CN101317623B - Preparation method for oil-in-water type soybean isolated protein - Google Patents
Preparation method for oil-in-water type soybean isolated protein Download PDFInfo
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Abstract
The invention discloses a preparation method for emulsion-typed soybean protein isolate, which comprises the steps as follows: pre-crushed low-temperature defatted soybean flour (DSF) is taken as raw material and mixed with deionized water with the water/material ratio of 5-20w/w, is heated to 40-60 DEG C under alkali condition and leached for 20-30min so as to lead the soybean meal to expand by absorbing water; Acalase alkaline protease is added to carry out the enzymolysis; the bottom residue is removed by centrifuging so as to obtain soybean proteolysis supernatant; furthermore, the soybean proteolysis supernatant is homogenized and acidity is adjusted to PH 4.5; the protein in the soybean proteolysis supernatant is deposited and centrifuged for 10-20min; the obtained deposition is mixed with the deionized water by the water/material ratio of 3-5w/w; the mixed solution is adjusted to neutral by adding NaOH and frozen dryly or sprayed dryly so as to obtain the emulsion-typed soybean protein isolate. The process of the invention is simple and reliable, which remarkably improves the emulsion performance of the soybean protein.
Description
Technical field
The present invention relates to the processing technique field of soybean protein, be specifically related to a kind of preparation method of oil-in-water type soybean isolated protein.
Background technology
There is abundant soybean protein resource (China's dregs of beans output reached 2,600 ten thousand tons in 2005) in China, and soybean protein is worth because of its better nutritivity and unique health care is favored.Yet; The soybean protein isolate product of China is essentially high gelling; Only for being applied to rely on import basically and be directed to the oil-in-water type functional soy albumen that contains the grease liquid beverage in the bowel lavage series products, this has greatly limited at soybean protein in Application in Food Industry.
Usually, commercial soy protein isolates be the defatted soybean meal powder through alkali carry, acid is heavy, through neutralization, sterilization, spray-drying makes then, the emulsibility difference is that the restriction soybean protein is containing the main cause that the grease liquid system is used.To the technological deficiency of soybean protein emulsibility difference, both at home and abroad the researcher has done more trial, wherein is treated to the master with protein-modified.The method of modifying of common protein comprises three types of physical modification, chemical modification and enzyme modifications.Chemical modification has reacts simply, is widely used and the obvious results characteristics, many such as byproduct of reaction but chemical modification exists, and is difficult in the final products remove, and nutritive value can receive shortcomings such as influence to a certain degree in course of reaction.Though the physical method modification is safe, modified effect is not remarkable, and a lot of physical modification technology is difficult to realize industrialization.Enzyme modification has safety, reliably reaches advantages such as efficient, is to improve protein function characteristic and a kind of effective method that increases its range of application.In order to get a desired effect, often be used between each method.From the angle of food security, the combination modification of physical modification and enzyme modification is more favored.From the domestic and international documents and materials of report in the past, physical modification often be arranged in enzymolysis modified before, to improve the enzymolysis sensitiveness of soybean protein.Yet because the globulin structure of soybean protein is compact, above-mentioned method of modifying is unsatisfactory.In addition, to improvement soybean protein emulsibility enzymolysis processing, the enzyme that goes out of heating immediately after general enzymolysis is accomplished, the protein thermal denaturation that heating is caused will further influence the functional characteristic of soybean protein.
Summary of the invention
To the objective of the invention is the above-mentioned deficiency that exists in the present prior art in order solving, a kind of preparation method of oil-in-water type soybean isolated protein to be provided.The present invention has significantly improved the emulsifiability of soybean protein isolate, and the average grain diameter of emulsion obviously reduces, and 60 days normal temperature leaves standstill the observation emulsion system to be stablized.
The present invention realizes through following technical scheme.
A kind of preparation method of oil-in-water type soybean isolated protein comprises the steps:
(1) the defatted soybean meal powder with precomminution is a raw material, presses ratio of water to material 5～20w/w and mixes with deionized water; Under alkali condition, being heated to temperature is 40～60 ℃, and lixiviate 20～30min makes the dregs of beans imbibition, adds the Acalase alkali protease and carries out enzymolysis, gets hydrolysate of soybean protein liquid;
(2) with 20-30 ℃ of hydrolysate of soybean protein liquid, the centrifugal 10-20min of 4000-6000g removes bottom residues, gets the hydrolysate of soybean protein supernatant, and the hydrolysate of soybean protein supernatant is carried out homogenization processing;
(3) with step (2) obtain through the hydrolysate of soybean protein supernatant acid adjustment of homogeneous to pH4.5; At 20～30 ℃; Albumen in the centrifugal 10～20min deposition of the 4000-6000g hydrolysate of soybean protein supernatant, gained deposition are pressed ratio of water to material 3～5w/w and are mixed with deionized water, after hydro-oxidation sodium transfers to neutrality; Freeze drying or spray-drying get oil-in-water type soybean isolated protein.
Among the above-mentioned preparation method, the pH value of the said alkali condition of step (1) is 7.5～9.5.
Among the above-mentioned preparation method, under said temperature, add the Acalase alkali protease in the step (1), behind enzymolysis to the degree of hydrolysis 0.4%～5%, enzymolysis liquid is cooled to 20 ℃-30 ℃.
Among the above-mentioned preparation method, the said homogenization processing of step (2) adopts high pressure homogenizer to carrying out homogeneous, and homogenization pressure is 20～30MPa, and the homogeneous number of times is 1～2 time.
The present invention compared with prior art has following advantage and beneficial effect:
1, the present invention's interpolation alkali protease on the technology of normal temperature alkali condition extraction albumen carries out enzymolysis modified; Omitted the enzyme process of going out that heats; Utilization rationally is high-pressure homogeneous subsequently to make the protease inactivation with dry run, has effectively avoided heating the protein transition sex change that enzyme caused of going out.
2, the control enzymolysis process that the present invention adopted can promote the stripping of albumen in the dregs of beans on the one hand, improves the protein recovery of dregs of beans, can improve the emulsification function characteristic of soybean protein on the other hand.
3, the present invention only with water as medium, to add chemical reagent few, safe, help the reduction of suitability for industrialized production and production cost.
4 compare with the sample that non-modified is handled; The emulsifiability of the soybean protein isolate product of modification of the present invention is particularly outstanding; Can form the littler emulsion of particle diameter with oil; Emulsifying activity and emulsion stability significantly improve, and corresponding emulsion is the system stable uniform in longer standing time, is not separated.
The specific embodiment
Be described further below in conjunction with the embodiment specific embodiments of the invention.
The coherent detection evaluation method that relates to effect of the present invention:
1, Kai Shiding nitrogen: an amount of protein sample adds the concentrated sulfuric acid and catalyst digests on digesting, on azotometer, absorbs with the boric acid that adds indicator subsequently, uses the watery hydrochloric acid titration again.Adopt Kai Shiding nitrogen determination protein utilization rate, protein recovery %=(centrifugal back albumen supernatant protein content/dregs of beans protein content) * 100%
2, size distribution: soybean protein isolate 4g, soybean oil 40g, deionized water 156g, 50 ℃ of aquation 30min, twice preparation of 30MPa homogeneous emulsion.Take Ma Erwen 2000 particles distribution instruments to carry out granulometry, the emulsion sample is pressed mass ratio usefulness deionized water dilution in 1: 1000, and analytical model is a common-mode.
3, emulsibility is measured: get 15mL concentration and be 0.1% protein solution (being dissolved in the phosphate buffer of 0.2mol/LpH7.0), add the pure soybean oil of 5mL, under 10000rpm, room temperature, stir 1min, respectively after stirring 0,10min takes a sample from the bottom.With 100 times of 0.1% (W/V) SDS dilutions, measure the light absorption value A at 500nm place
500, be blank with SDS solution.Light absorption value with zero moment is represented emulsibility (EAI), or representes with emulsification vitality index EAI:
EAI＝2×(2.303A
500)×N×10
-4/φLC
In the formula: EAI is the emulsification area (m of every gram protein
2/ g); N is an extension rate; φ is the shared mark of oil phase in the system, and oil phase accounts for 0.25 in this test; C is the concentration (g/mL) of protein; L is colorimetric pool optical path (1cm).
Emulsion stability (ES) is represented with stable emulsifying index (ESI):
ESI＝A
0×ΔT/ΔA＝A
0×ΔT/(A
0-A
t)
In the formula: A
0, the light absorption value in 0 moment; A
t, the light absorption value of t during the moment; Δ T, the time difference; Extinction value difference in the Δ A, Δ T.
4, leave standstill observation: above-mentioned emulsion adds 2/10000ths Sodium azide solution, and mixing is got the 10ml sample and is statically placed in the 10ml scale test tube, observes phase separation, and breast is analysed rate %=(stillness of night height/10) * 100%
Embodiment one
Degreasing low temperature soy meal powder 100kg mixes with deionization 1500kg water, adjustment pH value 8.0 left and right sides lixiviate 30min, and centrifugation is removed residue and is obtained the albumen leaching liquor; Albumen leaching liquor acid adjustment deposition, the centrifugation deposition, deposition adds water neutralisation, spray-drying or freeze drying.Protein utilization rate is 80.12%; The emulsion average grain diameter is 4.312 μ m; The numerical value of EAI and ESI is respectively 27.82 and 12.69; Left standstill in 60 days and observe breast to analyse rate be 4.8%.
Embodiment two
Degreasing low temperature soy meal powder 100kg mixes with deionization 1500kg water, adjustment pH value 8.0 left and right sides lixiviate 30min, and centrifugation is removed residue and is obtained the albumen leaching liquor; The albumen leaching liquor carries out high-pressure homogeneous processing, homogenization pressure 25MPa, and the homogeneous number of times is for once; Albumen leaching liquor acid adjustment deposition after the homogenization processing, the centrifugation deposition, deposition adds water neutralisation, spray-drying or freeze drying.Protein utilization rate is 80.5%; The emulsion average grain diameter is 4.297 μ m; The numerical value of EAI and ESI is respectively 33.45 and 12.46; Left standstill in 60 days and observe breast to analyse rate be 4.6%.
Embodiment three
Degreasing low temperature soy meal powder 100kg mixes with deionization 500kg water, is heated to 50 ℃, adjustment pH value 7.5 lixiviate 20min; Add the Acalase alkali protease; Behind enzymolysis to the degree of hydrolysis 1%, be cooled to 20 ℃, the centrifugal 10min separation of 6000g is removed residue and is got protein enzymatic hydrolyzate; With protein enzymatic hydrolyzate 25MPa homogeneous number of times 2 times; Adjustment albumen leaching liquor pH value to 4.5,30 ℃, the centrifugal 20min precipitation separation of 4000g; Gained deposition press ratio of water to material 1: 5w/w and is mixed with deionized water, and after hydro-oxidation sodium transferred to neutrality, freeze drying or spray-drying must oil-in-water type soybean isolated proteins.The protein utilization rate of dregs of beans is 84.32%; The emulsion average grain diameter is 3.354 μ m; The numerical value of EAI and ESI is respectively 35.12 and 17.35; Left standstill in 60 days and observe breast to analyse rate be 0.
Embodiment four
Degreasing low temperature soy meal powder 100kg mixes with deionization 1500kg water, is heated to 40 ℃, and adjustment pH value 8.5 lixiviate 25min add the Acalase alkali protease, are cooled to 25 ℃ behind enzymolysis to the degree of hydrolysis 0.4%; The centrifugal 15min of 5000g removes residue and gets protein enzymatic hydrolyzate; With protein enzymatic hydrolyzate 25MPa homogeneous number of times 1 time; Adjustment albumen leaching liquor pH value to 4.5,30 ℃, the centrifugal 20min precipitation separation of 4000g; The gained deposition is pressed ratio of water to material 1: 4w/w mix with deionized water, after hydro-oxidation sodium transferred to neutrality, freeze drying or spray-drying got oil-in-water type soybean isolated protein.The protein utilization rate of dregs of beans is 84.32%; The emulsion average grain diameter is 3.597 μ m; The numerical value of EAI and ESI is respectively 36.54 and 18.12; Left standstill in 60 days and observe breast to analyse rate be 0.
Embodiment five
Degreasing low temperature soy meal powder 100kg mixes with deionized water 2000kg, is heated to 60 ℃, and adjustment pH value 9.0 lixiviate 30min add the Acalase alkali protease, and enzymolysis to degree of hydrolysis 3.5% is cooled to 30 ℃; The centrifugal 20min of 4000g removes residue and gets protein enzymatic hydrolyzate; After protein enzymatic hydrolyzate 30MPa homogeneous number of times is 2 times; Adjustment albumen leaching liquor pH value to 4.5,30 ℃, the centrifugal 20min precipitation separation of 4000g; The gained deposition is pressed ratio of water to material 1: 3w/w mix with deionized water, after hydro-oxidation sodium transferred to neutrality, freeze drying or spray-drying got oil-in-water type soybean isolated protein.The protein utilization rate of dregs of beans is 83.92%; The emulsion average grain diameter is 3.617 μ m; The numerical value of EAI and ESI is respectively 35.72 and 17.48; Left standstill in 60 days and observe breast to analyse rate be 0.
Embodiment six
Degreasing low temperature soy meal powder 100kg mixes with deionized water 1000kg, is heated to 55 ℃, and adjustment pH value 8.0 lixiviate 30min add the Acalase alkali protease, and enzymolysis to degree of hydrolysis 5% is cooled to 20 ℃; The centrifugal 17min of 4000g removes residue and gets protein enzymatic hydrolyzate; After protein enzymatic hydrolyzate 26MPa homogeneous number of times is 1 time; Adjustment albumen leaching liquor pH value to 4.5,30 ℃, the centrifugal 20min precipitation separation of 4000g; The gained deposition is pressed ratio of water to material 1: 3w/w mix with deionized water, after hydro-oxidation sodium transferred to neutrality, freeze drying or spray-drying got oil-in-water type soybean isolated protein.The protein utilization rate of dregs of beans is 84.32%; The emulsion average grain diameter is 3.517 μ m; The numerical value of EAI and ESI is respectively 35.85 and 18.01; Left standstill in 60 days and observe breast to analyse rate be 0.
Show that to sum up the correlation technique of homogeneous is to improve protein utilization rate and the effective means of improving the Emulsifying Properties of Soy Protein Isolate ability behind the first enzymolysis; Simple homogenization processing DeGrain; The relevant treatment of homogeneous behind elder generation's enzymolysis can improve protein utilization rate on the one hand, can form the littler emulsion of average grain diameter on the other hand, and emulsifying activity and emulsion stability significantly improve, and leaves standstill the observation system in 60 days to stablize.
Claims (4)
1. the preparation method of an oil-in-water type soybean isolated protein is characterized in that comprising the steps:
(1) the defatted soybean meal powder with precomminution is a raw material, presses ratio of water to material 5～20w/w and mixes with deionized water; Under alkali condition, being heated to temperature is 40～60 ℃, and lixiviate 20min～30min makes the dregs of beans imbibition, adds the Acalase alkali protease and carries out enzymolysis, gets hydrolysate of soybean protein liquid; The pH value of said alkali condition is 7.5～9.5, under said temperature, adds the Acalase alkali protease, behind enzymolysis to the degree of hydrolysis 0.4%～5%, enzymolysis liquid is cooled to 20～30 ℃;
(2) with 20～30 ℃ of hydrolysate of soybean protein liquid, the centrifugal 10～20min of 4000～6000g removes bottom residues, gets the hydrolysate of soybean protein supernatant, and the hydrolysate of soybean protein supernatant is carried out homogenization processing; Said homogenization processing adopts high pressure homogenizer that the hydrolysate of soybean protein supernatant is carried out homogeneous, and homogenization pressure is 20～30MPa, and the homogeneous number of times is 1～2 time;
(3) with step (2) obtain through the hydrolysate of soybean protein supernatant acid adjustment of homogeneous to pH4.5; At 20～30 ℃; Albumen in the centrifugal 10～20min deposition of the 4000-6000g hydrolysate of soybean protein supernatant, gained deposition are pressed ratio of water to material 3～5w/w and are mixed with deionized water, after hydro-oxidation sodium transfers to neutrality; Freeze drying or spray-drying get oil-in-water type soybean isolated protein.
2. preparation method according to claim 1 is characterized in that the said ratio of water to material of step (1) is 10～15w/w, and said temperature is 45～55 ℃, and the pH value of said alkali condition is 7.5～9.5.
3. preparation method according to claim 2 is characterized in that said pH value is 7.8～8.2.
4. preparation method according to claim 3 is characterized in that said homogenization pressure is 25MPa, and the homogeneous number of times is 1 time.
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