CN111499771A - Method for simultaneously extracting grease, protein and polysaccharide in rice bran by three-phase extraction technology - Google Patents
Method for simultaneously extracting grease, protein and polysaccharide in rice bran by three-phase extraction technology Download PDFInfo
- Publication number
- CN111499771A CN111499771A CN202010313749.7A CN202010313749A CN111499771A CN 111499771 A CN111499771 A CN 111499771A CN 202010313749 A CN202010313749 A CN 202010313749A CN 111499771 A CN111499771 A CN 111499771A
- Authority
- CN
- China
- Prior art keywords
- rice bran
- protein
- polysaccharide
- phase
- simultaneously extracting
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 60
- 235000009566 rice Nutrition 0.000 title claims abstract description 60
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 48
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 48
- 150000004676 glycans Chemical class 0.000 title claims abstract description 43
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 43
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 43
- 238000000605 extraction Methods 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims abstract description 27
- 239000004519 grease Substances 0.000 title claims abstract description 23
- 240000007594 Oryza sativa Species 0.000 title 1
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 claims abstract description 72
- 241000209094 Oryza Species 0.000 claims abstract description 59
- 239000002002 slurry Substances 0.000 claims abstract description 26
- 150000003839 salts Chemical class 0.000 claims abstract description 13
- 239000007853 buffer solution Substances 0.000 claims abstract description 10
- 239000007787 solid Substances 0.000 claims abstract description 4
- 239000012071 phase Substances 0.000 claims description 47
- 239000000203 mixture Substances 0.000 claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 26
- 239000012153 distilled water Substances 0.000 claims description 25
- 238000007670 refining Methods 0.000 claims description 24
- 238000001035 drying Methods 0.000 claims description 18
- 150000002632 lipids Chemical class 0.000 claims description 16
- 238000002156 mixing Methods 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 10
- 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 9
- 238000011033 desalting Methods 0.000 claims description 8
- 238000001704 evaporation Methods 0.000 claims description 8
- 230000000887 hydrating effect Effects 0.000 claims description 8
- 239000012074 organic phase Substances 0.000 claims description 8
- 239000002994 raw material Substances 0.000 claims description 8
- 238000004064 recycling Methods 0.000 claims description 8
- 238000007873 sieving Methods 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 8
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 5
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 5
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 5
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 2
- UEGPKNKPLBYCNK-UHFFFAOYSA-L magnesium acetate Chemical compound [Mg+2].CC([O-])=O.CC([O-])=O UEGPKNKPLBYCNK-UHFFFAOYSA-L 0.000 claims description 2
- 239000011654 magnesium acetate Substances 0.000 claims description 2
- 235000011285 magnesium acetate Nutrition 0.000 claims description 2
- 229940069446 magnesium acetate Drugs 0.000 claims description 2
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 2
- 235000011152 sodium sulphate Nutrition 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims 1
- 238000001291 vacuum drying Methods 0.000 claims 1
- 238000009777 vacuum freeze-drying Methods 0.000 claims 1
- 238000001556 precipitation Methods 0.000 abstract description 4
- 230000008901 benefit Effects 0.000 abstract description 3
- 235000019198 oils Nutrition 0.000 abstract description 3
- 239000006184 cosolvent Substances 0.000 abstract description 2
- 238000005185 salting out Methods 0.000 abstract description 2
- 238000000926 separation method Methods 0.000 abstract description 2
- 239000003921 oil Substances 0.000 abstract 2
- 235000015112 vegetable and seed oil Nutrition 0.000 abstract 1
- 239000008158 vegetable oil Substances 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 32
- 239000010410 layer Substances 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 229920002521 macromolecule Polymers 0.000 description 3
- 238000003260 vortexing Methods 0.000 description 3
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 235000021329 brown rice Nutrition 0.000 description 2
- 239000000287 crude extract Substances 0.000 description 2
- 235000019621 digestibility Nutrition 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 description 1
- GJJVAFUKOBZPCB-UHFFFAOYSA-N 2-methyl-2-(4,8,12-trimethyltrideca-3,7,11-trienyl)-3,4-dihydrochromen-6-ol Chemical compound OC1=CC=C2OC(CCC=C(C)CCC=C(C)CCC=C(C)C)(C)CCC2=C1 GJJVAFUKOBZPCB-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- BHEOSNUKNHRBNM-UHFFFAOYSA-N Tetramethylsqualene Natural products CC(=C)C(C)CCC(=C)C(C)CCC(C)=CCCC=C(C)CCC(C)C(=C)CCC(C)C(C)=C BHEOSNUKNHRBNM-UHFFFAOYSA-N 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- GVJHHUAWPYXKBD-UHFFFAOYSA-N d-alpha-tocopherol Natural products OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 1
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N dodecahydrosqualene Natural products CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000000774 hypoallergenic effect Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000012044 organic layer Substances 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- -1 phytosterol Chemical compound 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 229940031439 squalene Drugs 0.000 description 1
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 229930003799 tocopherol Natural products 0.000 description 1
- 229960001295 tocopherol Drugs 0.000 description 1
- 239000011732 tocopherol Substances 0.000 description 1
- 235000010384 tocopherol Nutrition 0.000 description 1
- 229930003802 tocotrienol Natural products 0.000 description 1
- 239000011731 tocotrienol Substances 0.000 description 1
- 235000019148 tocotrienols Nutrition 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 230000002087 whitening effect Effects 0.000 description 1
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
-
- 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
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B1/00—Production of fats or fatty oils from raw materials
- C11B1/02—Pretreatment
- C11B1/04—Pretreatment of vegetable raw material
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B3/00—Refining fats or fatty oils
- C11B3/12—Refining fats or fatty oils by distillation
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Sustainable Development (AREA)
- Materials Engineering (AREA)
- Polymers & Plastics (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention relates to the technical field of extracting and separating biomacromolecules from vegetable oil, in particular to a method for simultaneously extracting grease, protein and polysaccharide from rice bran by a three-phase extraction technology. Under the combined action of salting out, plasma precipitation, cosolvent precipitation, etc., solid salt, tert-butanol and rice bran slurry are added, and fully mixed to form three phases, the upper phase is reduced pressure evaporated to obtain oil, the middle phase is dissolved in buffer solution, and then dialyzed and dried to obtain refined protein, and the lower phase is dialyzed and dried to obtain refined polysaccharide. The invention provides a simple, efficient, economic and green three-phase extraction technology, realizes the simultaneous extraction of oil, protein and polysaccharide in rice bran, and has high product yield and good quality. The method can directly act on the rice bran slurry, can be operated at room temperature, and has the advantages of high separation efficiency of target products, wide application range, easy expansion and the like.
Description
Technical Field
The invention relates to the technical field of simultaneous extraction of grease, protein and polysaccharide from rice bran, in particular to a method for simultaneously separating and extracting grease, protein and polysaccharide from rice bran by a three-phase extraction technology.
Background
The rice bran is an important byproduct in the rice processing process, is obtained in the brown rice whitening process, and accounts for about 8-10% of the weight of the brown rice. Rice bran is a good source of oil, protein and carbohydrates, in addition to some valuable nutrients, antioxidants, vitamins and minerals. The rice bran contains about 12% to 18% of oil and fat, which contains a large amount of nutrients beneficial to health, such as tocopherol, tocotrienol, phytosterol, squalene, oryzanol, phospholipids, and the like. The protein content of the rice bran is about 10% -16%, the protein efficiency ratio of the rice bran protein is 2.0-2.5, the casein is 2.5, and the protein digestibility of the rice bran protein is more than 90%. It has been widely recognized because of its hypoallergenic and nutritional properties, as well as its good biological value and digestibility. The rice bran polysaccharide has good physiological activity, and has good physiological health promotion and tumor prevention effects.
Three-phase extraction is a method for extracting, purifying and concentrating macromolecules under the combined action of salting out, plasma precipitation, cosolvent precipitation and the like. The principle of three-phase extraction is to mix the crude extract or suspension with solid salt and organic solvent to form three phases, the upper phase is organic layer mainly containing pigment, lipid and other substances with small polarity, the middle phase is protein layer, and the lower phase is water layer mainly containing some water-soluble substances. Compared with the traditional technology, the extraction time of the three-phase extraction is short, the three-phase extraction can directly act on a crude extract, the operation can be carried out at room temperature, and the method has the advantages of high separation efficiency of target products, wide application range, cost benefit, easiness in expansion and the like. In addition, the organic solvent used in the three-phase extraction is not flammable, as compared to the solvents used in conventional extraction methods, such as n-hexane, methanol, or ethanol. Three-phase extraction can be defined as a simple, efficient, economical, green (clean) bioseparation process for extracting and purifying macromolecules of interest (biologically active substances).
Therefore, the rice bran is taken as a research object, the three-phase extraction technology is applied to the extraction of macromolecular substances, and the oil, protein and polysaccharide in the rice bran are simultaneously extracted by the three-phase extraction technology, so that a theoretical basis is provided for industrial production.
Disclosure of Invention
The invention aims to provide a simple, efficient, economical and green method for simultaneously extracting grease, protein and polysaccharide from rice bran by a three-phase extraction technology.
The technical scheme of the invention is as follows:
a method for simultaneously extracting grease, protein and polysaccharide from rice bran by a three-phase extraction technology comprises the following steps:
(1) preparing raw materials: sieving testa oryzae with 60-100 mesh sieve, and mixing with distilled water at a ratio of 1 (10-30) to obtain testa oryzae slurry;
(2) constructing a three-phase system: adding 10-50% (w/v) ammonium sulfate into the rice bran slurry, slightly swirling, and adjusting pH of the mixture to 3-7; then slowly adding tert-butyl alcohol according to the ratio of the slurry to the tert-butyl alcohol of 1:0.5-1:2.5 (v/v); gently stirring the mixture for 0.5-2.5 h on a magnetic stirrer at 30-70 ℃, and centrifuging the mixture for 20min at 5000r/min after fully mixing to form clear three phases;
(3) refining the grease: collecting the upper organic phase, evaporating and recovering in vacuum, collecting a lipid sample, and recycling tert-butyl alcohol;
(4) refining protein by dissolving the intermediate phase in Tris-HCl buffer solution of 20-100 mmol/L and pH 7-9, hydrating at 4 deg.C overnight, dialyzing in distilled water to remove salt, collecting supernatant, concentrating, and drying to obtain rice bran protein;
(5) refining polysaccharide: dialyzing the lower layer solution in distilled water, desalting, concentrating, and drying to obtain testa oryzae polysaccharide.
The specific implementation mode is as follows:
the first embodiment is as follows:
a method for simultaneously extracting grease, protein and polysaccharide from rice bran by a three-phase extraction technology comprises the following steps:
(1) preparing raw materials: sieving rice bran with 60 mesh sieve, and mixing with distilled water at a ratio of 1:15 to obtain rice bran slurry;
(2) constructing a three-phase system: adding 15% (w/v) sodium sulfate to the rice bran slurry, and gently vortexing to adjust the pH of the mixture to 4; then slowly adding tert-butyl alcohol according to the ratio of the slurry to the tert-butyl alcohol of 1:0.5 (v/v); gently stirring the mixture on a magnetic stirrer at 40 ℃ for 0.5h, fully mixing, and centrifuging the mixture at 5000r/min for 20min to form clear three phases;
(3) refining the grease: collecting the upper organic phase, evaporating and recovering in vacuum, collecting a lipid sample, and recycling tert-butyl alcohol;
(4) refining protein by dissolving the intermediate phase in Tris-HCl buffer solution of 20 mmol/L and pH 7, hydrating at 4 deg.C overnight, dialyzing in distilled water to remove salt, collecting supernatant, concentrating, and drying to obtain rice bran protein;
(5) refining polysaccharide: dialyzing the lower layer solution in distilled water, desalting, concentrating, and drying to obtain testa oryzae polysaccharide.
The second embodiment is as follows:
a method for simultaneously extracting grease, protein and polysaccharide from rice bran by a three-phase extraction technology comprises the following steps:
(1) preparing raw materials: sieving rice bran with 80 mesh sieve, and mixing with distilled water at a ratio of 1:20 to obtain rice bran slurry;
(2) constructing a three-phase system: adding 20% (w/v) ammonium sulfate to the rice bran slurry, and gently swirling to adjust the pH of the mixture to 5; then slowly adding tert-butyl alcohol according to the ratio of the slurry to the tert-butyl alcohol of 1:1 (v/v); gently stirring the mixture on a magnetic stirrer at 30 ℃ for a certain time of 1h, fully mixing, and centrifuging the mixture at 5000r/min for 20min to form clear three phases;
(3) refining the grease: collecting the upper organic phase, evaporating and recovering in vacuum, collecting a lipid sample, and recycling tert-butyl alcohol;
(4) refining protein by dissolving the intermediate phase in Tris-HCl buffer solution of 50 mmol/L and pH 8, hydrating at 4 deg.C overnight, dialyzing in distilled water to remove salt, collecting supernatant, concentrating, and drying to obtain rice bran protein;
(5) refining polysaccharide: dialyzing the lower layer solution in distilled water, desalting, concentrating, and drying to obtain testa oryzae polysaccharide.
The third concrete implementation mode:
a method for simultaneously extracting grease, protein and polysaccharide from rice bran by a three-phase extraction technology comprises the following steps:
(1) preparing raw materials: sieving testa oryzae with 100 mesh sieve, and mixing with distilled water at a ratio of 1:25 to obtain testa oryzae slurry;
(2) constructing a three-phase system: adding 30% (w/v) ammonium sulfate to the rice bran slurry, and gently swirling to adjust the pH of the mixture to 5; then slowly adding tert-butyl alcohol according to the ratio of the slurry to the tert-butyl alcohol of 1:1.5 (v/v); gently stirring the mixture on a magnetic stirrer at 50 ℃ for a certain time of 1.5h, fully mixing, and centrifuging the mixture at 5000r/min for 20min to form clear three phases;
(3) refining the grease: collecting the upper organic phase, evaporating and recovering in vacuum, collecting a lipid sample, and recycling tert-butyl alcohol;
(4) refining protein by dissolving the intermediate phase in Tris-HCl buffer solution of 100 mmol/L and pH 9, hydrating at 4 deg.C overnight, dialyzing in distilled water to remove salt, collecting supernatant, concentrating, and drying to obtain rice bran protein;
(5) refining polysaccharide: dialyzing the lower layer solution in distilled water, desalting, concentrating, and drying to obtain testa oryzae polysaccharide.
The fourth concrete implementation mode:
a method for simultaneously extracting grease, protein and polysaccharide from rice bran by a three-phase extraction technology comprises the following steps:
(1) preparing raw materials: sieving rice bran with 60 mesh sieve, and mixing with distilled water at a ratio of 1:20 to obtain rice bran slurry;
(2) constructing a three-phase system: adding 40% (w/v) magnesium sulfate to the rice bran slurry, and gently vortexing to adjust the pH of the mixture to 5; then slowly adding tert-butyl alcohol according to the ratio of the slurry to the tert-butyl alcohol of 1:2 (v/v); gently stirring the mixture for 2h at 60 ℃ on a magnetic stirrer, fully mixing, and centrifuging the mixture for 20min at 5000r/min to form clear three phases;
(3) refining the grease: collecting the upper organic phase, evaporating and recovering in vacuum, collecting a lipid sample, and recycling tert-butyl alcohol;
(4) refining protein by dissolving the intermediate phase in Tris-HCl buffer solution of 20 mmol/L and pH 7, hydrating at 4 deg.C overnight, dialyzing in distilled water to remove salt, collecting supernatant, concentrating, and drying to obtain rice bran protein;
(5) refining polysaccharide: dialyzing the lower layer solution in distilled water, desalting, concentrating, and drying to obtain testa oryzae polysaccharide.
The fifth concrete implementation mode:
a method for simultaneously extracting grease, protein and polysaccharide from rice bran by a three-phase extraction technology comprises the following steps:
(1) preparing raw materials: sieving rice bran with 80 mesh sieve, and mixing with distilled water at a ratio of 1:25 to obtain rice bran slurry;
(2) constructing a three-phase system: adding 40% (w/v) ammonium sulfate to the rice bran slurry, and gently swirling to adjust the pH of the mixture to 4; then slowly adding tert-butyl alcohol according to the ratio of the slurry to the tert-butyl alcohol of 1:2.5 (v/v); gently stirring the mixture for a certain time of 2.5h at 30 ℃ on a magnetic stirrer, fully mixing, and centrifuging the mixture for 20min at 5000r/min to form clear three phases;
(3) refining the grease: collecting the upper organic phase, evaporating and recovering in vacuum, collecting a lipid sample, and recycling tert-butyl alcohol;
(4) refining protein by dissolving the intermediate phase in Tris-HCl buffer solution of 50 mmol/L and pH 8, hydrating at 4 deg.C overnight, dialyzing in distilled water to remove salt, collecting supernatant, concentrating, and drying to obtain rice bran protein;
(5) refining polysaccharide: dialyzing the lower layer solution in distilled water, desalting, concentrating, and drying to obtain testa oryzae polysaccharide.
The sixth specific implementation mode:
a method for simultaneously extracting grease, protein and polysaccharide from rice bran by a three-phase extraction technology comprises the following steps:
(1) preparing raw materials: sieving testa oryzae with 100 mesh sieve, and mixing with distilled water at a ratio of 1:30 to obtain testa oryzae slurry;
(2) constructing a three-phase system: adding 50% (w/v) magnesium acetate to the rice bran slurry, and gently vortexing to adjust the pH of the mixture to 3; then slowly adding tert-butyl alcohol according to the ratio of the slurry to the tert-butyl alcohol of 1:2 (v/v); gently stirring the mixture on a magnetic stirrer at 40 ℃ for a certain time of 1h, fully mixing, and centrifuging the mixture at 5000r/min for 20min to form clear three phases;
(3) refining the grease: collecting the upper organic phase, evaporating and recovering in vacuum, collecting a lipid sample, and recycling tert-butyl alcohol;
(4) refining protein by dissolving the intermediate phase in Tris-HCl buffer solution of 100 mmol/L and pH 9, hydrating at 4 deg.C overnight, dialyzing in distilled water to remove salt, collecting supernatant, concentrating, and drying to obtain rice bran protein;
(5) refining polysaccharide: dialyzing the lower layer solution in distilled water, desalting, concentrating, and drying to obtain testa oryzae polysaccharide.
Claims (8)
1. A method for simultaneously extracting grease, protein and polysaccharide from rice bran is characterized in that the grease, protein and polysaccharide in the rice bran are simultaneously separated and extracted by a three-phase extraction technology, and the method comprises the following steps:
(1) preparing raw materials: sieving testa oryzae with 60-100 mesh sieve, and mixing with distilled water at a certain proportion to obtain testa oryzae slurry;
(2) constructing a three-phase system: adding a certain amount of solid salt into the rice bran slurry, slightly swirling, and adjusting the pH of the mixture to 3-7; then slowly adding tert-butyl alcohol according to the proportion; gently stirring the mixture on a magnetic stirrer at a certain temperature for a certain time, fully mixing, and centrifuging the mixture at 5000r/min for 20min to form clear three phases;
(3) refining the grease: collecting the upper organic phase, evaporating and recovering in vacuum, collecting a lipid sample, and recycling tert-butyl alcohol;
(4) and (3) refining the protein: dissolving the intermediate phase in Tris-HCl buffer solution, hydrating at 4 ℃ overnight, dialyzing in distilled water to remove salt, taking supernatant, concentrating and drying to obtain rice bran protein;
(5) refining polysaccharide: dialyzing the lower layer solution in distilled water, desalting, concentrating, and drying to obtain testa oryzae polysaccharide.
2. The method for simultaneously extracting lipids, proteins and polysaccharides from rice bran according to claim, wherein: in the step (1), the rice bran and the distilled water are mixed according to the proportion 1 (10-30).
3. The method for simultaneously extracting lipids, proteins and polysaccharides from rice bran according to claim 1, wherein: the type of the solid salt in the step (2) is any one of ammonium sulfate, sodium sulfate, magnesium acetate and dipotassium hydrogen phosphate, and the addition amount of the salt is 10-50% (w/v).
4. The method for simultaneously extracting lipids, proteins and polysaccharides from rice bran according to claim 1, wherein: the ratio of the slurry to the tertiary butanol in the step (2) is 1:0.5-1:2.5 (v/v).
5. The method for simultaneously extracting lipids, proteins and polysaccharides from rice bran according to claim 1, wherein: the stirring temperature of the mixture in the step (2) is 30-70 ℃.
6. The method for simultaneously extracting lipids, proteins and polysaccharides from rice bran according to claim 1, wherein: the stirring time of the mixture in the step (2) is 0.5h-2.5 h.
7. The method for simultaneously extracting lipids, proteins and polysaccharides from rice bran according to claim 1, wherein the concentration of Tris-HCl buffer solution in the step (4) is 20-100 mmol/L, and the pH is 7-9.
8. The method for simultaneously extracting lipids, proteins and polysaccharides from rice bran according to claim 1, wherein: the drying of the refined protein and the refined polysaccharide in the step (4) and the step (5) can be drying, vacuum drying and freeze drying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010313749.7A CN111499771A (en) | 2020-04-20 | 2020-04-20 | Method for simultaneously extracting grease, protein and polysaccharide in rice bran by three-phase extraction technology |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010313749.7A CN111499771A (en) | 2020-04-20 | 2020-04-20 | Method for simultaneously extracting grease, protein and polysaccharide in rice bran by three-phase extraction technology |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111499771A true CN111499771A (en) | 2020-08-07 |
Family
ID=71871188
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010313749.7A Pending CN111499771A (en) | 2020-04-20 | 2020-04-20 | Method for simultaneously extracting grease, protein and polysaccharide in rice bran by three-phase extraction technology |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111499771A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111650319A (en) * | 2020-06-30 | 2020-09-11 | 广东省测试分析研究所（中国广州分析测试中心） | Gas-liquid multiphase extraction-gas chromatography-mass spectrometry analysis method for determining total chemical composition of peanut oil |
| CN115518087A (en) * | 2022-07-08 | 2022-12-27 | 李玉山 | Comprehensive extraction and separation method of active ingredients of ginkgo |
| CN116284465A (en) * | 2022-08-05 | 2023-06-23 | 西北农林科技大学 | Preparation method of eucommia ulmoides polysaccharide and application of eucommia ulmoides polysaccharide prepared by preparation method in anti-depression |
| CN116616344A (en) * | 2023-07-26 | 2023-08-22 | 山东蒙天乳业有限公司 | Low-sugar lactobacillus beverage and preparation method thereof |
| CN116874627A (en) * | 2023-07-11 | 2023-10-13 | 广东省科学院动物研究所 | Stemona tuberosa polysaccharide and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101560439A (en) * | 2009-05-25 | 2009-10-21 | 韶关学院 | Method for extracting teaseed oil from tea seeds by adopting three-phase separation method |
| CN108342247A (en) * | 2018-04-11 | 2018-07-31 | 安徽德运粮油有限公司 | A kind of method of three phase separation aqueous enzymatic extraction rice bran oil |
| CN110885384A (en) * | 2019-11-11 | 2020-03-17 | 江苏大学 | Method for efficiently separating active polysaccharide from fresh okra |
-
2020
- 2020-04-20 CN CN202010313749.7A patent/CN111499771A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101560439A (en) * | 2009-05-25 | 2009-10-21 | 韶关学院 | Method for extracting teaseed oil from tea seeds by adopting three-phase separation method |
| CN108342247A (en) * | 2018-04-11 | 2018-07-31 | 安徽德运粮油有限公司 | A kind of method of three phase separation aqueous enzymatic extraction rice bran oil |
| CN110885384A (en) * | 2019-11-11 | 2020-03-17 | 江苏大学 | Method for efficiently separating active polysaccharide from fresh okra |
Non-Patent Citations (2)
| Title |
|---|
| SUPHAT PHONGTHAI等: "Preparation of rice bran protein isolates using three-phase partitioning and its properties", 《FOOD AND APPLIED BIOSCIENCE JOURNAL》 * |
| 鞠兴荣等: "三相分离法提取米糠油的初步研究", 《中国粮油学报》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111650319A (en) * | 2020-06-30 | 2020-09-11 | 广东省测试分析研究所（中国广州分析测试中心） | Gas-liquid multiphase extraction-gas chromatography-mass spectrometry analysis method for determining total chemical composition of peanut oil |
| CN111650319B (en) * | 2020-06-30 | 2022-07-08 | 广东省测试分析研究所（中国广州分析测试中心） | Gas-liquid multiphase extraction-gas chromatography-mass spectrometry analysis method for determining total chemical composition of peanut oil |
| CN115518087A (en) * | 2022-07-08 | 2022-12-27 | 李玉山 | Comprehensive extraction and separation method of active ingredients of ginkgo |
| CN116284465A (en) * | 2022-08-05 | 2023-06-23 | 西北农林科技大学 | Preparation method of eucommia ulmoides polysaccharide and application of eucommia ulmoides polysaccharide prepared by preparation method in anti-depression |
| CN116874627A (en) * | 2023-07-11 | 2023-10-13 | 广东省科学院动物研究所 | Stemona tuberosa polysaccharide and preparation method and application thereof |
| CN116616344A (en) * | 2023-07-26 | 2023-08-22 | 山东蒙天乳业有限公司 | Low-sugar lactobacillus beverage and preparation method thereof |
| CN116616344B (en) * | 2023-07-26 | 2023-10-03 | 山东蒙天乳业有限公司 | Low-sugar lactobacillus beverage and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111499771A (en) | Method for simultaneously extracting grease, protein and polysaccharide in rice bran by three-phase extraction technology | |
| US20220080333A1 (en) | Ultrasonic composite acidic water extraction method for cordyceps polysaccharide and cordycepin in cordyceps militaris | |
| CN107858393B (en) | Method for extracting protein polypeptide from walnut meal | |
| CA2115571A1 (en) | Process for extracting lipids with a high proportion of long chain, highly unsaturated fatty acids | |
| US20080069922A1 (en) | Oil Seed Processing | |
| JP2017521084A (en) | Method for extracting soluble protein from microalgal biomass | |
| CN110699411B (en) | Preparation method of eggshell membrane polypeptide | |
| CN1765921A (en) | Separated soybean protein preparation process and prepared soybean small peptide and process | |
| CN101390564A (en) | Production method of corn separation protein | |
| CN106754834A (en) | A kind of preparation technology of high activity papain | |
| CN114286829A (en) | Compositions and methods for preparing hemicellulose products from coffee grounds | |
| CN108101980B (en) | Preparation method of high-purity phycocyanin | |
| JP4662846B2 (en) | Alcohol lowering agent | |
| US6228993B1 (en) | Soy isoflavone concentrate process and product | |
| Otero et al. | A process for the complete fractionation of baker's yeast | |
| KR20170105497A (en) | Method for fractionating components of a biomass of protein-rich microalgae | |
| CN101117641B (en) | Method for preparing secoisolariciresinol diglucoside | |
| JPH06315369A (en) | Treatment of distillation waste liquor of 'shochu' | |
| CN107467639A (en) | A kind of coproduction prepares oat beta glucan nutrient powder and the method for dietary fiber micro powder | |
| RU2815687C1 (en) | Method of producing complex of polysaccharides from rapeseed cake with coextracted protein | |
| CN110651886A (en) | Preparation method and application of macadamia nut protein | |
| RU2815553C1 (en) | Method of producing protein isolate from rapeseed cake | |
| RU2233104C1 (en) | Method for complex reprocessing of brown algae for producing of iodine-containing and polysaccharide products | |
| CN106801081A (en) | A kind of method that activated protein is extracted from rice bran | |
| RU2681442C1 (en) | Protein-vitamin complex from cedar cake obtaining method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200807 |
|
| RJ01 | Rejection of invention patent application after publication |