Further, in the step S3, the enzymolysis time is 1-3h.
Further, in the step S2, the mesh number of the sieving is 400-600 mesh.
Further, in the step L2, the heating temperature is 80-100 ℃, and the heating time is 12-20h.
The invention has the following beneficial technical effects:
according to the selenium polypeptide extraction method disclosed by the invention, the selenium polypeptide is obtained through degreasing of selenium-enriched broken rice, resin adsorption and desorption and enzymolysis, and the extraction efficiency of the obtained selenium polypeptide is high; and the purity is high, the selenium content of the selenium polypeptide is high, and the extraction effect is good. This is mainly because the adsorption resin of the present invention has a similar structure to selenoprotein and has a good specific surface area and porosity, thereby efficiently adsorbing selenoprotein.
Detailed Description
The present invention is further illustrated below with reference to specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
The selenium-enriched rice flour is purchased from a selenium-enriched rice flour processing plant in Ji-shou city, and crushed into particles with smaller particle size after being purchased;
the immobilized enzymolysis tank is 50L and purchased from Anhui Chenxianruida mechanical Limited company.
The HP20 resin of the present invention was purchased from Mitsubishi chemical in Japan.
Example 1
A method for extracting selenium polypeptide, which comprises the following steps:
s1, soaking selenium-enriched crushed rice in n-hexane, degreasing for 4 hours, removing a solvent, naturally airing, and adding water to obtain a mixed stock solution;
s2, homogenizing the mixed stock solution by using a homogenizer, sieving the homogenized stock solution by using a 200-mesh sieve, taking 30ml of obtained filtrate, passing through a column (with the diameter of 10cm and the height of 1 m) filled with modified porous adsorption resin, washing the adsorbed modified porous adsorption resin with water for a plurality of times, desorbing the adsorbed modified porous adsorption resin by using an ethanol solution (50 wt%) and removing the ethanol solution by reduced pressure distillation to obtain crude protein;
s3, injecting the crude protein into an immobilized enzymolysis tank, carrying out enzymolysis by alkaline protease under the conditions of pH=8 and 40 ℃, centrifuging the obtained crude product at a high speed by using a centrifuge, taking supernatant, and finally freeze-drying the supernatant to obtain the product selenium polypeptide;
the preparation method of the modified porous adsorption resin comprises the following steps:
l1, adding styrene, divinylbenzene, acrylic acid and amino acrylic ester according to the specified amount of substances (styrene: divinylbenzene: acrylic acid: amino acrylic ester=1:5:0.2:0.2, n/n/n/n), and reacting for 6 hours at 90 ℃ under the reaction condition of a catalytic amount of BPO and solvent water to obtain an intermediate product;
l2, soaking 50g of the intermediate product in 50ml of 1, 2-dichloroethane, then adding 30g of dimethoxymethane and 20g of catalyst ferric trichloride, reacting at 90 ℃ for 6 hours, removing the solvent, and washing with ethanol to obtain the modified porous adsorption resin.
Example 2
A method for extracting selenium polypeptide, which comprises the following steps:
s1, soaking selenium-rich crushed rice in petroleum ether, degreasing for 4 hours, removing a solvent, naturally airing, and adding water to obtain a mixed stock solution;
s2, homogenizing the mixed stock solution by using a homogenizer, sieving the homogenized stock solution by using a 200-mesh sieve, taking 30ml of obtained filtrate, passing through a column filled with modified porous adsorption resin (with the diameter of 12cm and the height of 1.2 m), washing the adsorbed modified porous adsorption resin with water for a plurality of times, desorbing by using ethanol solution (50 wt%) and removing the ethanol solution by vacuum distillation to obtain crude protein;
s3, injecting the crude protein into an immobilized enzymolysis tank, carrying out enzymolysis by alkaline protease under the conditions of pH=9 and 50 ℃, centrifuging the obtained crude product at a high speed by using a centrifuge, taking supernatant, and finally freeze-drying the supernatant to obtain the product selenium polypeptide;
the preparation method of the modified porous adsorption resin comprises the following steps:
l1, adding styrene, divinylbenzene, acrylic acid and amino acrylic ester according to the specified amount of substances (styrene: divinylbenzene: acrylic acid: amino acrylic ester=1:1:0.2:0.1, n/n/n/n), and reacting for 6 hours at 95 ℃ under the reaction condition of a catalytic amount of BPO and solvent water to obtain an intermediate product;
l2, soaking 50g of the intermediate product in 50ml of 1, 2-dichloroethane, then adding 30g of dimethoxymethane and 25g of catalyst ferric trichloride, reacting at 90 ℃ for 6 hours, removing the solvent, and washing with ethanol to obtain the modified porous adsorption resin.
Example 3
A method for extracting selenium polypeptide, which comprises the following steps:
s1, soaking selenium-rich crushed rice in petroleum ether, degreasing for 8 hours, removing a solvent, naturally airing, and adding water to obtain a mixed stock solution;
s2, homogenizing the mixed stock solution by using a homogenizer, sieving the homogenized stock solution by using a 200-mesh sieve, taking 30ml of obtained filtrate, passing through a column filled with modified porous adsorption resin (with the diameter of 12cm and the height of 1.2 m), washing the adsorbed modified porous adsorption resin with water for a plurality of times, desorbing the adsorbed modified porous adsorption resin by using ethanol solution (70 wt%) and removing the ethanol solution by vacuum distillation to obtain crude protein;
s3, injecting the crude protein into an immobilized enzymolysis tank, carrying out enzymolysis by alkaline protease under the conditions of pH=8.5 and temperature of 45 ℃, centrifuging the obtained crude product at a high speed by using a centrifuge, taking supernatant, and finally freeze-drying the supernatant to obtain the product selenium polypeptide;
the preparation method of the modified porous adsorption resin comprises the following steps:
l1, adding styrene, divinylbenzene, acrylic acid and amino acrylic ester according to the specified amount of substances (styrene: divinylbenzene: acrylic acid: amino acrylic ester=1:3:0.1:0.1, n/n/n/n), and reacting for 4 hours at 90 ℃ under the reaction condition of a catalytic amount of BPO and solvent water to obtain an intermediate product;
l2, soaking 50g of the intermediate product in 50ml of 1, 2-dichloroethane, then adding 35g of dimethoxymethane and 25g of catalyst ferric trichloride, reacting at 90 ℃ for 8 hours, removing the solvent, and washing with ethanol to obtain the modified porous adsorption resin.
Example 4
A method for extracting selenium polypeptide, which comprises the following steps:
s1, soaking selenium-enriched crushed rice in n-hexane, degreasing for 4 hours, removing a solvent, naturally airing, and adding water to obtain a mixed stock solution;
s2, homogenizing the mixed stock solution by using a homogenizer, sieving the homogenized stock solution by using a 200-mesh sieve, taking 40ml of obtained filtrate, passing through a column filled with modified porous adsorption resin (with the diameter of 12cm and the height of 1.5 m), washing the adsorbed modified porous adsorption resin with water for a plurality of times, desorbing the adsorbed modified porous adsorption resin by using ethanol solution (70 wt%) and removing the ethanol solution by vacuum distillation to obtain crude protein;
s3, injecting the crude protein into an immobilized enzymolysis tank, carrying out enzymolysis by alkaline protease under the conditions of pH=8 and 40 ℃, centrifuging the obtained crude product at a high speed by using a centrifuge, taking supernatant, and finally freeze-drying the supernatant to obtain the product selenium polypeptide;
the preparation method of the modified porous adsorption resin comprises the following steps:
l1, adding styrene, divinylbenzene, acrylic acid and amino acrylic ester according to the specified amount of substances (styrene: divinylbenzene: acrylic acid: amino acrylic ester=1:3:0.2:0.2, n/n/n/n), and reacting for 8 hours at 90 ℃ under the reaction condition of a catalytic amount of BPO and solvent water to obtain an intermediate product;
l2, soaking 50g of the intermediate product in 50ml of 1, 2-dichloroethane, then adding 30g of dimethoxymethane and 25g of catalyst ferric trichloride, reacting at 90 ℃ for 6 hours, removing the solvent, and washing with ethanol to obtain the modified porous adsorption resin.
Comparative example 1
The preparation method and the raw materials of the selenium polypeptide extraction method of comparative example 1 are the same as those of the selenium polypeptide extraction method of example 1, and the only difference is that the porous adsorption resin adopts HP20 resin.
Comparative example 2
The preparation method and the raw materials of the extraction method of the selenium polypeptide in the comparative example 2 are the same as those of the selenium polypeptide in the embodiment 1, and the only difference is that the synthetic monomer of the porous adsorption resin does not contain acrylic acid monomer, and styrene/divinylbenzene/amino acrylic acid ester=1:5:0.2 and n/n/n.
Comparative example 3
The preparation method and the raw materials of the extraction method of the selenium polypeptide in the comparative example 2 are the same as those of the selenium polypeptide in the embodiment 1, and the only difference is that the synthetic monomer of the porous adsorption resin does not contain amino acrylic ester monomer, and styrene and divinylbenzene are respectively acrylic acid=1:5:0.2 and n/n/n.
Test case
The selenium polypeptides prepared by the extraction method preparation method of the selenium polypeptides described in the embodiment 1 and the comparative examples 1-3 were subjected to the following tests.
Wherein,,
calculating the extraction rate of selenium polypeptide: according to the formula:
selenium polypeptide extraction = (mass of product collected in modified porous adsorption resin/mass of selenium-enriched broken rice flour) = 100%.
The results obtained are shown in Table 1.
Table 1: comparative data relating to example 1 and comparative examples 1 to 3
From the above data, the extraction method of selenium polypeptide adopted in example 1 has slightly better extraction rate than that of comparative example 1, mainly because the adsorption resin of the invention has similar structure with selenoprotein, and has slightly lower average pore diameter than HP20 resin, but has good specific surface area, thereby having high-efficiency adsorption on selenoprotein. The data fully prove that the modified porous adsorption resin designed by us has important significance in the aspect of selenium polypeptide extraction, is beneficial to breaking the monopoly pattern of the current European and American across-the-country enterprises, and has wide industrialized prospect.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (9)
1. The extraction method of the selenium polypeptide is characterized by comprising the following steps of:
s1, degreasing selenium-enriched crushed rice with a solvent, removing the solvent, and adding water to obtain a mixed stock solution;
s2, homogenizing the mixed stock solution, sieving, passing the obtained filtrate through a modified porous adsorption resin, washing the adsorbed modified porous adsorption resin with water, desorbing with an alcohol solution, and removing the alcohol solution to obtain crude protein;
s3, injecting the crude protein into an immobilized enzymolysis tank for enzymolysis, centrifuging the obtained crude product, taking supernatant, and drying the supernatant to obtain the selenium polypeptide;
the preparation method of the modified porous adsorption resin comprises the following steps:
l1, polymerizing styrene, divinylbenzene, acrylic acid and amino acrylic ester by free radicals to obtain an intermediate product;
and L2, soaking the intermediate product in 1, 2-dichloroethane, then adding dimethoxymethane and a catalyst ferric trichloride, heating for reaction, and removing the solvent to obtain the modified porous adsorption resin.
2. The method of claim 1, wherein in step S1, the solvent is selected from n-hexane and petroleum ether.
3. The method of claim 1, wherein in step S3, the enzyme used for the enzymolysis is one or more selected from the group consisting of protease, cellulase and lipase.
4. The method of claim 1, wherein in step S2, the alcoholic solution is selected from ethanol solution, propanol solution, butanol solution or hexanol solution.
5. The method of claim 1, wherein in step L1, the molar ratio of styrene, divinylbenzene, acrylic acid, and aminoacrylate is 1:1:1:1-1:5:0.2:0.2.
6. The method for extracting selenium polypeptide according to claim 1, wherein the enzymolysis conditions in the immobilized enzymolysis tank are: the pH is 8-10, and the temperature is 40-50 ℃.
7. The method of claim 1, wherein in step S3, the enzyme is performed for 1-3 hours.
8. The method of claim 1, wherein in step S2, the mesh size of the sieve is 400-600 mesh.
9. The method of claim 1, wherein in step L2, the heating is performed at 80-100 ℃ for 12-20 hours.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118027135A (en) * | 2024-04-11 | 2024-05-14 | 黑龙江八一农垦大学 | Method for extracting selenium polypeptide from selenium-enriched rice |
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|---|---|---|---|---|
| GB1221918A (en) * | 1968-07-24 | 1971-02-10 | Du Pont | Ethylene-amino acrylate copolymers |
| US6075105A (en) * | 1996-08-26 | 2000-06-13 | Xerox Corporation | Polymerization processes and resin particles formed thereby |
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