CN101130800A - Method for preparing antihypertensive peptide by enzymolysis of hairtail chine - Google Patents
Method for preparing antihypertensive peptide by enzymolysis of hairtail chine Download PDFInfo
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- CN101130800A CN101130800A CNA2007100710547A CN200710071054A CN101130800A CN 101130800 A CN101130800 A CN 101130800A CN A2007100710547 A CNA2007100710547 A CN A2007100710547A CN 200710071054 A CN200710071054 A CN 200710071054A CN 101130800 A CN101130800 A CN 101130800A
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Abstract
The present invention discloses a method for preparing peptide for reducing blood pressure by utilizing zymolysis of hairtail bone. Said method includes the following steps: pulverizing hairtail bone, adding water and uniformly stirring them, regulating its pH value to 2.0-4.0 (generally by using hydrochloric acid), heating to 40deg.C-50deg.C, retaining for 10-20 min, then adding acid protease, continuously thermo-insulating and continuously stirring to make zymolysis reaction for 4-8hr, then quickly heating zymolyte to 90-100deg.C, retaining for 10-15min, quickly cooling to room temperature and stopping reaction, making centrifugalization, taking out supernatant fluid, filtering, vacuum concentration, freeze-drying so as to obtain powdered peptide for reducing blood pressure.
Description
Technical field
The invention belongs to technical field of bioengineering, relate to medicine biological technique or functional foodstuff process for processing technology, relate in particular to and a kind ofly utilize the hairtail spine to prepare the method for blood pressure lowering peptide with enzymolysis process.
Background technology
Zinc metallopeptidase Zace1 (ACE) is played the part of important role in the blood pressure regulation process.It can make the Angiotensin decapeptide precursor conversion of non-activity is Angiotensin, thereby makes elevation of blood pressure.It is a kind of in the angiotensin converting enzyme inhibitor (ACEI) that Zinc metallopeptidase Zace1 (ACE) suppresses bioactive peptide, reaches the effect that brings high blood pressure down by the activity that suppresses ACE.The inhibitor of the synthetic peptide medicament of number of chemical as ACE arranged at present.The shortcoming of the synthetic Altace Ramipril of this class is that action time is not long, and blood pressure rebounds easily, and medicine is discharged the damage that causes renal function easily through kidney.
From natural protein, extract and have the functional peptides of trophicity and physiologically active, and with these functional peptides be applied in the middle of protective foods and the enriched food not only can full use food and food-processing in high-quality protein, and can effectively improve the added value of the protein raw material of wide material sources, promote the scientific and technological content of traditional food simultaneously greatly.
Food protein has the edible safety height, has no side effect through the blood pressure lowering peptide that enzymolysis produces, and particularly it has hypotensive effect to the hyperpietic, but to the advantage that the blood pressure normal people does not have hypotensive effect, is subjected to extensive concern in recent years.People have successfully obtained ace inhibitory peptide from krill, tuna, sardines, stripped tuna, corn, soybean, vinasse, silkworm chrysalis, casein at present.But above-mentioned research separation method cost height, the cost of large-scale production is also higher, and raw material mostly is people's life food.In the nineties later stage, both at home and abroad mainly be centered around proteinic degree of hydrolysis, molecular weight size, free aminoacid content, biological enzyme recycling and to the control of bitter taste and removal etc. about the research of proteolysis peptide.
Hairtail (Trichiurus japonicus) belongs to Perciformes, Trichiuridae, hairtail genus, popular name hairtail, opotism fish, tooth band, leukorrhea fish etc., be the warm water fishes of cluster, be distributed widely in marine sites such as China, Korea, Japan, Indonesia, Philippines, India, African eastern bank and Red sea in bottom.According to statistics, this fingerling is the highest in the catch of China, accounts for the 70%-80% of world's fish catch of the same race, occupies 1 of the single fingerling fishery harvesting of Marine Fishery of China flow control; And East Sea district hairtail catch is the highest, accounts for the 80%-90% of China with catch by species, is the most important object of fishing for of East Sea district fish production.
Traditional medicine thinks, hairtail is warm in nature, flavor is sweet, can warm up stomach, damp skin, tonifying Qi, nourish blood, fitness, it is edible that suitable weakness due to chronic disease, deficiency of blood dizziness, shortness of breath and fatigue, food are won the people of thin, malnutritive people and xerosis cutis less." food Chinese medicine is with just square " cloud: hairtail. enriching yin. nourishing the liver.The acute chronic enteritis steamed wheaten foods can improve symptom.Often edible hairtail. have the effect of help the five internal organs.
The hairtail spine is the main tankage after the hairtail processing, is commonly called as hairtail steck.It is mainly used in fodder industry at present, still belongs to blank for utilization that utilizes the hairtail spine and deep processing.
Summary of the invention
The invention provides a kind of is the method for feedstock production to the inhibited blood pressure lowering peptide of Zinc metallopeptidase Zace1 (ACE) with the hairtail spine.
A kind of method of preparing antihypertensive peptide by enzymolysis of hairtail chine, comprise the steps: to stir evenly with adding water after the pulverizing of hairtail spine, the adjusting pH value is 2.0-4.0 (generally using hydrochloric acid), temperature is risen to 40-50 ℃, keep adding aspartic protease (acid fungal protease after 10-20 minute, the effective PH of this enzyme is 2.0-6.0, temperature is 40-55 ℃, the typical application scope is baking, protein processing, fruit juice is produced and drinks production, it can strengthen the hydrolysis of protein fermentation media, fish, the processing of meat hydrolyzate and whole fish, it also can hydrolyzed soy etc. plant protein, processing soluble proteins product), continues also continuous stirring of insulation and carried out enzyme digestion reaction 4-8 hour, then enzymolysis solution is warming up to 90-100 ℃ rapidly, keep after 10-15 minute, be cooled to the room temperature termination reaction rapidly, get supernatant liquid filtering after centrifugal, vacuum concentration, lyophilize obtains the powdery blood pressure lowering peptide.
As preferably, after pulverizing, the hairtail spine stirs evenly with water, and the adjusting pH value is 2.5-2.8, behind the adjusting pH value temperature is risen to 45 ℃ (vigor of 45 ℃ of enzymes reaches highest level, and is the most favourable to enzymolysis).
When described hairtail spine added water and stirs evenly, the quality of hairtail spine and the volume ratio of water were 1: 2-7g/ml; Preferred 1: 4.22g/ml.
As preferably, temperature is risen to 40-50 ℃, keep adding aspartic protease after 20 minutes.
Described centrifugal under the condition of 2500-3000r/min centrifugal 20-30 minute, preferred centrifugal under the condition of 3000r/min centrifugal 30 minutes.
As preferably, the time of described enzyme digestion reaction is 6 hours.Reaction times is short, and enzymolysis is not thorough; Overlong time, the amino acid of generation increases.
The quality of described aspartic protease (enzyme activity is 5000U/g) is the 0.5-2% of albumen weight in the raw material hairtail spine.Enzyme concentration is too much, and reaction is not had too big promotion, wastes enzyme again; Enzyme-added very few, substrate utilization is incomplete.With 1% of albumen weight in the raw material hairtail spine is the best.
The advantage of the inventive method is:
1. the prepared blood pressure lowering peptide of the present invention is that the hairtail spine gets through biological enzyme hydrolysis, have and suppress the active effect of ACE, the hyperpietic is had hypotensive effect, safely, have no side effect, can play a role in health care to the normotensive, normal arterial pressure is not had influence.
2. raw material sources of the present invention are abundant, and are cheap, and process technical transform of the present invention can effectively improve the economic value added of this aquatic products processing tankage of hairtail spine, promote the development of fish processing industry that important meaning is arranged to continuing.
3. product of the present invention can be used as medicine, protective foods, food, foodstuff additive, medicament synergistic agent etc., and craft science is reasonable, and is simple to operate, has stronger industrializing implementation.
Embodiment
Embodiment 1
Take by weighing the hairtail spine (moisture 76.22% after the homogenate, protein content 12.17%, Ningbo City, Zhejiang Province superstar marine organisms Products Co., Ltd provides) 10kg, adding water 42kg, to stir evenly the back be that the HCl adjusting pH value of 6N is 2.5-2.8 with volumetric molar concentration, temperature is elevated to 45 ℃, keep after 20 minutes, 0.12% of hairtail spine weight aspartic protease 0.012kg (enzyme activity is 5000U/g) in the adding raw material continues insulation and continuous stirring was carried out enzyme digestion reaction 6 hours.Adopt enzymolysis solution to be warming up to 90-100 ℃ rapidly then, keep being cooled to room temperature rapidly and coming termination reaction after 10-15 minute.Under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 3000r/min centrifugal 30 minutes, supernatant liquor 30kg.With common filter paper filtering, then utilize tubular fibre polysulphones hyperfiltration membrane (molecular weight cut-off 10KDa) that the coarse filtration enzymolysis solution is carried out ultrafiltration, operating pressure 0.2MPa, working temperature obtains film and sees through liquid 23kg less than 40 ℃.Utilize vacuum decker that ultrafiltration is seen through enzymolysis solution and be concentrated into 3kg, carry out lyophilize, obtain having the peptide powder 0.45kg of antihypertensive function, peptide content is 91.95%, and the product yield is 4.5%, the tint lemon yellow exquisiteness, utilize vitro detection ACE to suppress active CUSHMAN method, record its IC
50Be 0.826mg/ml.
Embodiment 2
Take by weighing the hairtail spine (moisture 76.22% of pulverizing, protein content 12.17%, Ningbo City, Zhejiang Province superstar marine organisms Products Co., Ltd provides) 25kg, adding water 50kg, to stir evenly the back be that the HCl adjusting pH value of 6N is 3.0 with volumetric molar concentration, temperature is elevated to 40 ℃, keep after 20 minutes, 0.12% of hairtail spine weight aspartic protease 0.03kg (enzyme activity is 5000U/g) in the adding raw material continues insulation and continuous stirring was carried out enzyme digestion reaction 3 hours.Adopt enzymolysis solution to be warming up to 90-100 ℃ rapidly then, keep being cooled to room temperature rapidly and coming termination reaction after 10-15 minute.Under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 3000r/min centrifugal 20 minutes, supernatant liquor 42.4kg.With common filter paper filtering, then utilize tubular fibre polysulphones hyperfiltration membrane (molecular weight cut-off 10KDa) that the coarse filtration enzymolysis solution is carried out ultrafiltration, operating pressure 0.2MPa, working temperature obtains film and sees through liquid 35.3kg less than 40 ℃.Utilize vacuum decker that ultrafiltration is seen through enzymolysis solution and be concentrated into 4kg, carry out lyophilize, obtain having the peptide powder 0.69kg of antihypertensive function, peptide content is 76.42%, and the product yield is 2.76%, color and luster oyster white exquisiteness, utilize vitro detection ACE to suppress active CUSHMAN method, record its IC
50Be 4.831mg/ml.
Embodiment 3
Take by weighing the hairtail spine (moisture 76.22% of pulverizing, protein content 12.17%, Ningbo City, Zhejiang Province superstar marine organisms Products Co., Ltd provides) 15kg, adding water 90kg, to stir evenly the back be that the HCl adjusting pH value of 6N is 3.0 with volumetric molar concentration, temperature is elevated to 50 ℃, 0.06% of hairtail spine weight aspartic protease 0.009kg (enzyme activity is 5000U/g) in the adding raw material continues insulation and continuous stirring was carried out enzyme digestion reaction 4 hours.Adopt enzymolysis solution to be warming up to 90-100 ℃ rapidly then, keep being cooled to room temperature rapidly and coming termination reaction after 10-15 minute.Under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 2500r/min centrifugal 30 minutes, supernatant liquor 70.3kg.With common filter paper filtering, then utilize tubular fibre polysulphones hyperfiltration membrane (molecular weight cut-off 10KDa) that the coarse filtration enzymolysis solution is carried out ultrafiltration, operating pressure 0.2MPa, working temperature obtains film and sees through liquid 62.9kg less than 40 ℃.Utilize vacuum decker that ultrafiltration is seen through enzymolysis solution and be concentrated into 5kg, carry out lyophilize, obtain having the peptide powder 0.43kg of antihypertensive function, peptide content is 80.76%, and the product yield is 2.87%, color and luster oyster white exquisiteness, utilize vitro detection ACE to suppress active CUSHMAN method, record its IC
50Be 3.794mg/ml.
Claims (10)
1. the method for a preparing antihypertensive peptide by enzymolysis of hairtail chine, comprise the steps: to stir evenly with adding water after the pulverizing of hairtail spine, the adjusting pH value is 2.0-4.0, temperature is risen to 40-50 ℃, keep adding aspartic protease after 10-20 minute, continuing also continuous stirring of insulation carried out enzyme digestion reaction 4-8 hour, then enzymolysis solution is warming up to 90-100 ℃ rapidly, keep after 10-15 minute, be cooled to the room temperature termination reaction rapidly, get supernatant liquid filtering, vacuum concentration, lyophilize after centrifugal, obtain the powdery blood pressure lowering peptide.
2. the method for claim 1 is characterized in that: described hairtail spine is pulverized the back and is stirred evenly with water, and the adjusting pH value is 2.5-2.8.
3. the method for claim 1 is characterized in that: after regulating pH value temperature is risen to 45 ℃.
4. the method for claim 1, it is characterized in that: the quality of described hairtail spine and the volume ratio of water are 1: 2-7g/ml.
5. method as claimed in claim 4 is characterized in that: the quality of described hairtail spine and the volume ratio of water are for shining 1: 4.22g/ml.
6. the method for claim 1 is characterized in that: temperature is risen to 40-50 ℃, keep adding aspartic protease after 20 minutes.
7. the method for claim 1 is characterized in that: described centrifugal under the condition of 2500-3000r/min centrifugal 20-30 minute.
8. method as claimed in claim 7 is characterized in that: described centrifugal under the condition of 3000r/min centrifugal 30 minutes.
9. the method for claim 1, it is characterized in that: the time of described enzyme digestion reaction is 6 hours.
10. the method for claim 1, it is characterized in that: the quality of described aspartic protease is the 0.5-2% of albumen weight in the raw material hairtail spine.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100710547A CN101130800A (en) | 2007-09-01 | 2007-09-01 | Method for preparing antihypertensive peptide by enzymolysis of hairtail chine |
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| CNA2007100710547A CN101130800A (en) | 2007-09-01 | 2007-09-01 | Method for preparing antihypertensive peptide by enzymolysis of hairtail chine |
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| CN101130800A true CN101130800A (en) | 2008-02-27 |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101906455A (en) * | 2010-07-22 | 2010-12-08 | 东北农业大学 | Preparation method of egg white protein assistant antihypertensive peptides |
| CN102115491A (en) * | 2010-11-24 | 2011-07-06 | 河南工业大学 | Method for producing antihypertensive peptide by taking egg yolk as raw material |
| CN102115490A (en) * | 2010-11-24 | 2011-07-06 | 河南工业大学 | Method for producing blood pressure depressed peptide by using egg white as raw material |
| CN103014105A (en) * | 2012-06-11 | 2013-04-03 | 四川农业大学 | Method for preparing antihypertensive peptide by using porcine thighbones |
| WO2014194530A1 (en) * | 2013-06-08 | 2014-12-11 | 广州世优生物科技有限公司 | Method for preparing ace inhibitory peptides from hairtail leftovers |
| CN104694615A (en) * | 2015-03-03 | 2015-06-10 | 江南大学 | Method for preparing antihypertensive peptides by utilizing probiotic fermented marine organism processing leftovers |
| CN109206479A (en) * | 2017-09-18 | 2019-01-15 | 北京中医药大学 | A kind of vinegar beans glutelin source Antihypertensive Peptides and its application |
| CN113508828A (en) * | 2021-07-08 | 2021-10-19 | 泉州师范学院 | Preparation method of mackerel antihypertensive active peptide and application of mackerel antihypertensive active peptide in bread |
| CN114451505A (en) * | 2022-02-21 | 2022-05-10 | 福建农林大学 | Anti-fatigue beverage added with glycoprotein |
-
2007
- 2007-09-01 CN CNA2007100710547A patent/CN101130800A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101906455A (en) * | 2010-07-22 | 2010-12-08 | 东北农业大学 | Preparation method of egg white protein assistant antihypertensive peptides |
| CN101906455B (en) * | 2010-07-22 | 2013-07-31 | 东北农业大学 | Preparation method of egg white protein assistant antihypertensive peptides |
| CN102115491A (en) * | 2010-11-24 | 2011-07-06 | 河南工业大学 | Method for producing antihypertensive peptide by taking egg yolk as raw material |
| CN102115490A (en) * | 2010-11-24 | 2011-07-06 | 河南工业大学 | Method for producing blood pressure depressed peptide by using egg white as raw material |
| CN103014105A (en) * | 2012-06-11 | 2013-04-03 | 四川农业大学 | Method for preparing antihypertensive peptide by using porcine thighbones |
| WO2014194530A1 (en) * | 2013-06-08 | 2014-12-11 | 广州世优生物科技有限公司 | Method for preparing ace inhibitory peptides from hairtail leftovers |
| CN104694615A (en) * | 2015-03-03 | 2015-06-10 | 江南大学 | Method for preparing antihypertensive peptides by utilizing probiotic fermented marine organism processing leftovers |
| CN109206479A (en) * | 2017-09-18 | 2019-01-15 | 北京中医药大学 | A kind of vinegar beans glutelin source Antihypertensive Peptides and its application |
| CN109206479B (en) * | 2017-09-18 | 2021-10-29 | 北京中医药大学 | Vinegar bean gluten source antihypertensive peptide and application thereof |
| CN113508828A (en) * | 2021-07-08 | 2021-10-19 | 泉州师范学院 | Preparation method of mackerel antihypertensive active peptide and application of mackerel antihypertensive active peptide in bread |
| CN114451505A (en) * | 2022-02-21 | 2022-05-10 | 福建农林大学 | Anti-fatigue beverage added with glycoprotein |
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Open date: 20080227 |