CN113754716A - Technological method for extracting mulberry leaf protein by membrane method - Google Patents
Technological method for extracting mulberry leaf protein by membrane method Download PDFInfo
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- CN113754716A CN113754716A CN202111186028.5A CN202111186028A CN113754716A CN 113754716 A CN113754716 A CN 113754716A CN 202111186028 A CN202111186028 A CN 202111186028A CN 113754716 A CN113754716 A CN 113754716A
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- 238000000034 method Methods 0.000 title claims abstract description 45
- 235000008708 Morus alba Nutrition 0.000 title claims abstract description 36
- 240000000249 Morus alba Species 0.000 title claims abstract description 36
- 239000012528 membrane Substances 0.000 title claims abstract description 35
- 101710138460 Leaf protein Proteins 0.000 title claims abstract description 24
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 21
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 21
- 238000001035 drying Methods 0.000 claims abstract description 18
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 14
- 238000003756 stirring Methods 0.000 claims abstract description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 12
- 239000008367 deionised water Substances 0.000 claims abstract description 6
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 6
- 239000007788 liquid Substances 0.000 claims abstract description 5
- 238000002791 soaking Methods 0.000 claims abstract description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims description 17
- 239000002699 waste material Substances 0.000 claims description 12
- 239000000706 filtrate Substances 0.000 claims description 9
- 229920002239 polyacrylonitrile Polymers 0.000 claims description 6
- 238000009833 condensation Methods 0.000 claims description 5
- 230000005494 condensation Effects 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 5
- 229920002492 poly(sulfone) Polymers 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000000287 crude extract Substances 0.000 claims description 4
- 239000004695 Polyether sulfone Substances 0.000 claims description 3
- 229920006393 polyether sulfone Polymers 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 239000012535 impurity Substances 0.000 claims description 2
- 238000005057 refrigeration Methods 0.000 claims description 2
- 238000007873 sieving Methods 0.000 claims description 2
- 238000009210 therapy by ultrasound Methods 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims description 2
- 238000000967 suction filtration Methods 0.000 claims 2
- 239000012141 concentrate Substances 0.000 claims 1
- 238000000605 extraction Methods 0.000 abstract description 4
- 238000009826 distribution Methods 0.000 abstract description 2
- 239000006228 supernatant Substances 0.000 abstract 4
- 238000005119 centrifugation Methods 0.000 abstract 1
- 238000004140 cleaning Methods 0.000 abstract 1
- 230000014759 maintenance of location Effects 0.000 abstract 1
- 239000002994 raw material Substances 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 17
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 6
- 238000000751 protein extraction Methods 0.000 description 5
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 230000010874 maintenance of protein location Effects 0.000 description 4
- 235000019750 Crude protein Nutrition 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 238000000409 membrane extraction Methods 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000005185 salting out Methods 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 108010064851 Plant Proteins Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 235000006486 human diet Nutrition 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 235000021118 plant-derived protein Nutrition 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 230000003946 protein process Effects 0.000 description 1
- 229940116540 protein supplement Drugs 0.000 description 1
- 235000005974 protein supplement Nutrition 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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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
-
- 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
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Water Supply & Treatment (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a process method for extracting mulberry leaf protein by a membrane method, which comprises the following specific experimental steps of cleaning and drying mulberry leaves, crushing the mulberry leaves, soaking the mulberry leaves in deionized water according to a certain material-liquid ratio, stirring, centrifuging, collecting supernatant, adding water into the centrifuged filter residue, stirring, centrifuging, discarding the filter residue, and combining the obtained supernatant with the supernatant obtained after the first centrifugation. And (3) carrying out ultrafiltration on the supernatant through an ultrafiltration membrane to obtain a protein concentrated solution, then carrying out cold drying, and carrying out cold drying to obtain the mulberry leaf protein. The method is simple and convenient to operate, wide in raw material distribution, low in cost and environment-friendly. The retention rate of the obtained protein is as high as more than 97%, the extraction rate is 4%, and the purity of the obtained leaf protein is about 30%.
Description
Technical Field
The invention relates to the technical field of extracting protein from mulberry leaves, in particular to a technological method for extracting mulberry leaf protein by a membrane method.
Background
The mulberry has wide planting area and even distribution in China, a certain amount of mulberry leaves are exported to foreign countries every year, and in addition, a large amount of mulberry leaves cause waste. The leaf protein plant leaf protein is one of the largest green renewable protein resources in the world, contains trace elements required by a human body, does not contain cholesterol, and has great economic value. The mulberry leaves are rich in nutrition, have high protein content, especially have the highest content of glutamic acid and aspartic acid, and can be used as important protein supplement for human diet. In conclusion, the mulberry leaf protein is a plant protein resource with great development value, and can be widely applied along with the continuous improvement of the production process and the continuous improvement of the extraction rate of the mulberry leaf protein, so that a road for a mulberry leaf high-purity protein process which is simple to operate and environment-friendly is explored, and great economic benefits and social benefits are achieved.
The existing method for extracting the mulberry leaf protein comprises a heating method, an acid-base precipitation method, an acid-heating method, an alkali-heating method, a salting-out method, an organic solvent method and the like, and the methods have a plurality of defects. The yield of the protein by the direct heating method is not high, and the protein is easy to denature due to overhigh temperature; the pH value is not easy to control in the acid-base precipitation operation process, and waste liquid generated by using a large amount of acid or base is not environment-friendly; the salting-out method generally needs ammonium sulfate removal treatment by adding ammonium sulfate, so that the extraction process of the mulberry leaf protein becomes more complicated; the organic solvent method uses a large amount of organic solvent, and has problems such as difficulty in recovery and residue.
Disclosure of Invention
The invention aims to solve the problems of mulberry leaf resource waste and the defects of the technology. We invent a method for extracting mulberry leaf protein by a membrane method to solve the problems. The method has the advantages of low cost, simple operation, low energy consumption, environment-friendly and nuisanceless production process, high purity and green and nontoxic products. The invention adopts the following process method to achieve the aim.
(1) Washing impurities on the surface of fresh mulberry leaves, drying at 60 ℃, crushing and sieving with a 40-60-mesh sieve;
(2) according to a certain material-liquid ratio: mulberry leaf powder: deionized water 1: soaking and stirring the materials for 15-30 g/ml, performing ultrasonic treatment for 10-30 min, performing water bath, placing the materials in a refrigerator for refrigeration for 10-30 min at 20-40 ℃, centrifuging, and collecting centrifuged juice and waste residues respectively;
(3) adding deionized water into the waste residue, stirring, centrifuging again to obtain juice, discarding the waste residue, combining the juice obtained in the two steps, passing the juice through a gauze, centrifuging the obtained juice at a high speed of 4000-5000 r for 10 mm, and removing the waste residue to obtain a crude extract of the protein.
(4) Performing membrane separation on the centrifuged juice to obtain a mulberry leaf protein concentrated solution;
(5) and (3) carrying out cold drying on the concentrated solution to obtain a leaf protein solid, wherein the cold drying condition is that the condensation temperature is as follows: the vacuum degree is 24Pa at-57 to-30 ℃.
Wherein, the membrane material for membrane separation in the process (4) can be a polyacrylonitrile membrane, a polysulfone membrane and a polyethersulfone membrane, and is preferably a polyacrylonitrile membrane.
Wherein, in the process (4), the molecular weight cut-off of the ultrafiltration membrane in the ultrafiltration process is as follows: WMCO: 10000-300000, preferably polyacrylonitrile membrane (WMCO: 100000).
The invention has the beneficial effects that: the process method has the biggest characteristics of low cost, simple operation, low energy consumption, environment-friendly and pollution-free production process, suitability for large-scale production, high yield and green and nontoxic products. The mulberry leaf powder is added with water twice, stirred and centrifuged to increase the dissolution rate of protein, and the crude extract is centrifuged for 10min at 5000r and then ultrafiltered by an ultrafiltration membrane, so that suspended particles can be better removed to prevent the membrane from being blocked, and the purity of a product obtained by intercepting through the ultrafiltration membrane can be improved.
Drawings
FIG. 1 is a process flow chart of the mulberry leaf protein extraction method by a membrane method.
Detailed Description
The present invention will be further described with reference to the following specific operation examples, it should be noted that the examples are only illustrative of the extraction process of mulberry leaf protein of the present invention, and do not represent the whole patent of the present invention, and are not to be understood as limiting the scope of the present invention.
Preprocessing mulberry leaves: respectively weighing 3 parts of 4g of mulberry leaf powder according to a material-liquid ratio of 1: adding deionized water 20(g/ml), mixing, stirring, ultrasonic treating for 15min, stirring at 40 deg.C and 400r for 20min in water bath, cooling, centrifuging at 3000r, collecting juice, adding water 30ml into waste residue, mixing, stirring, filtering with gauze, mixing the filtrate with the juice obtained at the last time, centrifuging at 5000r, discardingMixing the three obtained juices, reserving 4ml, and measuring protein concentration by Coomassie brilliant blue method and recording as CCrude extract. The calculation formulas of the protein retention rate, the protein purity and the protein extraction rate are as follows:
EXAMPLE 1 Membrane extraction of Mulberry leaf protein (Polyacrylonitrile membrane)
Ultrafiltering 50ml of the pretreated juice with polyacrylonitrile ultrafiltration membrane, pressurizing with nitrogen in ultrafiltration cup at 0.6MPa, stirring at room temperature at 1000r, collecting 33ml of filtrate, and measuring protein concentration C by Coomassie brilliant blue methodFiltrateAnd carrying out cold drying after intercepting to obtain a protein concentrated solution, wherein the condensation temperature is-44 ℃, and the vacuum degree is 24 Pa. The mass M of the crude protein obtained after the cold drying for 48 hoursCold dryingThe protein retention rate, protein purity and protein extraction rate are shown in table 1.
Example 2 Membrane method for extracting Mulberry leaf protein (polysulfone membrane)
Taking 50ml of the pretreated juice, adopting a polysulfone ultrafiltration membrane, pressurizing the ultrafiltration cup with nitrogen at 0.6MPa, stirring at room temperature of 1000r, collecting 35ml of filtrate, and measuring the protein concentration C by adopting a Coomassie brilliant blue methodFiltrateAnd carrying out cold drying after intercepting to obtain a protein concentrated solution, wherein the condensation temperature is-44 ℃, and the vacuum degree is 24 Pa. The mass M of the crude protein obtained after the cold drying for 48 hoursCold dryingThe protein retention rate, protein purity and protein extraction rate are shown in table 1.
EXAMPLE 3 Membrane extraction of Mulberry leaf protein (polyethersulfone Membrane)
Collecting the above pretreated juice, subjecting 50ml of the juice to polysulfone ultrafiltration membrane, pressurizing with nitrogen in ultrafiltration cup at 0.6MPa, stirring at room temperature for 1000r, and collecting34ml of filtrate, and protein concentration C was measured by Coomassie brilliant blue methodFiltrateAnd carrying out cold drying after intercepting to obtain a protein concentrated solution, wherein the condensation temperature is-44 ℃, and the vacuum degree is 24 Pa. The mass M of the crude protein obtained after the cold drying for 48 hoursCold dryingThe protein retention rate, protein purity and protein extraction rate are shown in table 1.
TABLE 1
Claims (3)
1. A process for extracting mulberry leaf protein by a membrane method is characterized by comprising the following specific steps
(1) Washing impurities on the surface of fresh mulberry leaves, drying at 40-60 ℃, crushing and sieving with a 40-70 mesh sieve;
(2) according to a certain material-liquid ratio: mulberry leaf powder: deionized water 1: soaking and stirring the materials for 15-30 g/ml, performing ultrasonic treatment for 10-30 min, performing water bath, placing the materials in a refrigerator for refrigeration for 10-30 min at 20-50 ℃, centrifuging, and collecting centrifuged juice and waste residues respectively;
(3) adding deionized water into the waste residue, stirring, centrifuging again to obtain juice, discarding the waste residue, combining the juice obtained in the two steps, performing suction filtration by using gauze, and centrifuging 3000 r-5000 r of the filtrate obtained after suction filtration for 10 min; removing waste residues to obtain a crude extract of the protein;
(4) ultrafiltering the centrifuged juice to obtain folium Mori protein concentrate, collecting the filtrate, and measuring protein content;
(5) and (3) carrying out cold drying on the concentrated solution to obtain leaf protein powder, wherein the cold drying condition is that the condensation temperature is as follows: the vacuum degree is 24Pa at-57 to-30 ℃.
2. The method for extracting mulberry leaf protein by using the membrane method as claimed in claim, wherein the membrane for membrane separation is polyacrylonitrile membrane, polysulfone membrane and polyethersulfone membrane.
3. The method for extracting mulberry leaf protein by using a membrane method of claim, wherein the molecular weight cut-off of the ultrafiltration membrane in the ultrafiltration process is as follows: WMCO: 10000-300000.
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| CN202111186028.5A CN113754716A (en) | 2021-10-12 | 2021-10-12 | Technological method for extracting mulberry leaf protein by membrane method |
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| CN202111186028.5A CN113754716A (en) | 2021-10-12 | 2021-10-12 | Technological method for extracting mulberry leaf protein by membrane method |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103387599A (en) * | 2013-08-08 | 2013-11-13 | 河北考力森生物科技有限公司 | Technology for extracting protein of folium mori |
| CN103509097A (en) * | 2013-10-11 | 2014-01-15 | 中国科学院烟台海岸带研究所 | Extraction process method for plant leaf protein |
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2021
- 2021-10-12 CN CN202111186028.5A patent/CN113754716A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103387599A (en) * | 2013-08-08 | 2013-11-13 | 河北考力森生物科技有限公司 | Technology for extracting protein of folium mori |
| CN103509097A (en) * | 2013-10-11 | 2014-01-15 | 中国科学院烟台海岸带研究所 | Extraction process method for plant leaf protein |
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