CN100398660C - Method for producing compound bioactive peptide utilizing casein enzyme process - Google Patents
Method for producing compound bioactive peptide utilizing casein enzyme process Download PDFInfo
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- CN100398660C CN100398660C CNB2005101019010A CN200510101901A CN100398660C CN 100398660 C CN100398660 C CN 100398660C CN B2005101019010 A CNB2005101019010 A CN B2005101019010A CN 200510101901 A CN200510101901 A CN 200510101901A CN 100398660 C CN100398660 C CN 100398660C
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Abstract
The present invention provides a compound bioactive peptide producing method by using a casein enzyme method. In the present invention, casein is used for preparing enzymolysis substrate, compound enzyme-trypsin and neutral proteinase are used for enzymolysis, and the degree of hydrolysis is controlled at 18% to 20%; then enzyme killing operation and vacuum pressure reducing concentration are carried out on hydrolyzate, and spray-drying is carried out on concentrated solution; consequently, white-milky powdery compound active peptide finished products are obtained. The product produced by the present invention contains the compound bioactive peptide with high activity and can be used as an additive of health foods.
Description
Technical field
The present invention relates to a kind of method of utilizing casein enzyme process to produce compound bioactive peptide.
Background technology
With enzymolysis process is the common method that obtains the biological activity peptide product by proteolytic enzyme protolysate.CN1208565 discloses a kind of " milk protein active peptide preparation method ", and CN1101471C discloses a kind of " preparation method of active peptide of soybean protein ", and these methods are raw material with fresh milk or skim-milk or soybean, and production cost is higher, and technology is also complicated.
Summary of the invention
The objective of the invention is to propose a kind of wide material sources, inexpensive casein food grade raw material of utilizing, with the novel method of strong, the safe compound bioactive peptide product of prozyme solution production biological activity.
Processing step of the present invention is as follows:
(1) preparation enzymolysis substrate: with casein food grade with 4～6 times of pure water swellings to the casein food grade quality, transfer pH to 8.0-9.0 with NaOH again, then solution is put to heat in 50～60 ℃ of water baths and be stirred to casein food grade and dissolved, use HCl adjust pH to 7.0～7.3 again, with pure water solution is settled at last that the casein food grade mass concentration is 6～10% in the solution, is required enzymolysis substrate;
(2) in the enzymolysis substrate, add trypsinase and neutral protease, place 50～55 ℃ of water-baths to carry out enzyme digestion reaction, tryptic add-on be in the enzymolysis substrate casein food grade quality 0.8～1.5%, the add-on of neutral protease be in the enzymolysis substrate casein food grade quality 1.0～2.0%;
(3) when the degree of hydrolysis of hydrolyzed solution reaches 18%～20%, enzyme goes out;
(4) hydrolyzed solution that goes out behind the enzyme concentrates through vacuum decompression, and concentrated solution is spray-dried, promptly gets in vain～oyster white powdery compound bioactive peptide finished product.
The present invention is special, and the selection process step that proposes is as follows:
(1) preparation enzymolysis substrate: with casein food grade with 4 times of pure water swellings to the casein food grade quality, be the NaOH accent pH to 8.0-9.0 of 6mol/L again with concentration, then solution is put to heat in 60 ℃ of water baths and be stirred to casein food grade and dissolved, be the HCl adjust pH to 7.2 of 1mol/L again with concentration, with pure water solution is settled at last that the casein food grade mass concentration is 8% in the solution, is required enzymolysis substrate;
(2) in the enzymolysis substrate, add trypsinase and neutral protease, place 55 ℃ of water-baths to carry out enzyme digestion reaction 4～6 hours, tryptic add-on be in the enzymolysis substrate casein food grade quality 1%, the add-on of neutral protease be in the enzymolysis substrate casein food grade quality 1.5%;
(3) when the degree of hydrolysis of hydrolyzed solution reaches 18%～20%, enzyme goes out;
(4) hydrolyzed solution that goes out behind the enzyme concentrates through vacuum decompression, and concentrated solution is spray-dried, promptly gets in vain～oyster white powdery composite reactive peptide finished product.
In above-mentioned processing step, when (2) step process finishes,, need measure the degree of hydrolysis of hydrolyzed solution for reaching the condition that enters next processing step, when the degree of hydrolysis of measuring reaches 18%～20%, enter next step again.The mensuration of degree of hydrolysis can adopt following method: use Kai Shi trace nitriding to measure the substrate solution total nitrogen content, use two indicator formol titration to measure the amino nitrogen content of hydrolysis substrate solution hydrolysis front and back respectively, calculate degree of hydrolysis by following formula again:
The present invention utilizes casein food grade to pass through enzymolysis process, controls suitable degree of hydrolysis, and the product of producing contains active high compound bioactive peptide.The present invention is because in enzymolysis process, adopt combined-enzyme method, promptly carry out complex enzyme hydrolysis, microbiology class proteolytic enzyme-neutral protease wherein with trypsinase and neutral protease, it is strong to have hydrolysis ability, efficient height, and inexpensive characteristic, and animal proteinoid enzyme-trypsinase has the strong characteristics of hydrolysate biological activity, the combination of these two kinds of proteolytic enzyme, the complementary synergy of two enzyme different qualities, it is low to have solved use single enzymolysis efficient preferably, the difficult problem that cost is high.Hydrolysate behind concentrate drying, safe, the human or animal's that biological activity is strong immunostimulant can be used as the additive of heath food.
Technical scheme of the present invention, its rich raw material sources, cheapness, technology is simple, and mild condition is free from environmental pollution, is convenient to suitability for industrialized production.
Embodiment
(1) preparation of enzymolysis substrate:
Casein food grade 80g, adding distil water 320ml swelling is transferred pH to 8.0-9.0 with 6mol/L NaOH, heats to 60 ℃ and makes the casein food grade dissolving, transfers pH to 7.2 with 1mol/L HCl again, is settled to 1000ml at last.
(2) enzymolysis:
Trypsinase 0.8g, neutral protease 1.2g is dissolved in (the limit edged slowly is stirred to even) in the substrate solution for preparing equably.Enzymolysis solution is in 6 hours (reaction process is often stirred hydrolyzate or stirred automatically 50 times/minute) of 55 ℃ of water bath with thermostatic control reactions.
When hydrolysis finishes, degree of hydrolysis is measured:
Measure the substrate total nitrogen with micro-nitriding.When hydrolysis finishes, use Kai Shi trace nitriding to measure the substrate solution total nitrogen content, use two indicator formol titration to measure the content of hydrolysis front and back amino nitrogen in the substrate solution respectively, calculate degree of hydrolysis by following formula again.
When the degree of hydrolysis of hydrolyzed solution is 18～20%, be qualified, can enter next step.
(3) enzyme that goes out: temperature of reaction is risen to 80 ℃, keep the enzyme that went out in 20 minutes and live, cooling again.
(4) hydrolyzed solution that goes out behind the enzyme concentrates through vacuum decompression, and concentrated solution is spray-dried, promptly gets in vain～oyster white powdery composite reactive peptide finished product.
Claims (3)
1. method of utilizing casein enzyme process to produce compound bioactive peptide is characterized in that processing step is as follows:
(1) preparation enzymolysis substrate: with casein food grade with 4～6 times of pure water swellings to the casein food grade quality, transfer pH to 8.0-9.0 with NaOH again, then solution is put to heat in 50～60 ℃ of water baths and be stirred to casein food grade and dissolved, use HCl adjust pH to 7.0～7.3 again, with pure water solution is settled at last that the casein food grade mass concentration is 6～10% in the solution, is required enzymolysis substrate;
(2) in the enzymolysis substrate, add trypsinase and neutral protease, place 50～55 ℃ of water-baths to carry out enzyme digestion reaction, tryptic add-on be in the enzymolysis substrate casein food grade quality 0.8～1.5%, the add-on of neutral protease be in the enzymolysis substrate casein food grade quality 1.0～2.0%;
(3) when the degree of hydrolysis of hydrolyzed solution reaches 18%～20%, enzyme goes out;
(4) hydrolyzed solution that goes out behind the enzyme concentrates through vacuum decompression, and concentrated solution is spray-dried, promptly gets in vain～oyster white powdery composite reactive peptide finished product.
2. the method for utilizing casein enzyme process to produce compound bioactive peptide according to claim 1 is characterized in that described processing step is:
(1) preparation enzymolysis substrate: with casein food grade with 4 times of pure water swellings to the casein food grade quality, be the NaOH accent pH to 8.0-9.0 of 6mol/L again with concentration, then solution is put to heat in 60 ℃ of water baths and be stirred to casein food grade and dissolved, be the HCl adjust pH to 7.2 of 1mol/L again with concentration, with pure water solution is settled at last that the casein food grade mass concentration is 8% in the solution, is required enzymolysis substrate;
(2) in the enzymolysis substrate, add trypsinase and neutral protease, place 55 ℃ of water-baths to carry out enzyme digestion reaction 4～6 hours, tryptic add-on be in the enzymolysis substrate casein food grade quality 1%, the add-on of neutral protease be in the enzymolysis substrate casein food grade quality 1.5%;
(3) when the degree of hydrolysis of hydrolyzed solution reaches 18%～20%, enzyme goes out;
(4) hydrolyzed solution that goes out behind the enzyme concentrates through vacuum decompression, and concentrated solution is spray-dried, promptly gets in vain～oyster white powdery composite reactive peptide finished product.
3. the method for utilizing casein enzyme process to produce compound bioactive peptide according to claim 1 and 2, it is characterized in that when (2) step process finishes, earlier the degree of hydrolysis of hydrolyzed solution is measured, when the degree of hydrolysis of measuring reaches 18%～20%, enter next step again, the measuring method of degree of hydrolysis is as follows: measure the substrate solution total nitrogen content with Kai Shi trace nitriding, measure amino nitrogen content before and after the hydrolysis of hydrolysis substrate solution respectively with two indicator formol titration, calculate degree of hydrolysis by following formula again:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CNB2005101019010A CN100398660C (en) | 2005-12-07 | 2005-12-07 | Method for producing compound bioactive peptide utilizing casein enzyme process |
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| CNB2005101019010A CN100398660C (en) | 2005-12-07 | 2005-12-07 | Method for producing compound bioactive peptide utilizing casein enzyme process |
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| CN1814777A CN1814777A (en) | 2006-08-09 |
| CN100398660C true CN100398660C (en) | 2008-07-02 |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102150741B (en) * | 2011-03-07 | 2013-08-21 | 马忠仁 | Method for preparing lactalbumin hydrolysate by utilizing yak milk |
| CN105010755A (en) * | 2015-06-18 | 2015-11-04 | 广东碧德生物科技有限公司 | Biological active peptide preparation for aquatic products as well as preparation method and application of biological active peptide preparation |
| CN110904179A (en) * | 2019-12-10 | 2020-03-24 | 临夏州华安生物制品有限责任公司 | Production method of high-content and low-bitter casein phosphopeptide |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1378863A (en) * | 2002-04-29 | 2002-11-13 | 徐佳立 | Active casein phosphopeptide and its preparing process |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1378863A (en) * | 2002-04-29 | 2002-11-13 | 徐佳立 | Active casein phosphopeptide and its preparing process |
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