CN1896267A - Preparation of depressor peptide by silkworm chrysalis - Google Patents
Preparation of depressor peptide by silkworm chrysalis Download PDFInfo
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- CN1896267A CN1896267A CN 200610052094 CN200610052094A CN1896267A CN 1896267 A CN1896267 A CN 1896267A CN 200610052094 CN200610052094 CN 200610052094 CN 200610052094 A CN200610052094 A CN 200610052094A CN 1896267 A CN1896267 A CN 1896267A
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- silkworm chrysalis
- blood pressure
- enzymolysis solution
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Abstract
The present invention discloses a method of preparing antihypertensive peptide from silkworm pupa which includes the following steps: the defatted dry powder of silkworm pupa or silkworm pupa protein powder is mixed with water at the mass ratio of 1:6-7, the pH value is adjusted to 6.0-6.5, then neutral proteinase is added for hydrolysis at 50-55DEG C for 120-200 minutes, the result hydrolysate is antihypertensive peptide. The weight of the said neutral proteinase is 0.9-1.1% of the defatted dry powder of silkworm pupa or silkworm pupa protein powder. The method in the present invention that utilizes silkworm pupa to prepare antihypertensive peptide has lower cost and better effect.
Description
Technical field
The invention belongs to technical field of bioengineering, be specifically related to utilize silkworm chrysalis to prepare the method for blood pressure lowering peptide.
Background technology
Zinc metallopeptidase Zace1 (ACE) is played the part of important role in the blood pressure regulation process.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 these functional peptides are applied in 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 step-down 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, 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.
Silkworm chrysalis (Silkworm pupa) is the pupa of Bombycidae insect bombyx mori (Bombyx moil L.), is a kind of cuisines of high nutrition, also is a kind of Chinese medicine, has the effect with taste, wind-damp dispelling, long yang-energy, just obtains widespread use since ancient times.45.5 ten thousand tons of cocoons, about 60,000 tons of raw silk were produced in the whole nation in 2000.Just estimated to reach 550,000 tons to cocoon yield in 2005 in " Tenth Five-year plan of silk silk fabric industry " of calendar year 2001 China national economic and commercial committee issue, raw silk output reaches 6.2 ten thousand tons; By 2015, cocoon yield will be above 600,000 tons, and the raw silk annual production will reach more than 80,000 tons.Usually 1 ton of raw silk of reel silk from cocoons system can get 1.5 tons of dried silkworm chrysalises, and silkworm chrysalis output is considerable.While is along with the silk industry further developing. and the output of silkworm chrysalis will be bigger, therefore develops the silkworm chrysalis resource and have great importance and wide prospect.
At present domestic less for utilizing food proteins to prepare the blood pressure lowering peptide report, adopt filature not meet generation after extensive, the cheap protein source of this raw material of dried silkworm chrysalis of degreasing prepares the technology of blood pressure lowering peptide.
Summary of the invention
At the deficiencies in the prior art part, the invention provides low, the effective silkworm chrysalis that utilizes of a kind of cost and produce the method for blood pressure lowering peptide.
The present invention is for reaching above purpose, be to realize: a kind of method of utilizing silkworm chrysalis to produce blood pressure lowering peptide is provided by such technical scheme, may further comprise the steps: with the dried dried silkworm chrysalis meal of degreasing or pupa albumen powder and water according to 1: 6～7 weight ratio mixed after, regulating the pH value earlier is 6.0～6.5, add neutral protease again and carry out enzyme digestion reaction, the gained enzymolysis solution is blood pressure lowering peptide; The weight of described neutral protease is 0.9%～1.1% of dried dried silkworm chrysalis meal of degreasing or pupa albumen powder, 50 ℃～55 ℃ of enzyme digestion reaction temperature, 120～200 minutes enzyme digestion reaction time.
As a kind of improvement that utilizes the method that silkworm chrysalis produces blood pressure lowering peptide of the present invention: regulate after pH value is 6.0～6.5, be warming up to 50～55 ℃ and kept this temperature 18～22 minutes earlier, and then the adding neutral protease carries out enzyme digestion reaction.
As the further improvements in methods of utilizing silkworm chrysalis to produce blood pressure lowering peptide of the present invention: after the enzyme digestion reaction time arrives, the enzyme digestion reaction product is warming up to 85 ℃ and kept this temperature 10～15 minutes rapidly, is cooled to 0～30 ℃ more rapidly with enzymolysis reaction; The centrifugal then enzymolysis solution that promptly gets.
As the further improvements in methods of utilizing silkworm chrysalis to produce blood pressure lowering peptide of the present invention: adopt gac to the enzymolysis solution processing of decolouring, activated carbon dosage is the 5-30% of enzymolysis solution gross weight, 55 ℃ of temperature; Carry out again getting the coarse filtration enzymolysis solution after coarse filtration is handled.
As the further improvements in methods of utilizing silkworm chrysalis to produce blood pressure lowering peptide of the present invention: under the working temperature of the operating pressure of 0.1～0.2Mpa and 0～45 ℃, adopt ultra-filtration membrane to carry out uf processing the coarse filtration enzymolysis solution, the ultrafiltration enzymolysis solution; Again this ultrafiltration enzymolysis solution is carried out vacuum concentration, lyophilize successively.
As the further improvements in methods of utilizing silkworm chrysalis to produce blood pressure lowering peptide of the present invention: this ultra-filtration membrane molecular weight cut-off is 30 kilodaltons (Kda).
As the further improvements in methods of utilizing silkworm chrysalis to produce blood pressure lowering peptide of the present invention: the enzyme activity 1.5AU/g of neutral protease.
The method of utilizing silkworm chrysalis to produce blood pressure lowering peptide of the present invention has following advantage:
1. the prepared blood pressure lowering peptide of the present invention is that the dried silkworm chrysalis of degreasing 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 extensive, cheap, and process technical transform of the present invention can effectively improve the added value of this by product of Silk Mill's silkworm chrysalis, promote the development of silkworm 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 industrial implementation.
Embodiment
Embodiment 1,
Take by weighing the dried dried silkworm chrysalis meal of degreasing (moisture 1.2%, protein content 66.23% are bought in the Zhejiang silk one factory) 50kg after the pulverizing, adding water 320kg, to stir evenly the back be 6.0～6.5 with the HCl adjusting pH value that is 2N with concentration; Temperature with whole reaction system is elevated to 50 ℃～55 ℃ then, and is incubated 20 minutes; Add neutral protease (enzyme activity is 1.5AU/g) 0.5kg then, continue also continuous stirring of insulation and carried out enzyme digestion reaction 150 minutes.150 minutes time is warming up to 85 ℃ rapidly with this enzyme digestion reaction product, and is incubated 10～15 minutes after arriving; Be cooled to room temperature (promptly 0～30 ℃) then rapidly and come enzymolysis reaction.
Under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 3000r/min centrifugal 20 minutes, supernatant liquor 220kg, be enzymolysis solution; The throw out of centrifugal gained can be reused for participate in above-mentioned enzyme digestion reaction (be about to this throw out be used as be temperature be elevated to 50 ℃～55 ℃ and be incubated the raw material of gained after 20 minutes).
Decoloration active carbon with 22kg decoloured 2 hours to this enzymolysis solution, and temperature is controlled at 55 ℃, and the destainer of gained gets the coarse filtration enzymolysis solution through 3 layers of fine and close gauze bag coarse filtration.Utilize tubular fibre polysulphones hyperfiltration membrane (molecular weight cut-off 30KDa) that the coarse filtration enzymolysis solution is carried out ultrafiltration, operating pressure 0.1～0.2MPa, 0～40 ℃ of working temperature obtains ultrafiltration enzymolysis solution 123kg.Utilize vacuum decker that this ultrafiltration enzymolysis solution is concentrated into 30kg, carry out lyophilize, obtain having the peptide powder 1.85kg of antihypertensive function, peptide content is 88.21%, and the product yield is 3.7%, and color and luster white is fine and smooth, utilize vitro detection ACE to suppress active CUSHMAN method, record its IC
50Be 1.24mg/ml.
Embodiment 2,
Take by weighing the dried dried silkworm chrysalis meal of the degreasing (moisture 1.2% after the pulverizing, protein content 66.23%, buy in Zhejiang silk one factory) 10kg, utilize 1.0% NaOH to extract albumen, 60 ℃ of temperature, extraction time is 4 hours, under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 3000r/min centrifugal 20 minutes, supernatant liquor concentration is that the HCl of 2N is reduced to 4.6 with the pH value, precipitates centrifugal postlyophilization and obtains pupa albumen powder 2.38kg, protein content 96.4%.
Get above-mentioned pupa albumen powder 2kg, adding water 12kg, to stir evenly the back be that the HCl adjusting pH value of 2N is 6.0～6.5 with concentration; Temperature with whole reaction system is elevated to 50 ℃～55 ℃ then, is incubated after 20 minutes, adds 20g neutral protease (enzyme activity is 1.5AU/g), continues also continuous stirring of insulation and carries out enzyme digestion reaction 120～200 minutes.The reaction times that sets is warming up to 85 ℃ rapidly with this enzyme digestion reaction product after arriving, and is incubated 10～15 minutes; Be cooled to room temperature then rapidly and come enzymolysis reaction.
Under 4 ℃ of conditions of J-6M type large capacity refrigerated centrifuge (BECKMAN company) 3000r/min centrifugal 20 minutes, supernatant liquor 4.2kg supernatant liquor, be enzymolysis solution; The throw out of centrifugal gained can be reused for participates in above-mentioned enzyme digestion reaction.
Decoloration active carbon with 350g decoloured 2 hours 55 ℃ of temperature to above-mentioned enzymolysis solution; The destainer of gained gets the coarse filtration enzymolysis solution through 3 layers of fine and close gauze bag coarse filtration.Utilize tubular fibre polysulphones hyperfiltration membrane (molecular weight cut-off 30KDa) that the coarse filtration enzymolysis solution is carried out ultrafiltration, operating pressure 0.1～0.2MPa, working temperature 0～40 obtains ultrafiltration enzymolysis solution 1800ml.Utilize vacuum decker that the ultrafiltration enzymolysis solution is concentrated into 600ml, carry out lyophilize, obtain having the peptide powder 126g of antihypertensive function, peptide content is 92.12%, the product yield is 6.3%, color and luster white is fine and smooth, utilizes vitro detection ACE to suppress active CUSHMAN method, and recording its IC50 is 1.02mg/ml.
At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Claims (7)
1, a kind of method of utilizing silkworm chrysalis to produce blood pressure lowering peptide, it is characterized in that may further comprise the steps: with the dried dried silkworm chrysalis meal of degreasing or pupa albumen powder and water according to 1: 6～7 weight ratio mixed after, regulating the pH value earlier is 6.0～6.5, add neutral protease again and carry out enzyme digestion reaction, the gained enzymolysis solution is blood pressure lowering peptide; The weight of described neutral protease is 0.9%～1.1% of dried dried silkworm chrysalis meal of degreasing or pupa albumen powder, 50 ℃～55 ℃ of enzyme digestion reaction temperature, 120～200 minutes enzyme digestion reaction time.
2, the method for utilizing silkworm chrysalis to produce blood pressure lowering peptide according to claim 1 is characterized in that: regulate after pH value is 6.0～6.5, be warming up to 50～55 ℃ and kept this temperature 18～22 minutes earlier, and then the adding neutral protease carries out enzyme digestion reaction.
3, the method for utilizing silkworm chrysalis to produce blood pressure lowering peptide according to claim 1 and 2, it is characterized in that: after the described enzyme digestion reaction time arrives, the enzyme digestion reaction product is warming up to 85 ℃ and kept this temperature 10～15 minutes rapidly, is cooled to 0～30 ℃ more rapidly with enzymolysis reaction; The centrifugal then enzymolysis solution that promptly gets.
4, the method for utilizing silkworm chrysalis to produce blood pressure lowering peptide according to claim 3 is characterized in that: adopt gac to the processing of decolouring of described enzymolysis solution, activated carbon dosage is the 5-30% of enzymolysis solution gross weight, 55 ℃ of temperature; Carry out again getting the coarse filtration enzymolysis solution after coarse filtration is handled.
5, the method for utilizing silkworm chrysalis to produce blood pressure lowering peptide according to claim 4 is characterized in that: under the working temperature of the operating pressure of 0.1～0.2Mpa and 0～45 ℃, adopt ultra-filtration membrane to carry out uf processing described coarse filtration enzymolysis solution, the ultrafiltration enzymolysis solution; Again described ultrafiltration enzymolysis solution is carried out vacuum concentration, lyophilize successively.
6, the method for utilizing silkworm chrysalis to produce blood pressure lowering peptide according to claim 5 is characterized in that: described ultra-filtration membrane molecular weight cut-off is 30 kilodaltons.
7, the method for utilizing silkworm chrysalis to produce blood pressure lowering peptide according to claim 6 is characterized in that: the enzyme activity 1.5AU/g of described neutral protease.
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| CN2006100520942A CN1896267B (en) | 2006-06-23 | 2006-06-23 | Preparation of depressor peptide by silkworm chrysalis |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101411405B (en) * | 2008-11-26 | 2011-07-20 | 中国农业大学 | Insect tissue zymolyte and use thereof in artificial culture of entomopathogenic nematodes |
| RU2446711C1 (en) * | 2011-03-29 | 2012-04-10 | Игорь Иванович Агапов | Functional biologically active product (versions) and its production method (versions) |
| CN101701240B (en) * | 2009-11-12 | 2012-05-23 | 常州康和生物技术有限公司 | Method for preparing antihypertensive peptide by utilizing combined enzyme enzymolysis silkworm chrysalis protein |
| CN103190524A (en) * | 2013-04-02 | 2013-07-10 | 江苏科技大学 | Method for improving silkworm chrysalis protein functional characteristics using ultrasonic wave coordinated with enzymolysis technology and product and application thereof |
| CN103540636A (en) * | 2013-09-29 | 2014-01-29 | 浙江省农业科学院 | Method for preparing blood fat reducing peptide by use of silkworm chrysalis |
| CN106281644A (en) * | 2016-08-12 | 2017-01-04 | 安徽未来农业发展有限公司 | A kind of chlorella Camellia oil promoting infants growth and development and preparation method thereof |
| CN107828840A (en) * | 2017-11-02 | 2018-03-23 | 林峰 | Silkworm chrysalis active peptide, composition and preparation method |
| CN108192940A (en) * | 2018-02-10 | 2018-06-22 | 海盐县凌特生物科技有限公司 | The preparation method of silkworm chrysalis collagen |
| CN108409854A (en) * | 2018-02-10 | 2018-08-17 | 海盐县凌特生物科技有限公司 | A kind of small-molecular-weight silkworm chrysalis collagen |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1032868C (en) * | 1990-10-30 | 1996-09-25 | 辽宁师范大学 | Method for compound zymohydrolysis of silkworm chrysalis protein |
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2006
- 2006-06-23 CN CN2006100520942A patent/CN1896267B/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101411405B (en) * | 2008-11-26 | 2011-07-20 | 中国农业大学 | Insect tissue zymolyte and use thereof in artificial culture of entomopathogenic nematodes |
| CN101701240B (en) * | 2009-11-12 | 2012-05-23 | 常州康和生物技术有限公司 | Method for preparing antihypertensive peptide by utilizing combined enzyme enzymolysis silkworm chrysalis protein |
| RU2446711C1 (en) * | 2011-03-29 | 2012-04-10 | Игорь Иванович Агапов | Functional biologically active product (versions) and its production method (versions) |
| CN103190524A (en) * | 2013-04-02 | 2013-07-10 | 江苏科技大学 | Method for improving silkworm chrysalis protein functional characteristics using ultrasonic wave coordinated with enzymolysis technology and product and application thereof |
| CN103190524B (en) * | 2013-04-02 | 2014-06-04 | 江苏科技大学 | Method for improving silkworm chrysalis protein functional characteristics using ultrasonic wave coordinated with enzymolysis technology and product and application thereof |
| CN103540636A (en) * | 2013-09-29 | 2014-01-29 | 浙江省农业科学院 | Method for preparing blood fat reducing peptide by use of silkworm chrysalis |
| CN103540636B (en) * | 2013-09-29 | 2015-05-20 | 浙江省农业科学院 | Method for preparing blood fat reducing peptide by use of silkworm chrysalis |
| CN106281644A (en) * | 2016-08-12 | 2017-01-04 | 安徽未来农业发展有限公司 | A kind of chlorella Camellia oil promoting infants growth and development and preparation method thereof |
| CN107828840A (en) * | 2017-11-02 | 2018-03-23 | 林峰 | Silkworm chrysalis active peptide, composition and preparation method |
| CN108192940A (en) * | 2018-02-10 | 2018-06-22 | 海盐县凌特生物科技有限公司 | The preparation method of silkworm chrysalis collagen |
| CN108409854A (en) * | 2018-02-10 | 2018-08-17 | 海盐县凌特生物科技有限公司 | A kind of small-molecular-weight silkworm chrysalis collagen |
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