CN102578589A - Preparation method and application of sea cucumber polypeptide chelated active calcium - Google Patents
Preparation method and application of sea cucumber polypeptide chelated active calcium Download PDFInfo
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- CN102578589A CN102578589A CN2012100588777A CN201210058877A CN102578589A CN 102578589 A CN102578589 A CN 102578589A CN 2012100588777 A CN2012100588777 A CN 2012100588777A CN 201210058877 A CN201210058877 A CN 201210058877A CN 102578589 A CN102578589 A CN 102578589A
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Abstract
The invention discloses a preparation method and an application of sea cucumber polypeptide chelated active calcium. The preparation method comprises the following steps: softening sea cucumber through repeatedly boiling the sea cucumber by purified water, crushing the softened sea cucumber into sea cucumber particles with the diameter of 1-2mm, preparing a sea cucumber protoplasm from the sea cucumber particles, water and a composite protease, carrying out enzymatic hydrolysis for 120-180min at 45-65DEG C to obtain a sea cucumber enzymatic hydrolyzate, calcining oyster shell for 3h at 300-900DEG C, crushing the calcined oyster shell into 80-100 meshes to obtain oyster shell active calcium, mixing the sea cucumber enzymatic hydrolyzate with the oyster shell active calcium in a glass dish according to a weight ratio of 100:6-12, stirring for 10-30min, allowing the obtained product to stand for 30-40min, and drying to obtain the sea cucumber polypeptide chelated active calcium. The method has the advantages of simple preparation, convenient production, low production cost, and realization of a large amount of sea cucumber polypeptides, and the finally obtained sea cucumber polypeptide chelated active calcium has the effects of islet cell secretion improvement, and reduction of contents of glycosylated serum proteins, urea nitrogen and creatinine in blood.
Description
Technical field
The present invention relates to the production of sea cucumber polypeptide preparation, be specifically related to a kind of preparation method and application of sea cucumber polypeptide sequestering activity calcium.
Background technology
The islet cells hyposecretion causes in the blood saccharification haemocyanin, urea nitrogen and creatinine content higher, and sugar tolerance reduces, metabolism disorder of blood lipid; Blood sugar and blood fat raise, liver, decreased renal function.Sea cucumber contains abundant collagen, saponin and mucopolysaccharide, has raising immunity, and is antitumor, reducing blood lipid, and anticoagulation, delaying decrepitude of skin reduces the effect of high blood uric acid.Publication number is CN101514354, and name is called the application for a patent for invention of a kind of sea cucumber polypeptide and preparation method and application, fresh stichopus japonicus homogenate is disclosed after; With pepsin, trypsin digestion, add ethanol in the supernatant and leave standstill, the centrifuging and taking supernatant; Drying obtains the sea cucumber polypeptide bullion; Obtain sea cucumber polypeptide through chromatography then, sea cucumber polypeptide has increasing leukocyte, promotes the effect of bone marrow cell hyperplasia.Granted publication number is CN1951402; Name is called the patent of invention of the production method of a kind of sea cucumber polypeptide calcium, zinc albumen, then disclose sea cucumber milk with trypsin digestion after, add BIOLOGICAL CALCIUM and/or zinc; Drying obtains sea cucumber polypeptide calcium, zinc albumen; Have supplement calcium, improve bone, effect such as radioresistance, detoxifcation, but above-mentioned sea cucumber polypeptide or to contain in sea cucumber polypeptide calcium and/or the zinc sea cucumber polypeptide purity low, impurity such as sea cucumber polysaccharide and selenka are many; Also have residue of pesticide also more, the chelating degree is low between sea cucumber polypeptide and calcium, the zinc.
Summary of the invention
Technical problem to be solved by this invention provides a kind of preparation method of sea cucumber polypeptide sequestering activity calcium; This preparation method is convenient for production; Sea cucumber polypeptide purity is high, and kind is more, and molecular weight is than homogeneous; The sea cucumber polypeptide sequestering activity calcium that obtains has the secretion that improves islet cells, reduces the effect of saccharification haemocyanin, urea nitrogen and creatinine content in the blood.
The present invention solves the problems of the technologies described above the technical scheme that is adopted: a kind of preparation method of sea cucumber polypeptide sequestering activity calcium, and its step is following:
A, impurity elimination are outwelled boiling water after going the sea cucumber behind the internal organ to put into to fill the pot of pure water to boil, and add after pure water boils again; Outwell boiling water again; Change pure water repeatedly altogether and boil 4～5 times, obtain softening sea cucumber, will soften sea cucumber again and be ground into the sea cucumber granules that diameter is 1～2mm; Change pure water so repeatedly and boil and can fully remove salt marsh and agricultural chemicals, sea cucumber polysaccharide and selenka are removed in dissolving, and the sea cucumber bating effect is good, help pulverizing, and help enzymolysis;
B, add entry at sea cucumber granules by weight the ratio of 1:2～5; Add compound protease again, the weight ratio of said compound protease and said sea cucumber granules is 2～3:100, stirs; Hydrolysis temperature is under 45～65 ℃, and enzymolysis 120～180min obtains sea cucumber hydrolysate; Said compound protease is that stomach or trypsase and papain and neutral proteinase are formulated by weight 6:4:3, and the activity of each enzyme all is 10-20 million international units/g zymoprotein; Use this compound protease, enzymolysis under these conditions, the sea cucumber polypeptide kind that obtains is many, molecular weight is than homogeneous, and hydrolysis result is better;
C, oyster shell is cleaned up, at 300～900 ℃ of calcining 3h, cooling is ground into 80～100 orders, obtains oyster shell active calcium;
D, above-mentioned oyster shell active calcium is packed in the glass dish; Add above-mentioned sea cucumber hydrolysate again, the weight ratio of said sea cucumber hydrolysate and said oyster shell active calcium is 100:6～12, stirs 10～30min; Leave standstill behind 30～40min dryly, obtain sea cucumber polypeptide sequestering activity calcium; To calcium activated, the chelating degree is high, and uniformity is better with the basic chelating of the method sea cucumber polypeptide.
Hydrolysis temperature is 52 ℃ among the step b, enzymolysis time 177min, and the weight ratio of said compound protease and said sea cucumber granules is 2.5:100.
The sea cucumber polypeptide sequestering activity calcium that this method obtains is used in preparation improves the medicine of islet cells secretion, adds the necessary pharmaceutically assistant agent of approval, can be processed into tablet, pill, granule, capsule, microcapsules, oral liquid etc.
Compared with prior art, the invention has the advantages that: a kind of preparation method of sea cucumber polypeptide sequestering activity calcium, through boiling softening repeatedly with pure water to sea cucumber; Be ground into the sea cucumber granules that diameter is 1～2mm then, sea cucumber granules and water and compound protease are made into sea cucumber magma, and 45～65 ℃ of temperature; Enzymolysis 120～180min obtains sea cucumber hydrolysate, with 300～900 ℃ of calcinings of oyster shell 3h, is ground into 80～100 orders simultaneously; Obtain oyster shell active calcium, in glassware, sea cucumber hydrolysate and oyster shell active calcium are by weight mixing for 100:6～12; Stir 10～30min, leave standstill 30～40min drying, obtain sea cucumber polypeptide chelate oyster calcium activated.This method preparation is simple, and convenient for production, production cost is lower; Can fully remove salt marsh and agricultural chemicals, remove sea cucumber polysaccharide and selenka, the sea cucumber polypeptide purity that enzymolysis obtains is just high; Compound protease makes the sea cucumber polypeptide kind more, and molecular weight is than homogeneous, and the basic chelating of sea cucumber polypeptide is to calcium activated; The sea cucumber polypeptide sequestering activity calcium that obtains at last has the secretion that improves islet cells, reduces the effect of saccharification haemocyanin, urea nitrogen and creatinine content in the blood, can improve sugar tolerance; Improve blood lipid level, protection kidney, protection vascular endothelial cell etc.
The specific embodiment
Below in conjunction with embodiment the present invention is described in further detail.
Embodiment 1
A kind of preparation method of sea cucumber polypeptide sequestering activity calcium; Internal organ are removed in fresh sea cucumber impurity elimination, put into and outwell boiling water after the pot that fills pure water boils, add again and outwell boiling water again after pure water boils; So boil repeatedly altogether 4～5 times with pure water; Obtain softening sea cucumber, will soften sea cucumber again and be ground into the sea cucumber granules that diameter is 1～2mm, add 15 kg of water at 5 kilograms of sea cucumber granules; Add 0.125 kilogram of the compound protease (wherein the activity of each enzyme all is 10-20 million international units/g zymoprotein) that is mixed with by weight 6:4:3 by pepsin and papain and neutral proteinase again; Stir and process sea cucumber magma, under 52 ℃ of hydrolysis temperatures, enzymolysis 177min obtains sea cucumber hydrolysate; Simultaneously oyster shell is cleaned up, at 300～900 ℃ of calcining 3h, cooling is ground into 80～100 orders, obtains oyster shell active calcium; Then oyster shell active calcium is packed in the glass dish, add sea cucumber hydrolysate again, 100 kilograms of sea cucumber hydrolysate and 10 kilograms of oyster shell active calcium mixing chelatings stir 10-30min, and standing and drying 30-40min obtains sea cucumber polypeptide sequestering activity calcium.
Embodiment 2
Basic identical with embodiment 1, different just 5 kilograms of sea cucumber granules add 10 kilograms in water, add 0.1 kilogram of compound protease, hydrolysis temperature is 65 ℃, enzymolysis time is 120min, 100 kilograms of sea cucumber hydrolysate and 6 kilograms of oyster shell active calcium mixing chelatings.
Embodiment 3
Basic identical with embodiment 1; Different is that fresh sea cucumber is substituted by waterishlogged Stichopus japonicus, and 5 kilograms of sea cucumber granules add 50 kilograms in water, add 0.15 kilogram of compound protease; Pepsin is substituted by trypsase in the compound protease; Hydrolysis temperature is 45 ℃, and enzymolysis time is 180min, 100 kilograms of sea cucumber hydrolysate and 12 kilograms of oyster shell active calcium mixing chelatings.
Test Example
70 of SD rats, male and female half and half are divided into dosage G2 group and administration low dosage G3 group in blank control group, model group, administration high dose G1 group, the administration, model group rats by intraperitoneal injection STZ (100mg ﹒ kg at random
-1) carrying out modeling, administration G1, G2 and G3 group are pressed 10mgkg in advance respectively
-1D
-1, 6 mgkg
-1D
-1With 3 mgkg
-1D
-1Dosage is irritated the sea cucumber polypeptide sequestering activity calcium 60 days that embodiment 1 is stated on the Weishang, then lumbar injection STZ (100mg ﹒ kg
-1), blank group not medication is not injected STZ; Get blood through the tail vein in (blank is not injected, and gets blood simultaneously) the 3rd day behind the injection STZ, survey saccharification haemocyanin, urea nitrogen and creatinine content in the rat serum, obtain result as shown in table 1.
Table 1: sea cucumber polypeptide chelating calcium to rat blood serum biochemical indicator influence (
P<0.01)
| Group | Urea nitrogen (mmol/L) | Creatinine (μ mol/L) | Saccharification haemocyanin (mmol/L) |
| The blank group | 5.66±0.36 | 40.78±2.37 | 2.16±0.26 |
| Model group | 12.13±0.08 | 65.10±4.38 | 3.08±0.09 |
| Administration G1 group | 7.36±0.52 | 45.56±2.67 | 2.49±0.06 |
| Administration G2 group | 7.86±1.45 | 49.14±4.97 | 2.65±0.02 |
| Administration G3 group | 9.59±0.55 | 51.08±3.12 | 2.63±0.13 |
Can find out that from table 1 the administration group of taking sea cucumber polypeptide sequestering activity calcium explains that than each item biochemical indicator much less of model group the sea cucumber polypeptide sequestering activity calcium that the present invention obtains can reduce urea nitrogen, creatinine and saccharification serum albumin content in the blood.The people can increase the immunity of body if according to taking polypeptide chelate calcium activated 50-100mg/kg that the present invention obtains, ate 45 days continuously every day; Improve the secretion of islet cells; Improve sugar tolerance, reduce urea nitrogen and creatinine in the blood, improve blood lipid level and protection vascular endothelial cell; Prevention metabolism disorder of blood lipid etc. have certain protective role to kidney.In like manner, embodiment 2,3 sea cucumber polypeptide chelating calcium agent all have urea nitrogen, creatinine and the effect of saccharification haemocyanin that reduces in the blood, do not enumerate one by one at this.
Claims (3)
1. the preparation method of a sea cucumber polypeptide sequestering activity calcium is characterized in that step is following:
A, impurity elimination, go sea cucumber behind the internal organ to put into the pot that fills pure water to boil after; Outwell boiling water, add again and outwell boiling water after pure water boils, change pure water repeatedly altogether and boil 4～5 times; Obtain softening sea cucumber, will soften sea cucumber again and be ground into the sea cucumber granules that diameter is 1～2mm;
B, add entry at sea cucumber granules by weight the ratio of 1:2～5; Add compound protease again; The weight ratio of said compound protease and said sea cucumber granules is 2～3:100; Stirring, is under 45～65 ℃ at hydrolysis temperature, and enzymolysis 120～180min obtains sea cucumber hydrolysate; Said compound protease is that pepsin or trypsase and papain and neutral proteinase are formulated by weight 6:4:3, and the activity of said pepsin or trypsase and papain and neutral proteinase all is 10-20 million international units/g zymoprotein;
C, oyster shell is cleaned up, at 300～900 ℃ of calcining 3h, cooling is ground into 80～100 orders, obtains oyster shell active calcium;
D, above-mentioned oyster shell active calcium is packed in the glassware; Add above-mentioned sea cucumber hydrolysate again, the weight ratio of said sea cucumber hydrolysate and said oyster shell active calcium is 100:6～12, stirs 10～30min; Leave standstill behind 30～40min dryly, obtain sea cucumber polypeptide sequestering activity calcium.
2. the preparation method of a kind of sea cucumber polypeptide sequestering activity calcium as claimed in claim 1 is characterized in that hydrolysis temperature is 52 ℃ among the step b, enzymolysis time 177min, and the weight ratio of said compound protease and said sea cucumber granules is 2.5:100.
3. the sea cucumber polypeptide sequestering activity calcium that obtains of the preparation method of the described a kind of sea cucumber polypeptide sequestering activity calcium of claim 1 is used in preparation improves the medicine of islet cells secretion.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103829277A (en) * | 2012-11-23 | 2014-06-04 | 秦德文 | Novel functional food and making method thereof |
| CN104770766A (en) * | 2015-04-29 | 2015-07-15 | 郭中河 | Sea cucumber and oyster albumen powder preparation method |
| CN105671114A (en) * | 2016-02-25 | 2016-06-15 | 大连工业大学 | Preparation method of sea cucumber ovum peptide-calcium chelate |
| CN109355340A (en) * | 2018-11-23 | 2019-02-19 | 胜田(福清)食品有限公司 | A kind of preparation method with high thermal stability sea cucumber antioxidation chelation peptide |
| CN115153016A (en) * | 2021-04-06 | 2022-10-11 | 大连鑫玉龙海洋生物种业科技股份有限公司 | Sea cucumber polypeptide chelated zinc oral liquid and preparation method thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103829277A (en) * | 2012-11-23 | 2014-06-04 | 秦德文 | Novel functional food and making method thereof |
| CN104770766A (en) * | 2015-04-29 | 2015-07-15 | 郭中河 | Sea cucumber and oyster albumen powder preparation method |
| CN104770766B (en) * | 2015-04-29 | 2018-04-27 | 中食和畅生物科技有限公司 | A kind of preparation method of sea cucumber Oyster Protein powder |
| CN105671114A (en) * | 2016-02-25 | 2016-06-15 | 大连工业大学 | Preparation method of sea cucumber ovum peptide-calcium chelate |
| CN105671114B (en) * | 2016-02-25 | 2019-12-20 | 大连工业大学 | Preparation method of sea cucumber egg peptide calcium chelate |
| CN109355340A (en) * | 2018-11-23 | 2019-02-19 | 胜田(福清)食品有限公司 | A kind of preparation method with high thermal stability sea cucumber antioxidation chelation peptide |
| CN115153016A (en) * | 2021-04-06 | 2022-10-11 | 大连鑫玉龙海洋生物种业科技股份有限公司 | Sea cucumber polypeptide chelated zinc oral liquid and preparation method thereof |
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Application publication date: 20120718 |