CN101824453A - Two-stage preparation technology of high activity casein antioxidant peptides - Google Patents
Two-stage preparation technology of high activity casein antioxidant peptides Download PDFInfo
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- CN101824453A CN101824453A CN200910071495A CN200910071495A CN101824453A CN 101824453 A CN101824453 A CN 101824453A CN 200910071495 A CN200910071495 A CN 200910071495A CN 200910071495 A CN200910071495 A CN 200910071495A CN 101824453 A CN101824453 A CN 101824453A
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Abstract
The invention discloses a two-stage preparation technology of high activity casein antioxidant peptides, comprising the following steps of firstly, preparing aqueous solution of casein based on the mass volume ratio of 4-6 percent (W/V), regulating a pH value of the solution to 5.0-8.0, adding papain, keeping the solution under the temperature of 40-50 DEG C to react for 1-5h, heating for 20m under the temperature of 100 DEG C to inactivate enzyme, and obtaining the casein antioxidant peptides by freeze drying or spray drying supermate after centrifugation; secondly, preparing aqueous solution of the casein antioxidant peptides based on the mass volume ratio of 30-70 percent (W/V), reacting for 1-10h under the temperature of 13-47 DEG C after adding the papain, heating for 20m under the temperature of 100 DEG C to inactivate the enzyme, and obtaining the high activity casein antioxidant peptides by freeze drying or spray drying the solution. The activity of the prepared high activity casein antioxidant peptides is enhanced by 8 times, and the high activity casein antioxidant peptides can be used for food processing and health care product production.
Description
Technical field
The two-step approach that the present invention relates to the high activity casein anti-oxidation peptide prepares technology, relate to the casein is main raw material, utilize papoid to be catalyzer, by the further processing that the first step casein hydrolysis prepares the casein antioxidant peptide and the second step casein antioxidant peptide, two-step approach prepares the high activity casein anti-oxidation peptide.Prepared high activity casein anti-oxidation peptide can be applied to the production of food-processing and healthcare products.
Background technology
Anti-oxidation peptide is to handle by the enzymic hydrolysis of natural protein, thus the protein hydrolystate of preparing with anti-oxidant activity.Casein is one of proteinic main source in the anti-oxidation peptide preparation.Since anti-oxidation peptide have molecular weight little, easily digest and assimilate, safe and reliable, characteristics such as do not cause allergic reaction, so anti-oxidation peptide has good prospect in many fields such as food-processing, healthcare products productions.
The general preparation of anti-oxidation peptide mainly is to reach by proteinic direct enzymic hydrolysis, and the separation that the activity of anti-oxidation peptide can be by peptide, purification etc. are handled and further improved, but treatment technology complexity, operational difficulty.The present invention is a basic raw material with casein, papoid, and two-step approach prepares the high activity casein anti-oxidation peptide.The first step adopts papoid catalysis casein hydrolysis, the preparation casein antioxidant peptide; Second step utilized papoid that casein antioxidant peptide is handled once more, preparation high activity casein anti-oxidation peptide.Two-step approach prepares the high activity casein anti-oxidation peptide that, do not need the isolated or purified of peptide to handle, and the activity of product improves.Therefore, the two-step approach of high activity casein anti-oxidation peptide prepares technology, can be used as the production that a kind of new technique means is applied to the high activity casein anti-oxidation peptide.
Summary of the invention
The objective of the invention is with casein and papoid is basic raw material, the first step is prepared casein antioxidant peptide by the caseic hydrolysis of papoid catalysis, second step utilized papoid that casein antioxidant peptide is handled once more, thereby prepared a kind of high activity casein anti-oxidation peptide.
Purpose of the present invention is achieved through the following technical solutions:
(1) preparation method of casein antioxidant peptide:
(1) casein is carried out swelling with distilled water, constantly stir simultaneously and dissolve fully up to casein, the preparation mass volume ratio is the casein solution of 4%～6% (W/V);
(2) use 1molL
-1NaOH solution the pH value of casein solution is transferred to 5.0～8.0;
(3) add papoid in the casein solution of the intact pH value of above-mentioned adjusting, the addition of papoid is every gram casein 300U～600U enzyme activity unit;
(4) casein solution of above-mentioned adding papoid is put into 40 ℃～50 ℃ water-baths are heated, hydrolysis, agitator stirs, and mixing speed is that per minute 30 changes～90 and changes, and hydrolysis was carried out 1 hour～5 hours;
(5) use 1molL
-1HCl solution transfers to 4.6 with the casein solution pH value of hydrolysis, is warming up to 100 ℃ of heating then and carries out enzyme-deactivating in 20 minutes;
(6) casein solution with said hydrolyzed leaves heart processing 20 minutes with per minute 5000, discards residue, collects supernatant liquor;
(7) resulting supernatant liquor lyophilize or spraying drying.The shelf Heating temperature of freeze drier is 20 ℃ during lyophilize, takes out the lyophilize thing when loft drier pressure is reduced to 6 handkerchiefs～8 handkerchiefs, obtains casein antioxidant peptide.During spraying drying, flow obtains casein antioxidant peptide for per hour 150 kilograms, inlet temperature are that 160 ℃～200 ℃, temperature out are 70 ℃～80 ℃.
In the first step preparation, the mass volume ratio of casein solution is 5% (W/V) preferably, the pH value of solution preferably is adjusted to 6.5, the preferably every gram casein of papain dosage 500U enzyme activity unit, preferably 45 ℃ of temperature of reaction, mixing speed preferably per minute 60 changes hydrolysis time preferably 2 hours.
(2) preparation method of high activity casein anti-oxidation peptide:
(1) with the casein antioxidant peptide dissolved in distilled water that obtains, the preparation mass volume ratio is the casein antioxidant peptide solution of 30～70% (W/V);
(2) add papoid in above-mentioned casein antioxidant peptide solution, the addition of papoid is every gram casein antioxidant peptide 300U～600 enzyme activity units;
(3) the casein antioxidant peptide solution of above-mentioned adding papoid is stirred under 13 ℃～47 ℃ condition handle, stirring velocity is that per minute 60 changes～100 and changes, and the treatment time is 1 hour～10 hours; Be warming up to 100 ℃ of heating then and carried out enzyme-deactivating in 20 minutes;
(4) lyophilize of casein antioxidant peptide solution or the spraying drying after will handling.The shelf Heating temperature of freeze drier is 20 ℃ during lyophilize, when loft drier pressure is reduced to 6 handkerchiefs～8 handkerchiefs, takes out the lyophilize thing, obtains the high activity casein anti-oxidation peptide.During spraying drying, flow obtains the high activity casein anti-oxidation peptide for per hour 150 kilograms, inlet temperature are that 160 ℃～200 ℃, temperature out are 70 ℃～80 ℃.
In the preparation of second step, the mass volume ratio of casein antioxidant peptide is 50% (W/V) preferably, the preferably every gram casein antioxidant peptide of papain dosage 500U enzyme activity unit, preferably 30 ℃ of treatment temps, mixing speed preferably per minute 80 changes preferably 5.6 hours treatment time.
The present invention is a basic raw material with casein, papoid, and two-step approach prepares the high activity casein anti-oxidation peptide.The antioxidant peptide active of the present invention's preparation is higher, has removing 2, the advantage that bitter diazanyl (DPPH) free radical of 2-phenylbenzene-1-ability is significantly increased.
Embodiment
For understanding the present invention better, be elaborated below in conjunction with embodiment, but the scope of protection of present invention is not limited to the scope that embodiment represents.
Embodiment 1
(1) casein is carried out swelling with distilled water, constantly stir simultaneously and dissolve fully up to casein, the preparation mass volume ratio is the casein solution of 5% (W/V);
(2) use 1molL
-1NaOH solution the pH value of casein solution is transferred to 6.5;
(3) add papoid in the casein solution of the intact pH value of above-mentioned adjusting, the addition of papoid is every gram casein 500U enzyme activity unit;
(4) casein solution of above-mentioned adding papoid is put into 45 ℃ of water-baths are heated, hydrolysis, agitator stirs, and mixing speed is that per minute 60 changes, and hydrolysis was carried out 2 hours;
(5) use 1molL
-1HCl solution transfers to 4.6 with the casein solution pH value of hydrolysis, is warming up to 100 ℃ of heating then and carries out enzyme-deactivating in 20 minutes;
(6) casein solution with said hydrolyzed leaves heart processing 20 minutes with per minute 5000, discards residue, collects supernatant liquor;
(7) resulting supernatant liquor lyophilize or spraying drying.The shelf Heating temperature of freeze drier is 20 ℃ during lyophilize, takes out the lyophilize thing when loft drier pressure is reduced to 6 handkerchiefs～8 handkerchiefs, obtains casein antioxidant peptide.During spraying drying, flow obtains casein antioxidant peptide for per hour 150 kilograms, inlet temperature are that 160 ℃～200 ℃, temperature out are 70 ℃～80 ℃.
(8) with the casein antioxidant peptide dissolved in distilled water that obtains, the preparation mass volume ratio is the casein antioxidant peptide solution of 50% (W/V);
(9) add papoid in above-mentioned casein antioxidant peptide solution, the addition of papoid is every gram casein antioxidant peptide 500U enzyme activity unit;
(10) the casein antioxidant peptide solution of above-mentioned adding papoid is stirred under 30 ℃ condition handle, stirring velocity is that per minute 80 changes, and the treatment time is 5.6 hours; Be warming up to 100 ℃ of heating then and carried out enzyme-deactivating in 20 minutes;
(11) lyophilize of casein antioxidant peptide solution or the spraying drying after will handling.The shelf Heating temperature of freeze drier is 20 ℃ during lyophilize, when loft drier pressure is reduced to 6 handkerchiefs～8 handkerchiefs, take out the lyophilize thing, during spraying drying, flow obtains the high activity casein anti-oxidation peptide for per hour 150 kilograms, inlet temperature are that 160 ℃～200 ℃, temperature out are 70 ℃～80 ℃.
The high activity casein anti-oxidation peptide that adopts two-step approach of the present invention to prepare, method (the NsimbaRY of employing Nsimba etc., Kikuzaki H, Konishi Y.Antioxidant activity of various extracts and fractions of Chenopodiumquinoa and Amaranthus spp.seeds.Food Chemistry, 2008,106 (2): 760-766) carry out anti-oxidant activity and measure, obtain it and remove 2, the result of bitter diazanyl (DPPH) free radical of 2-phenylbenzene-1-ability is: TEAC (Trolox resistance to oxidation of equal value) is 90mmolg
-1, half of DPPH free radical presses down rate (IC
50) be 0.2mgmL
-1And under identical condition, the casein antioxidant peptide that a stage enzymatic hydrolysis method is prepared, the method for employing Nsimba etc. is carried out anti-oxidant activity mensuration, obtains its result who removes DPPH free radical ability to be: TEAC is 10mmolg
-1, half of DPPH free radical presses down rate (IC
50) be 1.8mgmL
-1The activity of high activity casein anti-oxidation peptide improves 8 times.
Claims (7)
1. the two-step approach of high activity casein anti-oxidation peptide prepares technology, it is characterized in that: utilize casein and papoid two-step approach to prepare the high activity casein anti-oxidation peptide.
2. the first step preparation method of high activity casein anti-oxidation peptide according to claim 1 is characterized in that:
(1) casein is carried out swelling with distilled water, constantly stir simultaneously and dissolve fully up to casein, the preparation mass volume ratio is the casein solution of 4%～6% (W/V);
(2) use 1molL
-1NaOH solution the pH value of casein solution is transferred to 5.0～8.0;
(3) add papoid in the casein solution of the intact pH value of above-mentioned adjusting, the addition of papoid is every gram casein 300U～600U enzyme activity unit;
(4) casein solution of above-mentioned adding papoid is put into 40 ℃～50 ℃ water-baths are heated, hydrolysis, agitator stirs, and mixing speed is that per minute 30 changes～90 and changes, and hydrolysis was carried out 1 hour～5 hours;
(5) use 1molL
-1HCl solution transfers to 4.6 with the casein solution pH value of hydrolysis, is warming up to 100 ℃ of heating then and carries out enzyme-deactivating in 20 minutes;
(6) casein solution with said hydrolyzed leaves heart processing 20 minutes with per minute 5000, discards residue, collects supernatant liquor;
(7) resulting supernatant liquor lyophilize or spraying drying.The shelf Heating temperature of freeze drier is 20 ℃ during lyophilize, takes out the lyophilize thing when loft drier pressure is reduced to 6 handkerchiefs～8 handkerchiefs, obtains casein antioxidant peptide.During spraying drying, flow obtains casein antioxidant peptide for per hour 150 kilograms, inlet temperature are that 160 ℃～200 ℃, temperature out are 70 ℃～80 ℃.
3. the first step preparation method according to claim 2 is characterized in that: the mass volume ratio of described casein solution is 5% (W/V) preferably, the preferably every gram casein of the addition of described papoid 500U enzyme activity unit.
4. the first step preparation method according to claim 2 is characterized in that: the pH value of described solution preferably is adjusted to 6.5, preferably 45 ℃ of described temperature of reaction, described hydrolysis time preferably 2 hours.
5. second one step preparation method of high activity casein anti-oxidation peptide according to claim 1 is characterized in that:
(1) with the casein antioxidant peptide dissolved in distilled water that obtains, the preparation mass volume ratio is the casein antioxidant peptide solution of 30%～70% (W/V);
(2) add papoid in above-mentioned casein antioxidant peptide solution, the addition of papoid is every gram casein antioxidant peptide 300U～600 enzyme activity units;
(3) the casein antioxidant peptide solution of above-mentioned adding papoid is stirred under 13 ℃～47 ℃ condition handle, stirring velocity is that per minute 60 changes～100 and changes, and the treatment time is 1 hour～10 hours; Be warming up to 100 ℃ of heating then and carried out enzyme-deactivating in 20 minutes;
(4) lyophilize of casein antioxidant peptide solution or the spraying drying after will handling.The shelf Heating temperature of freeze drier is 20 ℃ during lyophilize, when loft drier pressure is reduced to 6 handkerchiefs～8 handkerchiefs, takes out the lyophilize thing, obtains the high activity casein anti-oxidation peptide.During spraying drying, flow obtains the high activity casein anti-oxidation peptide for per hour 150 kilograms, inlet temperature are that 160 ℃～200 ℃, temperature out are 70 ℃～80 ℃.
6. second one step preparation method according to claim 5 is characterized in that: the mass volume ratio of described casein antioxidant peptide is 50% (W/V) preferably, the preferably every gram casein of the addition of described papoid 500U enzyme activity unit.
7. second one step preparation method according to claim 5 is characterized in that: preferably 30 ℃ of described treatment temps, preferably 5.6 hours described treatment time.
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| CN200910071495A CN101824453A (en) | 2009-03-05 | 2009-03-05 | Two-stage preparation technology of high activity casein antioxidant peptides |
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| CN200910071495A CN101824453A (en) | 2009-03-05 | 2009-03-05 | Two-stage preparation technology of high activity casein antioxidant peptides |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102787156A (en) * | 2012-08-30 | 2012-11-21 | 甘肃华羚生物技术研究中心 | Method for preparing antioxidant peptides from yak cheese protein |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992013964A1 (en) * | 1991-02-06 | 1992-08-20 | Novo Nordisk A/S | Protein preparations |
| CN1784145A (en) * | 2003-05-05 | 2006-06-07 | 荷兰联合利华有限公司 | Hydrolysed casein product comprising tripeptides ipp and/ or vpp |
-
2009
- 2009-03-05 CN CN200910071495A patent/CN101824453A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1992013964A1 (en) * | 1991-02-06 | 1992-08-20 | Novo Nordisk A/S | Protein preparations |
| CN1784145A (en) * | 2003-05-05 | 2006-06-07 | 荷兰联合利华有限公司 | Hydrolysed casein product comprising tripeptides ipp and/ or vpp |
Non-Patent Citations (2)
| Title |
|---|
| 杨倩: "大米类蛋白反应条件优化及其产物的氨基酸组成分析", 《中国粮油学报》 * |
| 鹿波: "固定化酶的制备与酪蛋白合成类蛋白的研究", 《中国优秀硕士学位论文全文数据库-工程科技I辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102787156A (en) * | 2012-08-30 | 2012-11-21 | 甘肃华羚生物技术研究中心 | Method for preparing antioxidant peptides from yak cheese protein |
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Application publication date: 20100908 |