CN104946713B - A kind of hypoglycemic peptide preparation method and applications based on sea cucumber albumen - Google Patents
A kind of hypoglycemic peptide preparation method and applications based on sea cucumber albumen Download PDFInfo
- Publication number
- CN104946713B CN104946713B CN201510369021.5A CN201510369021A CN104946713B CN 104946713 B CN104946713 B CN 104946713B CN 201510369021 A CN201510369021 A CN 201510369021A CN 104946713 B CN104946713 B CN 104946713B
- Authority
- CN
- China
- Prior art keywords
- sea cucumber
- added
- albumen
- supernatant
- hypoglycemic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The present invention discloses a kind of hypoglycemic peptide preparation method and applications based on sea cucumber albumen, and preparation method includes the following steps：Sea cucumber powder is taken, deionized water is added, heats in a water bath；Homogenate, obtains sea cucumber homogenate；PH value is adjusted to 7.0~10.0, protease I is added, after carrying out 2~4h of enzyme digestion reaction, proteinase II is added into the reaction system, continues 2~4h of enzymolysis, terminates reaction, obtains sea cucumber protein hydrolyzate；Inactivate protease I and proteinase II therein；Centrifugation removes impurity after cooling, retains supernatant；Be added 95% ethyl alcohol, centrifuging and taking supernatant after standing adjusts supernatant pH value to 6.0~8.0, and by the supernatant by the ultrafiltration membrane that molecular weight is 3000~10000Da that shuts off, collect ultrafiltrate to get.The sea cucumber protein peptides prepared using the method for the present invention can play good hypoglycemic lipid-lowering effect to diabetes and hyperlipidemia patient, and edible safety is high.
Description
Technical field
The present invention relates to aquatic products deep processing and biotechnology, more particularly to a kind of hypoglycemic drop based on sea cucumber albumen
Lipopeptid preparation method and applications.
Background technology
With social progress and economic development, prodigious change, diabetes and hyperlipemia has occurred in people's lives mode
The incidence of the metabolic diseases such as disease increases year by year.According to International Diabetes Federation (International Diabetes
Federation, IDF) statistics, end 2013, global diabetes number has reached 3.82 hundred million.It expects 2035, the whole world will
There are 5.92 hundred million people to be perplexed by diabetes.Currently, China has become the most country of diabetic's number, patient numbers are
Close to 100,000,000.Long term hyperglycemia can induce body fat metabolic disorder, lead to dyslipidemia, increase the illness wind of angiocardiopathy
Danger.Data are announced according to the World Health Organization (World Health Organization, WHO) to show, the whole world in 2008 is about
17300000 people die of angiocardiopathy.Cut-off 2012, China's cardiovascular patient number are 2.9 hundred million, and there are about 3,500,000 every year
People dies of angiocardiopathy.Currently, the method for the treatment of diabetes includes insulin injection and oral hypoglycemic drug, but these are controlled
Treatment method easily causes the adverse reactions such as hypoglycemia, hepar damnification, lactic acidosis and gastrointestinal discomfort.For controlling for hyperlipidemia
It treats, common stanin fat-reducing medicament, take for a long time also can generate toxic side effect to human body.Therefore, actively in research food
Functions of reducing sugar and reducing fat ingredient, safe, natural, efficient hypoglycemic peptide product of the exploitation in food have become sugared from now on
The pith of the metabolic diseases non-drug therapy such as urine disease and hyperlipidemia.Such as Chinese patent 201210462662.1, Wu
Neutral proteinase hydrolysis seaweed is utilized at industry etc., prepares the seaweed polypeptide with hypoglycemic and hypotensive activity.
Sea cucumber is food and drug resource important in ocean.Wall of sea cucumber Stichopus japonicus be sea cucumber mainly eat (medicine) use position, contain
Abundant collagen, glutinous polysaccharide and saponin(e, wherein sea cucumber protein content account for its 80% or more body wall dry weight.Sea cucumber albumen is rich
Sequence containing multiple biological activities releasably comes out under albumen enzyme effect appropriate, becomes with anti-oxidant, immunological regulation, drop
The peptide fragment of the biological activities such as blood pressure and hypoglycemic.In recent years, the activity of sea cucumber protein hydrolysate has obtained widely
Research.Li Wei etc. is disclosed with immunological regulation in Chinese patent 201310215842.4, is improved sleep and radiation-resisting functional
Sea cucumber fruit drink；Liu Kai etc. is prepared in Chinese patent 201410114474.9 using neutral proteinase hydrolysis sea cucumber albumen
Have effects that the sea cucumber protein peptides for promoting postoperative wound to heal.
Currently, the patented technology about enzyme hydrolytic sea cucumber albumen mostly uses greatly the enzymolysis mould of one one-step hydrolysis of single enzyme or complex enzyme
Formula, this digestion pattern are unfavorable for the hydrolysis of sea cucumber albumen, are also unsuitable for the release of bioactive sequences in sea cucumber albumen.Although
King, which is contained in Chinese patent 201310278840.X, utilizes alkali protease and immobilised enzymes (including neutral proteinase and pawpaw egg
White enzyme) fractional hydrolysis sea cucumber albumen, but in order to improve the enzymolysis efficiency of immobilised enzymes, the preliminary water of alkali protease need to be filtered to remove
The insoluble matter in liquid is solved, this process can lose the unhydrolysed sea cucumber albumen in part.Further, since in enzymolysis process lye use
And the inherent fishy smell of marine organisms, it can make sea cucumber protein peptides obtained that there is bad smell.Wang Qiukuan etc. is Chinese special
Using resin anion (R.A.) to the progress deodorant of sea cucumber protein peptides in profit 201210053085.0, this method is cumbersome and time-consuming, efficiency
It is low, it is unfavorable for industrialized production；Wu Qinglin in Chinese patent 201110449621.4 using activated carbon to sea cucumber protein peptides into
Row de-tastes processing, and this processing method can lose the functional component in polypeptide and reduce its biological activity.And it uses micro-
Encapsulated technology can not only cover the bad smell of sea cucumber protein peptides, but also will not lose the functional component in polypeptide.
The degree of hydrolysis of sea cucumber albumen is improved using double enzyme fractional hydrolysis patterns, preparing has the active sea cucumber egg of hypoglycemic
It white peptide and promotes its product Quality Research using microencapsulation and not yet finds.Therefore, a kind of controllable enzyme process preparation is developed
High activity, the method for flavorful sea cucumber albumen hypoglycemic peptide have important theory significance and application value.
Invention content
In view of the above shortcomings of the prior art, the present invention provides a kind of hypoglycemic peptide preparation method based on sea cucumber albumen
And its application, the hypoglycemic peptide activity height prepared using this method, excellent in flavor.
The technical scheme is that such realization：
A kind of hypoglycemic peptide preparation method based on sea cucumber albumen, includes the following steps：
1) it takes new fresh sea cucumber to clean, remove internal organs, is crushed after freeze-drying, obtain sea cucumber powder；Sea cucumber powder is taken, by 1:20~1:
Deionized water is added in 30 weight ratio, and 15~30min is then heated in 75~95 DEG C of water-baths.Homogenate power 1500~
Under the conditions of 3500r/min, it is homogenized 5~10min, obtains sea cucumber homogenate；
2) regulating step 1) the sea cucumber homogenate pH value to 7.0~10.0, protease I is added, make sea cucumber albumen and
Protease I carries out enzyme digestion reaction under the conditions of 40~60 DEG C；The protease I is alkali protease or trypsase；
3) step 2) enzyme digestion reaction carries out after 2~4h, keeps the temperature of reaction system of the enzyme digestion reaction constant, to institute
It states and proteinase II is added in reaction system, continue 2~4h of enzymolysis, terminate reaction, obtain sea cucumber protein hydrolyzate；
4) the sea cucumber protein hydrolyzate obtained by step 3) is warming up to 75~95 DEG C, keeps 15~30min, inactivation therein
Protease I and proteinase II；Centrifugation removes impurity after cooling, retains supernatant；The proteinase II is compound fertilizer production, wood
Any one in melon protease and neutral proteinase；
5) 95% ethyl alcohol is added into supernatant obtained by step 4), its final volume is made to reach 60~80%, 4 DEG C of total system
Centrifuging and taking supernatant after 10~15h of standing, adjusts the supernatant pH value to 6.0~8.0, and by the supernatant by shutting off
Molecular weight is the ultrafiltration membrane of 3000~10000Da, collects ultrafiltrate, obtains sea cucumber albumen hypoglycemic peptide.
A kind of hypoglycemic peptide preparation method based on sea cucumber albumen further includes the steps that following microencapsulation processing：
6) microencapsulation wall material is soluble in water, wall material solution is made；
7) wall material solution obtained by step 6) is added to the sea cucumber albumen hypoglycemic that step 5) obtains by a certain percentage
In peptide, 0.5~2h is stirred at room temperature, obtains mixed solution；
8) mixed solution obtained by step 7) is spray-dried, obtains microencapsulated protein polypeptide.
The addition of protease I is that 200~800KU protease I is added per 100g sea cucumber powders in the step 2).Wherein,
1U is defined as the amount of 1 μ g tyrosine of caseinhydrolysate generation per minute at 40 DEG C, KU 1000U.
The addition of proteinase II is that 200~800KU proteinase IIs are added per 100g sea cucumber powders in the step 3).Its
In, 1U is defined as the amount of 1 μ g tyrosine of caseinhydrolysate generation per minute at 40 DEG C, KU 1000U.
It is food-grade sodium carbonate, sodium bicarbonate, disodium hydrogen phosphate, carbonic acid that the lye used in pH value is adjusted in the step 2)
Any one in potassium, saleratus, dipotassium hydrogen phosphate or potassium hydroxide solution；It is food-grade breast to adjust the acid solution used in pH value
Acid or citric acid solution in any one.
Wall material in the step 7) is the whey protein concentrate of protein content >=80%.
Wall material in the step 7) is the mixture of the whey protein concentrate of sodium alginate and protein content >=80%,
Whey protein concentrate is 1 with sodium alginate mixed weight ratio:10~1:35.
Sea cucumber albumen hypoglycemic peptide and the mass ratio of wall material are 1 in the step 7):1~1:5.
The mass concentration of wall material can be molten according to the quality of polypeptide in polypeptide solution, and mixing in the wall material solution
The dry matter content of liquid determines.
The hypoglycemic peptide of sea cucumber albumen is in food source made from a kind of hypoglycemic peptide preparation method based on sea cucumber albumen
Application in property hypoglycemic health food.
Beneficial effects of the present invention：
1. the present invention can effectively improve the degree of hydrolysis of sea cucumber albumen, technique letter using the digestion pattern of double enzyme fractional hydrolysis
It is single easy, it is convenient for industrialized production；
2. the microencapsulation used in the present invention can effectively cover the bad smell of sea cucumber protein peptides, and not
Influence the prototype structure and biological activity of polypeptide；
3. the sea cucumber protein peptides edible safety of use the method for the present invention preparation is high, without side-effects, good absorption effect, to sugar
Urine disease and hyperlipidemia patient can play good hypoglycemic lipid-lowering effect, can play a role in health care to healthy population.
Specific implementation mode
The following is a clear and complete description of the technical scheme in the embodiments of the invention, it is clear that described embodiment
Only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, the common skill in this field
All other embodiment that art personnel are obtained without making creative work belongs to the model that the present invention protects
It encloses.
A kind of hypoglycemic peptide preparation method based on sea cucumber albumen, includes the following steps：
1) it takes new fresh sea cucumber to clean, remove internal organs, is crushed after freeze-drying, obtain sea cucumber powder；Sea cucumber powder is taken, by 1:20~1:
Deionized water is added in 30 weight ratio, and 15~30min is then heated in 75~95 DEG C of water-baths.Homogenate power 1500~
Under the conditions of 3500r/min, it is homogenized 5~10min, obtains sea cucumber homogenate；
2) regulating step 1) the sea cucumber homogenate pH value to 7.0~10.0, protease I is added, make sea cucumber albumen and
Protease I carries out enzyme digestion reaction under the conditions of 40~60 DEG C；The protease I is alkali protease or trypsase；
3) step 2) enzyme digestion reaction carries out after 2~4h, keeps the temperature of reaction system of the enzyme digestion reaction constant, to institute
It states and proteinase II is added in reaction system, continue 2~4h of enzymolysis, terminate reaction, obtain sea cucumber protein hydrolyzate；
4) the sea cucumber protein hydrolyzate obtained by step 3) is warming up to 75~95 DEG C, keeps 15~30min, inactivation therein
Protease I and proteinase II；Centrifugation removes impurity after cooling, retains supernatant；The proteinase II is compound fertilizer production, wood
Any one in melon protease and neutral proteinase；
5) 95% ethyl alcohol is added into supernatant obtained by step 4), its final volume is made to reach 60~80%, 4 DEG C of total system
Centrifuging and taking supernatant after 10~15h of standing, adjusts the supernatant pH value to 6.0~8.0, and by the supernatant by shutting off
Molecular weight is the ultrafiltration membrane of 3000~10000Da, collects ultrafiltrate, obtains sea cucumber albumen hypoglycemic peptide.
6) microencapsulation wall material is soluble in water, wall material solution is made；
7) wall material solution obtained by step 6) is added to the sea cucumber albumen hypoglycemic that step 5) obtains by a certain percentage
In peptide, 0.5~2h is stirred at room temperature, obtains mixed solution；
8) mixed solution obtained by step 7) is spray-dried, obtains microencapsulated protein polypeptide.
Embodiment 1
The hypoglycemic peptide preparation method based on sea cucumber albumen that the present embodiment provides a kind of, includes the following steps：
1) it takes new fresh sea cucumber to clean, remove internal organs, is crushed after freeze-drying, obtain sea cucumber powder；10g sea cucumber powders are weighed, are added
200mL deionized waters heat 20min in 90 DEG C of water-bath.Homogenate power is homogenized 5min under the conditions of being 2000r/min, uses carbon
Acid sodium solution adjusts pH to 9.0.Alkali protease 20KU is added, under 60 DEG C of water bath conditions, then hydrolysis 3h is directly added into wood
Melon protease 3 0KU continues to hydrolyze 3h, and it is 45.2% to measure enzymolysis liquid degree of hydrolysis at this time.The sea cucumber protein enzymatic hydrolyzate of gained heats up
To 85 DEG C, 15min, enzyme deactivation are kept.Supernatant is taken after 4000r/min centrifugations 15min, and 95% ethyl alcohol, which is added, makes its final volume be
80% (volume ratio), 4 DEG C of standing 10h.Centrifuging and taking supernatant adjusts supernatant pH and passes through the molecule that shuts off to 7.0, and by supernatant
Amount is the ultrafiltration membrane of 10000Da, collects ultrafiltrate, obtains sea cucumber albumen hypoglycemic peptide solution 15g, wherein content of peptides is
6.8g。
2) 13.6g whey protein concentrate is weighed, 150mL water is added, makes it completely dissolved.Then, it adds it to described
In sea cucumber albumen hypoglycemic peptide solution, 2h, spray-dried obtained microencapsulation polypeptide powder is stirred at room temperature.
Degree of hydrolysis can measure by the following method：
The sample of 0.5mL enzyme deactivations is drawn, cooling is simultaneously transferred in 25mL volumetric flasks, with 1% lauryl sodium sulfate
(SDS) solution constant volume is stood.Take the SDS solution of 0.125mL enzymolysis liquids and 1mL phosphate buffers (pH 8.2,0.2mol/L) and
Trinitrobenzene sulfonic acid method (TNBS) solution oscillation mixing of the 0.1% of 1mL, is protected from light 60min, reaction is finished in 50 DEG C of water-baths
Later, reaction is terminated immediately with the HCl of 2mL 0.1mol/L, after being placed at room temperature for 30min, its absorbance value is measured under 420nm.
0~5.0 × 10-3mol/L L-Leus do standard curve.Degree of hydrolysis is calculated with following formula：
In formula, h：Indicate every gram of peptide bond mM number (meq/g protein) being cleaved in hydrolysate；
htot：Indicate peptide bond mM number total in every gram of material protein.
Embodiment 2
1) 10g sea cucumber powders are weighed, 250mL deionized waters are added, 30min is heated in 85 DEG C of water-bath.Being homogenized power is
It is homogenized 8min under the conditions of 1500r/min, pH to 7.0 is adjusted with lactic acid solution.Trypsase 80KU, 40 DEG C of water bath conditions are added
Under, then hydrolysis 3h is directly added into papain 30KU, continue to hydrolyze 3h, measures enzymolysis liquid degree of hydrolysis at this time and is
42.5%.The sea cucumber protein enzymatic hydrolyzate of gained is warming up to 85 DEG C, keeps 15min, enzyme deactivation.It is taken after 4000r/min centrifugations 15min
Clear liquid, 95% ethyl alcohol, which is added, makes its final volume be 60% (volume ratio), 4 DEG C of standing 12h.Centrifuging and taking supernatant adjusts supernatant
PH collects ultrafiltrate, obtains sea cucumber albumen hypoglycemic to 7.0, and by supernatant by the ultrafiltration membrane that molecular weight is 3000Da that shuts off
Lipid-loweringing peptide 13.16g, wherein content of peptides are 6.4g.
2) 12.8g whey protein concentrate and 0.36g sodium alginates are weighed, 150mL water is added, makes it completely dissolved.Then,
It adds it in the sea cucumber albumen hypoglycemic peptide solution, 2h, spray-dried obtained microencapsulation polypeptide powder is stirred at room temperature.
Degree of hydrolysis assay method is the same as embodiment 1.
Embodiment 3
1) 10g sea cucumber powders are weighed, 250mL deionized waters are added, 30min is heated in 75 DEG C of water-bath.Being homogenized power is
It is homogenized 5min under the conditions of 2000r/min, pH to 10.0 is adjusted with sodium bicarbonate solution.Alkali protease 40KU, 50 DEG C of water are added
Under the conditions of bath, then hydrolysis 3h is directly added into compound fertilizer production 80KU, continues to hydrolyze 3h, measures enzymolysis liquid at this time and hydrolyzes
Degree is 41.8%.The sea cucumber protein enzymatic hydrolyzate of gained is warming up to 75 DEG C, keeps 30min, enzyme deactivation.After 4000r/min centrifuges 15min
Supernatant is taken, 95% ethyl alcohol, which is added, makes its final volume be 70% (volume ratio), 4 DEG C of standing 10h.Centrifuging and taking supernatant, in adjusting
Clear liquid pH collects ultrafiltrate, obtains sea cucumber albumen to 8.0, and by supernatant by the ultrafiltration membrane that molecular weight is 8000Da that shuts off
Hypoglycemic peptide 15g, wherein content of peptides are 6.5g.
2) 75.0g whey protein concentrate is weighed, 150mL water is added, makes it completely dissolved.Then, it adds it to described
In sea cucumber albumen hypoglycemic peptide solution, 0.5h, spray-dried obtained microencapsulation polypeptide powder is stirred at room temperature.Degree of hydrolysis measurement side
Method is the same as embodiment 1.
Embodiment 4
1) 10g sea cucumber powders are weighed, 250mL deionized waters are added, 20min is heated in 95 DEG C of water-bath.Being homogenized power is
It is homogenized 8min under the conditions of 1500r/min, pH to 8.0 is adjusted with solution of potassium carbonate.Trypsase 30KU, 40 DEG C of water bath conditions are added
Under, then hydrolysis 2h is directly added into neutral proteinase 40KU, continue to hydrolyze 4h, measures enzymolysis liquid degree of hydrolysis at this time and is
39.8%.The sea cucumber protein enzymatic hydrolyzate of gained is warming up to 85 DEG C, keeps 15min, enzyme deactivation.It is taken after 4000r/min centrifugations 15min
Clear liquid, 95% ethyl alcohol, which is added, makes its final volume be 60% (volume ratio), 4 DEG C of standing 15h.Centrifuging and taking supernatant adjusts supernatant
PH collects ultrafiltrate, obtains sea cucumber albumen hypoglycemic to 7.0, and by supernatant by the ultrafiltration membrane that molecular weight is 10000Da that shuts off
Lipid-loweringing peptide 15.2g, wherein content of peptides are 6.5g.
2) 13.6g whey protein concentrate is weighed, 100mL water is added, makes it completely dissolved.Then, it adds it to described
In sea cucumber albumen hypoglycemic peptide solution, 1.5h, spray-dried obtained microencapsulation polypeptide powder is stirred at room temperature.Degree of hydrolysis measurement side
Method is the same as embodiment 1.
Embodiment 5
1) 10g sea cucumber powders are weighed, 250mL deionized waters are added, 15min is heated in 95 DEG C of water-bath.Being homogenized power is
It is homogenized 10min under the conditions of 1500r/min, pH to 8.0 is adjusted with potassium bicarbonate solution.Trypsase 30KU, 40 DEG C of water-baths are added
Under the conditions of, then hydrolysis 2h is directly added into compound fertilizer production 20KU, continue to hydrolyze 4h, measure enzymolysis liquid degree of hydrolysis at this time
It is 40.5%.The sea cucumber protein enzymatic hydrolyzate of gained is warming up to 85 DEG C, keeps 15min, enzyme deactivation.It is taken after 4000r/min centrifugations 15min
Supernatant, 95% ethyl alcohol, which is added, makes its final volume be 60% (volume ratio), 4 DEG C of standing 15h.Centrifuging and taking supernatant adjusts supernatant
Liquid pH collects ultrafiltrate to 7.0, and by supernatant by the ultrafiltration membrane that molecular weight is 6000Da that shuts off, and obtains sea cucumber albumen drop
Sugared lipid-loweringing peptide 15.6g, wherein content of peptides are 6.4g.
2) 16.2g whey protein concentrate is weighed, 250mL water is added, makes it completely dissolved.Then, it adds it to described
In sea cucumber albumen hypoglycemic peptide solution, 1h, spray-dried obtained microencapsulation polypeptide powder is stirred at room temperature.Degree of hydrolysis assay method
With embodiment 1.
Embodiment 6
1) 10g sea cucumber powders are weighed, 300mL deionized waters are added, 30min is heated in 85 DEG C of water-bath.Being homogenized power is
It is homogenized 5min under the conditions of 3500r/min, pH to 10.0 is adjusted with potassium hydroxide solution.Alkali protease 40KU, 50 DEG C of water are added
Under the conditions of bath, then hydrolysis 4h is directly added into neutral proteinase 50KU, continues to hydrolyze 2h, measures enzymolysis liquid degree of hydrolysis at this time and is
40.8%.The sea cucumber protein enzymatic hydrolyzate of gained is warming up to 90 DEG C, keeps 15min, enzyme deactivation.It is taken after 4000r/min centrifugations 15min
Clear liquid, 95% ethyl alcohol, which is added, makes its final volume be 60% (volume ratio), 4 DEG C of standing 12h.Centrifuging and taking supernatant adjusts supernatant
PH collects ultrafiltrate, obtains sea cucumber albumen hypoglycemic to 6.0, and by supernatant by the ultrafiltration membrane that molecular weight is 8000Da that shuts off
Lipid-loweringing peptide 15.6g, wherein content of peptides are 6.5g.
2) 14.6g whey protein concentrate and 1.46g sodium alginates are weighed, 200mL water is added, makes it completely dissolved.Then,
It adds it in the sea cucumber albumen hypoglycemic peptide solution, 1.5h, spray-dried obtained microencapsulation polypeptide is stirred at room temperature
Powder.Degree of hydrolysis assay method is the same as embodiment 1.
In above-described embodiment, it is food-grade sodium carbonate, sodium bicarbonate, disodium hydrogen phosphate, carbon to adjust the lye used in pH value
Any one in sour potassium, saleratus, dipotassium hydrogen phosphate or potassium hydroxide solution；It is food-grade to adjust the acid solution used in pH value
Any one in lactic acid or citric acid solution.
Comparative example 1
10g sea cucumber powders are weighed, 200mL deionized waters are added, 20min is heated in 90 DEG C of water-bath.Being homogenized power is
It is homogenized 5min under the conditions of 2000r/min, adjusts pH to 9.0.Alkali protease 30KU is added, under 55 DEG C of water bath conditions, hydrolyzes 6h,
Lye is continuously added in hydrolytic process maintains reaction system pH constant, measures enzymolysis liquid at this time and is hydrolyzed to 25.8%；The sea of gained
Ginseng protein enzymatic hydrolyzate is warming up to 85 DEG C, keeps 15min, enzyme deactivation.Supernatant is taken after 4000r/min centrifugations 15min, 95% second is added
Alcohol makes its final volume be 60% (volume ratio), 4 DEG C of standing 10h.Centrifuging and taking supernatant adjusts supernatant pH to 7.0, and by supernatant
Liquid collects ultrafiltrate, obtains sea cucumber albumen hypoglycemic peptide by the ultrafiltration membrane that molecular weight is 10000Da that shuts off.Degree of hydrolysis is surveyed
Method is determined with embodiment 1.
By above-described embodiment and comparative example it is found that the method for the present invention can effectively improve the degree of hydrolysis of sea cucumber albumen, and
And the bad flavor of sea cucumber protein peptides can be covered.It is simple for process described in the method for the present invention, it is conducive to industrialized production.
Test example 1
The hypoglycemic of sea cucumber albumen made from the hypoglycemic peptide preparation method based on sea cucumber albumen that the present embodiment provides a kind of
Application of the lipid-loweringing peptide in food-borne hypoglycemic health food.
1. experiment material and method
1.1 experiment material
1) sea cucumber protein peptides
Sea cucumber protein peptides made from embodiment 1.
2) experimental animal
C57BL/6 mouse, male, 10~12 week old, 50.
3) key instrument and reagent
Blood glucose meter, total cholesterol kit, triglyceride reagent box, high-density lipoprotein cholesterol kit and low-density
Lipoprotein cholesterol kit etc..
4) other materials
(ingredient includes 73% basal feed, 10% lard, 10% yolk powder, 1% cholesterol and 6% breast to high lipid food
Powder).
1.2 experimental method
1) foundation of obese model
After 50 C57BL/6 mouse were with basal feed adaptable fed 1 week, it is grouped at random according to weight.40
C57BL/6 mouse continuously feed high lipid food 12 weeks, establish obesity C57BL/6 mouse models.10 C57BL/6 mouse feedings are just
Normal feed, as Normal group.
2) experimental design
By 40 C57BL/6 obesity mices, it is divided into 4 groups, every group 10, is set to obese model control group, sea cucumber egg
White peptide low dose group, sea cucumber protein peptides middle dose group and sea cucumber protein peptides high dose group.Specific experiment animal packet and give medicament
Amount is as described in Table 1.
1 experimental animal of table is grouped and dosage
| Group | Feed for nursing | Gavage drug | Dosage (mg/kg b.w.) |
| Normal group | Chow diet | Distilled water | —— |
| Obese model control group | High lipid food | Distilled water | —— |
| Sea cucumber protein peptides low dose group | High lipid food | Sea cucumber protein peptides | 100 |
| Sea cucumber protein peptides middle dose group | High lipid food | Sea cucumber protein peptides | 200 |
| Sea cucumber protein peptides high dose group | High lipid food | Sea cucumber protein peptides | 400 |
3) feeding manner
Every group of 10 C57BL/6 mouse are raised for 1 mouse cage, and 5 groups of experiments are carried out at the same time, and are fed according to the design of table 1 different
Feed, normal water supply.
4) administering mode
Rat body weight is measured before daily administration, according to different groups of other different dosing dosage, is dissolved in the distillation of same volume
In water, it is administered in a manner of gavage.Daily timing gastric infusion is primary.Test continuous 6 weeks, during experiment mouse free water and
It ingests.
5) index determining
After experiment, fasting 12h measures weight.Broken end takes blood after anesthesia, collects serum, measures fasting blood in serum
Sugar, triglycerides (TC), total cholesterol (TG), high-density lipoprotein cholesterol (HDL-c) and low density lipoprotein cholesterol
(LDL-c)。
2. experimental result and analysis
Different disposal group C57BL/6 mouse weights and blood items biochemical indicator, as described in Table 2.
2 different disposal group C57BL/6 mouse weights of table and blood items biochemical indicator
Note：Numerical value is indicated in the form of mean+SD；Different letters represent significant difference, p<0.05.
Each dosage group mouse weight is substantially less than hyperlipidemia model group (p<0.05).Middle dose group and high dose group Mice Body
Weight and the not notable (p> of Normal group mouse weight difference;0.05).
Compared with hyperlipidemia model group, fasting blood-glucose, the TC contents of middle dose group and high dose group mouse are substantially less than high in fat
Model group (p<0.05)；Not notable (the p> of difference between middle dose group and high dose group;0.05).
Hyperlipidemia model group and each dosage group mouse TG contents are significantly higher than Normal group (p<0.05)；Each dosage group mouse
TG contents are substantially less than hyperlipidemia model group (p<0.05)；Not notable (the p> of difference between each dosage group;0.05).
Middle dose group and high dose group mouse HDL-c contents are significantly higher than hyperlipidemia model group (p<0.05), with normal control
Not notable (the p> of difference between group;0.05)；Not notable (the p> of difference between middle dose group and high dose group;0.05).
Hyperlipidemia model group and each dosage group mouse LDL-c contents are significantly higher than Normal group (p<0.05), middle dose group
It is substantially less than hyperlipidemia model group (p< with high dose group mouse LDL-c contents;0.05)；Difference between middle dose group and high dose group
Not significantly (p>0.05).
3. experiment conclusion
The sea cucumber protein peptides of 200mg/kg b.w. dosage, can effective obesity controlling C57BL/6 mouse weights increase, drop
Its low fasting blood-glucose and triglycerides, total cholesterol and low density lipoprotein cholesterol content in blood, increase its blood
In high-density lipoprotein cholesterol content.As it can be seen that taking sea cucumber protein peptides of the present invention for a long time to body blood glucose and blood
Fat has certain adjustment effect, has certain auxiliary curative effect to treatment hyperglycemia and hyperlipidemia.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
With within principle, any modification, equivalent replacement, improvement and so on should all be included in the protection scope of the present invention god.
Claims (7)
1. a kind of hypoglycemic peptide preparation method based on sea cucumber albumen, it is characterised in that include the following steps：
1）It takes new fresh sea cucumber to clean, remove internal organs, is crushed after freeze-drying, obtain sea cucumber powder；Sea cucumber powder is taken, by 1:20~1:30 weights
Deionized water is added in amount ratio, and 15~30 min are then heated in 75~95 DEG C of water-baths；In homogenate power 1500~3500
Under the conditions of r/min, 5~10 min are homogenized, sea cucumber homogenate is obtained；
2）Regulating step 1）The pH value of the sea cucumber homogenate is added protease I, makes sea cucumber albumen and albumen to 7.0~10.0
Enzyme I carries out enzyme digestion reaction under the conditions of 40~60 DEG C；The protease I is alkali protease or trypsase；
3）Step 2）Enzyme digestion reaction carries out after 2~4 h, keeps the temperature of reaction system of the enzyme digestion reaction constant, to described
Proteinase II is added in reaction system, continues 2~4 h of enzymolysis, terminates reaction, obtains sea cucumber protein hydrolyzate；
4）By step 3）The sea cucumber protein hydrolyzate of gained is warming up to 75~95 DEG C, keeps 15~30 min, inactivates egg therein
White enzyme I and proteinase II；Centrifugation removes impurity after cooling, retains supernatant；The proteinase II is compound fertilizer production, pawpaw
Any one in protease and neutral proteinase；
5）To step 4）95% ethyl alcohol is added in gained supernatant, its final volume is made to reach 60~80%, 4 DEG C of standings of total system
Centrifuging and taking supernatant after 10~15 h adjusts the supernatant pH value to 6.0~8.0, and by the supernatant by shutting off point
The ultrafiltration membrane that son amount is 3000~10000 Da, collects ultrafiltrate, obtains sea cucumber albumen hypoglycemic peptide；
6）Microencapsulation wall material is soluble in water, wall material solution is made；
7）By step 6）The wall material solution of gained is added to step 5 by a certain percentage）In obtained sea cucumber albumen hypoglycemic peptide,
0.5~2 h is stirred at room temperature, obtains mixed solution；
8）To step 7）The mixed solution of gained is spray-dried, and microencapsulated protein polypeptide is obtained；
The step 7）Middle sea cucumber albumen hypoglycemic peptide and the mass ratio of wall material are 1:1~1:5.
2. a kind of hypoglycemic peptide preparation method based on sea cucumber albumen according to claim 1, which is characterized in that described
Step 2）The addition of middle protease I is that 200 ~ 800KU protease I is added per 100g sea cucumber powders.
3. a kind of hypoglycemic peptide preparation method based on sea cucumber albumen according to claim 1, which is characterized in that described
Step 3）The addition of middle proteinase II is that 200 ~ 800KU proteinase IIs are added per 100g sea cucumber powders.
4. a kind of hypoglycemic peptide preparation method based on sea cucumber albumen according to claim 1, which is characterized in that described
Step 2）Lye used in middle adjusting pH value be food-grade sodium carbonate, sodium bicarbonate, disodium hydrogen phosphate, potassium carbonate, saleratus,
Any one in dipotassium hydrogen phosphate or potassium hydroxide solution；It is that food grade lactic acid or citric acid are molten to adjust the acid solution used in pH value
Any one in liquid.
5. a kind of hypoglycemic peptide preparation method based on sea cucumber albumen according to claim 1, which is characterized in that described
Step 7）In wall material be protein content >=80% whey protein concentrate.
6. a kind of hypoglycemic peptide preparation method based on sea cucumber albumen according to claim 1, which is characterized in that described
Step 7）In wall material be sodium alginate and protein content >=80% whey protein concentrate mixture, whey protein concentrate
It is 1 with sodium alginate mixed weight ratio:10~1:35.
7. sea cucumber albumen made from hypoglycemic peptide preparation method of claim 1 ~ 6 any one of them based on sea cucumber albumen
Application of the hypoglycemic peptide in food-borne hypoglycemic health food.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510369021.5A CN104946713B (en) | 2015-06-29 | 2015-06-29 | A kind of hypoglycemic peptide preparation method and applications based on sea cucumber albumen |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510369021.5A CN104946713B (en) | 2015-06-29 | 2015-06-29 | A kind of hypoglycemic peptide preparation method and applications based on sea cucumber albumen |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104946713A CN104946713A (en) | 2015-09-30 |
| CN104946713B true CN104946713B (en) | 2018-10-26 |
Family
ID=54161776
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510369021.5A Active CN104946713B (en) | 2015-06-29 | 2015-06-29 | A kind of hypoglycemic peptide preparation method and applications based on sea cucumber albumen |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104946713B (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105506046B (en) * | 2016-01-15 | 2019-02-26 | 中国农业大学 | Casein hypoglycemic peptide based on yak milk Qula and its preparation method and application |
| CN105524967B (en) * | 2016-03-02 | 2019-07-26 | 集美大学 | A kind of combined preparation process of sea cucumber polysaccharide and holothurian collagen polypeptide |
| CN105695545B (en) * | 2016-03-02 | 2019-05-31 | 集美大学 | A kind of preparation method of sea cucumber fucoidan and sea cucumber glycoprotein |
| CN107184966A (en) * | 2017-06-19 | 2017-09-22 | 青岛金海宝生物科技发展有限公司 | A kind of pharmaceutical composition for preventing and treating diabetes and its complication |
| CN107397226A (en) * | 2017-06-30 | 2017-11-28 | 宁德师范学院 | A kind of reducing blood lipid sea cucumber polypeptide microcapsules and preparation method thereof |
| CN110295215A (en) * | 2019-07-08 | 2019-10-01 | 哈尔滨工业大学（威海） | A kind of preparation method of anti-type-2 diabetes mellitus sea cucumber polypeptide |
| US20210301314A1 (en) * | 2020-03-26 | 2021-09-30 | Chenland Nutritionals Inc. | Preparation method and application of sea cucumber protein peptides |
| CN111821421B (en) * | 2020-08-13 | 2021-11-02 | 上海交通大学 | Intestinal slow-release bovine colostrum and sea cucumber peptide chewable tablet and preparation method thereof |
| CN112176015B (en) * | 2020-10-09 | 2022-04-15 | 山东省科学院生物研究所 | Efficient bionic preparation method of sea cucumber bioactive peptide |
| CN112266405B (en) * | 2020-10-27 | 2021-05-14 | 赵博 | Method for recovering protein from potato starch processing wastewater |
| CN114317658B (en) * | 2022-01-14 | 2023-08-18 | 完美（广东）日用品有限公司 | Sea buckthorn seed meal protein peptide composition and preparation method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1478421A (en) * | 2002-08-30 | 2004-03-03 | 大连轻工业学院 | Seacucumbus whole powder food and its preparation method |
| CN102907558A (en) * | 2012-11-16 | 2013-02-06 | 山东省海洋水产研究所 | Processing method of sea cucumber polypeptide |
| CN103320487A (en) * | 2013-07-04 | 2013-09-25 | 威海益合元生物科技有限公司 | Method for preparing trepang peptide by combined hydrolysis of soluble enzyme and immobilized enzyme |
| CN103421867A (en) * | 2012-05-14 | 2013-12-04 | 宁波餐之皇食品有限公司 | Preparation method of sea cucumber collagens |
| CN104263788A (en) * | 2014-09-25 | 2015-01-07 | 冯群力 | Preparation method of sea cucumber polypeptide |
-
2015
- 2015-06-29 CN CN201510369021.5A patent/CN104946713B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1478421A (en) * | 2002-08-30 | 2004-03-03 | 大连轻工业学院 | Seacucumbus whole powder food and its preparation method |
| CN103421867A (en) * | 2012-05-14 | 2013-12-04 | 宁波餐之皇食品有限公司 | Preparation method of sea cucumber collagens |
| CN102907558A (en) * | 2012-11-16 | 2013-02-06 | 山东省海洋水产研究所 | Processing method of sea cucumber polypeptide |
| CN103320487A (en) * | 2013-07-04 | 2013-09-25 | 威海益合元生物科技有限公司 | Method for preparing trepang peptide by combined hydrolysis of soluble enzyme and immobilized enzyme |
| CN104263788A (en) * | 2014-09-25 | 2015-01-07 | 冯群力 | Preparation method of sea cucumber polypeptide |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104946713A (en) | 2015-09-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104946713B (en) | A kind of hypoglycemic peptide preparation method and applications based on sea cucumber albumen | |
| CN105079615B (en) | A kind of pressed candy and preparation method thereof | |
| CN105595338A (en) | Food for special medical purpose applicable to diabetes and preparation method of food for special medical purpose applicable to diabetes | |
| CN109757734A (en) | A kind of polypeptide oral liquor and preparation method thereof | |
| CN103975991B (en) | Sorosis anthocyanidin health-caring biscuit | |
| JP4132635B2 (en) | Uninactivated enzyme-enhanced composition | |
| CN109123036A (en) | Soybean-marrow peptide composition and application | |
| CN105506046B (en) | Casein hypoglycemic peptide based on yak milk Qula and its preparation method and application | |
| CN102379391B (en) | Preparation of health food of pine pollen, glucan and natto | |
| CN105476014A (en) | Method for preparing biological zinc from oysters | |
| CN106031436A (en) | Nutritional preparation containing egg white and preparation method thereof | |
| CN103385889A (en) | Carbohydrate and electrolyte mixed injection and preparation method thereof | |
| CN102511858A (en) | Thickened red jujube pulp suitable for being taken by pregnant women and manufacturing technology of same | |
| CN1120908A (en) | Choice seafood extract product and its production process and uses | |
| CN1240297C (en) | Peptide milk and its preparation technology | |
| CN107699602A (en) | It is a kind of that industrial process and application rich in machine ferrous iron donkey-hide gelatin peptide are prepared by donkey-hide gelatin | |
| CN102342400A (en) | High-calcium xylitol | |
| CN110664984A (en) | Composition with effects of dispelling effects of alcohol and protecting liver and preparation method thereof | |
| CN1298229C (en) | Method for preparing milk pellets contg. micro sang, white fungus and oligose for face-beauty, toning-lungs and lowering blood-sugar | |
| CN118511995B (en) | Polysaccharide composition capable of improving sugar metabolism disorder and application thereof | |
| CN109275914A (en) | Black fungus polypeptide function edible composition and its preparation method and application | |
| CN108991358A (en) | A kind of auxiliary reducing blood lipid, it is hypoglycemic, alleviate gout health care food and production method | |
| CN108785411A (en) | A kind of Preparation method and use of the cherry powder of control diabetes | |
| CN106360230A (en) | Ferment powder solid beverage and preparation method thereof | |
| CN100370905C (en) | Method for preparing sour milk contg. micron sang, white fungus and oligose for face-beauty, toning lungs and lower blood-sugar |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20180910 Address after: 265600 Dongguan Road, Penglai, Yantai, Shandong 25 Applicant after: YANTAI YUANLIDE MARINE BIOLOGICAL CO., LTD. Address before: 265600 Dongguan Road, Penglai, Yantai, Shandong 25 Applicant before: Jiang Naiyi |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |