CN101366469A - Preparation method for high-efficiency soy protein frothing agent - Google Patents
Preparation method for high-efficiency soy protein frothing agent Download PDFInfo
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- CN101366469A CN101366469A CNA2008101959886A CN200810195988A CN101366469A CN 101366469 A CN101366469 A CN 101366469A CN A2008101959886 A CNA2008101959886 A CN A2008101959886A CN 200810195988 A CN200810195988 A CN 200810195988A CN 101366469 A CN101366469 A CN 101366469A
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Abstract
The invention discloses a method for preparing a high-efficiency soybean protein foaming agent, which belongs to the biotechnical field of food. Soybean protein isolate is taken as a raw material, Alcalase is adopted and hydrolyzed first, then protein hydrolysate is ultrafiltered to obtain a protein peptide fragment with the molecular weight of more than 10kDa, the protein peptide fragment is recombined through the crosslinking of TGase, and a product after the crosslinking is condensed and freeze-dried to obtain the soybean protein foaming agent. Compared with untreated soybean protein isolate, the foaming ability and the foam stability of a product prepared by the method are greatly improved, the foaming ability of the product is improved by between 250 and 350 percent, the foam stability is improved by between 50 and 60 percent, and the foaming ability and the foam stability of the product are equivalent to those of egg white protein. The invention prepares the high-efficiency protein foaming agent by modifying the soybean protein isolate by an enzyme method, can forcefully promote the broad application of soybean protein in modern food processing, greatly improve the use value of the soybean protein, and has good economic benefit.
Description
Technical field
The invention provides a kind of preparation method of high-efficiency soy protein frothing agent, relate to the transformation of soybean protein structure and functional character, belong to technical field of food biotechnology.
Background technology
Soybean protein is the human important source of nutrition and the raw material of industry, aboundresources, best in quality, cheap, do not contain cholesterol, have higher nutritive value, and soybean protein has preferably and relevant functional characteristic such as foaming characteristic, emulsibility and gellings such as the hobby of food, processability.But the protein of natural premium properties such as various functional characters and animal sources albumen such as casein, parapeptone, albumen is compared, and sizable gap is still arranged, and causes its application in modern food processing to be subjected to certain limitation.
Soybean protein isolate is compared with albumen has relatively poor foaming characteristic, and its application in this respect is also far away not as good as albumen.Albumen is quite general in the application that cake etc. bakes in the based food as foaming agent.But, contain more cholesterol in the albumen, and the price of albumen is far above soybean protein, therefore, foaming characteristic and the foam stability of improving soybean protein have great social significance and economic implications.
For soybean protein structure of modification and the research that further improves functional character is the focus of research both at home and abroad always.Present soy proteinaceous method of modifying mainly contains physical modification, chemical modification, enzyme modification and bioengineering modification, and enzyme modification comes into one's own further because condition is gentle relatively.At present, many researchers use single or compound protease is hydrolyzed to soybean protein isolate, in the hope of improving the foaming characteristic of albumen, complex enzyme for hydrolyzing soybean protein as patent CN101011101A (improving the method for protein foamability) adopts the bent protease of rice, trypsase and papain to form has improved 83.3% before the foaming characteristic ratio is untreated.But people such as Sara, Amro and Elfadil studies show that, though enzymatic hydrolysis can significantly improve the foamability of soybean protein, also can reduce its foam stability simultaneously.In general, the raising bubbling character should improve its foamability and improve its foam stability again, and for protein, it is more meaningful to improve its foam stability.There have report to show that employing transglutaminase (TGase) carries out to be crosslinked, can significantly improve the foam stability of protein.
About the improvement of soybean protein foaming property, the most of research of forefathers concentrates on only to be used protease hydrolytic or only uses TGase crosslinked, and foamability and foam stability are not significantly improved simultaneously.Therefore, by of the influence of control molecular weight for the soybean protein foaming characteristic, earlier with soybean separation proteolysis, hydrolyzate separates according to molecular-weight gradation, it is crosslinked to adopt TGase that it is carried out again, to realize more purposively protein being carried out modification, increases substantially the foaming characteristic and the foam stability of soybean protein isolate, for the structure and the functional character remodeling method of soybean protein provides new approaches, widen its application in modern food processing.
Summary of the invention
Purpose of the present invention: be exactly the preparation method who proposes a kind of high-efficiency soy protein frothing agent.With the soybean protein isolate is raw material, adopt zymyhydrolyzed protein, and pass through hyperfiltration process, from protein hydrolyzate, obtain the protein peptides section of the above molecular weight of 10kDa, crosslinked through enzyme process again, obtain the soy protein frothing agent product of high foaming characteristic and foam stability, improve the use value of soybean protein, to widen the application of soybean protein in modern food processing.
Technical scheme of the present invention: with the soybean protein isolate is raw material, adopt alkali protease Alcalase with proteolysis earlier, from protein hydrolyzate, obtain the protein peptides section of the above molecular weight of 10kDa then by hyperfiltration process, by transglutaminase TGase the protein peptides section is carried out crosslinked reorganization again, product after crosslinked obtains the soy protein frothing agent product through concentrated, freeze drying.The inventive method specifically may further comprise the steps:
1. the hydrolysis of soybean protein isolate
Soybean protein isolate adds the water stirring and dissolving, be made into the protein solution of 80g/L (8%w/v), adjust pH value to 8.0 with 1mol/LNaOH solution, temperature remains on 55 ℃, adds the alkali protease Alcalase (0.2% of liquor capacity amount 0.2%, v/v) reaction (during keep protein solution pH value 8.0) that is hydrolyzed, when protein hydrolysis degree reaches 3%, cessation reaction, hydrolyzate heats the 15min enzyme that goes out in boiling water bath, through centrifugal (5000r/min, 20min) back collection supernatant.The soybean protein hydrolysis degree is defined as the ratio of the peptide bond that is hydrolyzed in the protein.
2. the ultrafiltration of soybean protein hydrolysis liquid
Using molecular cut off is that 3% soybean protein hydrolysis liquid carries out ultrafiltration as the polysulphone super-filter membrane of 10kDa with protein hydrolysis degree, part is held back in collection, obtain the protein peptides section of molecular weight greater than 10kDa, the foamability that records this Partial Protein is compared with soybean protein isolate and has been improved 250%～300%, and foam stability is compared with soybean protein isolate and descended 90%～98%.
3. the enzyme process of soybean protein hydrolysis liquid (molecular weight is greater than 10kDa) is crosslinked
The molecular weight that obtains with ultrafiltration is a substrate greater than the soybean protein hydrolysis liquid of 10kDa, substrate polypeptide content is 50g/L (5%w/v), (0.3%～0.5%w/v) TGase carries out cross-linking reaction to add 3～5g/L in the substrate solution, reaction temperature is 40～45 ℃, pH6.0～6.5, behind reaction 2～4h, place in 80 ℃ of water-baths the insulation 30min enzyme that goes out, in 25 ℃ of water-baths, cool off then, cross-linking products concentrates through vacuum, obtain the foaming agent product after the freeze drying, the foamability that records crosslinked afterproduct is compared with soybean protein isolate and has been improved 250%～350%, and foam stability is compared with soybean protein isolate and improved 50%～60%.
Described vacuum concentration is that 45～50 ℃ vacuum concentrates, and described freeze drying processing is the freeze drying under-50～-55 ℃ of temperature.
The enzyme of the alkali protease Alcalase that hydrolysis the is used unit that lives is 400000u/mL, and the enzyme of the crosslinked used transglutaminase unit that lives is 100u/g.
The foamability of soybean protein and the mensuration of foam stability are to carry out as follows: the soy bean proteinous soln 100mL of 1% (w/v) is placed 500mL graduated cylinder (bore 6cm, height is 20cm) in, use the rotating speed homogeneous 40s of emulsify at a high speed homogenizer with 17500r/min, amount to 2min continuous 3 times, liquid level Bing Ji Suan liquor capacity is V behind the record homogeneous
0(mL), leave standstill behind the 30min record liquid level and to count the Suan liquor capacity be V
30min(mL).The computing formula of foamability of soybean protein (FC) and foam stability (FS) is respectively:
Beneficial effect of the present invention: the preparation method who the invention provides a kind of high-efficiency soy protein frothing agent.Pass through enzyme modification, the foaming characteristic and the foam stability of soybean protein isolate have been increased substantially, for the structure and the functional character remodeling method of soybean protein provides new approaches, improved the use value of soybean protein, widen the application of soybean protein in modern food processing, had good market prospects and economic benefit.
The soy protein frothing agent of the present invention's preparation, foamability and foam stability and albumen are suitable, and have bigger price advantage than albumen, can substitute albumen and be widely used in the food processing, and soy protein frothing agent is natural, nutrition, and edible safety is good.
The technology of the present invention is not only applicable to soybean protein isolate, also can be the structure of some other vegetable proteins such as zein, peanut protein and the transformation of functional character and offer reference, this is for efficiently utilizing plant protein resource to produce high value added product and promoting agro based economic development all to have actively and important practical sense.
The specific embodiment
Comparative examples
Accurately taking by weighing soybean protein isolate 40g, add the water stirring and dissolving, be made into the protein solution of 8% (w/v), is 47.3% without the foamability of the soybean protein isolate of enzymatic treatment, and foam stability is 62.3%.
Embodiment 1
Accurately take by weighing soybean protein isolate 40g, add the water stirring and dissolving, be made into the protein solution of 8% (w/v), adjust pH value to 8.0 with 1mol/L NaOH solution, temperature remains on 55 ℃, (0.2%, the v/v) reaction (during keep protein solution pH value 8.0) that is hydrolyzed is when protein hydrolysis degree reaches 3% to add alkali protease Alcalase, cessation reaction, hydrolyzate heats the 15min enzyme that goes out in boiling water bath, (5000r/min, 20min) supernatant is collected in the back through centrifugal.
Using molecular cut off is that 3% soybean protein hydrolysis liquid carries out ultrafiltration as the polysulphone super-filter membrane of 10kDa with protein hydrolysis degree, part is held back in collection, obtain the protein peptides section of molecular weight greater than 10kDa, the foamability that records this Partial Protein is 165.6%, compare with soybean protein isolate (47.3%) and to have improved 250%, and the foam stability that records this Partial Protein is 1.3%, compares with soybean protein isolate (62.3%) to have descended 98%.
The molecular weight that obtains with ultrafiltration is a substrate greater than the soybean protein hydrolysis liquid of 10kDa, substrate polypeptide content is 5% (w/v), the TGase that adds 0.3% (w/v) in the substrate solution carries out cross-linking reaction, reaction temperature is 40 ℃, pH6.0, behind the reaction 2h, place in 80 ℃ of water-baths the insulation 30min enzyme that goes out, in 25 ℃ of water-baths, cool off then, cross-linking products concentrates through vacuum, obtain the foaming agent product after the freeze drying, the foamability that records crosslinked afterproduct is 165.8%, compare with soybean protein isolate (47.3%) and to have improved 250%, recording the cross-linking products foam stability is 93.4%, compares with soybean protein isolate (62.3%) and has improved 50%.
Embodiment 2
Accurately take by weighing soybean protein isolate 50g, add the water stirring and dissolving, be made into the protein solution of 8% (w/v), adjust pH value to 8.0 with 1mol/L NaOH solution, temperature remains on 55 ℃, (0.2%, the v/v) reaction (during keep protein solution pH value 8.0) that is hydrolyzed is when protein hydrolysis degree reaches 3% to add alkali protease Alcalase, cessation reaction, hydrolyzate heats the 15min enzyme that goes out in boiling water bath, (5000r/min, 20min) supernatant is collected in the back through centrifugal.
Using molecular cut off is that 3% soybean protein hydrolysis liquid carries out ultrafiltration as the polysulphone super-filter membrane of 10kDa with protein hydrolysis degree, part is held back in collection, obtain the protein peptides section of molecular weight greater than 10kDa, the foamability that records this Partial Protein is 175%, compare with soybean protein isolate (47.3%) and to have improved 270%, and foam stability is 2.5%, compares with soybean protein isolate (62.3%) to have descended 96%.
The molecular weight that obtains with ultrafiltration is a substrate greater than the soybean protein hydrolysis liquid of 10kDa, substrate polypeptide content is 5% (w/v), the TGase that adds 0.4% (w/v) in the substrate solution carries out cross-linking reaction, reaction temperature is 40 ℃, pH6.5, behind the reaction 3h, place in 80 ℃ of water-baths the insulation 30min enzyme that goes out, in 25 ℃ of water-baths, cool off then, cross-linking products concentrates through vacuum, obtain the foaming agent product after the freeze drying, the foamability that records crosslinked afterproduct is 184.5%, compare with soybean protein isolate (47.3%) and to have improved 290%, foam stability is 95.3%, compares with soybean protein isolate (62.3%) and has improved 53%.
Embodiment 3
Accurately take by weighing soybean protein isolate 50g, add the water stirring and dissolving, be made into the protein solution of 8% (w/v), adjust pH value to 8.0 with 1mol/L NaOH solution, temperature remains on 55 ℃, (0.2%, the v/v) reaction (during keep protein solution pH value 8.0) that is hydrolyzed is when protein hydrolysis degree reaches 3% to add alkali protease Alcalase, cessation reaction, hydrolyzate heats the 15min enzyme that goes out in boiling water bath, (5000r/min, 20min) supernatant is collected in the back through centrifugal.
Using molecular cut off is that 3% soybean protein hydrolysis liquid carries out ultrafiltration as the polysulphone super-filter membrane of 10kDa with protein hydrolysis degree, part is held back in collection, obtain the protein peptides section of molecular weight greater than 10kDa, the foamability that records this Partial Protein is 172.6%, compare with soybean protein isolate (47.3%) and to have improved 265%, and foam stability is 3.1%, compares with soybean protein isolate (62.3%) to have descended 95%.
The molecular weight that obtains with ultrafiltration is a substrate greater than the soybean protein hydrolysis liquid of 10kDa, substrate polypeptide content is 5% (w/v), the TGase that adds 0.4% (w/v) in the substrate solution carries out cross-linking reaction, reaction temperature is 45 ℃, pH6.5, behind the reaction 3.5h, place in 80 ℃ of water-baths the insulation 30min enzyme that goes out, in 25 ℃ of water-baths, cool off then, cross-linking products concentrates through vacuum, obtain the foaming agent product after the freeze drying, the foamability that records crosslinked afterproduct is 203.4%, compare with soybean protein isolate (47.3%) and to have improved 330%, foam stability is 96.6%, compares with soybean protein isolate (62.3%) and has improved 55%.
Embodiment 4
Accurately take by weighing soybean protein isolate 60g, add the water stirring and dissolving, be made into the protein solution of 8% (w/v), adjust pH value to 8.0 with 1mol/L NaOH solution, temperature remains on 55 ℃, (0.2%, the v/v) reaction (during keep protein solution pH value 8.0) that is hydrolyzed is when protein hydrolysis degree reaches 3% to add alkali protease Alcalase, cessation reaction, hydrolyzate heats the 15min enzyme that goes out in boiling water bath, (5000r/min, 20min) supernatant is collected in the back through centrifugal.
Using molecular cut off is that 3% soybean protein hydrolysis liquid carries out ultrafiltration as the polysulphone super-filter membrane of 10kDa with protein hydrolysis degree, part is held back in collection, obtain the protein peptides section of molecular weight greater than 10kDa, the foamability that records this Partial Protein is 179.7%, compare with soybean protein isolate (47.3%) and to have improved 280%, and foam stability is 5%, compares with soybean protein isolate (62.3%) to have descended 92%.
The molecular weight that obtains with ultrafiltration is a substrate greater than the soybean protein hydrolysis liquid of 10kDa, substrate polypeptide content is 5% (w/v), the TGase that adds 0.5% (w/v) in the substrate solution carries out cross-linking reaction, reaction temperature is 45 ℃, pH6.0, behind the reaction 4h, place in 80 ℃ of water-baths the insulation 30min enzyme that goes out, in 25 ℃ of water-baths, cool off then, cross-linking products concentrates through vacuum, obtain the foaming agent product after the freeze drying, the foamability that records crosslinked afterproduct is 212.9%, compare with soybean protein isolate (47.3%) and to have improved 350%, foam stability is 99.7%, compares with soybean protein isolate (62.3%) and has improved 60%.
Embodiment 5
Accurately take by weighing soybean protein isolate 80g, add the water stirring and dissolving, be made into the protein solution of 8% (w/v), adjust pH value to 8.0 with 1mol/L NaOH solution, temperature remains on 55 ℃, (0.2%, the v/v) reaction (during keep protein solution pH value 8.0) that is hydrolyzed is when protein hydrolysis degree reaches 3% to add alkali protease Alcalase, cessation reaction, hydrolyzate heats the 15min enzyme that goes out in boiling water bath, (5000r/min, 20min) supernatant is collected in the back through centrifugal.
Using molecular cut off is that 3% soybean protein hydrolysis liquid carries out ultrafiltration as the polysulphone super-filter membrane of 10kDa with protein hydrolysis degree, part is held back in collection, obtain the protein peptides section of molecular weight greater than 10kDa, the foamability that records this Partial Protein is 189.2%, compare with soybean protein isolate (47.3%) and to have improved 300%, and foam stability is 6.1%, compares with soybean protein isolate (62.3%) to have descended 90%.
The molecular weight that obtains with ultrafiltration is a substrate greater than the soybean protein hydrolysis liquid of 10kDa, substrate polypeptide content is 5% (w/v), the TGase that adds 0.5% (w/v) in the substrate solution carries out cross-linking reaction, reaction temperature is 40 ℃, pH6.5, behind the reaction 3h, place in 80 ℃ of water-baths the insulation 30min enzyme that goes out, in 25 ℃ of water-baths, cool off then, cross-linking products concentrates through vacuum, obtain the foaming agent product after the freeze drying, the foamability that records crosslinked afterproduct is 193.9%, compare with soybean protein isolate (47.3%) and to have improved 310%, foam stability is 97.8%, compares with soybean protein isolate (62.3%) and has improved 57%.
Claims (3)
1. the preparation method of a high-efficiency soy protein frothing agent, it is characterized in that with the soybean protein isolate being raw material, adopt alkali protease Alcalase with proteolysis earlier, from protein hydrolyzate, obtain the protein peptides section of the above molecular weight of 10kDa then by hyperfiltration process, by transglutaminase the protein peptides section is carried out crosslinked reorganization again, product after crosslinked obtains the soy protein frothing agent product through concentrated, freeze drying, and step is:
(1) hydrolysis of soybean protein isolate
Soybean protein isolate adds the water stirring and dissolving, be made into the protein solution of 80g/L, adjust pH value to 8.0 with 1mol/L NaOH solution, temperature remains on 55 ℃, the alkali protease Alcalase that adds liquor capacity amount 0.2% reaction that is hydrolyzed, during keep protein solution pH value 8.0, when protein hydrolysis degree reaches 3%, cessation reaction, hydrolyzate heat the 15min enzyme that goes out in boiling water bath, collect supernatant behind the centrifugal 20min of 5000r/min;
(2) ultrafiltration of soybean protein hydrolysis liquid
Using molecular cut off is that 3% soybean protein hydrolysis liquid carries out ultrafiltration as the polysulphone super-filter membrane of 10kDa with protein hydrolysis degree, part is held back in collection, obtain the protein peptides section of molecular weight greater than 10kDa, the foamability that records this Partial Protein is compared with soybean protein isolate and has been improved 250%～300%, and foam stability is compared with soybean protein isolate and descended 90%～98%;
(3) molecular weight is crosslinked greater than the enzyme process of 10kDa soybean protein hydrolysis liquid
The molecular weight that obtains with ultrafiltration is a substrate greater than the soybean protein hydrolysis liquid of 10kDa, substrate polypeptide content is 50g/L, the transglutaminase that adds 3～5g/L in the substrate solution carries out cross-linking reaction, reaction temperature is 40～45 ℃, pH6.0～6.5, behind reaction 2～4h, place in 80 ℃ of water-baths the insulation 30min enzyme that goes out, in 25 ℃ of water-baths, cool off then, cross-linking products concentrates through vacuum, obtain the foaming agent product after the freeze drying, the foamability that records crosslinked afterproduct is compared with soybean protein isolate and has been improved 250%～350%, and foam stability is compared with soybean protein isolate and improved 50%～60%.
2. the preparation method of a kind of high-efficiency soy protein frothing agent according to claim 1 is characterized in that: described vacuum concentration is that 45～50 ℃ vacuum concentrates, and it is freeze drying under-50～-55 ℃ of temperature that described freeze drying is handled.
3. the preparation method of a kind of high-efficiency soy protein frothing agent according to claim 1 is characterized in that: the enzyme of the alkali protease Alcalase that hydrolysis the is used unit that lives is 400000u/mL, and the enzyme of the crosslinked used transglutaminase unit that lives is 100u/g.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102669722A (en) * | 2012-04-24 | 2012-09-19 | 苏州欧福蛋业有限公司 | Making method of highly-beaten liquid egg white |
| CN104544283A (en) * | 2014-12-30 | 2015-04-29 | 中国农业科学院农产品加工研究所 | High-foaming egg white powder and preparation method thereof |
| CN106082752A (en) * | 2016-06-15 | 2016-11-09 | 中原工学院 | A kind of preparation method of enzyme crosslinking mixed hydrolysis albumen cement blowing agent |
| CN106912519A (en) * | 2015-12-25 | 2017-07-04 | 河南工业大学 | It is a kind of to add the method that albumen dispensing improves characteristics of cake |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1377590A (en) * | 2001-03-30 | 2002-11-06 | 吴炳新 | Method for producing soybean protein hydrolyate and use |
| CN100489115C (en) * | 2006-02-08 | 2009-05-20 | 沈阳农业大学 | Double-enzyme process for modified preparation of injection type soybean protein isolate |
| CN1944662A (en) * | 2006-10-18 | 2007-04-11 | 江南大学 | Preparing and application method for foam active protein of beer |
| CN101011101A (en) * | 2007-02-07 | 2007-08-08 | 东北农业大学 | Method of modifying protein foamability |
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2008
- 2008-09-09 CN CN2008101959886A patent/CN101366469B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102669722A (en) * | 2012-04-24 | 2012-09-19 | 苏州欧福蛋业有限公司 | Making method of highly-beaten liquid egg white |
| CN102669722B (en) * | 2012-04-24 | 2014-12-03 | 苏州欧福蛋业有限公司 | Making method of highly-beaten liquid egg white |
| CN104544283A (en) * | 2014-12-30 | 2015-04-29 | 中国农业科学院农产品加工研究所 | High-foaming egg white powder and preparation method thereof |
| CN106912519A (en) * | 2015-12-25 | 2017-07-04 | 河南工业大学 | It is a kind of to add the method that albumen dispensing improves characteristics of cake |
| CN106082752A (en) * | 2016-06-15 | 2016-11-09 | 中原工学院 | A kind of preparation method of enzyme crosslinking mixed hydrolysis albumen cement blowing agent |
| CN106082752B (en) * | 2016-06-15 | 2017-11-24 | 中原工学院 | A kind of preparation method of enzyme crosslinking mixed hydrolysis albumen cement blowing agent |
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