CN1748714A - Method for preparing biological active substance of siphon-worm and its products - Google Patents
Method for preparing biological active substance of siphon-worm and its products Download PDFInfo
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- CN1748714A CN1748714A CNA2004100663487A CN200410066348A CN1748714A CN 1748714 A CN1748714 A CN 1748714A CN A2004100663487 A CNA2004100663487 A CN A2004100663487A CN 200410066348 A CN200410066348 A CN 200410066348A CN 1748714 A CN1748714 A CN 1748714A
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- sipunculoidea
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Abstract
The preparation process of bioactive substance of sipunculan includes the following steps: washing sipunculan and beating to obtain water solution; centrifuging to separate supernatant as sipunculan liquid and precipitate; adding water to the precipitate to form water solution and enzymolysis to obtain enzymolyzed liquid. The preparation process has the advantages of maintaining the bioactive substance, simple operation, low cost and no secondary pollution. The prepared sipunculan liquid and the enzymolyzed sipunculan liquid contains high-F functional peptide oligomer, polysaccharide, free amino acids and other active matters, and has antifatigue, anoxia-resisting, heat resisting, immune regulating and other biological regulating functions. The sipunculan liquid and the enzymolyzed sipunculan liquid may be further prepared into oral liquid, capsule, granule, tablet, etc. for easy taking.
Description
Technical field
The present invention relates to a kind of marine product manufactured goods and processing method thereof, specifically a kind of is that raw material carries out the preparation method of bioactive substance and the series of products that obtained by this preparation method thereof with the Sipunculoidea.
Background technology
Sipunculoidea is commonly called as HAIQU, sandworm, Hai Ding, native Radix Crotalariae szemoensis etc., is a kind of euryhaline organism, and not only meat is tender and crisp for it, delicious flavour, and has higher medicinal, food therapy value, can regulate the multiple function of body.Put down in writing its edible and medical value in the multiple pharmacopeia of China, Sipunculoidea is cold in nature, sweet in the mouth, salty, have nourishing YIN to lower pathogenic fire, lung heat clearing tonify deficiency, invigorate blood circulation and keep fit and function such as the kidney invigorating skin care, can treat that osteopyrexia and fever, night sweat due to yin deficiency, the deficiency of the lung are breathed with cough, disease such as abundant expectoration uncomfortable in chest and women's milk in puerperal rareness, to treatment pulmonary tuberculosis cough, neurasthenia, infantile spleen deficiency and deficiency of the kidney and nocturia is frequent etc. that disease all produces effect, so the title of " Cordyceps " in the ocean is arranged.Because Sipunculoidea is a kind of nutritious, therefore trace element such as rich in proteins, several amino acids and calcium, phosphorus, ferrum and can regulate the multiple composition of taurine of brain function also has the good reputation of " seabeach sausage ", favored by domestic and international consumer.
At present very single to the eating method of this animal, mainly be directly edible.But because its nutrient substance mostly is macromolecule greatly, be not easy directly to be absorbed by human body and utilize, particularly Functional Polypeptides and polysaccharide isoreactivity material do not dissociate out fully, make human body can not effectively transform and utilize its nutrient substance, both increased production cost of products, and reduced the effect of product again, utilization of nutrients is not high, but also contain certain toxin, easily human body is produced certain harm.
The disclosed ocean of Chinese invention patent application CN1430975A Sipunculoidea effective component extracts and method thereof are that to select the ocean Sipunculoidea for use be raw material, use the saline solution extracting, the organic solvent fractional precipitation, and ultrafiltration obtains the active component extract after the lyophilization.The following shortcoming that this extracting method exists: 1, extracting method complexity, cost height.2, adopt organic solvent, have the latency of harmful to human.3, lose the existing nutrient substance of Sipunculoidea easily.
Summary of the invention
Technical problem to be solved by this invention is to overcome the above-mentioned defective of prior art and preparation method that a kind of biological active substance of siphon-worm is provided, make it can access natural bioactivity substance in the Sipunculoidea, transform the polymer substance in the Sipunculoidea, reach the purpose of harmless, efficient, low-cost preparation biological active substance of siphon-worm.
Another object of the present invention is to utilize containing of Sipunculoidea to enrich the enzymic hydrolysates of Functional Polypeptides, free amino acid, preparation has oral liquid, capsule, tablet and the electuary of the hydrolysate of certain special physiological function, to improve medicinal, the nutritional activities composition of product, instant edible, and fully improve its using value.
The present invention solves the problems of the technologies described above the technical scheme that is adopted:
The preparation method of biological active substance of siphon-worm may further comprise the steps:
(1), Sipunculoidea cleaned, smashed and produce certain density aqueous solution;
(2), centrifugal aqueous solution, be divided into supernatant and precipitation, supernatant is a sipunculids fluid;
(3), precipitation is added water make solution in different concentration and add enzyme and carry out enzymolysis, make enzyme hydrolyzate.
Main feature of the present invention is 2 points: the one, extract the initial condition dissolubility functional mass in the Sipunculoidea earlier, and the 2nd, enzyme hydrolysis.At first extract the natural active matter of Sipunculoidea such as oligopeptide, cyclic peptide enzyme, polypeptide, enzyme, terpene, alkaloid, polysaccharide, unsaturated fatty acid, nucleotide, free amino acid; these materials have multiple pharmacological function; be exactly because the effect of these natural active matters can be protected Sipunculoidea prior biological active substance like this as Chinese medicine in one's early years Sipunculoidea.Utilize zymolysis technique insoluble hydrolysate large protein then, improve and the powerful active component of conversion Sipunculoidea, the content of the oligomeric Functional Polypeptides of particularly high F value is convenient to these nutrient substance in the intravital conversion of people, promotes the absorption of human body to these functional components.
Above-mentioned enzyme can be used papain, trypsin, 1389 neutral protease and compound protease, particularly adopts compound protease, and hydrolysis effect is better.Compound protease can be commercial enzyme, can buy from the market, also can oneself produce.Compound protease contains cellulase, amylase and multiple protein enzyme.
The preparation of Sipunculoidea aqueous solution can utilize fresh individuality, also can utilize the individuality of other states.
A kind of the Sipunculoidea hydrotrope and enzyme hydrolyzate by method for preparing, this product contains rich functions peptide, polysaccharide and free amino acid, has human body immunity improving power, effect such as degeneration-resistant.
The use of the above-mentioned Sipunculoidea hydrotrope and enzyme hydrolyzate also can be made into oral liquid, capsule, tablet and electuary etc., as can be made into athlete's health product for convenience.Because it is similar that this enzyme hydrolyzate liquid similar with other is made the method for various preparations, therefore as long as in said hydrolyzed liquid, add respective substance, just can be made into oral liquid, capsule, tablet and electuary, the preparation method of these preparations is known, does not do detailed argumentation.
Compared with prior art, the invention has the advantages that and select two-step method for use, and with the polymer substance of protease hydrolysis Sipunculoidea, can keep original bioactive substance like this, can fully transform potential bioactive substance again, also without organic solvent, therefore easy and simple to handle, production cost is low, non-secondary pollution.The special compound protease that uses, its method is easier, hydrolysis time is shorter, active substance transforms manyly.The oligomeric Functional Polypeptides, polysaccharide and the free amino acid isoreactivity composition that contain more high F value in natural bioactivity substance that makes and the enzyme hydrolyzate, have better resisting fatigue, anoxia enduring, high temperature resistant, regulate multiple biological regulatory functions such as immunity, lactogenic.
The sipunculids fluid and the enzyme hydrolyzate that utilize said method to make can further be processed into oral liquid, capsule, electuary, tablet etc., and be edible more convenient like this, takes better effects if.
The specific embodiment
Below in conjunction with embodiment the present invention is described in further detail.
Embodiment 1:
The preparation method of biological active substance of siphon-worm, step is as follows:
(1), fresh Sipunculoidea water cleans up, after the Sipunculoidea that cleans up was pulverized with meat grinder, according to 1: 2-3 concentration doubly added water, and the reuse fiberizer further is milled to certain density aqueous solution;
(2), to get the Sipunculoidea aqueous solution that makes in the step 1 centrifugal, supernatant is sipunculids fluid, but its vacuum drying or lyophilization;
(3), get precipitate in the step 2 and add entry and make solution in different concentration and add protease again and carry out enzymolysis, obtain enzymolysis solution.
The different Sipunculoidea concentration and the addition of enzyme obtain different results, the condition of enzymolysis and the results are shown in Table 1 to table 8.
Identical enzyme addition and different Sipunculoidea concentration see Table 9 to table 10 with the result of variations of enzymolysis time.
As can be seen, the enzymatic hydrolysis condition of sipunculids fluid is preferably from table 1 to table 10:
The concentration of Sipunculoidea is 10-60% (weight), and concentration is the percentage ratio that Sipunculoidea accounts for whole aqueous solution;
The addition of enzyme is for being 2 * 10
3IU/kg-1.4 * 10
6IU/kg (Sipunculoidea);
Temperature is selected the serviceability temperature of composite zymoprotein enzyme for use, is generally 35-70 ℃, heats up gradually;
Enzymolysis time 1-6h.
Embodiment 2:
By sipunculids fluid that obtains among the embodiment 1 and Sipunculoidea enzymolysis solution, add the lactose tabletting and make tablet, have the function of human body immunity improving power; Sipunculids fluid and Sipunculoidea enzymolysis solution filtrate 1000: Mel 25: the Fructus Jujubae water cooking liquid filters 10: sugared 70 fills become oral liquid.
Embodiment 3
By the sipunculids fluid that obtains among the embodiment 1 and enzymolysis solution directly or add behind the lactose spray drying to make powder encapsulated, the concrete ratio of lactose can be decided by the polysaccharide and the amino acid whose content of required product.
Embodiment 4
Concentrated by sipunculids fluid that obtains among the embodiment 1 and enzymolysis solution, as inserts tabletting sugar coating, tablet is made in polishing with starch.
Embodiment 5
Electuary is made in adding lactose or beta cyclodextrin pelletize in sipunculids fluid that obtains in by embodiment 1 and the enzymolysis solution.
Embodiment 6
Will by sipunculids fluid that obtains among the embodiment 1 and enzymolysis solution add the L-carnitine again according to embodiment 2, embodiment 3, embodiment 4, embodiment 5 method system oral liquids, do capsule, electuary and film-making, this series products is athletic health product.
Enzymatic hydrolysis condition and experimental result:
Amino acid content (mg/ml) behind the table 1 Sipunculoidea neutral proteinase hydrolysis
| Substrate/enzyme concentration | 1.3×10 4IU/kg | 1.1×10 5IU/kg | 2.0×10 5IU/kg | 3.0×10 5IU/kg |
| 10％ 20％ 40％ 60％ | 3.841 3.899 3.778 3.808 | 4.121 4.173 3.767 3.713 | 4.553 4.202 4.123 3.825 | 4.985 4.274 3.961 4.145 |
The content (mg/ml) of polysaccharide behind the table 2 Sipunculoidea neutral proteinase hydrolysis
| Substrate/enzyme concentration | 1.3×10 4IU/kg | 1.1×10 5IU/kg | 2.0×10 5IU/kg | 3.0×10 5IU/kg |
| 10％ 20％ 40％ 60％ | 84.45 66.43 72.95 96.15 | 158.45 125.58 117.88 110.95 | 217.91 134.70 136.71 124.17 | 241.04 191.09 165.45 144.78 |
Amino acid content (mg/ml) after the table 3 Sipunculoidea trypsin hydrolyzing
| Substrate/enzyme concentration | 1.3×10 5IU/kg | 3.8×10 5IU/kg | 6.3×10 5IU/kg | 8.8×10 5IU/kg |
| 10％ 20％ 40％ 60％ | 4.078 3.611 3.508 3.186 | 4.208 3.943 4.177 4.033 | 4.812 4.017 4.274 3.938 | 5.633 4.317 4.371 4.439 |
Polyoses content (mg/ml) after the table 4 Sipunculoidea trypsin hydrolyzing
| Substrate/enzyme concentration | 1.3×10 5IU/kg | 3.8×10 5IU/kg | 6.3×10 5IU/kg | 8.8×10 5IU/kg |
| 10％ 20％ 40％ 60％ | 65.29 73.04 73.28 79.24 | 84.45 83.17 83.19 85.32 | 89.07 76.56 75.59 72.10 | 81.80 75.68 62.71 81.88 |
Amino acid content (mg/ml) after the table 5 Sipunculoidea papain hydrolysis
| Substrate/enzyme concentration | 9.5×10 3IU/kg | 7.6×10 4IU/kg | 1.5×10 5IU/kg |
| 10％ 20％ 40％ 60％ | 2.826 2.589 2.309 2.193 | 3.646 3.151 2.914 2.953 | 3.949 3.367 2.968 2.849 |
The content (mg/ml) of polysaccharide after the table 6 Sipunculoidea papain hydrolysis
| Substrate/enzyme concentration | 9.5×10 3IU/kg | 7.6×10 4IU/kg | 1.5×10 5IU/kg |
| 10％ 20％ 40％ 60％ | 120.79 106.4 115.71 | 148.54 125.65 111.22 | 136.64 119.61 114.12 |
Amino acid content (mg/ml) after the enzyme hydrolysis of table 7 Sipunculoidea compound protein
| Substrate/enzyme concentration | 2×10 4IU/kg | 6×10 4IU/kg | 1×10 4IU/kg | 1.4×10 5IU/kg... |
| 10％ 20％ 40％ 60％ | 11.571 9.395 5.063 11.947 | 13.558 12.374 11.542 13.530 | 19.567 14.678 15.117 12.357 | 22.309 16.404 14.579 14.012 |
The content (mg/ml) of polysaccharide after the enzyme hydrolysis of table 8 Sipunculoidea compound protein
| Substrate/enzyme concentration | 2×10 4IU/kg | 6×10 4IU/kg | 1×10 4IU/kg | 1.4×10 5IU/kg... |
| 10％ 20％ 40％ 60％ | 138.65 74.11 95.34 50.63 | 90.19 104.03 82.90 60.38 | 124.31 169.65 100.17 76.69 | 150.39 151.19 100.21 81.41 |
The relation of amino acid content (mg/ml) and enzymolysis time in the table 9 Sipunculoidea compound protein enzyme hydrolyzate:
The enzyme addition is: 5 * 10
4IU/kg, temperature adopts gradually and heats up
| Temperature/time concentration of substrate | 10％ | 20％ | 40％ | 60％ |
| 35℃-70℃/2h 35℃-70℃/4h 35℃-70℃/6h | 11.108 19.359 13.023 | 9.528 18.306 12.311 | 6.926 15.066 12.064 | 5.783 12.846 12.098 |
The relation of polyoses content (mg/ml) and enzymolysis time in the table 10 Sipunculoidea compound protein enzyme hydrolyzate:
The enzyme addition is: 5 * 10
4IU/kg, temperature adopts gradually and heats up
| Temperature/time concentration of substrate | 10％ | 20％ | 40％ | 60％ |
| 35-70℃/2h 35-70℃/4h 35-70℃/6h | 110.19 156.45 121.03 | 99.45 163.23. 132.25 | 96.23 144.56 133.56 | 96.05 139.02 135.23 |
Claims (8)
1, the preparation method of biological active substance of siphon-worm is characterized in that: may further comprise the steps:
(1), Sipunculoidea cleaned, smashed and produce certain density aqueous solution;
(2), centrifugal aqueous solution, be divided into supernatant and precipitation, supernatant is a sipunculids fluid;
(3), precipitation is added water make solution in different concentration and add enzyme and carry out enzymolysis, make enzyme hydrolyzate.
2, the preparation method of biological active substance of siphon-worm as claimed in claim 1 is characterized in that: described enzyme is a compound protease.
3, the preparation method of biological active substance of siphon-worm as claimed in claim 1 or 2 is characterized in that: described enzymatic hydrolysis condition is: the content of Sipunculoidea is 10-60% (weight), and the enzyme addition is 2 * 10
3IU/kg-1.4 * 10
6IU/kg (Sipunculoidea), hydrolysis temperature are the serviceability temperature of compound protease, are 35-70 ℃, and adopt intensification gradually, and enzymolysis time is 1-6h.
4, sipunculids fluid and the enzymolysis solution that obtains by the preparation method of claim 1 or 2 or 3 biological active substance of siphon-worm.
5, biological active substance of siphon-worm oral liquid is characterized in that: the sipunculids fluid and the enzymolysis solution that are obtained by claim 4 add Mel, Fructus Jujubae water cooking liquid, sugared fill, perhaps add Mel, Fructus Jujubae water cooking liquid, sugar and the fill of L-carnitine.
6, biological active substance of siphon-worm capsule is characterized in that: sipunculids fluid that is obtained by claim 4 and enzymolysis solution add lactose and form, and perhaps add lactose and the L-carnitine forms.
7, biological active substance of siphon-worm tablet is characterized in that: sipunculids fluid that is obtained by claim 4 or enzymolysis solution add starch and form, and perhaps add starch, the L-carnitine forms.
8, biological active substance of siphon-worm electuary is characterized in that: sipunculids fluid that is obtained by claim 4 or enzymolysis solution add lactose or the beta cyclodextrin pelletize is made, and perhaps add to add lactose again behind the L-carnitine or the beta cyclodextrin pelletize is made.
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| CNA2004100663487A CN1748714A (en) | 2004-09-14 | 2004-09-14 | Method for preparing biological active substance of siphon-worm and its products |
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|---|---|---|---|
| CNA2004100663487A CN1748714A (en) | 2004-09-14 | 2004-09-14 | Method for preparing biological active substance of siphon-worm and its products |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102153666A (en) * | 2011-02-22 | 2011-08-17 | 中国人民解放军海军医学研究所 | Sipunculidae polysaccharide and preparation method thereof and application thereof to anti-fatigue function |
| CN101381411B (en) * | 2008-09-28 | 2011-12-14 | 宁波大学 | Sipunculid collagen protein and preparation method thereof |
| CN101210261B (en) * | 2006-12-29 | 2011-12-14 | 宁波大学 | Method for preparing sipunculid polypeptide and application thereof |
| CN103169081A (en) * | 2013-03-22 | 2013-06-26 | 薛命雄 | Sandworm polypeptide enteric capsule |
| CN103330204A (en) * | 2013-07-03 | 2013-10-02 | 广西壮族自治区海洋研究所 | Sipunculus nudus polysaccharide oral liquid |
| CN103483462A (en) * | 2013-09-23 | 2014-01-01 | 宁波大学 | Preparation method for phascolosoma esculenta polysaccharide and application of phascolosoma esculenta polysaccharide in treating septicemia |
| CN104146233A (en) * | 2014-04-30 | 2014-11-19 | 浙江省海洋水产养殖研究所 | Natural seasoner prepared from sipunculus nudus and preparation method of natural seasoner |
| CN104473185A (en) * | 2014-12-27 | 2015-04-01 | 广西壮族自治区海洋研究所 | Effervescent tablet containing vitamin C and sipunculusnudus enzymatic protein |
| CN105918461A (en) * | 2016-04-08 | 2016-09-07 | 宁波大学 | Low-calorific-value yogurt containing phascolosoma esculenta polysaccharide and preparation method thereof |
| CN112825986A (en) * | 2021-01-20 | 2021-05-25 | 中国热带农业科学院农产品加工研究所 | Sipunculus nudus polypeptide beverage composition, beverage prepared from Sipunculus nudus polypeptide beverage composition and preparation method of beverage |
| CN116144725A (en) * | 2023-04-19 | 2023-05-23 | 青岛大学附属医院 | Application of marine organism polypeptide in preparing anesthetic auxiliary drug |
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2004
- 2004-09-14 CN CNA2004100663487A patent/CN1748714A/en active Pending
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101210261B (en) * | 2006-12-29 | 2011-12-14 | 宁波大学 | Method for preparing sipunculid polypeptide and application thereof |
| CN101381411B (en) * | 2008-09-28 | 2011-12-14 | 宁波大学 | Sipunculid collagen protein and preparation method thereof |
| CN102153666A (en) * | 2011-02-22 | 2011-08-17 | 中国人民解放军海军医学研究所 | Sipunculidae polysaccharide and preparation method thereof and application thereof to anti-fatigue function |
| CN102153666B (en) * | 2011-02-22 | 2013-02-27 | 中国人民解放军海军医学研究所 | Sipunculidae polysaccharide and preparation method thereof and application thereof to anti-fatigue function |
| CN103169081B (en) * | 2013-03-22 | 2014-12-10 | 薛命雄 | Sandworm polypeptide enteric capsule |
| CN103169081A (en) * | 2013-03-22 | 2013-06-26 | 薛命雄 | Sandworm polypeptide enteric capsule |
| CN103330204A (en) * | 2013-07-03 | 2013-10-02 | 广西壮族自治区海洋研究所 | Sipunculus nudus polysaccharide oral liquid |
| CN103483462A (en) * | 2013-09-23 | 2014-01-01 | 宁波大学 | Preparation method for phascolosoma esculenta polysaccharide and application of phascolosoma esculenta polysaccharide in treating septicemia |
| CN103483462B (en) * | 2013-09-23 | 2015-12-23 | 宁波大学 | A kind of preparation method of Phascolosoma polysaccharide and application |
| CN104146233A (en) * | 2014-04-30 | 2014-11-19 | 浙江省海洋水产养殖研究所 | Natural seasoner prepared from sipunculus nudus and preparation method of natural seasoner |
| CN104146233B (en) * | 2014-04-30 | 2015-09-09 | 浙江省海洋水产养殖研究所 | Natural flavouring utilizing siphon-worm to prepare and preparation method thereof |
| CN104473185A (en) * | 2014-12-27 | 2015-04-01 | 广西壮族自治区海洋研究所 | Effervescent tablet containing vitamin C and sipunculusnudus enzymatic protein |
| CN105918461A (en) * | 2016-04-08 | 2016-09-07 | 宁波大学 | Low-calorific-value yogurt containing phascolosoma esculenta polysaccharide and preparation method thereof |
| CN112825986A (en) * | 2021-01-20 | 2021-05-25 | 中国热带农业科学院农产品加工研究所 | Sipunculus nudus polypeptide beverage composition, beverage prepared from Sipunculus nudus polypeptide beverage composition and preparation method of beverage |
| CN116144725A (en) * | 2023-04-19 | 2023-05-23 | 青岛大学附属医院 | Application of marine organism polypeptide in preparing anesthetic auxiliary drug |
| CN116144725B (en) * | 2023-04-19 | 2023-06-20 | 青岛大学附属医院 | Application of marine organism polypeptide in preparing anesthetic auxiliary drug |
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