CN111758967A - Giant salamander collagen peptide composition and application thereof - Google Patents
Giant salamander collagen peptide composition and application thereof Download PDFInfo
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- CN111758967A CN111758967A CN202010682106.XA CN202010682106A CN111758967A CN 111758967 A CN111758967 A CN 111758967A CN 202010682106 A CN202010682106 A CN 202010682106A CN 111758967 A CN111758967 A CN 111758967A
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- giant salamander
- enzymolysis
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/30—Artificial sweetening agents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/06—Enzymes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/16—Inorganic salts, minerals or trace elements
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/175—Amino acids
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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Abstract
The invention relates to the technical field of giant salamander deep processing, and particularly relates to a giant salamander collagen peptide composition and application thereof. The composition provided by the invention comprises the following components in percentage by mass (1-50): 1 giant salamander collagen peptide and mangosteen pericarp extract. The giant salamander collagen peptide is obtained by sequentially carrying out three-stage enzymolysis on a giant salamander raw material from which fat and non-collagen are removed by using alkaline protease, neutral protease and flavourzyme, and then filtering and refining an enzymolysis product. The average molecular weight of the giant salamander collagen peptide is below 1000Da, the protein content is above 95%, the ash content is below 2%, the giant salamander collagen peptide and the mangosteen pericarp extract can be cooperated with each other, and after the giant salamander collagen peptide and the mangosteen pericarp extract are compounded, the giant salamander collagen peptide has an obvious lipid-lowering effect, can effectively reduce fat deposition, and assists obese people to recover weight.
Description
Technical Field
The invention relates to the technical field of giant salamander deep processing, and particularly relates to a composition containing giant salamander collagen peptide, application and a preparation thereof.
Background
Giant salamanders (Andrias davidianus) are amphibians with tails in the giant salamander family and the giant salamander genus, also called giant salamanders, are amphibians with high nutritional values, and are known as 'underwater ginseng'. Collagen is a fibrous protein, the main constituent of which is amino acid, and collagen peptide and by-product amino acid are produced after hydrolysis. In recent years, collagen peptide has been increasingly used in various skin care products, health care products, medicines and foods because of its remarkable biological functional activity. The skin and the meat of the giant salamander both contain abundant collagen. Besides various physiological active functions of general collagen peptide such as water replenishing, water locking, oxidation resistance, aging resistance, tumor resistance, blood pressure reduction, gastric mucosa, bone and ligament health care, hair loss prevention, hair growth promotion and the like, the giant salamander collagen peptide also has the special effects of repairing epithelial cells, promoting wound healing, protecting liver injury and enhancing immunity.
The preparation method of the giant salamander collagen peptide mainly comprises acid hydrolysis, alkali hydrolysis and an enzymolysis method. At present, the preparation of giant salamander collagen peptide is more concerned about the molecular weight of hydrolysate peptide so as to obtain collagen peptide with smaller molecular weight and favorable absorption of organisms, and the content of active peptide with specific function is less concerned. Patent application CN105018555A discloses a preparation method of giant salamander skin collagen peptide, which comprises the steps of taking giant salamander skin as a raw material, degreasing, removing black skin and foreign protein, carrying out high-pressure treatment, adding alkaline protease for enzymolysis, decoloring by using activated carbon, centrifuging, microfiltration by using a filter membrane, ultrafiltration by using an ultrafiltration membrane, and nanofiltration by using a nanofiltration membrane to obtain filtrate, concentrating and freeze-drying the filtrate to obtain giant salamander skin collagen peptide powder, wherein the molecular weight of the prepared collagen peptide is small, and more than 90% of the molecular weight is less than 1000 daltons. Patent application CN111172226A discloses collagen peptides for intestinal absorption and extraction method and application thereof. Cleaning giant salamander skin, removing fishy smell, soaking in NaOH solution, and washing with water; carrying out enzymolysis on the treated giant salamander skin, then carrying out enzyme deactivation treatment, then carrying out centrifugal filtration and collecting filter residues; and sequentially carrying out microfiltration, ultrafiltration and sodium filtration on the filter residue, concentrating, freeze-drying and crushing to obtain the collagen peptide for intestinal absorption.
The polypeptide product composition of the giant salamander collagen after enzymolysis is very complex, wherein the polypeptide product composition contains functional active peptide with known or unknown sequence, but also contains a large amount of peptides without target effect or even any substantial effect, and the existence of the peptides can form a competitive relationship with the absorption and utilization of the functional active peptide and is not beneficial to the absorption and utilization of the functional active peptide. On the other hand, active peptides having specific functions often include a plurality of peptides having different amino acid compositions and different molecular weights, and are difficult to distinguish by some common characteristics. Although collagen peptide with small molecular weight can be obtained by proper single enzymolysis or compound enzymolysis in the prior art, the content and the activity of the peptide with specific function are difficult to ensure.
Disclosure of Invention
The invention aims to provide a giant salamander collagen peptide composition with excellent lipid-lowering function, and the invention also aims to provide a preparation containing the composition.
The invention provides a giant salamander collagen peptide composition which comprises the following components in percentage by mass (1-50): 1 giant salamander collagen peptide and mangosteen pericarp extract.
The invention carries out long-term and deep research on the deep processing method of the giant salamander collagen and the functions of the processed product. The invention aims to obtain giant salamander collagen peptide with the lipid-lowering function, and a large number of giant salamander collagen enzymolysis methods are tried, but the giant salamander collagen peptide with the ideal lipid-lowering effect is still difficult to obtain. The invention unexpectedly discovers that the composition obtained by compounding the giant salamander collagen peptide prepared by the specific enzymolysis process and the mangosteen pericarp extract according to the proportion has the remarkably improved lipid-lowering activity, the giant salamander collagen peptide and the mangosteen pericarp extract can have a synergistic effect, and the lipid-lowering activity of the compounded composition is far higher than that of the lipid-lowering activity of the giant salamander collagen peptide and the mangosteen pericarp extract when the giant salamander collagen peptide and the mangosteen pericarp extract are used.
In the giant salamander collagen peptide composition provided by the invention, the mass ratio of the giant salamander collagen peptide to the mangosteen pericarp extract is preferably (1-40): 1; more preferably (1-35): 1; more preferably (1-10): 1.
preferably, the giant salamander collagen peptide is prepared by a method comprising the following steps: the giant salamander skin and/or meat from which fat and non-collagen are removed is sequentially subjected to three-stage enzymolysis by alkaline protease, neutral protease and flavourzyme to prepare zymolyte.
The skin and/or meat of the giant salamander are/is used as raw materials, the three-stage enzymolysis method adopting the enzymolysis sequence can ensure that the giant salamander raw materials are subjected to full enzymolysis, high-content small molecular collagen peptide (less than 1000Da) is obtained, and more importantly, the zymolyte can generate a synergistic effect with the mangosteen skin extract, so that the lipid-lowering effect which is remarkably better than the effect of the skin and/or meat of the giant salamander on independent action is achieved.
In the three-stage enzymolysis, the reaction temperature of each stage of enzymolysis is not higher than 60 ℃.
In the enzymolysis process, reaction conditions such as enzymolysis temperature and the like are generally selected according to the optimal reaction conditions of the enzyme, however, the invention unexpectedly finds that even though the alkaline protease can exert excellent enzymolysis activity under the temperature condition of higher than 60 ℃, the synergistic effect of the zymolyte obtained by enzymolysis under the temperature condition of higher than 60 ℃ and the mangosteen pericarp extract is reduced.
The activities of alkaline protease, neutral protease and flavourzyme are comprehensively considered, and the enzymolysis temperature of each section is preferably controlled to be 40-60 ℃.
In the three-stage enzymolysis process, the pH value of each stage of enzymolysis is preferably controlled to be 6-9. The giant salamander collagen hydrolysate obtained under the condition has better synergistic effect with the mangosteen pericarp extract.
Specifically, the three-stage enzymolysis comprises the following steps:
(1) alkaline protease enzymolysis: adding alkaline protease, wherein the mass ratio of the alkaline protease to the giant salamander skin and/or meat without fat and non-collagen is 0.1-0.5%, the enzymolysis temperature is 55-60 ℃, and the enzymolysis time is 0.5-1.5 h;
(2) carrying out enzymolysis by neutral protease: the mass ratio of the added neutral protease to the skin and/or meat of the giant salamander without the non-collagen protein is 0.1-0.5%, the enzymolysis temperature is 45-50 ℃, and the enzymolysis time is 1-2.5 h;
(3) carrying out enzymolysis on flavourzyme: the mass ratio of the added flavourzyme to the giant salamander skin and/or meat without the non-collagen protein is 0.1-0.5%, the enzymolysis temperature is 45-55 ℃, and the enzymolysis time is 0.3-3 h.
In the three-stage enzymolysis process, after the last stage of enzymolysis is finished and the temperature is reduced, protease for the next stage of enzymolysis can be directly added for enzymolysis.
The mangosteen pericarp extract is an ethanol extract of mangosteen pericarp. The invention finds that the ethanol extract of the mangosteen pericarp can better cooperate with the giant salamander collagen peptide prepared by enzymolysis, and better lipid-lowering effect is achieved.
Preferably, the mangosteen pericarp extract is obtained by leaching with 40-60% ethanol water solution by volume percentage and drying the leaching solution.
In order to be more beneficial to enzymolysis, before the enzymolysis of the alkaline protease, the giant salamander skin and/or meat from which fat and non-collagen are removed is/are preferably treated at the temperature of 90-100 ℃ for 0.5-1 h. The high-temperature treatment can lead the collagen in the giant salamander raw material to be properly denatured, which is beneficial to the enzymolysis reaction.
The removal of non-collagen from giant salamander raw material can be carried out by conventional methods, such as: soaking in alkali solution. Preferably, soaking the fat-removed giant salamander skin and/or meat in 0.05-0.1M alkali liquor for 3.5-4.5 h, and removing non-collagen to obtain the fat-removed giant salamander skin and/or meat.
Specifically, before three-stage enzymolysis, the pretreatment method of the giant salamander raw material comprises the following steps: after the giant salamander whole fish is cut into pieces and is subjected to skin, meat and fat separation treatment, soaking the skin and/or meat of the giant salamander in 0.05-0.1M NaOH solution for 3.5-4.5 h to remove non-collagen, wherein the material-liquid ratio (mass-volume ratio g: ml) is 1 (5-10); then washing the mixture by deionized water to be neutral, draining, and mixing the mixture according to a material-liquid ratio (mass-volume ratio g: ml) of 1: (1-3) adding water, heating to 90-100 ℃, and keeping the temperature for 0.5-1 h.
After the pretreatment, the giant salamander raw material can be more fully subjected to enzymolysis (especially small molecule active peptide which is beneficial to reducing blood fat and has synergistic effect with the mangosteen pericarp extract).
In the preparation process of the giant salamander collagen peptide, after pretreatment and three-stage enzymolysis of the giant salamander raw material are finished, a proper amount of acid is added to adjust the pH of an zymolyte to be neutral, and then enzyme deactivation treatment is carried out.
The enzyme deactivation treatment described above may be carried out by methods conventional in the art, for example: and preserving the heat for 4-8 min at 85-90 ℃.
The alkaline protease, neutral protease and flavourzyme used in the present invention are all commonly used proteases in the art and commercially available.
Aiming at the characteristics of impurities such as peptide composition, ash content and the like of the zymolyte obtained by three-stage enzymolysis, the invention further provides a filtering and refining method of the adapted zymolyte, which specifically comprises the following steps:
and (3) after enzyme deactivation treatment, performing primary filtration, activated carbon adsorption, fine filtration and drying treatment on the zymolyte in sequence.
Wherein, the primary filtration comprises the filtration of a plate and frame filter which takes perlite as a filter aid. The perlite is matched with a plate and frame filter for filtering, so that macromolecular impurities in zymolyte can be well removed, and meanwhile, the adverse effect on the peptide activity of the zymolyte can not be generated.
The preferable mass ratio of the perlite to the zymolyte is 0.1-2%, and the pressure of the plate-and-frame filter is 1.5-2 MPa.
Further, after primary filtration, activated carbon adsorption treatment is carried out, and after the zymolyte after the activated carbon adsorption treatment is concentrated, fine filtration treatment is carried out.
Wherein the fine filtration sequentially comprises the steps of taking the pore volume of 0.3-1.0 cm3The filter is filtered by a diatomite filter with the aperture of 1-2 mu m, a paperboard filter with the filtering precision of 0.4-0.8 mu m and a polypropylene micropore folding filter element with the filtering precision of 0.22/0.22 mu m.
By adopting the filtering and refining method of primary filtering, activated carbon adsorption and fine filtering, the ash content of the giant salamander collagen peptide can be effectively reduced, the protein content and purity of the giant salamander collagen peptide are ensured, and adverse effects on the activity of the peptide are avoided.
The drying treatment can be spray drying or freeze drying, and the giant salamander collagen peptide powder is prepared by spray drying or freeze drying.
The giant salamander collagen peptide composition provided by the invention can also contain other components with lipid-lowering and other effects, such as resveratrol.
The invention provides a giant salamander collagen peptide composition which comprises the following components in parts by weight: 0.5-20 parts of mangosteen pericarp extract, 10-50 parts of giant salamander collagen peptide and 1-5 parts of resveratrol. In the composition, the addition of resveratrol can further improve the lipid-lowering function of the composition.
Specifically, the invention also provides a giant salamander collagen peptide composition, which is used for realizing more comprehensive lipid-lowering function and maintaining the balance of nutrition and metabolism of a body; the paint comprises the following components in parts by weight: 0.5-20 parts of mangosteen pericarp extract, 10-50 parts of giant salamander collagen peptide, 1-5 parts of resveratrol, 0.0003-0.001 part of biotin, 1-20 parts of plant dietary fiber, 1-15 parts of taurine, 0.0005-0.004 part of folic acid, 0.2-0.5 part of coenzyme Q10, 0.5-4 parts of beta-Nicotinamide Mononucleotide (NMN), 0.005-0.05 part of manganese, 0.01-0.15 part of iron, 0.0001-0.0011 part of selenium and 0.0005-0.015 part of copper. The composition can ensure nutrition supply of organism and reduce body weakness caused by blood lipid while exerting excellent blood lipid lowering function.
Wherein the plant dietary fiber can be inulin or fruit and vegetable powder, and can improve gastrointestinal function.
In addition, the manganese, iron, selenium and copper are provided in the form of manganese sulfate, ferrous sulfate, sodium selenite and copper sulfate. The amount is calculated by the mass of manganese, iron, selenium and copper.
The invention also provides application of the giant salamander collagen peptide-containing composition in preparation of products with lipid-lowering or weight-losing functions.
Preferably, the product is a food, a medicine, a health product, or the like.
The invention provides a giant salamander collagen peptide preparation which takes the giant salamander collagen peptide-containing composition as an active ingredient.
The giant salamander collagen peptide preparation can be a liquid preparation (oral liquid, aqueous solution and the like) or a solid preparation (powder, electuary, tablets and the like).
The giant salamander collagen peptide preparation further comprises an auxiliary material, and the auxiliary material comprises a seasoning auxiliary material.
Preferably, the seasoning auxiliary material comprises the following components in parts by weight: 2-7 parts of sour modifier, 0.1-50 parts of sweet modifier and 0.5-3 parts of natural flavoring agent.
Wherein, the sour flavoring agent is preferably one or more of citric acid and malic acid. The sweet flavoring agent is preferably one or more of xylitol, stevioside and erythritol. The sugar substitute component is used as sweet flavoring agent instead of sugar component, so that the oral preparation can achieve low sugar and no sugar. The natural flavoring agent is preferably one or more of fresh flower essential oil (such as rose essential oil and jasmine essential oil), fruit essential oil, fruit essence and concentrated fruit juice.
The auxiliary materials of the preparation also can comprise auxiliary materials required by preparing corresponding dosage forms. For example: when the preparation is powder or granules, the auxiliary material also comprises an anti-caking agent (such as silicon dioxide); when the preparation is a tablet, the auxiliary material also comprises a tabletting glidant (such as magnesium stearate). When the preparation is oral liquid or aqueous solution, the auxiliary material also comprises distilled pure water.
The invention also provides a preparation method of the preparation, which is a preparation method for a liquid preparation and comprises the following steps: after all the components are mixed evenly, a polyethersulfone filter membrane and/or a normal-temperature nanofiltration membrane are adopted for filtration.
Compared with the sterilization and preservation method by high-temperature sterilization and adding artificial preservatives (such as sorbate, benzoate and the like), the method disclosed by the invention has the advantages that the microorganisms are removed by adopting the filtering method, the preservation effect is excellent, the influence on the activity of active ingredients is less, the artificial preservatives are not required to be added, and the preservation and preservation performance of the liquid preparation can be improved to more than 365 days from 15-30 days.
The invention has the beneficial effects that:
(1) the giant salamander collagen peptide and the mangosteen pericarp extract in the composition provided by the invention have a remarkable synergistic effect, the lipid-lowering function of the composition is remarkably improved, and the composition can effectively reduce the accumulation of body fat such as visceral fat and the like when high-fat is taken.
(2) The giant salamander collagen peptide provided by the invention is rich in small molecule active peptide with the molecular weight below 1000Da, has the average molecular weight below 1000Da, and has higher absorption utilization rate; meanwhile, the product has high purity, the protein content is more than 95%, and the ash content is less than 2%.
(3) The giant salamander collagen peptide preparation provided by the invention does not contain additives such as artificial pigments, essences, solubilizers, deodorants, stabilizers and the like, and sugar substitutes are used for replacing sugar as sweet taste modifiers, so that the giant salamander collagen peptide preparation is more beneficial to the health of organisms; and long-time corrosion prevention and fresh keeping performance can be ensured without adding artificial preservatives.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
The experimental procedures used in the following examples are conventional unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified. Wherein the alkaline protease (Alcalase) is derived from Bacillus subtilis, CAS number is 9014-01-1, and enzyme activity is 2.4 AU/g; the neutral protease is derived from bacillus subtilis, the CAS number is 9014-01-1, and the enzyme activity is 10 ten thousand U/g; the Flavourzyme (Flavourzyme) is prepared by fermenting Aspergillus oryzae and adding flavourous substances, screening and compounding, the CAS number is 9014-01-1, and the enzyme activity is 500 LAPU/g.
In the following examples, the mangosteen pericarp extract was prepared by the following method:
1. physically separating mangosteen flesh and mangosteen peel;
2. drying and crushing the separated mangosteen pericarp;
3. extracting pulverized mangosteen pericarp with 50% ethanol water solution for 3 hr, freeze drying the extractive solution, and grinding into powder.
Example 1
The embodiment provides a giant salamander collagen peptide composition which comprises the following components in parts by weight: 20 parts of giant salamander collagen peptide and 20 parts of mangosteen pericarp extract.
The giant salamander collagen peptide is prepared by the following method:
1. pretreating giant salamander raw materials: after the giant salamander whole fish is cut into pieces and is subjected to skin, meat and fat separation treatment, soaking the skin and the meat for 4 hours by using 0.1mol/L sodium hydroxide solution to remove non-collagen, wherein the soaking material-liquid ratio (g: ml) is 1:8, and the NaOH solution is replaced once during the soaking period; washing the soaked product to be neutral by using deionized water, and draining for later use;
2. enzymolysis: putting the pretreated giant salamander fish raw material obtained in the step 1 into an enzymolysis tank, adding deionized water according to a feed-liquid ratio of 1:1.5 (g: ml), starting stirring, heating to 100 ℃, and slightly boiling and preserving heat for 60 min.
1) Cooling to 60 deg.C, adjusting pH to 8.0 with calcium hydroxide, adding alkaline protease 3 ‰ of the material mass, performing enzymolysis for 1 hr, and maintaining pH at 8.0-9.0 during enzymolysis;
2) cooling to 50 ℃, adding neutral protease with the mass of 3 per mill of the material under the natural pH, continuing enzymolysis for 2 hours, and maintaining the pH at 6.0-8.0 in the enzymolysis process;
3) adding flavourzyme with the mass of 5 per mill of the material, continuing enzymolysis for 1h at 50 ℃, keeping the pH value between 6.0 and 8.0 in the enzymolysis process, and adjusting the pH value to be neutral by using phosphoric acid after the enzymolysis is finished.
3. Enzyme deactivation: after the enzymolysis is finished, heating to 90 ℃ and preserving the temperature for 5 min; adding perlite with the quality of 0.5 percent of the zymolyte, and cooling to 60 ℃;
4. primary filtering: and (4) filtering the material obtained in the step (3) to a decoloring tank by using a plate and frame filter, wherein the pressure of the plate and frame filter is 2 Mpa.
5. And (3) decoloring: adjusting pH to 4.0-4.8 with phosphoric acid (measured at 60 deg.C), adding activated carbon at 1% of material ratio, stirring and maintaining the temperature for 30 min; after the decolorization is completed, the pH is adjusted to 6.0 with calcium hydroxide (pH needs to be unchanged within 15 minutes to prevent false pH caused by insufficient or uneven reaction), and then the mixture is filtered.
6. Concentration: the material is concentrated to the concentration of 45 plus or minus 1 percent (concentration meter at the discharge port of the concentration tower).
7. Fine filtering: passing the concentrated solution through a diatomite filter (pore volume is 0.3-1.0 cm)3The filter precision of the filter is 1-2 mu m, the paper board filter (the filter precision of 0.4 mu m) and the micropore folding filter element (the filter precision of 0.22/0.22 mu m polypropylene) enter a sterile tank.
8. Spray drying: the air inlet temperature is 210 ℃, the air outlet temperature is 85 ℃, the atomizer frequency is 50Hz, and the density requirement is as follows: 0.23-0.27 g/ml.
Example 2
The present example provides a giant salamander collagen peptide composition, which is different from example 1 only in step 2 of the preparation method of the giant salamander collagen peptide, wherein step 2 is specifically as follows:
enzymolysis: putting the pretreated giant salamander fish raw material obtained in the step 1 into an enzymolysis tank, adding deionized water according to a feed-liquid ratio of 1:1.5 (g: ml), starting stirring, heating to 100 ℃, and slightly boiling and preserving heat for 60 min.
1) Cooling to 60 deg.C, adjusting pH to 6.5 with calcium hydroxide, adding alkaline protease 1 ‰ of the material mass, performing enzymolysis for 1.5 hr, and maintaining pH at 6.0-8.0 during enzymolysis;
2) cooling to 50 deg.C, adding neutral protease with a mass of 1 ‰ of the material under natural pH, and continuing enzymolysis for 2.5 hr while maintaining pH at 6.0-8.0;
3) adding flavourzyme with the mass of 1 per mill of the material, continuing enzymolysis for 3 hours at 50 ℃, keeping the pH value between 6.0 and 8.0 in the enzymolysis process, and adjusting the pH value to be neutral by using phosphoric acid or calcium hydroxide after the enzymolysis is finished.
Example 3
The embodiment provides a giant salamander collagen peptide composition which comprises the following components in parts by weight: 20 parts of giant salamander collagen peptide and 10 parts of mangosteen pericarp extract.
The preparation method of the giant salamander collagen peptide is the same as that in example 1.
Example 4
The embodiment provides a composition, which consists of the following components in parts by weight: 5 parts of mangosteen pericarp extract, 25 parts of giant salamander collagen peptide and 2.5 parts of resveratrol.
Example 5
The embodiment provides an oral liquid which comprises the following components in parts by weight: 1.5 parts of mangosteen pericarp extract, 50 parts of giant salamander collagen peptide, 2.5 parts of resveratrol and 0.0005 part of biotin; 5 parts of inulin, 5 parts of taurine, 0.001 part of folic acid, 10.3 parts of coenzyme Q, 0.6 part of beta-nicotinamide mononucleotide, 0.01 part of manganese, 0.02 part of iron, 0.0002 part of selenium, 0.0015 part of copper, 500 parts of distilled water, 5 parts of citric acid, 0.2 part of stevioside and 1.5 parts of natural rose essential oil.
The embodiment also provides a preparation method of the oral liquid, which comprises the following steps: mixing the components according to the dosage ratio, and filtering with a polyethersulfone filter membrane to obtain the final product.
The oral liquid of the embodiment is not added with artificial preservative, and the quality guarantee period can reach more than 365 days at normal temperature.
Comparative example 1
This comparative example provides a composition which differs from example 1 only in step 2 of the process for preparing giant salamander collagen peptide, step 2 being specifically as follows:
enzymolysis: putting the pretreated giant salamander fish raw material obtained in the step 1 into an enzymolysis tank, adding deionized water according to a feed-liquid ratio of 1:1.5 (g: ml), starting stirring, heating to 100 ℃, and slightly boiling and preserving heat for 60 min.
1) Cooling to 50 ℃, adding neutral protease with the mass of 3 per mill of the material, performing enzymolysis for 2 hours, and maintaining the pH value to be 6.0-8.0 in the enzymolysis process;
2) heating to 60 deg.C, adjusting pH to 8.0 with calcium hydroxide, adding alkaline protease 3 ‰ of the material mass, and continuing enzymolysis for 1 hr while maintaining pH at 8.0-9.0;
3) adding flavourzyme with the mass of 5 per mill of the material, continuing enzymolysis for 1h at 50 ℃, keeping the pH value between 6.0 and 8.0 in the enzymolysis process, and adjusting the pH value to be neutral by using phosphoric acid after the enzymolysis is finished.
Comparative example 2
This comparative example provides a composition which differs from example 1 only in step 2 of the process for preparing giant salamander collagen peptide, step 2 being specifically as follows:
enzymolysis: putting the pretreated giant salamander fish raw material obtained in the step 1 into an enzymolysis tank, adding deionized water according to a feed-liquid ratio of 1:1.5 (g: ml), starting stirring, heating to 100 ℃, and slightly boiling and preserving heat for 60 min.
1) Cooling to 60 deg.C, adjusting pH to 8.0 with calcium hydroxide, adding alkaline protease 3 ‰ of the material mass, performing enzymolysis for 1 hr, and maintaining pH at 8.0-9.0 during enzymolysis;
2) cooling to 50 ℃, adding trypsin with the mass of 3 per mill of the material under the natural pH, continuing enzymolysis for 2 hours, and maintaining the pH at 6.0-8.0 in the enzymolysis process;
3) adding flavourzyme with the mass of 5 per mill of the material, continuing enzymolysis for 1h at 50 ℃, keeping the pH value between 6.0 and 8.0 in the enzymolysis process, and adjusting the pH value to be neutral by using phosphoric acid after the enzymolysis is finished.
Comparative example 3
The comparative example provides a composition consisting of the following components in parts by weight: 20 parts of giant salamander collagen peptide and 20 parts of lotus leaf extract.
The preparation method of the giant salamander collagen peptide is the same as that in example 1.
Comparative example 4
The comparative example provides a composition consisting of the following components in parts by weight: 20 parts of giant salamander collagen peptide and 20 parts of bitter gourd extract.
The preparation method of the giant salamander collagen peptide is the same as that in example 1.
Experimental example 1 quality test of giant salamander collagen peptide
The giant salamander collagen peptides of examples 1 and 2 and comparative examples 1 and 2 were mass analyzed, respectively. The results of the average molecular weight, protein content and ash content detection of the giant salamander collagen peptides are shown in table 1, and the results show that the average molecular weight, the protein content and the ash content of the giant salamander collagen peptides of examples 1 and 2 are below 1000Da, the protein content is above 95% and the ash content is below 2%.
TABLE 1 quality test of giant salamander collagen peptide
| Detecting the index | Example 1 | Example 2 | Comparative example 1 | Comparative example 2 |
| Average molecular weight (DAL) | 720 | 960 | 1500 | 850 |
| Protein (dry basis,%) | ≥95 | ≥95 | ≥95 | ≥95 |
| Ash (%) | ≤2 | ≤2 | ≤2 | ≤2 |
Experimental example 2 lipid-lowering Activity of giant salamander collagen peptide composition
1. The experimental method comprises the following steps: the experimental animal selects 5-week-old clean male Wistar rats with the body weight of about 110 g. A total of 55 rats were selected and individually fed with normal feed and drinking water for the first 2 weeks to suit the feeding environment. After 2 weeks 55 rats were randomized into 11 groups. Rats of group 1 were fed normal diet as a basic control group, rats of group 2 were fed high fat diet as a control group, rats of group 3 were fed high fat diet while feeding 0.2g/kg/day of hydrolyzed collagen peptide of giant salamander via normal drinking water, rats of group 4 were fed high fat diet while feeding 0.2g/kg/day of mangosteen pericarp extract via normal drinking water, rats of group 5 were fed high fat diet while feeding the composition of example 1 (the amount of the composition is such that 0.2g/kg/day of hydrolyzed collagen peptide of giant salamander and 0.2g/kg/day of mangosteen pericarp extract) via normal drinking water, rats of group 6 were fed high fat diet while feeding the composition of example 2 (the amount of the composition is such that 0.2g/kg/day of hydrolyzed collagen peptide of giant salamander, 0.2g/kg/day of mangosteen pericarp extract), 7 th group of rats were fed with high-fat feed while feeding the composition of example 3 via normal drinking water (the amount of the composition was such that 0.2g/kg/day of giant salamander hydrolyzed collagen peptide and 0.1 g/kg/day of mangosteen pericarp extract), 8 th group of rats were fed with high-fat feed while feeding the composition of comparative example 1 via normal drinking water (the amount of the composition was such that 0.2g/kg/day of giant salamander hydrolyzed collagen peptide and 0.2g/kg/day of mangosteen pericarp extract), 9 th group of rats were fed with high-fat feed while feeding the composition of comparative example 2 via normal drinking water (the amount of the composition was such that 0.2g/kg/day of giant salamander hydrolyzed collagen peptide and 0.2g/kg/day of mangosteen pericarp extract), rats of group 10 were fed high-fat diet while feeding the composition of comparative example 3 via normal drinking water (the amount of the composition was such that the hydrolyzed collagen peptide of giant salamander was 0.2g/kg/day, and the lotus leaf extract was 0.2 g/kg/day). Rats of group 11 were fed high-fat diet while feeding the composition of comparative example 4 via normal drinking water (the amount of the composition was such that 0.2g/kg/day of hydrolyzed collagen peptide of giant salamander and 0.2g/kg/day of extract of Momordica charantia). Groups 11 of rats were housed for a total of 10 weeks as described above.
The normal diet and high fat diet formulations are shown in table 2.
TABLE 2 Normal and high fat diet formulas
| Feed composition | Normal feed (g/kg) | High fat feed (g/kg) |
| Wheat flour | 340 | 150 |
| Corn starch | 250 | 200 |
| Rice flour | - | 300 |
| Soybean powder | 140 | 100 |
| Fish meat powder | 160 | 100 |
| Fat | 70 | 200 |
Within 24 hours of the end of feeding, all rats were sacrificed by carbon dioxide asphyxiation and the body weight of each rat was measured immediately and perirenal and perianal fat weights of all specimens were measured after decapitating.
2. The experimental results are as follows: the fat index is calculated based on the perirenal fat weight, the perianal fat weight as a percentage of the rat body weight, and the total weight of the perirenal fat weight and the perianal fat weight as a percentage of the rat body weight. The results of the fat index measurements (averaged for each group) for 11 rats are shown in table 3.
TABLE 3 fat index test results
Statistical results of the rate of change of fat index of rats in other experimental groups compared to the basic control group (group 1) are shown in Table 4.
TABLE 4 fat Change Rate statistics
As can be seen from the experimental results in tables 3 and 4, the perirenal fat index, perianal fat index and total fat index of the rats in the control group (high fat intake) were significantly increased compared to those of the rats in the basic control group (normal diet), indicating that the high fat intake resulted in a significant increase in body fat of the rats, causing obesity. Compared with the rats in the basic control group, the rats fed with the high-fat feed and the giant salamander hydrolyzed collagen peptide have the increase degrees of 73%, 15% and 46% in the kidney, perianal and total fat indexes, which indicates that the addition of the giant salamander hydrolyzed collagen peptide in the feed can inhibit the increase of body fat of the rats caused by high-fat intake, and has a certain effect on preventing obesity. Compared with rats fed with high-fat feed, giant salamander hydrolyzed collagen peptide and mangosteen pericarp extract, rats fed with the high-fat feed, giant salamander hydrolyzed collagen peptide and the mangosteen pericarp extract have significantly reduced or unchanged perianal fat index, and have no change observed in perianal fat index, so that the total fat index is significantly reduced, which shows that the simultaneous feeding of the giant salamander hydrolyzed collagen peptide and the mangosteen pericarp extract has a very significant inhibition effect on the increase of body fat of the rats caused by high fat intake, and proves that the compatibility of the giant salamander hydrolyzed collagen peptide and the mangosteen pericarp extract has very excellent effects of reducing body fat and preventing obesity.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. The giant salamander collagen peptide composition is characterized by comprising the following components in percentage by mass (1-50): 1 giant salamander collagen peptide and mangosteen pericarp extract.
2. The composition of claim 1, wherein the giant salamander collagen peptide is prepared by a method comprising the steps of: the giant salamander skin and/or meat from which fat and non-collagen are removed is sequentially subjected to three-stage enzymolysis by alkaline protease, neutral protease and flavourzyme to prepare zymolyte.
3. The composition of claim 2, wherein in the three-stage enzymolysis, the reaction temperature of each stage of enzymolysis is not higher than 60 ℃; preferably 40-60 ℃;
preferably, in the three-stage enzymolysis process, the pH of each stage of enzymolysis is controlled to be 6-9.
4. The composition according to claim 2 or 3, wherein the alkaline protease is used in an amount of 0.1-0.5% by mass of the giant salamander skin and/or meat from which fat and non-collagen are removed, the enzymolysis temperature is 55-60 ℃, and the enzymolysis time is 0.5-1.5 h;
the mass ratio of the neutral protease to the giant salamander skin and/or meat without fat and non-collagen is 0.1-0.5%, the enzymolysis temperature is 45-50 ℃, and the enzymolysis time is 1-2.5 h;
the mass ratio of the amount of the flavor protease to the giant salamander skin and/or meat from which fat and non-collagen are removed is 0.1-0.5%, the enzymolysis temperature is 45-55 ℃, and the enzymolysis time is 0.3-3 h.
5. The composition according to any one of claims 1 to 4, wherein the mangosteen pericarp extract is an ethanol extract of mangosteen pericarp;
preferably, the mangosteen pericarp extract is obtained by leaching with 40-60% ethanol water solution by volume percentage and drying the leaching solution.
6. The composition according to any one of claims 2 to 5, wherein the giant salamander skin and/or meat from which fat and non-collagen are removed is treated at 90 to 100 ℃ for 0.5 to 1 hour before the enzymolysis by the alkaline protease;
preferably, soaking the fat-removed giant salamander skin and/or meat in 0.05-0.1M alkali liquor for 3.5-4.5 h, and removing non-collagen to obtain the fat-removed and non-collagen-removed giant salamander skin and/or meat.
7. The composition as claimed in any one of claims 2 to 6, wherein the enzymatic hydrolysate is subjected to enzyme deactivation treatment, and then subjected to primary filtration, activated carbon adsorption, fine filtration and drying treatment in sequence;
the primary filtration comprises filtration of a plate and frame filter taking perlite as a filter aid;
the fine filtration sequentially comprises the steps of taking the pore volume of 0.3-1.0 cm3Filtering by a diatomite filter with the aperture of 1-2 mu m, filtering by a paperboard filter with the filtering precision of 0.4-0.8 mu m, and filtering by a polypropylene micropore folding filter element with the filtering precision of 0.22/0.22 mu m;
preferably, in the primary filtration, the mass ratio of the perlite to the zymolyte is 0.1% -2%, and the pressure of the plate and frame filter is 1.5-2 Mpa.
8. The composition according to any one of claims 1 to 7, wherein the composition comprises the following components in parts by weight: 0.5-20 parts of mangosteen pericarp extract, 10-50 parts of giant salamander collagen peptide and 1-5 parts of resveratrol;
or, the composition comprises the following components in parts by weight: 0.5-20 parts of mangosteen pericarp extract, 10-50 parts of giant salamander collagen peptide, 1-5 parts of resveratrol, 0.0003-0.001 part of biotin, 1-20 parts of plant dietary fiber, 1-15 parts of taurine, 0.0005-0.004 part of folic acid, 0.2-0.5 part of coenzyme Q10, 0.5-4 parts of beta-nicotinamide mononucleotide, 0.005-0.05 part of manganese, 0.01-0.15 part of iron, 0.0001-0.0011 part of selenium and 0.0005-0.015 part of copper.
9. Use of the composition of any one of claims 1 to 8 for preparing a product with lipid-lowering or weight-reducing function, preferably the product is a food, a drug or a health product.
10. A giant salamander collagen peptide preparation which comprises the composition according to any one of claims 1 to 8 as an active ingredient;
preferably, the preparation further comprises a seasoning auxiliary material, wherein the seasoning auxiliary material comprises the following components in parts by weight: 2-7 parts of sour modifier, 0.1-50 parts of sweet modifier and 0.5-3 parts of natural flavoring agent;
more preferably, the sour flavor is one or more selected from the group consisting of citric acid and malic acid; the sweet flavoring agent is one or more selected from xylitol, stevioside and erythritol; the natural flavoring agent is one or more selected from fresh flower essential oil, fruit essence and concentrated fruit juice.
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