CN102827685A - Method for extracting soybean phospholipids through aqueous enzymatic method - Google Patents
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- CN102827685A CN102827685A CN2012103433034A CN201210343303A CN102827685A CN 102827685 A CN102827685 A CN 102827685A CN 2012103433034 A CN2012103433034 A CN 2012103433034A CN 201210343303 A CN201210343303 A CN 201210343303A CN 102827685 A CN102827685 A CN 102827685A
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Abstract
A method for extracting soybean phospholipids through an aqueous enzymatic method belongs to a plant phospholipid extracting and processing technology, and comprises the following steps that (1) soybeans are ground to be subjected to extrusion pretreatment to obtain extruded materials, the extruded materials are mixed with water to obtain a mixed liquid, alkaline protease is added into the mixed liquid for enzymolysis, and centrifugal separation is carried out after the enzymolysis to obtain free oil, emulsion, hydrolysate and residues; (2) the emulsion which is obtained in the step (1) is preheated, the pH value of the emulsion is regulated, and the emulsion is continuously heated, mixed, added with carboxylic acid hydrolase for enzymolysis, subjected to enzyme deactivation after the enzymolysis, added with water to hydrate, and centrifugally separated after the hydration to obtain supernatant; and (3) the supernatant which is obtained in step (2) is filtered through a micro-filtration membrane to obtain soybean phospholipids. By the method for extracting soybean phospholipids, the functionality of the soybean phospholipids can be better maintained, moreover, the required equipment is simple and is easy to operate, and the obtained soybean phospholipids have high purity and high yield.
Description
Technical field
The invention belongs to the extraction processing technology of plant phosphatide, relate generally to a kind of method of aqueous enzymatic extraction soybean phospholipid.
Background technology
At present, the preparation soybean phospholipid mainly contains silica gel or methods such as calcium salt absorption, supercritical extraction, SX, membrane sepn and gas antisolvent crystallisation.Method commonly used has solvent extration, supercritical extraction and membrane separation process.To the powdered soybean phospholipid development, start from the fifties in last century abroad, Japan's end of the seventies, existing patent report promptly was raw material with the concentrated soybean phospholipid, employing acetone is solvent, processes powdered soybean phospholipid through refining the purification.U.S. Riceland Food Company company the eighties because of succeeding in developing the flow model powdered soybean phospholipid.1998, U.S. ADM company held in the food exhibition at Frankfurt, Germany and shows that to the public ultrafiltration process prepares powder lecithin.China's powdered soybean phospholipid development starts from the sixties in last century, and owing to technology, equipment and technology still fail to make a breakthrough, the production cycle reaches 20 h ~ 30 h, and solvent consumption is big, poor product quality, does not also reach commercialization up to the end of the eighties.
The solvent extration advantage is that technology is simpler, and productive rate is higher, is easy to industriallization; But this method acetone, phosphatide consumption are big, and energy consumption is high, and acetone and acetone secondary metabolite are residual too high.Present domestic most soybean phospholipid products are not measured its acetone residual quantity.The residual too high maximum potential safety hazard of China's phospholipid prod that become of acetone and acetone secondary metabolite; Improve constantly with standard of living, the human consumer requires more strict to food safety.Particularly occur repeatedly today, should more pay close attention to there being potential safety hazard on the powdered soybean phospholipid quality product on the market in the food safety incident.
Supercritical CO
2The abstraction technique advantage is that percentage extraction is high, product does not contain dissolvent residual etc.; But it also has comparatively critical defect: (1) must under high pressure work, and equipment requirements is harsh, costs an arm and a leg; (2) treatment capacity is lower, is not easy to scale operation; (3) only can intermittently change production, be unfavorable for industrial applications.
Membrane separation technique is nearly ten years research focuses, is mainly used in fields such as fat degumming, depickling, decolouring, solvent recuperation.In preparation phosphatide field, mainly concentrate on the preparation food-level concentrated soybean phospholipid in recent years, preparation is then reported seldom for powder lecithin.Membrane separation technique is new and effective, the accurate stripping technique of succeeding in developing in 20th century.Advantages such as membrane separation technique has the separation efficiency height, equipment is simple, easy to operate and energy-conservation are widely used in each industrial circle.Membrane sepn can be divided into r-o-(RO, < 1 nm), nanofiltration (NF is about 1 nm), ultrafiltration (UF, 1 ~ 100 nm) and micro-filtration (MF, 0. 1 ~ 10 μ m) according to the difference of membrane pore size.R-o-is mainly used in the processing to water, and the molecular weight cut-off of nanofiltration is generally 10
2~ 10
3U, the molecular weight cut-off of ultrafiltration are 10
3~ 10
6U, micro-filtration is also referred to as millipore filtration, is a kind of secondary filter.Membrane separation technique can be divided into organic membrane and mineral membrane by mould material, and mostly the carrier of mineral membrane is porous ceramic materials, compares with the organic membrane material, and inorganic ceramic film has following many advantages in practical application.
At present, it is less both at home and abroad to be used for the report of evaporation concentration and solvent recuperation aspect about membrane sepn, and people such as external Kuk use the solvent in RO/NF film recovery ethanol and the cottonseed oil mixture, draw the aperture and be not more than 2nm, and aperture density is 10
12Individual/cm
2The RO/NF film be suitable for from mixture, reclaiming ethanol most; 7 RO/NF films of humans such as Koseoglu separate Oleum Gossypii semen with 5 UF films from normal hexane, ethanol, Virahol; Human 0.02 μ m inorganic ceramic membrane ultrafiltration soybean miscellas such as Chi-sheng Wu have 20% VT 18 to be held back by ultra-filtration membrane approximately, and solvent is not trapped.Selecting suitable membrane material and definite optimized process conditions is that membrane sepn is effective to oil prodution industry subject matter, also is one of focus of research at present.
Membrane separation process has plurality of advantages: (1) sepn process place normal temperature process is particularly suitable for heat-sensitive substance; (2) sepn process does not undergo phase transition, and energy consumption is relatively low; (3) because of being physical process, can keep the product function characteristic; (4) only make impellent with pressure, therefore device is easier, processing ease, for ease of maintenaince.
So far, there is not a kind of process method that can the recovery of phosphatide in the aqueous enzymatic extraction soybean oil process be come out.
Summary of the invention
Technical problem to be solved by this invention is the deficiency that overcomes above-mentioned prior art, and a kind of method of aqueous enzymatic extraction soybean phospholipid is provided.
Technical problem to be solved by this invention realizes through following technical scheme:
A kind of method of aqueous enzymatic extraction soybean phospholipid; This method may further comprise the steps: (1) is pulverized laggard capable expelling-expansion pretreatment with soybean and is obtained expanded material; Expanded material mixed with water obtain mixed solution; In mixed solution, add Sumizyme MP and carry out enzymolysis, spinning obtains free oil, milk sap, hydrolyzed solution and residue behind the enzymolysis; (2) milk sap that step (1) is obtained adds lemon acid for adjusting pH value to 2.16 after 50 ℃ of following preheatings, in 55 ℃ of water-baths behind the heating 1h; The pH value is adjusted to 5.5, continues heating 1h, and constantly stir; Add carboxylic acid lytic enzyme enzymolysis then, the enzyme that goes out behind the enzymolysis adds water hydratable 30-150min; Spinning obtains supernatant behind the hydration reaction, and described hydrolysis temperature is that 50-90 ℃, the addition and the milk sap mass ratio of carboxylic acid lytic enzyme are that 0.3-0.7:1, enzymolysis time are 1-5h; (3) both got soybean phospholipid after the supernatant that step (2) is obtained filters with microfiltration membrane, described microfiltration membrane aperture is that inorganic ceramic film, the micro-filtration pressure of 600-2000nm is that 0.1-0.5MPa, micro-filtration temperature are 20-60 ℃.
Described hydration time is 90min.
Described enzymolysis preferred parameter is: the addition of carboxylic acid lytic enzyme and milk sap mass ratio are that 0.6:1, hydrolysis temperature are that 80 ℃, enzymolysis time are 3h.
Described micro-filtration preferred parameter is: the microfiltration membrane aperture is that 800nm, micro-filtration temperature are that 50 ℃, micro-filtration pressure are 0.3 MPa.
The inventive method is on the basis of aqueous enzymatic method, in milk sap, adds the carboxylic acid lytic enzyme and makes wherein water-fast non-hydrated phosphatide be converted into the water-soluble hemolytic phosphatide of ability, makes the phosphorus content in the grease be less than or equal to 10 * 10
-6, after film is removed impurity excessively, isolate phosphatide; The soybean phospholipid that present method is extracted can keep it functional preferably, and present method required equipment is simple simultaneously, obtains phospholipid purity height, quality better.
Description of drawings
Fig. 1 overall process route map of the present invention;
The addition of Fig. 2 enzyme is to the influence of phosphatide decreasing ratio;
Fig. 3 preheating temperature is to the influence of phosphatide decreasing ratio;
Fig. 4 hydration time is to the influence of phosphatide decreasing ratio;
The influence of Fig. 5 enzymolysis time comparison phosphatide decreasing ratio;
Fig. 6 microfiltration membrane aperture is to the influence of phosphatide rejection;
Fig. 7 micro-filtration temperature is to the influence of phosphatide rejection;
Fig. 8 micro-filtration pressure is to the influence of phosphatide rejection.
Embodiment
Further describe the present invention below in conjunction with specific embodiment.
A kind of method of aqueous enzymatic extraction soybean phospholipid; This method may further comprise the steps: (1) is pulverized laggard capable expelling-expansion pretreatment with soybean and is obtained expanded material; Expanded material mixed with water obtain mixed solution; In mixed solution, add Sumizyme MP and carry out enzymolysis, spinning obtains free oil, milk sap, hydrolyzed solution and residue behind the enzymolysis; (2) milk sap that step (1) is obtained adds lemon acid for adjusting pH value to 2.16 after 50 ℃ of following preheatings, in 55 ℃ of water-baths behind the heating 1h; The pH value is adjusted to 5.5, continues heating 1h, and constantly stir; Add carboxylic acid lytic enzyme enzymolysis then, the enzyme that goes out behind the enzymolysis adds water hydratable 30-150min; Spinning obtains supernatant behind the hydration reaction, and described hydrolysis temperature is that 50-90 ℃, the addition and the milk sap mass ratio of carboxylic acid lytic enzyme are that 0.3-0.7:1, enzymolysis time are 1-5h; (3) both got soybean phospholipid after the supernatant that step (2) is obtained filters with microfiltration membrane, described microfiltration membrane aperture is that inorganic ceramic film, the micro-filtration pressure of 600-2000nm is that 0.1-0.5MPa, micro-filtration temperature are 20-60 ℃.
Described hydration time is 90min.
Described enzymolysis preferred parameter is: the addition of carboxylic acid lytic enzyme and milk sap mass ratio are that 0.6:1, hydrolysis temperature are that 80 ℃, enzymolysis time are 3h.
Described micro-filtration preferred parameter is: the microfiltration membrane aperture is that 800nm, micro-filtration temperature are that 50 ℃, micro-filtration pressure are 0.3 MPa.
The screening experiment of embodiment 1 phosphatide enzymolysis extraction process condition optimum parameter
1 materials and methods
1.1 material, reagent
| Soybean | Sky, Tianjin power chemical reagent ltd |
| Protex 6L enzyme | Denmark Novo company |
| The carboxylic acid lytic enzyme | Guangzhou one hundred silent bio tech ltd |
| NaOH | Sky, Tianjin power chemical reagent ltd |
| Sodium orthomolybdate | Sky, Tianjin power chemical reagent ltd |
| The sulfuric acid hydrazine solution | Sky, Tianjin power chemical reagent ltd |
1.2 key instrument equipment
| Spectrophotometer | Deyang City Ke Rui instrument company |
| Low speed centrifuge | Good Instr Ltd. in the section in the Anhui |
| The desk type high speed refrigerated centrifuge | Whizzer ltd is found in capital, Beijing |
| The cryogenic thermostat stirring reaction is bathed | Zhengzhou Greatwall Scientific Industrial & Trading Co., Ltd. |
1.3 experimental technique
1.3.1 technical process (see figure 1)
1.3.2 calculation formula
2 results and discussion
2.1 phosphatide removing process list factor condition is to the influence of phosphatide decreasing ratio
2.1.1 the addition of enzyme is to the influence of phosphatide decreasing ratio
80 ℃ of preheatings, adding 45% Hydrocerol A, to be transferred to pH be 2.16 with milk sap, in 55 ℃ of water-baths behind the heating 1h; Using the NaOH of 2mol/L is 5.5 with pH regulator, heating 1h, and constantly stir; Add carboxylic acid lytic enzyme enzymolysis 3h then, after the deactivation, aquation 60min; Investigate of the influence of the addition of enzyme, result such as Fig. 2 to the phosphatide decreasing ratio.Can find out that by Fig. 2 result the phosphatide decreasing ratio increases along with the increase of the addition of enzyme; When the addition of carboxylic acid lytic enzyme and milk sap mass ratio are 0.6:1; It is maximum that decreasing ratio reaches, so the addition and the milk sap mass ratio of the carboxylic acid lytic enzyme of optimizing are 0.6:1.
2.1.2 preheating temperature is to the influence of phosphatide decreasing ratio
After the milk sap preheating, adding 45% Hydrocerol A, to be transferred to pH be 2.16, in 55 ℃ of water-baths behind the heating 1h; Using the NaOH of 2mol/L is 5.5 with pH regulator, heating 1h, and constantly stir; Add carboxylic acid lytic enzyme enzymolysis 3h then, after the deactivation, aquation 60min; Investigate the influence of hydrolysis temperature, result such as Fig. 3 to the phosphatide decreasing ratio.Can find out temperature 80 ℃ the time by Fig. 3 result, it is maximum that the phosphatide decreasing ratio reaches, so preferred hydrolysis temperature is 80 ℃.
2.1.3 hydration time is to the influence of phosphatide decreasing ratio
80 ℃ of preheatings, adding 45% Hydrocerol A, to be transferred to pH be 2.16 with milk sap, in 55 ℃ of water-baths behind the heating 1h; Using the NaOH of 2mol/L is 5.5 with pH regulator, heating 1h, and constantly stir; Add carboxylic acid lytic enzyme enzymolysis 3h then, after the deactivation, hydration reaction; Investigate the influence of hydration time, result such as Fig. 4 to the phosphatide decreasing ratio.Can find out that by Fig. 4 result aquation 90min phosphatide decreasing ratio reaches maximum, so the hydration time of optimizing is 90min.
2.1.4 the influence of enzymolysis time comparison phosphatide decreasing ratio
80 ℃ of preheatings, adding 45% Hydrocerol A, to be transferred to pH be 2.16 with milk sap, in 55 ℃ of water-baths behind the heating 1h; Using the NaOH of 2mol/L is 5.5 with pH regulator, heating 1h, and constantly stir; Add carboxylic acid lytic enzyme enzymolysis then, after the deactivation, aquation 60min; Investigate the influence of enzymolysis time comparison phosphatide decreasing ratio, result such as Fig. 5.Can find out that by Fig. 5 result enzymolysis 3h phosphatide decreasing ratio reaches maximum, so the enzymolysis time of optimizing is 3h.
3 confirmatory experiments and simultaneous test
Under the top condition that single-factor analysis therapy is tried to achieve; Addition and the milk sap mass ratio that is the carboxylic acid lytic enzyme is that 0.6:1, hydrolysis temperature are that 80 ℃, hydration time are that 90min, enzymolysis time are to carry out 3 parallel laboratory tests under the 3h, and the MV of 3 parallel laboratory tests of phosphatide decreasing ratio is 97.67%.
4 experiment conclusion
Utilize the single factor analysis method that the processing parameter of the recovery method of phosphatide in the aqueous enzymatic extraction soybean oil process is optimized.Having set up corresponding mathematical model is that later pilot scale and suitability for industrialized production provided fundamental basis, and the addition that to have obtained optimum phosphatide removing process condition be the carboxylic acid lytic enzyme and milk sap mass ratio are that 0.6:1, hydrolysis temperature are that 80 ℃, hydration time are that 90min, enzymolysis time are 3h.Can know that through checking the MV of 3 parallel laboratory tests of decreasing ratio under optimum dephosphorization technology condition is about 97.67%.
The screening experiment of embodiment 2 membrane separation process condition optimum parameters
1 materials and methods
1.1 key instrument equipment
MF-50 inorganic ceramic film ultrafiltration apparatus, inorganic ceramic film.
1.2 experimental technique
1.3 calculation formula
In the formula, J is a permeation flux, L/m
2H; V is the penetrating fluid volume, L; A is a membrane area; m
2T is the time, h; R is phosphatide or greasy rejection; C
f, C
pBe respectively phosphatide or fat content in liquid stock and the penetrating fluid.
2 results and discussion
2.1 membrane separation process list factor condition is to the influence of the rejection of phosphatide
2.1.1 the microfiltration membrane aperture is to the influence of the rejection of phosphatide
Micro-filtration pressure 0.3 MPa, the micro-filtration temperature is 50 ℃, investigates the influence of microfiltration membrane aperture to rejection, the result sees Fig. 6.The rejection that can be found out phosphatide by Fig. 6 result increases along with the increase in microfiltration membrane aperture, and when aperture greater than 800nm the time, rejection begins to descend.So the membrane pore size of optimizing is 800nm.
2.1.2 the micro-filtration temperature is to the influence of the rejection of phosphatide
Micro-filtration pressure 0.3 MPa, the microfiltration membrane aperture is 800nm, investigates the influence of micro-filtration temperature to the rejection of phosphatide, the result sees Fig. 7.Can find out when temperature is 50 ℃ that by Fig. 7 result it is maximum that the rejection of phosphatide reaches, so the micro-filtration temperature of optimizing is 50 ℃.
2.1.3 micro-filtration pressure is to the influence of the rejection of phosphatide
The microfiltration membrane aperture is 800nm, and the micro-filtration temperature is 50 ℃, investigates the influence of micro-filtration pressure to rejection, and the result sees Fig. 8.The rejection that can be found out phosphatide by Fig. 8 result increases along with the increase of pressure, and when pressure was 0.3 MPa, it is maximum that the phosphatide rejection reaches, so the micro-filtration pressure of optimizing is 0.3 MPa.
3 confirmatory experiments
Under the top condition that single-factor analysis therapy is tried to achieve, promptly the microfiltration membrane aperture is 800nm, and the micro-filtration temperature is 50 ℃, and micro-filtration pressure is 0.3 MPa, carries out 3 parallel laboratory tests, and the MV of 3 parallel laboratory tests of rejection of phosphatide is for being 72.56%.
4 experiment conclusion
Utilize the single factor analysis method that the processing parameter of the recovery method of phosphatide in the aqueous enzymatic extraction soybean oil process is optimized.Having set up corresponding mathematical model is that later pilot scale and suitability for industrialized production provided fundamental basis, and to have obtained optimum membrane separation process condition be 800nm for the microfiltration membrane aperture, and the micro-filtration temperature is 50 ℃, and micro-filtration pressure is 0.3 MPa.The MV that can know 3 parallel laboratory tests of rejection of phosphatide under optimum membrane separation process condition through checking is about 72.56%.
Claims (4)
1. the method for an aqueous enzymatic extraction soybean phospholipid; This method may further comprise the steps: (1) is pulverized laggard capable expelling-expansion pretreatment with soybean and is obtained expanded material; Expanded material mixed with water obtain mixed solution; In mixed solution, add Sumizyme MP and carry out enzymolysis, spinning obtains free oil, milk sap, hydrolyzed solution and residue behind the enzymolysis; It is characterized in that: the milk sap that (2) obtain step (1) adds lemon acid for adjusting pH value to 2.16 after 50 ℃ of following preheatings, in 55 ℃ of water-baths behind the heating 1h; The pH value is adjusted to 5.5; Continue heating 1h, and constantly stir, add carboxylic acid lytic enzyme enzymolysis then; Enzyme goes out behind the enzymolysis; Add water hydratable 30-150min, spinning obtains supernatant behind the hydration reaction, and described hydrolysis temperature is that 50-90 ℃, the addition and the milk sap mass ratio of carboxylic acid lytic enzyme are that 0.3-0.7:1, enzymolysis time are 1-5h; (3) both got soybean phospholipid after the supernatant that step (2) is obtained filters with microfiltration membrane, described microfiltration membrane aperture is that inorganic ceramic film, the micro-filtration pressure of 600-2000nm is that 0.1-0.5MPa, micro-filtration temperature are 20-60 ℃.
2. the method for a kind of aqueous enzymatic extraction soybean phospholipid according to claim 1 is characterized in that described hydration time is 90min.
3. the method for a kind of aqueous enzymatic extraction soybean phospholipid according to claim 1 is characterized in that described enzymolysis preferred parameter is: the addition of carboxylic acid lytic enzyme and milk sap mass ratio are that 0.6:1, hydrolysis temperature are that 80 ℃, enzymolysis time are 3h.
4. the method for a kind of aqueous enzymatic extraction soybean phospholipid according to claim 1 is characterized in that described micro-filtration preferred parameter is: the microfiltration membrane aperture is that 800nm, micro-filtration temperature are that 50 ℃, micro-filtration pressure are 0.3 MPa.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106753747A (en) * | 2016-12-05 | 2017-05-31 | 东北农业大学 | A kind of method that enzyme process prepares soybean grease and nanoemulsions |
| CN107117668A (en) * | 2017-05-31 | 2017-09-01 | 成都振中电气有限公司 | A kind of effluent purifier for handling electroplating technology waste water and preparation method thereof |
| CN107319100A (en) * | 2017-08-20 | 2017-11-07 | 合肥信达膜科技有限公司 | A kind of method that soybean lecithin is extracted from soybean |
| CN114478621A (en) * | 2022-01-25 | 2022-05-13 | 海南乐孕生物科技有限公司 | Process for extracting enzymolysis soybean phospholipid |
| CN115353528A (en) * | 2022-08-17 | 2022-11-18 | 东北林业大学 | Novel green production process of powdered phospholipid |
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2012
- 2012-09-17 CN CN2012103433034A patent/CN102827685A/en active Pending
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| 李杨等: "水酶法制取大豆油的水解度对提油率影响机理研究", 《食品与发酵工业》 * |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106753747A (en) * | 2016-12-05 | 2017-05-31 | 东北农业大学 | A kind of method that enzyme process prepares soybean grease and nanoemulsions |
| CN107117668A (en) * | 2017-05-31 | 2017-09-01 | 成都振中电气有限公司 | A kind of effluent purifier for handling electroplating technology waste water and preparation method thereof |
| CN107319100A (en) * | 2017-08-20 | 2017-11-07 | 合肥信达膜科技有限公司 | A kind of method that soybean lecithin is extracted from soybean |
| CN114478621A (en) * | 2022-01-25 | 2022-05-13 | 海南乐孕生物科技有限公司 | Process for extracting enzymolysis soybean phospholipid |
| CN114478621B (en) * | 2022-01-25 | 2024-03-26 | 海南乐孕生物科技有限公司 | Process for extracting enzymatic soybean phospholipids |
| CN115353528A (en) * | 2022-08-17 | 2022-11-18 | 东北林业大学 | Novel green production process of powdered phospholipid |
| CN115353528B (en) * | 2022-08-17 | 2024-03-15 | 东北林业大学 | New green production process of powder phosphatide |
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Application publication date: 20121219 |