CN101343309A - Method for simultaneously separating soy protein and oil fat with inverse micelle abstraction technique - Google Patents
Method for simultaneously separating soy protein and oil fat with inverse micelle abstraction technique Download PDFInfo
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Abstract
Disclosed is a method for separating soybean protein and fat by the reversed micellar extraction, which comprises the following procedures: (1) reversed micellar solution is prepared; (2)a pretreated raw material of soybean is added into the reversed micellar solution to extract protein and fat; (3) electrolyte solution is added so that the protein is transferred into the aqueous phase from the reversed micellar solution to be separated, thereby realizing back extraction of the protein, and protein aqueous solution and mixed oil are obtained; (4) the obtained protein product and the fat product are processed. The method has the following main advantages: the fat and the protein are separated at the same time in the reversed micellar extraction system, so the production process is greatly simplified, and the production cost and the investment on the fixed assets are reduced; the method has the advantages of solvent extraction; in the process of extraction, the protein is surrounded by the water environment in the reversed micellar, and the environment is close to the environment in the cells with mild condition, so the protein is not denatured and can keep the biological activity of the protein, and the purity of the product is high; and no acid and alkali wastewater is discharged without pollution.
Description
Technical field
The present invention relates to a kind of soybean protein and greasy isolation technique, particularly a kind of novel soybean albumen and greasy high efficient separation technology---utilize inverse micelle abstraction technique soy protein isolate and greasy method simultaneously.
Background technology
Soybean is to cultivate one of crop the most widely in the world, and large-area plantation is all arranged all over the world.According to statistics, 2007, world soybean output was 22,159 ten thousand tons; China's soybean yields is about 1,440 ten thousand tons, and import volume is expected to be 3,300 ten thousand tons, and aggregate consumption is expected to be 4,706 ten thousand tons; Along with developing rapidly of modern agriculture, the output of soybean also increasing year by year, estimates that world soybean output in 2008 is 22,162 ten thousand tons.This provides abundant raw material for soybean protein and greasy research with using; Also effectively promoted developing rapidly of soybean protein and oil prodution industry thus, this just requires soybean protein and grease can not be confined to be used at present foodstuffs industry, fodder industry, even the raw material dregs of beans (cake) that will produce soybean protein is wasted as fertilizer, therefore, press for new high efficient separation technology and extract soybean protein, keep it active and functional, expand new Application Areas, elongate the chain of deep processing of farm products, improve additional value of farm products and farmers' income, also meet the industry policy of country simultaneously.
The main component of soybean has protein, fat, carbohydrate etc., and wherein protein content is about 40%, and lipid content is about 18%, and carbohydrate content is about 25%.Protein is the basic substance that constitutes tissue, is one of human essential nutritive ingredient; The not only amino acid contained kind of soybean protein is more complete, and the ratio of contained essential amino acid is higher, is a kind of desirable nutrient excellent product that can replace animal proteinum few in number; Because soybean protein has economy, especially have functional performances such as emulsifying property, tackiness, gel formation, foaminess simultaneously, in recent years, it has become the indispensable raw material of foodstuffs industry.The grease that is rich in the soybean is one of indispensable important nutritive ingredient in the food, and at first its major function just provides the heat of needed by human body; Secondly grease also provides lipid acid and the numerous food product local flavor and the making function of needed by human such as linoleic acid plus linolenic acid; In addition, grease also has very important purposes in industry such as light industry, chemical industry.Therefore, soybean protein and oil resource furtherd investigate and made full use of and important economy and social effect had.
At present, soyabean processing is the center based on grease generally both at home and abroad, under the condition that as far as possible satisfies the oil prodution industry requirement, just considers the utilization of protein resource that is:, and the grease in the raw material is extracted more than 99%, few waste; But 20% of not enough its Tot Prot of soybean protein that the present food-processing of China utilizes, and the main approach that utilizes of protein resource is to be used as feed, even as fertilizer; On the one hand, food protein is supplied with wretched insufficiency, on the other hand, causes the huge waste of protein resource.Basic reason is exactly to lack advanced soybean protein separation technology.At present, albumen and grease etc. are (as shown in Figure 1) of adopting proceed step by step in the separation soybean both at home and abroad, that is: at first adopting once leachings etc. technology is first oil extraction in the soybean is come out, is that raw material carries out proteic separation then with the defatted soybean meal, removes fiber etc.Because residual in the dregs of rice have a large amount of solvents, needs cause the protein denaturation in the dregs of rice thus through the high temperature precipitation, make the dregs of rice behind the precipitation mainly as feed, even as fertilizer sources, cause the very big waste of albumen, thereby reduce proteic use value and utilization ratio.Leach and low temperature desolventizing technology even adopt, protein denaturation is little in the dregs of rice, can further be used for producing protein concentrate or protein isolate, yet proteic separation is to carry out after system oil again, and promptly be proceed step by step separating of soybean oil, albumen and fiber etc.On the other hand, at present both at home and abroad soy protein isolate mainly adopts the traditional technology of alkali extraction and acid precipitation, and not only technology, equipment complexity, investment are big, and still have the Partial Protein sex change, thereby influence proteic use value; Alkali extraction and acid precipitation technology need be used a large amount of bronsted lowry acids and bases bronsted lowries simultaneously, and discharges the waste water (as shown in Figure 1) of nutritive substances such as containing sugar in a large number and soda acid, thus serious environment pollution.And this method requires raw material dregs of beans quality than uniformity, so that technological operation.China's soy material is wide in variety, mix purchase, causes control difficulty in the production, and unstable product quality and acid and alkali consumption amount are big, the cost height, isolated whey liquid does not reclaim with discharge of wastewater, and wherein low molecular weight protein is wasted, and soluble components is removed also not thorough.Therefore, along with the fast development of soybean protein and oil prodution industry, research is easy to industrialized high efficiency separation new technology has become the task of top priority.
The research worker explores new soybean protein separation technology constantly both at home and abroad.Inverse micelle abstraction be exactly a kind of new and effective isolation technique that under this background, grows up " Li Xiang village; He Gaohong; the new development [J] of inverse micelle abstraction protein technology. the water treatment skill is stated; 2005.31 (1) 7-12. ", for plant (soybean) albumen and greasy the separation simultaneously having opened up a new way with applications well prospect.
The eighties in 20th century, there is document to introduce the reverse micelle technology at home, and the work that conducts a research, Shi Hongqin and Lei Xia " Shi Hongqin; Lei Xia; the research [J] of Shen Zhong credit .AOT-phosphatide reverse micelle system extracting protein matter. chemical industry journal, 1990 (6): 702-708 " wait the people to study aspect the inverse micelle abstraction separation enzyme utilizing.Lu Qiang and Li Kuanhong " Lu Qiang; Li Kuanhong; Shi Yajun. extract new technology---inverse micelle abstraction [J] with isolated protein. modern chemical industry; 1993; (12): 37-39 " etc. the people with CTAB/ n-hexyl alcohol/octane inverse micelle abstraction separation system, the kinetics of research extracting and separating bovine serum albumin from the aqueous solution has been measured the apparent mass transfer coefficient under the multiple condition, thereby judges extraction and back extraction process controlled step separately.After this, the domestic report that begins to have inverse micelle abstraction amino acid separation etc., but reported mostly be some micromolecular albumen or enzymes, concentrate on cytopigment-C, bovine hemoglobin, serum albumin, α-Dian Fenmei and trypsinase etc.Gong Fuzhong etc. introduce two kinds of interfacial mass transfers of inverse micelle abstraction, promptly with the relevant mass transfer of dissolving with based on the mass transfer of liquid film.
These researchs all fully show, inverse micelle abstraction technique is as a new separation technology, having an enormous advantage aspect the separation purification biologically active substance, along with going deep into of research, inverse micelle abstraction technique very likely becomes the important method of biologically active substance separation, purification and is applied to industrial production.Simultaneously, inverse micelle abstraction technique separates, purification of protein institute obtains achievement and progress are indicating that also inverse micelle abstraction technique is applied to soybean protein and greasy IP prospecting is very wide, press for us and continue research in depth.
Summary of the invention
Purpose of the present invention just is being based on above-mentioned prior art and a kind of inverse micelle abstraction technique simultaneously soy protein isolate and greasy method utilized of providing specially.
The objective of the invention is to be achieved through the following technical solutions: inverse micelle abstraction technique while soy protein isolate and the greasy method utilized of the present invention may further comprise the steps:
(1) configuration inverse micellar solution;
(2) pretreated soy material is added in the inverse micellar solution, realize extraction albumen and grease wherein simultaneously;
(3) add electrolyte solution, isolate protein, realize proteic reextraction, obtain protein solution and mixing oil thereby make protein transfer to aqueous phase from reverse micelle;
(4) protein solution that separation is obtained and mixing oil are handled and are obtained protein product and oil and fat product.
In the present invention, inverse micellar solution is made up of succinate sodium 2-ethylhexyl (AOT)/octane-iso/damping fluid, wherein: the AOT addition is 60g/L～140g/L, and the damping fluid addition is 0.01L/L～0.10L/L (mol ratio W of control reverse micelle system water and tensio-active agent
04～21), regulate pH6.0～9.0; Damping fluid in the described inverse micellar solution can be sylvite, sodium salt and GaCl
2, MgCl
2Deng phosphate buffered saline buffer.
Electrolyte solution can be KCl-phosphate buffered saline buffer, MgCl
2-phosphate buffered saline buffer, NaCl-phosphate buffered saline buffer, GaCl
2-phosphate buffered saline buffer, NaNO
3-phosphate buffered saline buffer, KNO
3-phosphate buffered saline buffer, Na
2SO
4In-the phosphate buffered saline buffer any one, as: KCl-phosphate buffered saline buffer, MgCl
2-phosphate buffered saline buffer, NaCl-phosphate buffered saline buffer, GaCl
2-phosphate buffered saline buffer, NaNO
3-phosphate buffered saline buffer, KNO
3-phosphate buffered saline buffer, Na
2SO
4-phosphate buffered saline buffer etc.
In strip stage, the electrolyte solution amount and the inverse micellar solution equal-volume that add, described electrolyte solution can further be chosen as KCl-phosphate buffer soln or NaCl-phosphate buffer soln, wherein KCl in the damping fluid or NaCl concentration are 0.5mol/L～1.5mol/L, pH6.0～9.0,20 ℃～35 ℃ of temperature, time 1h～1.5h.
Pretreated soy material of the present invention refers to full fat soybean powder; The ratio that soy material adds in the inverse micellar solution is to add 10-60 gram soy material in every liter of inverse micellar solution.
25 ℃～60 ℃ of extraction temperature in the step of the present invention (2), extraction time 1h～2h.
Also can add the Alcalase Sumizyme MP in the inverse micelle abstraction process, adding proportion is 2.0% of a soy material quality, its objective is the characteristic of utilizing enzyme to have " superactivity " at reverse micelle system, and extraction process can carry out proteic enzyme modification simultaneously.
The present invention adopts the reverse micelle isolation technique as soybean protein and isolating technology of grease while, its major advantage:
(1) realizes that in the inverse micelle abstraction system grease separates simultaneously with proteinic, simplified soybean protein and grease production technological process widely, reduce production cost and investment in fixed assets;
(2) has solvent-extracted advantage;
(3) in extraction process, albumen is surrounded by water surrounding in the reverse micelle, and environment approaches intracellular environment, mild condition, and albumen is volatility not, thereby keeps its biological activity, the product purity height;
(4) extraction process does not need to add acid or alkali, and used tensio-active agent and solvent can recycle, and solves the difficult problem of a large amount of acidic and alkaline waste water contaminate environment of discharging in the conventional production methods;
(5) utilize enzyme to have the characteristic of " superactivity " at reverse micelle system, extraction process can carry out advantages such as proteic enzyme modification simultaneously.
Description of drawings
Fig. 1 is traditional soybean grease and protein production technical process.
Fig. 2 is inverse micelle abstraction soybean oil of the present invention and protein technical process.
Embodiment
The present invention is further described below in conjunction with embodiment (accompanying drawing), originally is not restriction the present invention:
The solution of the present invention is to use reverse micelle system that grease in the soybean is separated simultaneously with albumen.
At first prepare inverse micellar solution: reverse micelle system be tensio-active agent in non-polar organic solvent, concentration forms when surpassing threshold concentration has thermodynamically stable aggregate system.Used reverse micelle system is succinate sodium 2-ethylhexyl (AOT)/octane-iso/KCl-phosphoric acid salt reverse micelle system.Take by weighing a certain amount of AOT, add the octane-iso of respective volume according to concentration requirement.Magnetic agitation is dissolved AOT fully, treat that solution is transparent after, add an amount of KCl-phosphate buffer soln, the shaking table jolting, leave standstill under the room temperature on 12h or the whizzer with 3000r/min and separate 20min, solution if transparent be reverse micelle then, otherwise then be not (the bottom less water to the system influence not quite).
The AOT addition is 60g/L～140g/L, and KCl-phosphate buffer soln addition is 0.01L/L～0.10L/L (mol ratio W of control reverse micelle system water and tensio-active agent
04～21), regulate pH6.0～9.0, KCl concentration is 0.00mol/L～0.35mol/L.
During the preparation inverse micellar solution, the KCl-phosphate buffered saline buffer in the reverse micelle system can be by MgCl
2-phosphate buffered saline buffer, NaCl-phosphate buffered saline buffer etc. substitute, the MgCl that adds during the collection of when just preparing and back
2The pH of-phosphate buffered saline buffer, NaCl-phosphate buffered saline buffer, MgCl
2Or difference such as NaCl concentration.
Separating step is with reference to shown in Figure 2, and the committed step processing parameter is as follows:
The inverse micelle abstraction step: the raw material add-on is 10g/L～60g/L, 25 ℃～60 ℃ of extraction temperature, extraction time 1h～2h.
Strip stage: add isopyknic electrolyte solution KCl-phosphate buffer soln or NaCl-phosphate buffer soln, KCl in the damping fluid or NaCl concentration are 0.5mol/L～1.5mol/L, pH6.0～9.0,20 ℃～35 ℃ of temperature, time 1h～1.5h.
Embodiment 1
80g is joined in the AOT inverse micellar solution of 4L 80g/L through pretreated full fat soybean powder, control condition, the damping fluid in the inverse micellar solution is NaNO
3-phosphate buffered saline buffer, control condition W
09, NaNO
3Concentration 0.05mol/L regulates pH 8.0, and 45 ℃ of extraction temperature are with the speed mechanical stirring 2h of 180r/min.Albumen forward extraction rate 60.0%.
Add equal-volume 1.0mol/L NaCl-phosphate buffered saline buffer, regulate pH value 7.7, with centrifugal 0.5h behind the velocity fluctuation 1.5h of 300r/min, static layering obtains the upper strata mixing oil, lower floor's protein solution.Collection rate 87.1% behind the albumen.Oil extraction rate 90.0%.
Generally, the protein extracting rate is 52.3%, and the oil extraction rate is 90.0%.The soybean protein that the foaminess of soybean protein, emulsifying property and emulsifying stability are produced than alkali extraction and acid precipitation method is good.
Embodiment 2
It is in the inverse micellar solution of 120g/L that 0.3g is joined 20mL AOT concentration through pretreated full fat soybean powder, and the damping fluid in the inverse micellar solution is the KCl-phosphate buffered saline buffer, control condition W
018, KCl concentration 0.05mol/L regulates pH7.5, and 40 ℃ of extraction temperature are with the velocity fluctuation 1h of 180r/min.Albumen forward extraction rate 64.6%.
Add equal-volume 1.0mol/L KCl-phosphate buffered saline buffer, regulate pH value 7.0, with centrifugal 15min behind the velocity fluctuation 2h more than the 250r/min, static layering obtains the upper strata mixing oil, lower floor's protein solution in vibrator.Collection rate 89.0% behind the albumen.Oil extraction rate 86.0%.Generally, the protein extracting rate is 57.5%, and the oil extraction rate is 86.0%.
Embodiment 3
It is in the inverse micellar solution of 80g/L that 15g is joined 1L AOT concentration through pretreated full fat soybean powder, adds the 2.0%Alcalase Sumizyme MP simultaneously, control condition W
020, KCl concentration 0.10mol/L regulates pH 7.0,45 ℃ of extraction temperature, extraction 2h.Albumen forward extraction rate is 73.8%.
Add equal-volume 0.8mol/L KCl-phosphate buffered saline buffer, regulate pH value 8.0, with centrifugal 20min behind the velocity fluctuation 1.5h more than the 250r/min, static layering obtains the upper strata mixing oil, lower floor's protein solution in vibrator.Collection rate 82.8% behind the albumen.Oil extraction rate 87.5%.
Generally, the protein extracting rate is 61.1%, and the oil extraction rate is 86.0%.Add the extraction of Alcalase Sumizyme MP, protein hydrolysis degree is about 4%, and extraction yield increases.
Embodiment 4
It is in the inverse micellar solution of 60g/L that 1g is joined 100mL AOT concentration through pretreated full fat soybean powder, and the damping fluid in the inverse micellar solution is MgCl
2-phosphate buffered saline buffer, control condition W0 12, MgCl
2Concentration 0.15mol/L regulates pH6.5, and 40 ℃ of extraction temperature are with the velocity fluctuation 1.5h of 180r/min.Albumen forward extraction rate 61.4%.
Add equal-volume 1.5mol/L KCl-phosphate buffered saline buffer, regulate pH value 8.0, with centrifugal 15min behind the velocity fluctuation 3h more than the 250r/min, static layering obtains the upper strata mixing oil, lower floor's protein solution in vibrator.Collection rate 83.1% behind the albumen.Oil extraction rate 79.8%.
Generally, the protein extracting rate is 51.0%, and the oil extraction rate is 79.8%.
Claims (10)
1, a kind of inverse micelle abstraction technique soy protein isolate and greasy method simultaneously utilized is characterized in that: may further comprise the steps:
(1) configuration inverse micellar solution;
(2) pretreated soy material is added in the inverse micellar solution, realize extraction albumen and grease wherein simultaneously;
(3) add electrolyte solution, isolate protein, realize proteic reextraction, obtain protein solution and mixing oil thereby make protein transfer to aqueous phase from reverse micelle;
(4) protein solution that separation is obtained and mixing oil are handled and are obtained protein product and oil and fat product.
2, while soy protein isolate according to claim 1 and greasy method, it is characterized in that: inverse micellar solution is made up of succinate sodium 2-ethylhexyl (AOT)/octane-iso/damping fluid, wherein: the AOT addition is 60g/L～140g/L, and the damping fluid addition is 0.01L/L～0.10L/L (mol ratio W of control reverse micelle system water and tensio-active agent
04～21), regulate pH6.0～9.0.
3, while soy protein isolate according to claim 2 and greasy method is characterized in that: the damping fluid in the described inverse micellar solution can be sylvite, sodium salt and GaCl
2, MgCl
2Deng phosphate buffered saline buffer.
4, while soy protein isolate according to claim 1 and greasy method is characterized in that: the electrolyte solution that is added in step (3) can be KC1-phosphate buffered saline buffer, MgCl
2-phosphate buffered saline buffer, NaCl-phosphate buffered saline buffer, GaCl
2-phosphate buffered saline buffer, NaNO
3-phosphate buffered saline buffer, KNO
3-phosphate buffered saline buffer, Na
2SO
4In-the phosphate buffered saline buffer any one.
5, according to claim 1 or 4 described while soy protein isolate and greasy methods, it is characterized in that: in the reextraction process in step (3), the electrolyte solution amount of adding and inverse micellar solution equal-volume.
6, according to claim 1 or 4 described while soy protein isolate and greasy methods, it is characterized in that: at electrolyte solution described in the step (3) is KC1-phosphate buffer soln or NaC1-phosphate buffer soln, wherein KCl in the damping fluid or NaCl concentration are 0.5mol/L～1.5mol/L, pH6.0～9.0,20 ℃～35 ℃ of temperature, time 1h～1.5h.
7, while soy protein isolate according to claim 1 and 2 and greasy method is characterized in that: the ratio that soy material adds in the inverse micellar solution is to add 10-60 gram soy material in every liter of inverse micellar solution.
8, while soy protein isolate according to claim 1 and greasy method is characterized in that: 25 ℃～60 ℃ of extraction temperature, extraction time 1h～2h.
9, while soy protein isolate according to claim 1 and greasy method is characterized in that: be added with the Alcalase Sumizyme MP in the inverse micelle abstraction process, adding proportion is 2.0% of a soy material quality.
10, while soy protein isolate according to claim 1 and greasy method, it is characterized in that: pretreated soy material refers to full fat soybean powder.
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| CN101912030A (en) * | 2010-09-26 | 2010-12-15 | 河南工业大学 | Method for extracting soybean protein by using dodecyltrimethylammonium chloride reverse micellar solution |
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| CN101619099B (en) * | 2009-08-17 | 2012-07-11 | 南京财经大学 | Method for extracting rapeseed protein by reverse micelle |
| CN102599326A (en) * | 2012-03-23 | 2012-07-25 | 山东省农业科学院农产品研究所 | Backward extraction method for reversed micellar extraction of soybean protein |
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| CN101619098A (en) * | 2009-08-17 | 2010-01-06 | 南京财经大学 | Backward extraction method for extracting rapeseed protein by reverse micelle |
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| CN102533688B (en) * | 2012-03-07 | 2014-07-02 | 湖南大学 | Method for extracting and purifying lignin peroxidase by using reverse micelles |
| CN102533688A (en) * | 2012-03-07 | 2012-07-04 | 湖南大学 | Method for extracting and purifying lignin peroxidase by using reverse micelles |
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| CN103988975A (en) * | 2014-05-23 | 2014-08-20 | 东北农业大学 | Method for extracting protein from soybean meal by adopting ultrasonic-assisted reverse micelle extraction process |
| CN104004809A (en) * | 2014-05-23 | 2014-08-27 | 东北农业大学 | Method for preparing soybean oil and protein by using reverse micelle aqueous enzymatic method |
| CN104004811A (en) * | 2014-05-23 | 2014-08-27 | 东北农业大学 | Method for extracting soybean oil and proteins |
| CN105175485A (en) * | 2015-09-23 | 2015-12-23 | 江西省科学院应用化学研究所 | Method for synchronously extracting protein and oil from taxus chinenesis seeds |
| CN105175485B (en) * | 2015-09-23 | 2019-01-15 | 江西省科学院应用化学研究所 | A method of it is synchronous from Chinese yew seed to extract protein and grease |
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