CN105294822A - Process for extracting rapeseed protein - Google Patents
Process for extracting rapeseed protein Download PDFInfo
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- CN105294822A CN105294822A CN201410348962.6A CN201410348962A CN105294822A CN 105294822 A CN105294822 A CN 105294822A CN 201410348962 A CN201410348962 A CN 201410348962A CN 105294822 A CN105294822 A CN 105294822A
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Abstract
The invention relates to a process for extracting rapeseed protein. The process is simple, and has the advantages that the degree of protein denaturation is greatly reduced, the protein extraction rate is high, and the loss of a surfactant is remarkable reduced; besides, water resources are saved, and energy consumption in industrial production is lowered. In the process for extracting the rapeseed protein by use of reverse micelle, a method for backward extraction comprises the following steps: extracting the rapeseed protein by use of a reverse micelle solution to obtain a forward extraction solution; conducting vacuum concentration on the forward extraction solution to remove an organic solvent in the reverse micelle solution; then, adding a buffer solution for extraction and separation, so as to obtain an aqueous solution of the rapeseed protein. A method for extracting the rapeseed protein by use of the reverse micelle solution comprises the following steps: adding powder of rapeseed dregs into the reverse micelle solution; carrying out extraction under the ultrasonic action; then, carrying out solid-liquid separation to obtain the forward extraction solution. According to the process, vacuum concentration is conducted through rotary evaporation, and the water bath temperature is controlled to be 50-70 DEG C; the process is simple, lower in resource consumption, low in production cost, mild in operating condition, and high in protein extraction rate.
Description
Technical field
The present invention relates to a kind of technique extracting rape seed protein.
Background technology
Rape is a kind of oil crops that China's big area is produced, and output ranks first in the world.The by-product volume produced in fundamentals process is huge, only rapeseed meal output, every year just more than 1,000 ten thousand tons.Containing the crude protein of 35% ~ 42% in rapeseed meal, and in rape seed protein, the content of Methionin, Gelucystine and methionine(Met) is higher, and amino acid balance is better than soybean protein, is a kind of vegetable protein sources.
At present with vegetable seed or rapeseed meal for raw material, the method for producing rape seed protein mainly contains Aqueous phase, aqueous enzymatic extraction, organic solvent extraction, the method such as ultrafiltration, percolation process.Often protein extracting ratio is lower in the industrial production for Aqueous phase, also has waste water not easily to process, easily causes the problems such as environmental pollution; Although aqueous enzymatic extraction improves the extraction yield of rape seed protein, reduce some functional property with protein and nutritive value; Prepare the method for rape seed protein with the organic solvent such as alcohol, ketone, because of complex process, cost are high and solvent on problems such as the impacts of protein nutritive value; And containing the macromolecular substance such as protein, pectin in the feed liquid of ultrafiltration, percolation process production, ultra-filtration membrane face or fenestra are easy to contaminated, cause the processing power of ultra-filtration membrane to decline even can not work, and pollute after cleaning also quite numerous and diverse, cause ultra-filtration technique promotion and application to be industrially subject to great restriction.
Domestic and international investigator is not exploring new protein separation technology to section.The extraction of anti-glue card be exactly grow up in this context new and effective isolation technique [Li Xiang village, He Gaohong, the new development [J] of inverse micelle abstraction protein techniques. water technology, 2005.31 (1) 7-12.].Much research shows that inverse micelle abstraction technique is as a novel isolation technique, has very large advantage in separating-purifying biologically active substance.Reverse micelle is a kind of aggregate of the tensio-active agent nanoscale of spontaneous formation in organic solvent, in reverse micelle, the polar group part of tensio-active agent surrounds a polarity core, be called pond, this pond comprises polar group internal surface and the moisture wherein of tensio-active agent, and the ion etc. be dissolved in the water, there is hydrophilic biomacromolecule and just can be dissolved in the moisture in pond and be extracted by the form with micella.An important parameter of reverse micelle is its water content WO, the water namely dissolved in pond and the mol ratio of tensio-active agent, and it determines the size of reverse micelle.It is reported that bio-active substance enters inverse micellar solution is a kind of collaborative processes, namely the surfactant layer between macroscopical two-phase (organic phase and aqueous phase) interface is out of shape with contiguous protein generation electrostatic interaction, then the reverse micelle including bio-active substance is defined at two-phase interface, this reverse micelle diffuses into organic phase, thus achieving extracting and separating, this process is called forward extraction.
For reverse micelle method extracting method, obtain forward extraction by forward extraction technique liquid-solid so important, but from forward extraction liquid, target product is separated no less important, this process is referred to as rear extraction usually.Backward extraction method is in the past generally by anti-limb bundle solution and buffer solution contact, and the factor such as pH value, ionic strength of regulator solution, makes protein proceed to aqueous phase, extraction after realizing.
But adopting traditional backward extraction method, is generally large with buffered soln consumption, waste great lot of water resources, and the significant advantage making reverse micelle extract protein is covered, and also will evaporate large quantity of moisture, consume electric power in the later stage; And, in traditional rear extraction process, two-phase interface can produce a kind of white compound.This is because in rear extraction process, protein meeting and Action of Surfactant, deactivation, is deposited on the interface of two-phase.Like this, not only cause extraction rate of protein low, also cause a large amount of losses of tensio-active agent simultaneously.
Summary of the invention
The object of the invention is for overcoming the low shortcoming of traditional vegetable seed protein extracting method extraction rate of protein.
For achieving the above object, technical scheme of the present invention is as follows:
A kind of technique extracting rape seed protein is provided, it is characterized in that, comprise the following steps: extract rape seed protein with inverse micellar solution, obtain forward extraction liquid, forward extraction liquid, through vacuum concentration, adds buffered soln after the organic solvent in removing inverse micellar solution, extracting and separating, obtains the aqueous solution of rape seed protein; Extract rape seed protein with inverse micellar solution, the method obtaining forward extraction liquid is as follows:
Rapeseed meal powder is added in inverse micellar solution, 40 DEG C of extraction 80min under frequency 30KHz, power 120KW ul-trasonic irradiation, then carry out centrifugation 20min with the relative centrifugal force of 3200 × g, obtain forward extraction liquid, described inverse micellar solution is by succinate sodium 2-ethylhexyl, octane-iso and contain K
+buffered soln mix, in described inverse micellar solution, the mass volume ratio of succinate sodium 2-ethylhexyl and octane-iso is 0.02g/mL, W
0be 33, buffered soln is the Sodium phosphate dibasic-sodium hydroxide buffer solution of KCl, and pH value is 11, K
+concentration is 0.15mol.L
-1, the rapeseed meal powder added be 0.02g/mL with the mass volume ratio of inverse micellar solution.
After forward extraction liquid adds buffered soln after vacuum concentration, also add the aqueous ethanolic solution that volumetric concentration is 10%.
The method of described vacuum concentration is rotary evaporation, and bath temperature controls at 70 DEG C.
The buffered soln added after rotary evaporation is the buffered soln of KCl.
The pH value of buffer solution 7, K of the KCl added after rotary evaporation
+concentration is 1.Omol.L
-1.
The buffered soln of the KCl added after rotary evaporation is Sodium phosphate dibasic one sodium dihydrogen phosphate buffer of KCl.
The present invention selects suitable processing condition according to the characteristic of rape seed protein, adopts reverse micelle to extract forward extraction, rear extraction technique, significantly improves the extraction yield of protein in conjunction with ultrasonic technology, is the new way that the Efficient Development of rapeseed meal protein resource utilizes.Because rape seed protein is protected by tensio-active agent and water molecules, not with organic solvent exposure, therefore can not inactivation.After solid-liquid separation, gained solid phase is treated can obtain the byproducts such as fiber, makes full use of rapeseed meal.Adopt unique rear extraction technique, protein denaturation degree reduces greatly, and the loss of tensio-active agent significantly reduces; Not only after protein, extraction rate is high, and saving water resource, and reduction subsequent protein concentrates the energy consumption in link, and production cost reduces greatly.In a word, present invention process is simple, and economize on resources, production cost is low, and operational condition is gentle, and extraction rate of protein is high.
Embodiment
Below by embodiment, the invention will be further described.
EXAMPLE l:
(1) preparation of rapeseed meal powder:
Select 2008 annual output high-quality Double-low Rape Seed, on hulling machine, carry out heat treatment, process the choosing of laggard sector-style.Vegetable seed after shelling is pulverized on hammer Cyclone mill, after pulverizing, crosses 100 mesh sieves.On Soxhlet extraction device, at 50 DEG C, deoiling treatment is carried out to the rear vegetable seed of pulverizing with sherwood oil (30-60 DEG C of boiling range), be placed on ventilation after process and carry out precipitation, sherwood oil is volatilized completely.Air seasoning again, gained is the required rapeseed meal powder of test, and surveying its protein (N × 6.25 butt) content is 50.1%.
(2) configuration of inverse micellar solution
KCl buffer preparation: take 110.8g bis-hypophosphite monohydrate disodium hydrogen, 9.07g potassium primary phosphate, is all mixed with the solution of 1L, get 190mL and lOmL respectively to mix, then get 0.596g Repone K and add in above-mentioned mixed solution, make it dissolve completely, obtaining pH is 9, K
+concentration is 0.04mol.L
-1kCl buffered soln.
Take 3g Surfactant Aerosol OT, be placed in 250mL Erlenmeyer flask, add 50mL octane-iso, magnetic agitation makes tensio-active agent dissolve completely, after solution is transparent, adds KCI buffered soln, makes the W of inverse micellar solution
0value is 25.Then being placed in 40 DEG C of constant temperature oscillators to vibrate to solution and fully mix, then with the relative centrifugal force 20min of 1800 × g, if transparent, is inverse micellar solution, otherwise is not then (a small amount of water of bottom is little on system impact).The density of gained inverse micellar solution is 0.06g/mL.
(3) inverse micellar solution extracts rape seed protein, obtains forward extraction liquid:
Get above-mentioned inverse micellar solution 50mL, add 0.050g rapeseed meal powder in Erlenmeyer flask, be placed in ultrasonic cleaner, frequency 20KHz, power 1OOKW, extract 80min with 60 DEG C.Carry out centrifugation 15min with the relative centrifugal force of 3200 × g, gained supernatant liquor is forward extraction liquid.
Recording its protein (N × 6.25) content by GB/T-500915-2003 is 0.0203g, and the extraction yield of rape seed protein is 81.17%.
(4) vacuum concentration:
Above-mentioned forward extraction liquid is evaporated on the rotary evaporator, reclaims octane-iso, obtain solid substance.The temperature of rotary evaporation, vacuum tightness and rotating speed are controlled as 50 DEG C, 0.1Mpa and lOOr/s.
(5) add buffered soln, obtain rear extraction liquid, extracting and separating:
KCl buffer preparation: take 110.8g 12 hydration and to see sour disodium hydrogen, 9.07g potassium primary phosphate, is all mixed with the solution of 1L, get 190mL and lOmL respectively to mix, then get 11.133gKCl and add in above-mentioned mixed solution, make it dissolve completely, obtaining pH is 9, K
+concentration is 0.75mol.L
-1kCl buffered soln.
The above-mentioned K of 25mL is added in solid substance
+concentration is 0.75mol.L
-1kCl buffered soln, magnetic agitation makes remaining solid substance dissolve completely.Add the ethanol 1mL that concentration is 10% (v/v), be then placed in 30 DEG C of constant temperature oscillators and vibrate to solution and fully mix, then with the relative centrifugal force 20min of 1800 × g.
Take off a layer aqueous phase after separating funnel layering, survey protein (N × 6.25) content wherein for 0.0112g with GB/T-5009.5-2003 Kjeldahl determination, calculating percentage extraction after protein is 55.2%.
Embodiment 2:
(1) preparation of rapeseed meal powder:
Select 2008 annual output high-quality Double-low Rape Seed, on hulling machine, carry out heat treatment, process the choosing of laggard sector-style.Vegetable seed after shelling is pulverized on hammer Cyclone mill, after pulverizing, crosses 80 mesh sieves.On Soxhlet extraction device, at 50 DEG C, deoiling treatment is carried out to the rear vegetable seed of pulverizing with sherwood oil (30-60 DEG C of boiling range), be placed on ventilation after process and carry out precipitation, sherwood oil is volatilized completely.Air seasoning again, gained is the required rapeseed meal powder of test, and surveying its protein (N × 6.25, butt) content is 45.6%.
(2) preparation of inverse micellar solution
KCl buffer preparation: preparation 0.2mol.L
-1disodium phosphate soln and 0.1mol.L
-1the each 1L of citric acid solution, get 10.30mL and 9.70mL respectively and mix.Get 0.1193g Repone K again to add in above-mentioned mixed solution, make it dissolve completely, obtaining pH is 5, K
+concentration is 0.08mol.L
-1kCl buffered soln.
Take 3g Surfactant Aerosol OT, be placed in the Erlenmeyer flask of 250mL, add 50mL octane-iso, magnetic agitation makes tensio-active agent dissolve completely, after solution is transparent, adds above-mentioned KCl buffered soln, makes the W of inverse micellar solution
0value is 33.Then being placed in 30 DEG C of constant temperature oscillators to vibrate to solution and fully mix, then with the relative centrifugal force 15min of 2000 × g, if transparent, is inverse micellar solution, otherwise is not then (a small amount of water of bottom is little on system impact).
(3) inverse micellar solution extracts rape seed protein, obtains forward extraction liquid:
Get above-mentioned inverse micellar solution 50mL, add 0.501g rapeseed meal powder in Erlenmeyer flask, be placed in ultrasonic cleaner, frequency 40KHz, power 90KW, extract 120min with 20 DEG C.Carry out centrifugation lOmin with the relative centrifugal force of 3600 × g, gained supernatant liquor is forward extraction liquid.Recording its protein (N × 6.25) content by GB/T-5009.5-2003 is 0.1874g.The extraction yield of rape seed protein is 82.01%.
(4) vacuum concentration:
Above-mentioned forward extraction liquid is evaporated on the rotary evaporator, reclaims octane-iso, obtain solid substance.The temperature of rotary evaporation, vacuum tightness and rotating speed are controlled as 55 DEG C, 0.1Mpa and 90r/s.
(5) add buffered soln, obtain rear extraction liquid, extracting and separating:
KCl buffer preparation: preparation 0.2mol.L
-1disodium phosphate soln and 0.1mol.L
-1the each 1L of citric acid solution, get 19.45mL and 0.55mL respectively and mix.Get 1.86g Repone K again to add in above-mentioned mixed solution, make it dissolve completely, obtaining pH is 8.0, K
+concentration is 1.25mol.L
-1kCl buffered soln.
In solid substance, add 5mLKCl buffered soln, wherein potassium concentration is 1.25mol.L
-1, pH is 8.0, and magnetic agitation makes remaining solid substance dissolve completely.Add the ethanol 2mL that concentration is 10% (v/v), be then placed in 25 DEG C of constant temperature oscillators and vibrate to solution and fully mix, then with the relative centrifugal force 20min of 2400 × g.
Take off a layer aqueous phase after separating funnel layering, survey protein (N × 6.25) content wherein for 0.095g with GB/T-5009.5-2003 Kjeldahl determination, calculating percentage extraction after protein is 50.69%.
Claims (6)
1. extract a technique for rape seed protein, it is characterized in that, comprise the following steps: extract rape seed protein with inverse micellar solution, obtain forward extraction liquid, forward extraction liquid, through vacuum concentration, adds buffered soln after the organic solvent in removing inverse micellar solution, extracting and separating, obtains the aqueous solution of rape seed protein; Extract rape seed protein with inverse micellar solution, the method obtaining forward extraction liquid is as follows:
Rapeseed meal powder is added in inverse micellar solution, 40 DEG C of extraction 80min under frequency 30KHz, power 120KW ul-trasonic irradiation, then carry out centrifugation 20min with the relative centrifugal force of 3200 × g, obtain forward extraction liquid, described inverse micellar solution is by succinate sodium 2-ethylhexyl, octane-iso and contain K
+buffered soln mix, in described inverse micellar solution, the mass volume ratio of succinate sodium 2-ethylhexyl and octane-iso is 0.02g/mL, W
0be 33, buffered soln is the Sodium phosphate dibasic-sodium hydroxide buffer solution of KCl, and pH value is 11, K
+concentration is 0.15mol.L
-1, the rapeseed meal powder added be 0.02g/mL with the mass volume ratio of inverse micellar solution.
2. the technique of the extraction rape seed protein as described in claim l, is characterized in that, after forward extraction liquid adds buffered soln after vacuum concentration, also adds the aqueous ethanolic solution that volumetric concentration is 10%.
3. the technique extracting rape seed protein as claimed in claim 1 or 2, it is characterized in that, the method for described vacuum concentration is rotary evaporation, and bath temperature controls at 70 DEG C.
4. as the technique of the extraction rape seed protein of claim 3 spouse, it is characterized in that, the buffered soln added after rotary evaporation is the buffered soln of KCl.
5. the technique extracting rape seed protein as claimed in claim 4, is characterized in that, the pH value of buffer solution 7, K of the KCl added after rotary evaporation
+concentration is 1.Omol.L
-1.
6. the technique of extraction rape seed protein according to claim 5, is characterized in that, the buffered soln of the KCl added after rotary evaporation is Sodium phosphate dibasic one sodium dihydrogen phosphate buffer of KCl.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107751295A (en) * | 2017-11-14 | 2018-03-06 | 唐林元 | A kind of method that ship biscuit is prepared using rapeseed dregs |
| CN108314702A (en) * | 2018-04-24 | 2018-07-24 | 台州学院 | A kind of method of inverse micelle abstraction-impulse electric field combined separation purifying protein |
| CN109111497A (en) * | 2018-09-06 | 2019-01-01 | 武汉轻工大学 | A kind of method for producing of peony seeds albumen |
| CN112048012A (en) * | 2020-09-17 | 2020-12-08 | 南京财经大学 | Preparation method of rapeseed napin protein |
-
2014
- 2014-07-22 CN CN201410348962.6A patent/CN105294822A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107751295A (en) * | 2017-11-14 | 2018-03-06 | 唐林元 | A kind of method that ship biscuit is prepared using rapeseed dregs |
| CN108314702A (en) * | 2018-04-24 | 2018-07-24 | 台州学院 | A kind of method of inverse micelle abstraction-impulse electric field combined separation purifying protein |
| CN108314702B (en) * | 2018-04-24 | 2020-08-18 | 台州学院 | Method for separating and purifying protein by combining reverse micelle extraction and pulsed electric field |
| CN109111497A (en) * | 2018-09-06 | 2019-01-01 | 武汉轻工大学 | A kind of method for producing of peony seeds albumen |
| CN112048012A (en) * | 2020-09-17 | 2020-12-08 | 南京财经大学 | Preparation method of rapeseed napin protein |
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Application publication date: 20160203 |