CN103749939B - A kind of soybean protein isolate preparation method being rich in branched-chain amino acid - Google Patents
A kind of soybean protein isolate preparation method being rich in branched-chain amino acid Download PDFInfo
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- 239000006228 supernatant Substances 0.000 claims abstract description 7
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- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 6
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 4
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- QVDXUKJJGUSGLS-LURJTMIESA-N methyl L-leucinate Chemical compound COC(=O)[C@@H](N)CC(C)C QVDXUKJJGUSGLS-LURJTMIESA-N 0.000 claims description 6
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- KUGLDBMQKZTXPW-JEDNCBNOSA-N methyl (2s)-2-amino-3-methylbutanoate;hydrochloride Chemical compound Cl.COC(=O)[C@@H](N)C(C)C KUGLDBMQKZTXPW-JEDNCBNOSA-N 0.000 claims description 3
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- Coloring Foods And Improving Nutritive Qualities (AREA)
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Abstract
The invention discloses a kind of soybean protein isolate preparation method being rich in branched-chain amino acid, first prepare soybean protein isolate, stir, heating, adopt alkali protease to be hydrolyzed, in hydrolytic process, stablize pH value in prescribed limit with NaOH; By hydrolyzate through boiling water deactivation, add HCl adjust ph, centrifugal, get supernatant for subsequent use; Again the freeze-drying of supernatant vacuum freeze drier, porphyrize are placed in drier; Regulate pH value of solution, under Cys exists, add Papain Catalyzed reaction, enzyme concentration 2%; Reacted rear alcohol settling protein, centrifugal, protein crosses bag filter in rear centrifugal, and freeze drying is precipitated as end product.Compared with prior art, the present invention adopts plastein reaction method to be imported by branched-chain amino acid in the protein such as soybean protein, the obtained albuminoid product being rich in branched-chain amino acid, the object of supplementary branched-chain amino acid can be reached equally after edible, and product characteristics are stablized, good processability, without bad mouthfeel.
Description
Technical field
The present invention relates to a kind of soybean protein isolate preparation method, particularly relate to a kind of soybean protein isolate preparation method being rich in branched-chain amino acid.
Background technology
All the time, known comprise pick propylhomoserin, leucine, this 3 seed amino acid of isoleucine branched-chain amino acid have and increase the function of protein, or energy source when can be used as motion and playing an important role, develop various medicine, food and supplies etc. containing branched-chain amino acid and go on the market.
But; usually; free branched-chain amino acid or the small peptide production containing more branched-chain amino acid are normally become parenteral solution, particle or tablet with supplies, or are added to the product obtaining in food, beverage and be rich in branched-chain amino acid by existing medicine containing branched-chain amino acid, food.On the one hand, these products have the distinctive bitter taste stink deriving from branched-chain amino acid, amino acid free in addition or the less stable of little peptide, and product is in the reduction even change of shape of product of preserving, selling period easily causes branched-amino acid content.Meanwhile, branched-chain amino acid is owing to being hydrophobic amino acid, and the granular preparation therefore such as containing branched-chain amino acid is difficult to water affine, when swallowing in anhydrous situation or when taking together with water, has particle a large amount of residual problem waited in mouth of failing to swallow.Have researcher to utilize enzyme hydrolysis and utilize particular technology to carry out being separated to prepare the small peptide material being rich in branched-chain amino acid, but these little peptides do not have the functional characters such as matter structure, and certain bitter taste may be had, be unfavorable for applying in the food industry yet.
Summary of the invention
Object of the present invention is just to provide a kind of soybean protein isolate preparation method being rich in branched-chain amino acid to solve the problem.
The present invention is achieved through the following technical solutions above-mentioned purpose:
The present invention includes following steps:
(1) soybean protein isolate of 5% is prepared, stir, 80 DEG C of heating 10min, as storing solution, employing alkali protease is hydrolyzed, to enzyme amount 5%, pH8.5,55 DEG C of waters bath with thermostatic control hydrolysis 4h, in hydrolytic process with 0.2mol/L NaOH stablize pH value prescribed limit ± 0.05 in;
(2) after reaching hydrolysis time, by hydrolyzate through boiling water deactivation 10min, add the HCL adjust ph to 4.0 of 2mol/L, unhydrolysed albumen precipitation is got off, the centrifugal 20min of 3500r/min, gets supernatant for subsequent use;
(3) freeze-drying of supernatant vacuum freeze drier, porphyrize are placed in drier, for subsequent use;
(4) using step (1) to the product in step (3) as the substrate of plastein reaction, be mixed with the aqueous solution of concentration 30%, regulate pH value of solution=6.0, reaction temperature 37 DEG C, ratio between leucine methyl ester hydrochloride m and substrate m is 10:1, Papain Catalyzed reaction is added, enzyme concentration 2%, reaction time 24h under the Cys of 0.01mol/L exists;
(5) reacted rear use 80% alcohol settling protein, the centrifugal 30min of 3500r/min, protein cross 3500Da bag filter dialyse 4 days at 4 DEG C after centrifugal, freeze drying is precipitated as end product.
Improve as one, described step (1) can be taked as under type is replaced: the soybean protein isolate of preparation 5%, stir, 80 DEG C of heating 10min, as storing solution, adopt chymotrypsin to be hydrolyzed, to enzyme amount 5%, pH9.0,50 DEG C of waters bath with thermostatic control hydrolysis 6h, in hydrolytic process with 0.2mol/L NaOH stablize pH value prescribed limit ± 0.05 in.
Improve as one, described step (3) can be taked as under type is replaced: hydrolyzate thermal spray drying machine spraying dry, porphyrize are placed in drier, for subsequent use.
Improve as one, described step (4) can be taked as under type is replaced: using step (1) to the product in step (3) as the substrate of plastein reaction, be mixed with the aqueous solution of concentration 30%, regulate pH value of solution=6.0, reaction temperature 37 DEG C, the ratio between valine methyl ester hydrochloride m and substrate m is 10:1, adds Papain Catalyzed reaction under the Cys of 0.01mol/L exists, enzyme concentration 2%, reaction time 24h;
Improve as one, described step (4) can be taked as under type is replaced: using step (1) to the product in step (3) as the substrate of plastein reaction, be mixed with the aqueous solution of concentration 30%, regulate pH value of solution=7.0, reaction temperature 37 DEG C, ratio between leucine methyl ester hydrochloride m and substrate m is 10:1, alkali protease (Alcalase) catalytic reaction is added under 0.01mol/L Cys exists, enzyme concentration 2%, reaction time 24h.
Improve as one, described step (5) can be replaced in the following way: reacted rear employing 10% trichloroacetic acid precipitation protein, the centrifugal 30min of 3500r/min, protein cross 3500 bag filters dialyse 4 days at 4 DEG C after centrifugal, freeze drying is precipitated as end product.
Improve as one, described step (5) can be replaced in the following way: reacted rear boiling water bath heating 15 ~ 20min postprecipitation protein, the centrifugal 30min of 3500r/min, protein cross 3500Da bag filter dialyse 4 days at 4 DEG C after centrifugal, freeze drying is precipitated as end product.
Beneficial effect of the present invention is:
The present invention is a kind of soybean protein isolate preparation method being rich in branched-chain amino acid, compared with prior art, the present invention adopts plastein reaction method to be imported by branched-chain amino acid in the protein such as soybean protein, the obtained albuminoid product being rich in branched-chain amino acid, the object of supplementary branched-chain amino acid can be reached equally after edible, and product characteristics are stablized, good processability, without bad mouthfeel.
Detailed description of the invention
The invention will be further described below:
The present invention includes following steps:
(1) soybean protein isolate of 5% is prepared, stir, 80 DEG C of heating 10min, as storing solution, employing alkali protease is hydrolyzed, to enzyme amount 5%, pH 8.5,55 DEG C of waters bath with thermostatic control hydrolysis 4h, in hydrolytic process with 0.2mol/L NaOH stablize pH value prescribed limit ± 0.05 in;
(2) after reaching hydrolysis time, by hydrolyzate through boiling water deactivation 10min, add the HCL adjust ph to 4.0 of 2mol/L, unhydrolysed albumen precipitation is got off, the centrifugal 20min of 3500r/min, gets supernatant for subsequent use;
(3) freeze-drying of hydrolyzate vacuum freeze drier, porphyrize are placed in drier, for subsequent use;
(4) using step (1) to the product in step (3) as the substrate of plastein reaction, be mixed with the aqueous solution of concentration 30%, regulate pH value of solution=6.0, reaction temperature 37 DEG C, ratio between leucine methyl ester hydrochloride m and substrate m is 10:1, Papain Catalyzed reaction is added, enzyme concentration 2%, reaction time 24h under the Cys of 0.01mol/L exists;
(5) reacted rear use 80% alcohol settling protein, the centrifugal 30min of 3500r/min, protein cross 3500Da bag filter dialyse 4 days at 4 DEG C after centrifugal, freeze drying is precipitated as end product.
Embodiment 1: improve as one, described step (1) can be taked as under type is replaced: the soybean protein isolate of preparation 5%, stir, 80 DEG C of heating 10min, as storing solution, adopt chymotrypsin to be hydrolyzed, to enzyme amount 5%, pH9.0,50 DEG C of waters bath with thermostatic control hydrolysis 6h, in hydrolytic process with 0.2mol/LNaOH stablize pH value prescribed limit ± 0.05 in.
Embodiment 2: improve as one, described step (3) can be taked as under type is replaced: hydrolyzate thermal spray drying machine spraying dry, porphyrize are placed in drier, for subsequent use.
Embodiment 3: improve as one, described step (4) can be taked as under type is replaced: using step (1) to the product in step (3) as the substrate of plastein reaction, be mixed with the aqueous solution of concentration 30%, regulate pH value of solution=6.0, reaction temperature 37 DEG C, the ratio between valine methyl ester hydrochloride m and substrate m is 10:1, adds Papain Catalyzed reaction under the Cys of 0.01mol/L exists, enzyme concentration 2%, reaction time 24h;
Embodiment 4: improve as one, described step (4) can be taked as under type is replaced: using step (1) to the product in step (3) as the substrate of plastein reaction, be mixed with the aqueous solution of concentration 30%, regulate pH value of solution=7.0, reaction temperature 37 DEG C, m (leucine methyl ester hydrochloride): m (substrate)=10:1, alkali protease (Alcalase) catalytic reaction is added under 0.01mol/L Cys exists, enzyme concentration 2%, reaction time 24h.
Embodiment 5: improve as one, described step (5) can be replaced in the following way: reacted rear employing 10% trichloroacetic acid precipitation protein, the centrifugal 30min of 3500r/min, protein cross 3500 bag filters dialyse 4 days at 4 DEG C after centrifugal, freeze drying is precipitated as end product.
Embodiment 6: improve as one, described step (5) can be replaced in the following way: reacted rear boiling water bath heating 15 ~ 20min postprecipitation protein, the centrifugal 30min of 3500r/min, protein cross 3500Da bag filter dialyse 4 days at 4 DEG C after centrifugal, freeze drying is precipitated as end product.
For existing technology, the soluble problem of this technology (or advantage is as follows):
(1) utilize plastein reaction method to import in peptide chain by amino acid, and that cause properties of product problem unstable due to free amino acid can be solved, reduce bitter taste and peculiar smell that free amino acid causes simultaneously;
(2) the little peptide of the high branched-chain amino acid prepared from simple hydrolysis is different, and protein can be made to occur by plastein reaction crosslinked, produce the material with good rheological character, increase the viscosity of solution, these character are very useful in matter structure helping digestion product;
(3) this technology stability that the branched-chain amino acid of importing can be made to present prior art do not have;
(4) final products of this technology are compared with existing product, effectively can improve the utilization rate of protein, and be rich in physiologically active.
Claims (7)
1. be rich in a soybean protein isolate preparation method for branched-chain amino acid, it is characterized in that, comprise the following steps:
(1) soybean protein isolate of 5% is prepared, stir, 80 DEG C of heating 10min, as storing solution, employing alkali protease is hydrolyzed, to enzyme amount 5%, pH 8.5,55 DEG C of waters bath with thermostatic control hydrolysis 4h, in hydrolytic process with 0.2mol/L NaOH stablize pH value prescribed limit ± 0.05 in;
(2) after reaching hydrolysis time, by hydrolyzate through boiling water deactivation 10min, add the HCL adjust ph to 4.0 of 2mol/L, unhydrolysed albumen precipitation is got off, the centrifugal 20min of 3500r/min, gets supernatant for subsequent use;
(3) freeze-drying of supernatant vacuum freeze drier, porphyrize are placed in drier, for subsequent use;
(4) using step (1) to the product in step (3) as the substrate of plastein reaction, be mixed with the aqueous solution of concentration 30%, regulate pH value of solution=6.0, reaction temperature 37 DEG C, ratio between leucine methyl ester hydrochloride m and substrate m is 10:1, Papain Catalyzed reaction is added, enzyme concentration 2%, reaction time 24h under the Cys of 0.01mol/L exists;
(5) reacted rear use 80% alcohol settling protein, the centrifugal 30min of 3500r/min, protein cross 3500Da bag filter dialyse 4 days at 4 DEG C after centrifugal, freeze drying is precipitated as end product.
2. the soybean protein isolate preparation method being rich in branched-chain amino acid according to claim 1, it is characterized in that: described step (1) can be taked as under type is replaced: the soybean protein isolate of preparation 5%, stir, 80 DEG C of heating 10min, as storing solution, adopt chymotrypsin to be hydrolyzed, to enzyme amount 5%, pH9.0,50 DEG C of waters bath with thermostatic control hydrolysis 6h, in hydrolytic process with 0.2mol/L NaOH stablize pH value prescribed limit ± 0.05 in.
3. the soybean protein isolate preparation method being rich in branched-chain amino acid according to claim 1, it is characterized in that: described step (3) can be taked as under type is replaced: hydrolyzate thermal spray drying machine spraying dry, porphyrize are placed in drier, for subsequent use.
4. the soybean protein isolate preparation method being rich in branched-chain amino acid according to claim 1, it is characterized in that: described step (4) can be taked as under type is replaced: using step (1) to the product in step (3) as the substrate of plastein reaction, be mixed with the aqueous solution of concentration 30%, regulate pH value of solution=6.0, reaction temperature 37 DEG C, ratio between valine methyl ester hydrochloride m and substrate m is 10:1, Papain Catalyzed reaction is added under the Cys of 0.01mol/L exists, enzyme concentration 2%, reaction time 24h;
5. the soybean protein isolate preparation method being rich in branched-chain amino acid according to claim 1, it is characterized in that: described step (4) can be taked as under type is replaced: using step (1) to the product in step (3) as the substrate of plastein reaction, be mixed with the aqueous solution of concentration 30%, regulate pH value of solution=7.0, reaction temperature 37 DEG C, ratio between leucine methyl ester hydrochloride m and substrate m is 10:1, basic protein enzymic catalytic reaction is added under 0.01mol/LL-cysteine exists, enzyme concentration 2%, reaction time 24h.
6. according to the soybean protein isolate preparation method being rich in branched-chain amino acid wherein described in Claims 1-4, it is characterized in that: described step (5) can be taked as under type is replaced: reacted rear employing 10% trichloroacetic acid precipitation protein, the centrifugal 30min of 3500r/min, protein cross 3500Da bag filter dialyse 4 days at 4 DEG C after centrifugal, freeze drying is precipitated as end product.
7. according to the soybean protein isolate preparation method being rich in branched-chain amino acid wherein described in Claims 1-4, it is characterized in that: described step (5) can be replaced in the following way: reacted rear boiling water bath heating 15 ~ 20min postprecipitation protein, the centrifugal 30min of 3500r/min, protein cross 3500Da bag filter dialyse 4 days at 4 DEG C after centrifugal, freeze drying is precipitated as end product.
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| CN106819358A (en) * | 2016-12-22 | 2017-06-13 | 山东禹王生态食业有限公司 | A kind of soybean protein isolate processing method |
| CN109007238A (en) * | 2018-07-09 | 2018-12-18 | 东北农业大学 | A kind of method that cysteine combination transglutaminase modification prepares low irritability lactalbumin |
| CN109777849B (en) * | 2019-03-07 | 2022-08-19 | 中南林业科技大学 | Preparation method for extracting proteolysis polypeptide from debitterized peach kernel |
| CN112011585A (en) * | 2020-09-09 | 2020-12-01 | 武汉轻工大学 | Preparation method of pumpkin seed polypeptide |
| CN115444078B (en) * | 2022-08-31 | 2024-05-31 | 山东省海洋资源与环境研究院(山东省海洋环境监测中心、山东省水产品质量检验中心) | Amino acid balance method of non-grain vegetable protein based on enzymolysis hydration reaction |
| CN115886076B (en) * | 2022-11-26 | 2024-04-19 | 黑龙江飞鹤乳业有限公司 | Composition containing leucine protein reaction product and application of composition in resisting muscle attenuation |
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| CN103005142A (en) * | 2012-12-13 | 2013-04-03 | 内蒙古伊利实业集团股份有限公司 | Alpha-lactalbumin hydrolysate and preparation method and application thereof |
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| CN103005142A (en) * | 2012-12-13 | 2013-04-03 | 内蒙古伊利实业集团股份有限公司 | Alpha-lactalbumin hydrolysate and preparation method and application thereof |
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| 酶促水解大豆分离蛋白的研究;赵新淮等;《食品与发酵工业》;19941231(第5期);7-11 * |
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Inventor after: Jiang Bin Inventor after: Feng Zhibiao Inventor after: Liu Chunhong Inventor after: Li Dongmei Inventor before: Feng Zhibiao Inventor before: Jiang Bin Inventor before: Liu Chunhong Inventor before: Li Dongmei |
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Free format text: CORRECT: INVENTOR; FROM: FENG ZHIBIAO JIANG BIN LIU CHUNHONG LI DONGMEI TO: JIANG BIN FENG ZHIBIAO LIU CHUNHONG LI DONGMEI |
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Granted publication date: 20151021 |
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