CN105400857A - Method for extracting antioxidative peptide from degreased rice bran - Google Patents
Method for extracting antioxidative peptide from degreased rice bran Download PDFInfo
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Abstract
The invention discloses a method for extracting antioxidative peptide from degreased rice bran, belonging to the technical field of preparation of antioxidative peptide. The method comprises the following steps: with degreased rice bran as a raw material, cooking the degreased rice bran with water at first and then carrying out filtering with crocus cloth so as to obtain rice bran liquid; then adding a certain amount of degreased rice bran and distilled water, carrying out mixing and heat preservation, putting the obtained mixture into a constant-temperature water bath, adding protease and cellulase and carrying out a hydrolysis reaction; after enzyme killing, successively carrying out soaking with hydrochloric acid, washing with deionized water, soaking with a sodium hydroxide solution, washing with deionized water again, hydrolysis and high-speed centrifugation so as to obtain crude antioxidative peptide extract; and mixing the crude antioxidative peptide extract with absolute ethyl alcohol and the like, adding cellulose, carrying out hydrolysis, killing cellulase and then carrying out centrifugation so as to prepare the antioxidative peptide. The invention has the following beneficial effects: the preparation method is simple, and preparation cost of the preparation method is 5% lower than the preparation cost of other methods; and the problem of the low utilization rate of rice bran is overcome, production cost for the antioxidative peptide is decreased, and sustainable cyclic utilization of resources is realized.
Description
Technical field
The present invention relates to a kind of method extracting anti-oxidation peptide from defatted rice bran, belong to anti-oxidation peptide preparing technical field.
Background technology
Paddy is China the 1st generalized grain kind. produce about 2.06 hundred million t per year at present.Account for 42% of national total output of grain.Rice yield accounts for 37% of total output of grain in the world.Paddy will remove shell and account for the pericarp of gross weight about 10%, seed coat, perisperm, mush layer and embryo in the process being processed into polished rice, traditional rice bran i.e. existing national standards rice bran are mainly processed into by pericarp, seed coat, perisperm, aleurone layer and embryo, therefore a small amount of rice husk and a certain amount of dust and microorganism can be infiltrated in the course of processing, so can only feed be used for, it is the principal by product of paddy processing.Result of study both domestic and external and data show, are rich in various nutrient substance and physiologically active substance in rice bran.The developed countries such as the present U.S. have had edible rice bran to come out, China also has analogous products to be invented, namely apply modern food to process accurate stone roller technology the not beneficial food matter (rice husk, pericarp, seed coat, dust, microorganism etc.) in rice bran is precisely milled be separated with beneficial nutritive substance (the outer endosperm such as embryo, aleurone layer) of eating in the production plant of cleaning, rice bran can be classified as feed grade rice bran and food grade rice bran two portions by this isolation technique, wherein food grade rice bran accounts for 80% of rice bran gross weight, more than 90% of nutrition.Because although food grade rice bran only accounts for 6% of paddy weight, and accounting for about 60% of paddy nutrition, is thing under the stone roller in rice whitening process.The anticancer property of rice bran protein uniqueness, anti-lipid, the function such as anti-oxidant, becomes the bioactive peptide containing multiple physiologically active by proteolysis.Along with people are to the growing interest of antioxidant, utilize rice bran protein enzyme-squash techniqued to have antioxidation active peptides and have great significance.The rice bran processing and utilization rate of China is low, except few part is as extracting except the raw material of Rice pollard oil, most as feed, much goes out of use in addition, causes the huge waste of resource.Nowadays the antioxidant used usually based on chemical antioxidants, because safety issue is limited; Some natural antioxidants, because high in cost of production is restricted.Therefore from rice bran, find anti-oxidation peptide or other active substances with anti-oxidation characteristics there is significantly exploitation and prospect and realistic meaning.
Summary of the invention
Technical problem to be solved by this invention: low for current rice bran utilization ratio, majority is used as feed or abandons, cause the waste of resource, and due to the high drawback of the natural antioxidants cost produced now, providing a kind of is raw material with defatted rice bran, first and water boiling by it, rice bran liquid is filtered to obtain in crocus cloth, after adding a certain amount of defatted rice bran and distilled water mixing insulation again, put into water bath with thermostatic control, add proteolytic enzyme and cellulase hydrolysis reaction, carry out going out after ferment treatment, through salt acid soak, deionized water wash, sodium hydroxide solution soaks, deionization washs again, be hydrolyzed reaction again, high speed centrifugation is separated to obtain anti-oxidation peptide crude extract, after mixing with dehydrated alcohol etc., after adding cellulase hydrolysis, go out ferment treatment, the method of obtained anti-oxidation peptide after centrifugation.Preparation method of the present invention is simple, not only solves the problem that rice bran utilization ratio is low, decreases the production cost of anti-oxidation peptide, achieve the sustainable recycle of resource.
For solving the problems of the technologies described above, the present invention adopts technical scheme as described below to be:
(1) get 50 ~ 60mL water and put into ellipse circular bottom pot, add 20 ~ 30g defatted rice bran simultaneously, 100 ~ 105 DEG C are warming up to the speed program of 5 DEG C/min, boiling 30 ~ 40min, until by boiled for water to boiling, after water boiling, open wide pot cover, make mixture temperature in pot naturally cool to 35 ~ 40 DEG C, taking out and putting into aperture is that the crocus cloth of 0.1 ~ 0.3 μm filters, and obtains rice bran liquid;
(2) the rice bran liquid of above-mentioned gained is put into there-necked flask, respectively to wherein adding 30 ~ 50g defatted rice bran and 80 ~ 100mL distilled water, control temperature is 40 ~ 45 DEG C, is under the condition of 500 ~ 700r/min at rotating speed, stir 20 ~ 30min to mix, then be incubated 30 ~ 40min;
(3) after above-mentioned insulation completes, put it in water bath with thermostatic control, regulate temperature to be 48 ~ 50 DEG C, add the proteolytic enzyme of 1.5 ~ 1.8% and the cellulase of 3 ~ 5%, control rotating speed is 800 ~ 900r/min, and magnetic agitation mixes, then keep temperature-resistant, hydrolysis reaction 5 ~ 6h;
(4) after hydrolysis completes, ferment treatment 50 ~ the 60min that goes out is carried out under zymolyte being put into ultraviolet lamp, go out after ferment treatment, add salt acid soak 7 ~ 10min that 20 ~ 30mL massfraction is 30 ~ 40%, with deionized water wash to neutral, adding 20 ~ 30mL mass concentration is again soak 7 ~ 10min in the sodium hydroxide solution of 0.1 ~ 0.3mol/L, then with deionized water wash to neutral;
(5) again wash as adding 10 ~ 12g Potassium Persulphate in neutral zymolyte to above-mentioned, the proteolytic enzyme simultaneously adding 1.5 ~ 1.8% mixes, hydrolysis reaction 3 ~ 4h, after hydrolysis reaction completes, put into the centrifugal 12 ~ 14min of whizzer that rotating speed is 8000 ~ 12000r/min, be separated to obtain supernatant liquor, i.e. anti-oxidation peptide crude extract;
(6) the anti-oxidation peptide crude extract of above-mentioned gained is put into reaction vessel, add 5 ~ 7mL ethanol solution wherein, add 5 ~ 7g Sodium phosphate dibasic wherein simultaneously, it is 45 ~ 50 DEG C in temperature, rotating speed is under 500 ~ 700r/min condition, stirs 30 ~ 40min, adds 10 ~ 12g cellulase wherein while stirring with the speed of 1 ~ 2g/min, after interpolation, hydrolysis reaction 2 ~ 3h;
(7) after hydrolysis reaction completes, under zymolyte being put into ultraviolet lamp, carry out the ferment treatment 50 ~ 60min that goes out, after the ferment treatment that goes out, put into the centrifugal 12 ~ 14min of whizzer that rotating speed is 8000 ~ 12000r/min, be separated to obtain supernatant liquor, i.e. fine work anti-oxidation peptide.
The present invention is compared with additive method, and Advantageous Effects is:
(1) preparation method of the present invention is simple, and preparation cost is lower than additive method 5%;
(2) not only solve the problem that rice bran utilization ratio is low, decrease the production cost of anti-oxidation peptide, achieve the sustainable recycle of resource.
Embodiment
First get 50 ~ 60mL water and put into ellipse circular bottom pot, add 20 ~ 30g defatted rice bran simultaneously, 100 ~ 105 DEG C are warming up to the speed program of 5 DEG C/min, boiling 30 ~ 40min, until by boiled for water to boiling, after water boiling, open wide pot cover, make mixture temperature in pot naturally cool to 35 ~ 40 DEG C, taking out and putting into aperture is that the crocus cloth of 0.1 ~ 0.3 μm filters, and obtains rice bran liquid; Then the rice bran liquid of above-mentioned gained is put into there-necked flask, respectively to wherein adding 30 ~ 50g defatted rice bran and 80 ~ 100mL distilled water, control temperature is 40 ~ 45 DEG C, is under the condition of 500 ~ 700r/min at rotating speed, stir 20 ~ 30min to mix, then be incubated 30 ~ 40min; After above-mentioned insulation completes, put it in water bath with thermostatic control, regulate temperature to be 48 ~ 50 DEG C, add the proteolytic enzyme of 1.5 ~ 1.8% and the cellulase of 3 ~ 5%, control rotating speed is 800 ~ 900r/min, and magnetic agitation mixes, then keep temperature-resistant, hydrolysis reaction 5 ~ 6h; Again after hydrolysis completes, ferment treatment 50 ~ the 60min that goes out is carried out under zymolyte being put into ultraviolet lamp, go out after ferment treatment, add salt acid soak 7 ~ 10min that 20 ~ 30mL massfraction is 30 ~ 40%, with deionized water wash to neutral, adding 20 ~ 30mL mass concentration is again soak 7 ~ 10min in the sodium hydroxide solution of 0.1 ~ 0.3mol/L, then with deionized water wash to neutral; Again wash as adding 10 ~ 12g Potassium Persulphate in neutral zymolyte to above-mentioned, the proteolytic enzyme simultaneously adding 1.5 ~ 1.8% mixes, hydrolysis reaction 3 ~ 4h, after hydrolysis reaction completes, put into the centrifugal 12 ~ 14min of whizzer that rotating speed is 8000 ~ 12000r/min, be separated to obtain supernatant liquor, i.e. anti-oxidation peptide crude extract; The anti-oxidation peptide crude extract of above-mentioned gained is put into reaction vessel, add 5 ~ 7mL ethanol solution wherein, add 5 ~ 7g Sodium phosphate dibasic wherein simultaneously, it is 45 ~ 50 DEG C in temperature, rotating speed is under 500 ~ 700r/min condition, stirs 30 ~ 40min, adds 10 ~ 12g cellulase wherein while stirring with the speed of 1 ~ 2g/min, after interpolation, hydrolysis reaction 2 ~ 3h; Finally after hydrolysis reaction completes, under zymolyte being put into ultraviolet lamp, carry out the ferment treatment 50 ~ 60min that goes out, after the ferment treatment that goes out, put into the centrifugal 12 ~ 14min of whizzer that rotating speed is 8000 ~ 12000r/min, be separated to obtain supernatant liquor, i.e. fine work anti-oxidation peptide.
Example 1
First get 50mL water and put into ellipse circular bottom pot, add 20g defatted rice bran simultaneously, 100 DEG C are warming up to the speed program of 5 DEG C/min, boiling 30min, until by boiled for water to boiling, after water boiling, open wide pot cover, make mixture temperature in pot naturally cool to 35 DEG C, taking out and putting into aperture is that the crocus cloth of 0.1 μm filters, and obtains rice bran liquid; Then the rice bran liquid of above-mentioned gained is put into there-necked flask, respectively to wherein adding 30g defatted rice bran and 80mL distilled water, control temperature is 40 DEG C, is under the condition of 500r/min at rotating speed, stirs 20min and mixes, then be incubated 30min; After above-mentioned insulation completes, put it in water bath with thermostatic control, regulate temperature to be 48 DEG C, add the proteolytic enzyme of 1.5% and the cellulase of 3%, control rotating speed is 800r/min, and magnetic agitation mixes, and then keeps temperature-resistant, hydrolysis reaction 5h; Again after hydrolysis completes, the ferment treatment 50min that goes out is carried out under zymolyte being put into ultraviolet lamp, go out after ferment treatment, add the salt acid soak 7min that 20mL massfraction is 30%, with deionized water wash to neutral, adding 20mL mass concentration is again soak 7min in the sodium hydroxide solution of 0.1mol/L, then with deionized water wash to neutral; Again wash as adding 10g Potassium Persulphate in neutral zymolyte to above-mentioned, the proteolytic enzyme simultaneously adding 1.5% mixes, hydrolysis reaction 3h, after hydrolysis reaction completes, put into the centrifugal 12min of whizzer that rotating speed is 8000r/min, be separated to obtain supernatant liquor, i.e. anti-oxidation peptide crude extract; The anti-oxidation peptide crude extract of above-mentioned gained is put into reaction vessel, add 5mL ethanol solution wherein, add 5g Sodium phosphate dibasic wherein simultaneously, it is 45 DEG C in temperature, rotating speed is under 500r/min condition, stirs 30min, adds 10g cellulase wherein while stirring with the speed of 1g/min, after interpolation, hydrolysis reaction 2h; Finally after hydrolysis reaction completes, under zymolyte being put into ultraviolet lamp, carry out the ferment treatment 50min that goes out, after the ferment treatment that goes out, put into the centrifugal 12min of whizzer that rotating speed is 8000r/min, be separated to obtain supernatant liquor, i.e. fine work anti-oxidation peptide.Preparation method of the present invention is simple, and preparation cost is lower than additive method 5%; Not only solve the problem that rice bran utilization ratio is low, decrease the production cost of anti-oxidation peptide, achieve the sustainable recycle of resource.
Example 2
First get 55mL water and put into ellipse circular bottom pot, add 25g defatted rice bran simultaneously, 103 DEG C are warming up to the speed program of 5 DEG C/min, boiling 35min, until by boiled for water to boiling, after water boiling, open wide pot cover, make mixture temperature in pot naturally cool to 38 DEG C, taking out and putting into aperture is that the crocus cloth of 0.2 μm filters, and obtains rice bran liquid; Then the rice bran liquid of above-mentioned gained is put into there-necked flask, respectively to wherein adding 40g defatted rice bran and 90mL distilled water, control temperature is 43 DEG C, is under the condition of 600r/min at rotating speed, stirs 25min and mixes, then be incubated 35min; After above-mentioned insulation completes, put it in water bath with thermostatic control, regulate temperature to be 49 DEG C, add the proteolytic enzyme of 1.65% and the cellulase of 4%, control rotating speed is 850r/min, and magnetic agitation mixes, and then keeps temperature-resistant, hydrolysis reaction 5.5h; Again after hydrolysis completes, the ferment treatment 55min that goes out is carried out under zymolyte being put into ultraviolet lamp, go out after ferment treatment, add the salt acid soak 9min that 25mL massfraction is 35%, with deionized water wash to neutral, adding 25mL mass concentration is again soak 9min in the sodium hydroxide solution of 0.2mol/L, then with deionized water wash to neutral; Again wash as adding 11g Potassium Persulphate in neutral zymolyte to above-mentioned, the proteolytic enzyme simultaneously adding 1.7% mixes, hydrolysis reaction 3.5h, after hydrolysis reaction completes, put into the centrifugal 13min of whizzer that rotating speed is 10000r/min, be separated to obtain supernatant liquor, i.e. anti-oxidation peptide crude extract; The anti-oxidation peptide crude extract of above-mentioned gained is put into reaction vessel, add 6mL ethanol solution wherein, add 6g Sodium phosphate dibasic wherein simultaneously, it is 48 DEG C in temperature, rotating speed is under 600r/min condition, stirs 35min, adds 11g cellulase wherein while stirring with the speed of 1.5g/min, after interpolation, hydrolysis reaction 2.5h; Finally after hydrolysis reaction completes, under zymolyte being put into ultraviolet lamp, carry out the ferment treatment 55min that goes out, after the ferment treatment that goes out, put into the centrifugal 13min of whizzer that rotating speed is 10000r/min, be separated to obtain supernatant liquor, i.e. fine work anti-oxidation peptide.Preparation method of the present invention is simple, and preparation cost is lower than additive method 5%; Not only solve the problem that rice bran utilization ratio is low, decrease the production cost of anti-oxidation peptide, achieve the sustainable recycle of resource.
Example 3
First get 60mL water and put into ellipse circular bottom pot, add 30g defatted rice bran simultaneously, 105 DEG C are warming up to the speed program of 5 DEG C/min, boiling 40min, until by boiled for water to boiling, after water boiling, open wide pot cover, make mixture temperature in pot naturally cool to 40 DEG C, taking out and putting into aperture is that the crocus cloth of 0.3 μm filters, and obtains rice bran liquid; Then the rice bran liquid of above-mentioned gained is put into there-necked flask, respectively to wherein adding 50g defatted rice bran and 100mL distilled water, control temperature is 45 DEG C, is under the condition of 700r/min at rotating speed, stirs 30min and mixes, then be incubated 40min; After above-mentioned insulation completes, put it in water bath with thermostatic control, regulate temperature to be 50 DEG C, add the proteolytic enzyme of 1.8% and the cellulase of 5%, control rotating speed is 900r/min, and magnetic agitation mixes, and then keeps temperature-resistant, hydrolysis reaction 6h; Again after hydrolysis completes, the ferment treatment 60min that goes out is carried out under zymolyte being put into ultraviolet lamp, go out after ferment treatment, add the salt acid soak 10min that 30mL massfraction is 40%, with deionized water wash to neutral, adding 30mL mass concentration is again soak 10min in the sodium hydroxide solution of 0.3mol/L, then with deionized water wash to neutral; Again wash as adding 12g Potassium Persulphate in neutral zymolyte to above-mentioned, the proteolytic enzyme simultaneously adding 1.8% mixes, hydrolysis reaction 4h, after hydrolysis reaction completes, put into the centrifugal 14min of whizzer that rotating speed is 12000r/min, be separated to obtain supernatant liquor, i.e. anti-oxidation peptide crude extract; The anti-oxidation peptide crude extract of above-mentioned gained is put into reaction vessel, add 7mL ethanol solution wherein, add 7g Sodium phosphate dibasic wherein simultaneously, it is 50 DEG C in temperature, rotating speed is under 700r/min condition, stirs 40min, adds 12g cellulase wherein while stirring with the speed of 2g/min, after interpolation, hydrolysis reaction 3h; Finally after hydrolysis reaction completes, under zymolyte being put into ultraviolet lamp, carry out the ferment treatment 60min that goes out, after the ferment treatment that goes out, put into the centrifugal 14min of whizzer that rotating speed is 12000r/min, be separated to obtain supernatant liquor, i.e. fine work anti-oxidation peptide.Preparation method of the present invention is simple, and preparation cost is lower than additive method 5%; Not only solve the problem that rice bran utilization ratio is low, decrease the production cost of anti-oxidation peptide, achieve the sustainable recycle of resource.
Claims (1)
1. from defatted rice bran, extract a method for anti-oxidation peptide, it is characterized in that concrete extracting method is:
(1) get 50 ~ 60mL water and put into ellipse circular bottom pot, add 20 ~ 30g defatted rice bran simultaneously, 100 ~ 105 DEG C are warming up to the speed program of 5 DEG C/min, boiling 30 ~ 40min, until by boiled for water to boiling, after water boiling, open wide pot cover, make mixture temperature in pot naturally cool to 35 ~ 40 DEG C, taking out and putting into aperture is that the crocus cloth of 0.1 ~ 0.3 μm filters, and obtains rice bran liquid;
(2) the rice bran liquid of above-mentioned gained is put into there-necked flask, respectively to wherein adding 30 ~ 50g defatted rice bran and 80 ~ 100mL distilled water, control temperature is 40 ~ 45 DEG C, is under the condition of 500 ~ 700r/min at rotating speed, stir 20 ~ 30min to mix, then be incubated 30 ~ 40min;
(3) after above-mentioned insulation completes, put it in water bath with thermostatic control, regulate temperature to be 48 ~ 50 DEG C, add the proteolytic enzyme of 1.5 ~ 1.8% and the cellulase of 3 ~ 5%, control rotating speed is 800 ~ 900r/min, and magnetic agitation mixes, then keep temperature-resistant, hydrolysis reaction 5 ~ 6h;
(4) after hydrolysis completes, ferment treatment 50 ~ the 60min that goes out is carried out under zymolyte being put into ultraviolet lamp, go out after ferment treatment, add salt acid soak 7 ~ 10min that 20 ~ 30mL massfraction is 30 ~ 40%, with deionized water wash to neutral, adding 20 ~ 30mL mass concentration is again soak 7 ~ 10min in the sodium hydroxide solution of 0.1 ~ 0.3mol/L, then with deionized water wash to neutral;
(5) again wash as adding 10 ~ 12g Potassium Persulphate in neutral zymolyte to above-mentioned, the proteolytic enzyme simultaneously adding 1.5 ~ 1.8% mixes, hydrolysis reaction 3 ~ 4h, after hydrolysis reaction completes, put into the centrifugal 12 ~ 14min of whizzer that rotating speed is 8000 ~ 12000r/min, be separated to obtain supernatant liquor, i.e. anti-oxidation peptide crude extract;
(6) the anti-oxidation peptide crude extract of above-mentioned gained is put into reaction vessel, add 5 ~ 7mL ethanol solution wherein, add 5 ~ 7g Sodium phosphate dibasic wherein simultaneously, it is 45 ~ 50 DEG C in temperature, rotating speed is under 500 ~ 700r/min condition, stirs 30 ~ 40min, adds 10 ~ 12g cellulase wherein while stirring with the speed of 1 ~ 2g/min, after interpolation, hydrolysis reaction 2 ~ 3h;
(7) after hydrolysis reaction completes, under zymolyte being put into ultraviolet lamp, carry out the ferment treatment 50 ~ 60min that goes out, after the ferment treatment that goes out, put into the centrifugal 12 ~ 14min of whizzer that rotating speed is 8000 ~ 12000r/min, be separated to obtain supernatant liquor, i.e. fine work anti-oxidation peptide.
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Citations (5)
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| CN101012471A (en) * | 2007-02-07 | 2007-08-08 | 东北农业大学 | Method for extracting rice bran protein by complex enzyme |
| JP2008308445A (en) * | 2007-06-14 | 2008-12-25 | Gekkeikan Sake Co Ltd | Angiotensin converting enzyme-inhibitory peptide mixture and method for producing the same |
| CN102080118A (en) * | 2010-06-23 | 2011-06-01 | 东北农业大学 | Preparation method of rice bran antioxidant active protein peptide |
| CN102115690A (en) * | 2011-01-12 | 2011-07-06 | 武汉百信食品有限公司 | Method for comprehensively utilizing rice bran |
| CN102178151A (en) * | 2011-04-22 | 2011-09-14 | 淮南市楚丰工贸有限公司 | Method for co-producing rice bran dietary fiber and rice bran protein by utilizing defatted rice bran |
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2015
- 2015-12-12 CN CN201510916929.3A patent/CN105400857A/en active Pending
Patent Citations (5)
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|---|---|---|---|---|
| CN101012471A (en) * | 2007-02-07 | 2007-08-08 | 东北农业大学 | Method for extracting rice bran protein by complex enzyme |
| JP2008308445A (en) * | 2007-06-14 | 2008-12-25 | Gekkeikan Sake Co Ltd | Angiotensin converting enzyme-inhibitory peptide mixture and method for producing the same |
| CN102080118A (en) * | 2010-06-23 | 2011-06-01 | 东北农业大学 | Preparation method of rice bran antioxidant active protein peptide |
| CN102115690A (en) * | 2011-01-12 | 2011-07-06 | 武汉百信食品有限公司 | Method for comprehensively utilizing rice bran |
| CN102178151A (en) * | 2011-04-22 | 2011-09-14 | 淮南市楚丰工贸有限公司 | Method for co-producing rice bran dietary fiber and rice bran protein by utilizing defatted rice bran |
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Application publication date: 20160316 |