CN102146427A - Method for preparing swine blood active peptide by microwave-accelerated enzymolysis - Google Patents
Method for preparing swine blood active peptide by microwave-accelerated enzymolysis Download PDFInfo
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- CN102146427A CN102146427A CN2010106132580A CN201010613258A CN102146427A CN 102146427 A CN102146427 A CN 102146427A CN 2010106132580 A CN2010106132580 A CN 2010106132580A CN 201010613258 A CN201010613258 A CN 201010613258A CN 102146427 A CN102146427 A CN 102146427A
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Abstract
The invention discloses a method for preparing swine blood active peptide by microwave-accelerated enzymolysis, belonging to the field of finish and deep processing and comprehensive utilization of animal byproducts. The method comprises the steps of anticoagulation of swine blood, centrifugal separation, ultrasonic wall breaking, microwave-accelerated enzymolysis, secondary enzymolysis, inactivation, membrane filtration and spray drying. The product can be used as a raw material or additive for various foods. Compared with the traditional methods, the protein molecules move violently due to microwave-accelerated enzymolysis, the enzymolysis velocity can be obviously improved and the enzymolysis time is shortened. The method has simple process, low cost and environment friendliness and is suitable for industrial production of deep processing of swine blood.
Description
Technical field
The invention belongs to a kind of method of quick preparation pig blood bioactive peptide, belong to the technical field of deep processing of livestock and poultry byproduct and comprehensive utilization.
Background technology
China's pig blood resource is abundant, but pig blood is in China's serious waste phenomenon.Roughing at present becomes blood meal or blood bean curd, and intensive processing mainly concentrates on produces protoheme, SOD and benefit iron product etc.Protein content height in the pig blood, but process for processing bioactive peptide product.Bioactive peptide not only has trophic function and biological activity, but also has by the characteristics of enteron aisle rapid absorption.Present industrial protein powder mainly is to adopt acid-base method or enzymolysis process, reduces but be to use acid-base method that amino acid nutrient is worth, and adopts enzymolysis process, and enzymolysis process needed more than ten hour even the longer time.Microwave is baking, drying, blanching, application is all being arranged aspect thawing, and the research on assistance enzymolysis is also more and more.The microwave-assisted enzymolysis can shorten enzymolysis time greatly, enhances productivity, and is convenient to suitability for industrialized production.
Summary of the invention
The object of the invention provides a kind of microwave-assisted enzymolysis porcine haemoglobin, produces the method for bioactive peptide.This method technology is simple, and the enzyme digestion reaction time is short.Described method comprises: the anti-freezing of pig blood, centrifugation, supersonic wave wall breaking, microwave-assisted enzymolysis, secondary enzymolysis, the enzyme that goes out, membrane filtration, spraying drying.
1) the qualified fresh pig blood of quarantine adds the anti-freezing of 0.3%-0.5% trisodium citrate;
2) pig blood centrifugation: centrifugal use high speed freezing centrifuge, 1 ℃-4 ℃ of centrifuging temperatures, centrifugal rotational speed 2500rpm-4000rpm, centrifugation time 15min-20min, centrifugal after, hemocyte adds with volume 0.9% normal saline flushing 1-3 time;
3) the broken born of the same parents of ultrasonic wave: ultrasonic power is 200w-400w, and the time is 10min-20min, and intermittently than adopting 2: 1,1: 1,1: 2, operation is carried out at normal temperatures;
4) microwave-assisted enzymolysis: microwave-assisted is enzymolysis for the first time, microwave power is at 500w-700w, temperature is at 45 ℃-60 ℃, the time of enzymolysis is 10min-20min, concentration of substrate is 7%-10%, and the NaOH adjusting pH to 8.0-10.0 with 1M adds the phosphate buffered saline buffer that pH is 8.0-10.0 again, adding enzyme activity is the Alcalase restriction endonuclease of 2.4AU-A/g, and the addition of enzyme is the 2%-7% of substrate;
5) secondary enzymolysis: after the first time, enzymolysis finished, adding enzyme activity again was that 200000U/g food grade flavor protease carries out enzymolysis second time, and enzyme concentration is 2% of a substrate, enzymolysis 1min-3min, microwave power and temperature with the first time enzymolysis identical, debittering;
6) enzyme that goes out: the enzyme that in 90 ℃ of-100 ℃ of water-baths, goes out, the enzyme time of going out is 5min-10m;
7) membrane filtration: adopting the aperture is the film of 3000Da-10000Da, enzymolysis solution is filtered the filtering macromolecular substance;
8) spraying drying: spraying drying feed liquid flow 6L/h-8L/h, 75 ℃-85 ℃ of feed temperatures, intake air temperature are at 120 ℃-140 ℃, and outlet wind-warm syndrome degree is at 70 ℃-85 ℃, and spraying drying obtains pig blood bioactive peptide powder.
Embodiment
Embodiment
The separation of 1 hemocyte
The qualified fresh pig blood of quarantining adds 0.33% trisodium citrate anti-freezing, slowly stirs.Use the high speed freezing centrifuge separating blood, under 4 ℃ of conditions, the centrifugal 15min of rotating speed with 3000rpm pours out supernatant liquor, adds equal-volume 0.9% physiological saline recentrifuge, and above-mentioned cleaning step repeats twice, obtains more purified hemocyte.
2 supersonic wave wall breakings
Abolish the cell walls of hemocyte with ultrasonic wave, adjustment power is 400w, intermittently than 2: 1, adds the two volumes deionized water, ultrasonication 20min.The broken wall operation is carried out at normal temperatures.Measure above-mentioned solution with ultraviolet-visible spectrophotometer, absorbancy rises to 0.408 from 0.102, and shell-broken effect is remarkable.
3 microwave-assisted enzymolysis
The microwave-assisted enzymolysis adopts the method for substep enzymolysis.Substrate behind the supersonic wave wall breaking adds the solution of distilled water furnishing concentration 7%, regulates substrate pH to 9.0 with the NaOH of 1M, adds pH again and be 9.0 phosphate buffered saline buffer, and adding enzyme activity is the Alcalase restriction endonuclease of 2.4AU-A/g, and enzyme concentration is 6% of a substrate.The enzyme digestion reaction solution for preparing is poured into the microwave reaction instrument, and the setting microwave power is 500w, and controlled temperature is at 50 ℃, and behind the enzymolysis 11min, enzymolysis efficiency can reach 29.74%.The food grade flavor protease that adds 2% enzyme activity again and be 200000U/g carries out secondary enzymolysis, enzymolysis 2min, debittering.After enzyme digestion reaction is finished, with 90 ℃ of water-bath constant temperature 10min enzyme that goes out.At last enzymolysis solution is used 3000Da membrane filtration, filtering macromolecular substance.Using efficient liquid phase chromatographic analysis filtrate, is the bioactive peptide less than 1000Da more than 80%.
4 spraying dryings
Enzymolysis solution behind the membrane filtration carries out spraying drying, and the feed liquid flow is 6.5L/h, and feed temperature is 80 ℃, and intake air temperature is 130 ℃, and the air outlet temperature is 75 ℃, and spraying drying obtains pig blood bioactive peptide powder.
Claims (4)
1. the present invention relates to the method that a kind of microwave-assisted enzymolysis prepares pig blood bioactive peptide, it is characterized in that described method comprises the steps:
A) supersonic wave wall breaking: the ultrasonic wave hemocyte wall power that breaks is 200w-400w, and the time is 10min-20min, intermittently than adopting 2: 1,1: 1,1: 2, operates and carries out at normal temperatures;
B) microwave-assisted enzymolysis: the microwave-assisted enzymolysis carries out in two steps, and twice enzymolysis microwave power is identical with temperature, and microwave power is at 500w-700w, and temperature is carried out the enzymolysis second time in the first time after the enzymolysis end at 45 ℃-60 ℃ again;
C) membrane filtration: adopting the aperture is the film of 3000Da-10000Da, enzymolysis solution is filtered the filtering macromolecular substance;
D) spraying drying: the spraying drying of pig blood bioactive peptide, feed liquid flow are at 6L/h-8L/h, and feed temperature is at 70 ℃-85 ℃, and intake air temperature is at 120 ℃-140 ℃, and the air outlet temperature is at 70 ℃-85 ℃, and spraying drying obtains pig blood bioactive peptide powder.
2. according to the described supersonic wave wall breaking of claim 1a, it is characterized in that ultrasonic wave institute disruptive hemocyte is after centrifugation, add with volume 0.9% physiological saline washing 1-3 time.
3. according to the claim 1b described first time of enzymolysis, it is characterized in that substrate protein white matter concentration is 7%-10%, regulate substrate pH to 8.0-10.0 with 1MNaOH, add the pH8.0-10.0 phosphate buffered saline buffer again, adding enzyme activity is the Alcalase restriction endonuclease of 2.4AU-A/g, the addition of enzyme is the 2%-7% of substrate, and enzymolysis time is 9min-14min.
4. according to the claim 1b described second time of enzymolysis, it is characterized in that the enzyme that is added is that vigor is a 200000U/g food grade flavor protease, enzyme concentration is 2% of a substrate, and enzymolysis time is 1min-3min.
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| CN2010106132580A CN102146427A (en) | 2010-12-30 | 2010-12-30 | Method for preparing swine blood active peptide by microwave-accelerated enzymolysis |
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| CN2010106132580A CN102146427A (en) | 2010-12-30 | 2010-12-30 | Method for preparing swine blood active peptide by microwave-accelerated enzymolysis |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103952458A (en) * | 2014-04-30 | 2014-07-30 | 桂林军供生化技术开发有限公司 | Method for preparing active peptides in duck blood through microwave-assisted enzymolysis |
| CN106086140A (en) * | 2016-08-10 | 2016-11-09 | 柳江县渡庄生物科技有限公司 | A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide |
| CN107245422A (en) * | 2017-08-03 | 2017-10-13 | 重庆市药物种植研究所 | A kind of deer-blood wine and preparation method thereof |
| CN107418992A (en) * | 2017-09-19 | 2017-12-01 | 广东雅道生物科技有限公司 | A kind of bone peptide extracting method |
| CN115010803A (en) * | 2021-03-03 | 2022-09-06 | 内蒙古天奇生物科技有限公司 | Preparation of hemoglobin polypeptide rich in heme iron |
| CN117126909A (en) * | 2023-08-15 | 2023-11-28 | 深圳中科生物药业有限公司 | Method for preparing hemoglobin peptide from animal blood and prepared hemoglobin peptide |
-
2010
- 2010-12-30 CN CN2010106132580A patent/CN102146427A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103952458A (en) * | 2014-04-30 | 2014-07-30 | 桂林军供生化技术开发有限公司 | Method for preparing active peptides in duck blood through microwave-assisted enzymolysis |
| CN106086140A (en) * | 2016-08-10 | 2016-11-09 | 柳江县渡庄生物科技有限公司 | A kind of method that microwave-assisted and membrane filtration prepare fresh-water fishes hemepeptide |
| CN107245422A (en) * | 2017-08-03 | 2017-10-13 | 重庆市药物种植研究所 | A kind of deer-blood wine and preparation method thereof |
| CN107245422B (en) * | 2017-08-03 | 2021-04-13 | 重庆市药物种植研究所 | Deer blood wine and preparation method thereof |
| CN107418992A (en) * | 2017-09-19 | 2017-12-01 | 广东雅道生物科技有限公司 | A kind of bone peptide extracting method |
| CN115010803A (en) * | 2021-03-03 | 2022-09-06 | 内蒙古天奇生物科技有限公司 | Preparation of hemoglobin polypeptide rich in heme iron |
| CN117126909A (en) * | 2023-08-15 | 2023-11-28 | 深圳中科生物药业有限公司 | Method for preparing hemoglobin peptide from animal blood and prepared hemoglobin peptide |
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Application publication date: 20110810 |