CN113785860B - Preparation method and application of lentinan-fish scale collagen peptide conjugate - Google Patents
Preparation method and application of lentinan-fish scale collagen peptide conjugate Download PDFInfo
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- CN113785860B CN113785860B CN202110910927.9A CN202110910927A CN113785860B CN 113785860 B CN113785860 B CN 113785860B CN 202110910927 A CN202110910927 A CN 202110910927A CN 113785860 B CN113785860 B CN 113785860B
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- lentinan
- collagen peptide
- fish scale
- scale collagen
- fish
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- 108010035532 Collagen Proteins 0.000 title claims abstract description 104
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- 239000000863 peptide conjugate Substances 0.000 title claims abstract description 63
- 238000002360 preparation method Methods 0.000 title claims abstract description 32
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- 238000000034 method Methods 0.000 claims abstract description 28
- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 claims abstract description 24
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- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 claims abstract description 12
- HWTZYBCRDDUBJY-UHFFFAOYSA-N Rhynchosin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=CC(O)=C(O)C=C2O1 HWTZYBCRDDUBJY-UHFFFAOYSA-N 0.000 claims abstract description 12
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 claims abstract description 12
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- YLZOPXRUQYQQID-UHFFFAOYSA-N 3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-1-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]propan-1-one Chemical compound N1N=NC=2CN(CCC=21)CCC(=O)N1CCN(CC1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F YLZOPXRUQYQQID-UHFFFAOYSA-N 0.000 description 7
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- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
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- 241000233866 Fungi Species 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
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- 238000004090 dissolution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
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- 102000035122 glycosylated proteins Human genes 0.000 description 1
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- QYHFIVBSNOWOCQ-UHFFFAOYSA-N selenic acid Chemical compound O[Se](O)(=O)=O QYHFIVBSNOWOCQ-UHFFFAOYSA-N 0.000 description 1
- 229940000207 selenious acid Drugs 0.000 description 1
- MCAHWIHFGHIESP-UHFFFAOYSA-N selenous acid Chemical compound O[Se](O)=O MCAHWIHFGHIESP-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D13/00—Finished or partly finished bakery products
- A21D13/06—Products with modified nutritive value, e.g. with modified starch content
- A21D13/062—Products with modified nutritive value, e.g. with modified starch content with modified sugar content; Sugar-free products
-
- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D13/00—Finished or partly finished bakery products
- A21D13/06—Products with modified nutritive value, e.g. with modified starch content
- A21D13/064—Products with modified nutritive value, e.g. with modified starch content with modified protein content
-
- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D2/00—Treatment of flour or dough by adding materials thereto before or during baking
- A21D2/08—Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
- A21D2/14—Organic oxygen compounds
- A21D2/18—Carbohydrates
- A21D2/181—Sugars or sugar alcohols
-
- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D2/00—Treatment of flour or dough by adding materials thereto before or during baking
- A21D2/08—Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
- A21D2/24—Organic nitrogen compounds
-
- A—HUMAN NECESSITIES
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D2/00—Treatment of flour or dough by adding materials thereto before or during baking
- A21D2/08—Treatment of flour or dough by adding materials thereto before or during baking by adding organic substances
- A21D2/24—Organic nitrogen compounds
- A21D2/26—Proteins
- A21D2/268—Hydrolysates from proteins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
- C07K1/1072—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups
- C07K1/1077—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides by covalent attachment of residues or functional groups by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- Life Sciences & Earth Sciences (AREA)
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- Zoology (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Polymers & Plastics (AREA)
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- Materials Engineering (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
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- Biotechnology (AREA)
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- General Engineering & Computer Science (AREA)
- Toxicology (AREA)
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- Coloring Foods And Improving Nutritive Qualities (AREA)
- Meat, Egg Or Seafood Products (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a preparation method and application of lentinan-fish scale collagen peptide conjugate, wherein the preparation method comprises (1) washing fresh lentinus edodes, blanching, crushing, pulping to obtain lentinus edodes homogenate, treating with dynamic high-pressure microjet for three times, extracting lentinan from the lentinus edodes by water extraction and alcohol precipitation; (2) Removing impurities and decalcification from fish scales, adding deionized water for extraction, separating, concentrating, freeze-drying to obtain fish gelatin, and performing ultrasonic-assisted synergistic two-step enzymolysis to obtain fish scale collagen peptide; (3) Re-dissolving lentinan and fish scale collagen peptide, adding selenite and quercetin to prepare a solution, pretreating in an ultrasonic environment, spray-drying, treating in a strong magnetic field environment, and finally carrying out glycosylation under the condition of superheated steam. The lentinan and fish scale collagen peptide prepared by the method has high purity and rich nutrition; the biscuit manufactured by the invention meets the requirements of modern consumers on healthy diet, and simultaneously enriches the taste of the biscuit.
Description
Technical Field
The invention belongs to the technical field of food processing, and particularly relates to a preparation method and application of a lentinan-fish scale collagen peptide conjugate.
Background
The lentinus edodes is an edible fungus, has long history, good color and taste, high nutritive value, is rich in protein, mineral elements, vitamins and dietary fibers, has the effects of tonifying qi and nourishing body, promoting blood circulation and removing blood stasis, has both edible value and pharmacological value, and is a food with homology of medicine and food. The lentinan has high pharmacological value, including immunoregulation, anti-tumor, anti-aging, antioxidation, bacteriostasis, antivirus, and anti-radiation effects.
The fish scales are common byproducts in the processing of freshwater fish, are rich in nutrition and rich in protein, vitamins and microelements, but are often discarded as waste, so that the fish scales are waste of resources and cause certain pollution. Compared with collagen, the collagen peptide extracted from fish scales has the advantage that the collagen peptide is easier to be absorbed by human bodies due to the proper proportion of amino acid, dipeptide and polypeptide.
The dynamic high-pressure micro-jet is a novel food processing technology, and has remarkable physical modification effects, such as physicochemical properties, physiological activity and the like. The invention adopts dynamic high-pressure micro-jet treatment before lentinan extraction, and the effects of high pressure, high-speed impact, high-frequency vibration, cavitation shearing and the like lead lentinus edodes homogenate to be more uniform and finer and improve the lentinan extraction efficiency.
The ultrasonic wave is a mechanical wave, and can effectively improve the reaction efficiency due to the physical effect and the mechanical effect, and has mild condition and no pollution, and the ultrasonic wave is often used as an auxiliary means at present. The method has the advantages of mild enzymolysis process conditions, simple operation, high efficiency and environmental protection, is one of the common methods for hydrolyzing protein as peptide, adopts ultrasonic-assisted synergistic two-step enzymolysis process, and has good ratio of the obtained amino acid, dipeptide and polypeptide and high extraction efficiency.
The strong magnetic field sterilization is a cold sterilization mode, is different from the heating sterilization, can retain the primary color fragrance to a large extent, has fast propagation and high strength, can instantaneously sterilize, and has wide sterilization range. The lentinan-fish scale collagen peptide conjugate is treated by the strong magnetic field technology, so that the lentinan-fish scale collagen peptide conjugate can be rapidly sterilized, the original properties of the lentinan-fish scale collagen peptide conjugate are not influenced, the bacteriostasis of the treated lentinan-fish scale collagen peptide conjugate is greatly improved, and the shelf life of the lentinan-fish scale collagen peptide conjugate can be prolonged.
Glycosylation refers to the process of forming glycosylated proteins by cross-linking of reducing sugars with proteins/peptides, AGEs refers to post-substances generated during glycosylation, and AGEs may induce cross-linking of extracellular matrix in humans to affect normal function. The glycosylation reaction can lead the protein/peptide and the sugar to react to form a conjugate, but the generation of AGEs is often accompanied in the process, and the invention adds trace selenite and quercetin into the raw materials, so that the generation of the AGEs is well inhibited.
The biscuit is a common leisure food, has the advantages of good taste, long preservation period, convenience, portability and the like, and is widely popular with eaters. With the change of diet concept, consumers pay more and more attention to the nutrition of foods, and the biscuit with single raw materials cannot meet the demands, so that the development prospect of the nutritional biscuit is broad.
Disclosure of Invention
Aiming at the defects and the problems in the prior art, the invention aims to provide a preparation method of lentinan-fish scale collagen peptide conjugate and a preparation method of biscuits rich in the lentinan-fish scale collagen peptide conjugate, and the prepared lentinan and fish scale collagen peptide have high purity and rich nutrition. The invention combines ultrasonic pretreatment technology and strong magnetic field treatment means to carry out glycosylation on the lentinan and the fish scale collagen peptide conjugate in the superheated steam environment, and finally the prepared lentinan-fish scale collagen peptide conjugate not only maintains the flavor of the original product, but also makes the color and aroma more attractive due to glycosylation reaction. In addition, the invention also provides a preparation method of the lentinan-rich biscuits, and the lentinan-fish scale collagen peptide conjugate prepared by the preparation method is added on the basis of original preparation of the biscuits, so that the requirements of modern consumers on healthy diet are met, and meanwhile, the taste of the biscuits is enriched.
The invention is realized by the following technical scheme:
the invention provides a preparation method of lentinan-fish scale collagen peptide conjugate, which specifically comprises the following steps:
(1) Extraction of lentinan:
A. washing fresh lentinus edodes, and carrying out blanching, crushing and pulping treatment to obtain lentinus edodes homogenate;
B. adding water into the lentinus edodes homogenate, stirring, performing dynamic high-pressure microjet treatment for three times, wherein the lentinus edodes homogenate and water are 1:15-20, the dynamic high-pressure microjet pressure is 110-130Mpa, centrifuging, filtering, and collecting supernatant;
C. coarse extraction: alcohol precipitation is carried out according to the ratio of supernatant to absolute ethyl alcohol=1:1, standing is carried out for 12 hours at normal temperature, centrifugal filtration is carried out, and sediment is crude polysaccharide;
D. purifying: re-dissolving the crude polysaccharide, regulating pH to 3.0 with citric acid, mixing, refrigerating in refrigerator for 24 hr, centrifuging at 8000r/min for 10min, filtering to obtain water phase, and freeze drying to obtain lentinan;
(2) Preparation of fish scale collagen peptide
A. Removing impurities: soaking fish scales in 0.2mol/L NaOH solution for 30min, and then washing with deionized water to remove most of pigment, impurity protein and other impurities;
B. decalcification: soaking fish scales in citric acid solution for decalcification treatment, wherein the decalcification time is 1-2h, the solid-liquid ratio is 1:5-10, the concentration of the used citric acid is 0.25-0.5mol/L, and stirring is kept in the whole process;
C. extracting: adding 10 times of deionized water in volume parts into the decalcified fish scales, adjusting the pH value of the solution to 4.0, and heating at 55 ℃ for 3-5 hours for extraction;
D. separating and concentrating: extracting fish gelatin, filtering, concentrating the supernatant, and freeze-drying to obtain fish gelatin;
E. ultrasonic-assisted enzymolysis: the fish gelatin is placed in an ultrasonic environment, and is subjected to hydrolysis by alkaline protease and flavourzyme respectively after the fish gelatin is finished, wherein the enzymolysis conditions of the alkaline protease are as follows: pH7.5-8.5, temperature 50-55deg.C, and time 2-4 hr; the enzymolysis condition of the flavourzyme is that the pH is 6-6.5, the temperature is 45-55 ℃ and the time is 3-4h; ultrasonic power is 250W, and ultrasonic time is 20-25min, so that the fish scale collagen peptide is obtained.
(3) Preparation of lentinan-fish scale collagen peptide conjugate:
A. ultrasonic pretreatment: re-dissolving lentinan and fish scale collagen peptide in a ratio of 1:1, adding selenite and quercetin, adding lentinan, fish scale collagen peptide, selenite and quercetin=2000:1000:1-2:1-2, preparing a 1mg/ml solution, and treating the mixture with 150-200w ultrasonic power for 10-20min;
B. spray drying: spray drying the mixture in a spray tower after ultrasonic pretreatment, wherein the spray drying condition is that the air inlet temperature is 175-180 ℃ and the air outlet temperature is 170-180 ℃;
C. strong magnetic field treatment: placing the dried mixture in a strong magnetic field environment, and treating with 2-4T intensity;
D. superheated steam treatment: and (3) carrying out glycosylation on the solid powder under the condition of superheated steam, wherein the temperature of the superheated steam is 170 ℃, and the treatment time is 1-3min.
The second aspect of the invention provides the lentinan-fish scale collagen peptide conjugate prepared by the method.
The third aspect of the invention provides an application of the lentinan-fish scale collagen peptide conjugate, wherein the lentinan-fish scale collagen peptide conjugate is used in food.
Preferably, the food comprises biscuits rich in lentinan-fish scale collagen peptide conjugate, and the preparation method of the biscuits comprises the steps of mixing butter, erythritol and egg liquid, whipping, adding the lentinan-fish scale collagen peptide conjugate, baking soda, flour and salt, stirring to form dough, and baking after molding.
Based on the total weight of flour, the butter additive amount is 30-40%, the erythritol additive amount is 10-15%, the baking soda additive amount is 0.8%, the lentinan-fish scale collagen peptide conjugate additive amount is 8-12%, the salt additive amount is 0.5%, and the egg liquid additive amount is 30%; the flour uses mixed flour of low gluten flour and common flour, and the common flour is low gluten flour=1-3:1; heating and melting butter before adding, and beating egg liquid and erythritol for several times; the baking condition is that the upper layer temperature is 180-200deg.C, the lower layer temperature is 60-100deg.C, the time is 12-15min, and the biscuit turns off-white.
In a fourth aspect, the invention provides a lentinan-fish scale collagen peptide conjugate-enriched biscuit prepared by the method.
Compared with the prior art, the invention has the beneficial effects that:
1. in the preparation of lentinan, the invention ensures that lentinus edodes homogenate is more uniform by dynamic high-pressure micro-jet treatment, and the biological activity of the extracted lentinan is higher and the yield and purity are kept under the combined actions of high-speed impact, high-frequency vibration, instantaneous pressure drop, strong shearing and the like.
2. The invention extracts fish gelatin from fish scales, which are common byproducts in fish processing, but in fact the fish scales are rich in proteins, vitamins and various unsaturated fatty acids, and further extracts fish scale collagen peptide, and compared with collagen, the collagen peptide prepared by the invention is easier to be absorbed by human body.
3. When the fish scale collagen peptide is prepared, the invention adopts the ultrasonic-assisted synergistic two-step enzymolysis treatment mode, the physical and mechanical effects generated by ultrasonic waves can improve the reaction efficiency, the operation is simple, mild and pollution-free, and the two-step enzymolysis of alkaline protease and flavourzyme can lead the product to have better amino acid, dipeptide and polypeptide ratio.
4. The collagen peptide prepared from the fish scales has slight fishy smell, and the collagen peptide is glycosylated in the superheated steam environment by combining the ultrasonic pretreatment technology with the strong magnetic field treatment means, so that a reaction system formed by the glycosylation of lentinan is effectively utilized, and the fishy smell of the collagen peptide is reduced.
5. The lentinan-fish scale collagen peptide conjugate prepared by the invention has good antibacterial property through treating the product by a strong magnetic field.
6. The lentinan and the fish scale collagen peptide are two substances with high nutritive value, and the lentinan and the fish scale collagen peptide react through a glycosylation reaction means, so that the lentinan-fish scale collagen peptide conjugate is obtained, the prepared conjugate has both nutritive value and pharmacological value, the flavor of the original product is reserved, and the glycosylation reaction also makes the color and aroma more attractive.
7. The invention well inhibits the generation of AGEs by adding trace selenious acid and quercetin into the raw materials.
8. The basic formula of the biscuit is improved, and the heat of the biscuit is effectively reduced by using erythritol to replace white granulated sugar; the mixing and matching of the common flour and the low-gluten flour enhances the toughness of the dough and enables the dough to be easier to form.
9. The lentinan-fish scale collagen peptide conjugate prepared by the invention is added into a biscuit formula, so that the nutrition value of the biscuit is increased, the taste of the product is enriched, and the flavor of the product is increased.
Detailed Description
The invention will be further illustrated with reference to examples.
In a first aspect, the present invention provides a method for preparing a lentinan-fish scale collagen peptide conjugate, specifically:
(1) Washing fresh lentinus edodes, blanching, crushing and pulping to obtain lentinus edodes homogenate, carrying out dynamic high-pressure microjet treatment for three times, and extracting lentinan from the lentinus edodes by a water extraction and alcohol precipitation method;
(2) Removing impurities and decalcification from fish scales, adding deionized water for extraction, separating, concentrating, freeze-drying to obtain fish gelatin, and performing ultrasonic-assisted synergistic two-step enzymolysis to obtain fish scale collagen peptide;
(3) Re-dissolving the lentinan and the fish scale collagen peptide, adding selenite and quercetin to prepare a solution, pretreating in an ultrasonic environment, spray-drying, treating in a strong magnetic field environment, and finally carrying out glycosylation under the condition of superheated steam.
According to the invention, in step (1), the lentinus edodes homogenate is mixed with water=1:15-20, and the dynamic high-pressure micro-jet pressure is 110-130Mpa; in the step (1), the water extraction and alcohol precipitation method adopts absolute ethyl alcohol to carry out crude extraction, and alcohol precipitation is carried out for 12 hours according to the proportion of solution to absolute ethyl alcohol=1:1; in the step (1), the polysaccharide after the crude extraction is purified, the pH value is regulated to 3.0 by citric acid after the re-dissolution, the polysaccharide is refrigerated for 24 hours in a refrigerator after the uniform mixing, the polysaccharide is centrifuged for 10 minutes at 8000r/min, the water phase is obtained after the filtration, and the lentinan is obtained after the freeze drying.
According to the invention, in step (2), the decalcification time is 1: 1h, the solid-to-liquid ratio is 1:5-10, the concentration of citric acid used is 0.5mol/L, and the stirring is kept proper. According to the invention, in the step (2), 10 times of deionized water in volume parts is added into the decalcified fish scales, the pH value of the solution is regulated to 4.0, and the fish scales are heated for 3 to 5 hours at the temperature of 55 ℃ for extraction.
According to the invention, in the step (2), the parameter conditions of the ultrasonic wave are as follows: ultrasonic power is 250w, and ultrasonic time is 20-30min; in the step (2), the enzymes used for enzymolysis are alkaline protease and flavourzyme, and the enzymolysis conditions of the alkaline protease are as follows: pH7.5-8.5, temperature 50-55deg.C, time 2-4h, and enzymolysis conditions of flavourzyme are: the pH is 6-6.5, the temperature is 45-55 ℃ and the time is 3-4h;
according to the invention, in step (3), when preparing the solution, lentinan, fish scale collagen peptide, selenic acid and quercetin=2000:1000:2:1 are added; in the step (3), the ultrasonic pretreatment parameter conditions are as follows: the power is 150-200w, and the time is 10-20min; in the step (3), the spray drying parameter conditions are as follows; the air inlet temperature is 175-180 ℃, and the air outlet temperature is 170-180 ℃; in the step (3), the conditions of the strong magnetic field processing parameters are as follows: adopting 2-4T intensity; in the step (3), the superheated steam treatment parameter conditions are as follows: the superheated steam temperature is 170 ℃, and the treatment time is 1-3min.
In a second aspect, the present invention provides a lentinan-fish scale collagen peptide conjugate prepared by the above method.
In a third aspect, the present invention provides an application of the lentinan-fish scale collagen peptide conjugate, namely an additive for food, comprising a biscuit rich in the lentinan-fish scale collagen peptide conjugate, wherein the raw materials of the biscuit are the lentinan-fish scale collagen peptide conjugate, butter, erythritol, egg white, baking soda, flour and table salt.
Wherein, based on the total amount of flour, the butter addition amount is 40%, the erythritol addition amount is 10-15%, the baking soda addition amount is 0.8%, the lentinan-fish scale collagen peptide conjugate addition amount is 8-12%, the salt addition amount is 0.5%, and the egg liquid addition amount is 30%.
According to the present invention, a mixed flour of low gluten flour and ordinary flour is used as the flour, and ordinary flour: low gluten flour=1-3:1.
In a fourth aspect, the invention provides a preparation method of lentinan-enriched biscuits, which comprises the steps of mixing butter, erythritol and egg liquid, whipping, adding lentinan-fish scale collagen peptide conjugate, baking soda, flour and salt, stirring to form dough, and baking after molding.
According to the invention, butter needs to be heated and melted, and egg liquid and erythritol need to be added in three times of whipping; the dough enters a mould for molding, and the thickness after molding is preferably not more than 5 mm; the baking condition is that the upper layer temperature is 180-200deg.C, the lower layer temperature is 60-100deg.C, the time is 12-15min, and the biscuit should be beige at the end.
The following are several examples of the preparation of lentinan-fish scale collagen peptide conjugate and lentinan-fish scale collagen peptide conjugate-enriched biscuits, which provide examples with feasibility and superiority, but the invention is not limited thereto.
Example 1-1
(1) Extraction of lentinan
A. Washing fresh lentinus edodes, and carrying out blanching, crushing and pulping treatment to obtain lentinus edodes homogenate;
B. adding water into the lentinus edodes homogenate (lentinus edodes homogenate: water=1:15), stirring, performing dynamic high-pressure microjet treatment for three times, homogenizing under 110Mpa, centrifuging, filtering, and collecting supernatant;
C. coarse extraction: alcohol precipitation is carried out according to the proportion of solution to absolute ethyl alcohol=1:1, standing is carried out for 12 hours at normal temperature, centrifugal filtration is carried out, and sediment is crude polysaccharide;
D. purifying: re-dissolving the crude polysaccharide, regulating pH to 3.0 with citric acid, mixing, refrigerating in refrigerator for 24 hr, centrifuging at 8000r/min for 10min, filtering to obtain water phase, and freeze drying to obtain lentinan;
(2) Preparation of fish scale collagen peptide
A. Removing impurities: soaking fish scales in 0.2mol/LNaOH solution for 30min, and then washing with deionized water to remove most of pigment, foreign protein and other impurities;
B. decalcification: soaking fish scales in 0.5mol/L citric acid solution for decalcification treatment, wherein the decalcification time is 1h, the solid-liquid ratio is 1:5, and stirring properly;
C. extracting: adding 10 times of deionized water in volume parts into the decalcified fish scales, adjusting the pH value of the solution to 4.0, and heating at 55 ℃ for 3 hours for extraction;
D. separating and concentrating: extracting fish gelatin, filtering, concentrating the supernatant, and freeze-drying to obtain fish gelatin;
E. ultrasonic-assisted enzymolysis: placing fish gelatin in an ultrasonic environment, wherein the power is 250w, the time is 20min, and after the fish gelatin is ended, respectively hydrolyzing with alkaline protease and flavourzyme to obtain fish scale collagen peptide, wherein the enzymolysis conditions of the alkaline protease are as follows: pH7.5, 50 ℃ and 2 hours of enzymolysis time, and the enzymolysis conditions of the flavourzyme are as follows: pH6, temperature 50 ℃ and enzymolysis time 4h;
(3) Preparation of lentinan-fish scale collagen peptide conjugate
A. Compounding: the lentinan, the fish scale collagen peptide, the selenite and the quercetin=2000:1000:2:1 are compounded into a solution according to the mass portion;
B. ultrasonic pretreatment: the lentinan and the fish scale collagen peptide are compounded into a 1mg/ml solution, and the mixture is treated for 20min by adopting 150w ultrasonic power;
C. spray drying: spray drying the mixture in a spray tower after ultrasonic pretreatment, wherein the air inlet temperature is 175 ℃ and the air outlet temperature is 170 ℃;
D. strong magnetic field treatment: placing the dried mixture in a strong magnetic field environment, wherein the magnetic field strength is 2T;
E. superheated steam treatment: the solid powder mixture treated by the strong magnetic field is put under the condition of superheated steam for glycosylation at 170 ℃ for 1min;
examples 1 to 2
(1) Extraction of lentinan
A. Washing fresh lentinus edodes, and carrying out blanching, crushing and pulping treatment to obtain lentinus edodes homogenate;
B. adding water into the lentinus edodes homogenate (lentinus edodes homogenate: water=1:18), stirring, performing dynamic high-pressure microjet treatment for three times, homogenizing under 120Mpa, centrifuging, filtering, and collecting supernatant;
C. coarse extraction: alcohol precipitation is carried out according to the proportion of solution to absolute ethyl alcohol=1:1, standing is carried out for 12 hours at normal temperature, centrifugal filtration is carried out, and sediment is crude polysaccharide;
D. purifying: re-dissolving the crude polysaccharide, regulating pH to 3.0 with citric acid, mixing, refrigerating in refrigerator for 24 hr, centrifuging at 8000r/min for 10min, filtering to obtain water phase, and freeze drying to obtain lentinan;
(2) Preparation of fish scale collagen peptide
A. Removing impurities: soaking fish scales in 0.2mol/L NaOH solution for 30min, and then washing with deionized water to remove most of pigment, impurity protein and other impurities;
B. decalcification: soaking fish scales in 0.5mol/L citric acid solution for decalcification treatment for 1.5h with a solid-to-liquid ratio of 1:10, and maintaining proper stirring;
C. extracting: adding 10 times of deionized water in volume parts into the decalcified fish scales, adjusting the pH value of the solution to 4.0, and heating at 55 ℃ for 4 hours for extraction;
D. separating and concentrating: extracting fish gelatin, filtering, concentrating the supernatant, and freeze-drying to obtain fish gelatin;
E. ultrasonic-assisted enzymolysis: placing fish gelatin in an ultrasonic environment with the power of 250W for 25min, and respectively hydrolyzing with alkaline protease and flavourzyme after the completion of the process to obtain fish scale collagen peptide, wherein the enzymolysis conditions of the alkaline protease are as follows: the pH value is 8.0, the temperature is 55 ℃, the enzymolysis time is 3 hours, and the enzymolysis conditions of the flavourzyme are as follows: pH6.5, temperature 55 ℃ and enzymolysis time 3h;
(3) Preparation of lentinan-fish scale collagen peptide conjugate
A. Compounding: the lentinan, the fish scale collagen peptide, the selenite and the quercetin=2000:1000:1:1 are compounded into a solution according to the mass portion;
B. ultrasonic pretreatment: the lentinan and the fish scale collagen peptide are compounded into a 1mg/ml solution, and the mixture is treated for 15min by adopting 180w ultrasonic power;
C. spray drying: spray drying the mixture in a spray tower after ultrasonic pretreatment, wherein the air inlet temperature is 175 ℃ and the air outlet temperature is 170 ℃;
D. strong magnetic field treatment: placing the dried mixture in a strong magnetic field environment, wherein the magnetic field strength is 2T;
E. superheated steam treatment: the solid powder mixture treated by the strong magnetic field is put under the condition of superheated steam for glycosylation at the temperature of 170 ℃ for 2min;
examples 1 to 3
(1) Extraction of lentinan
A. Washing fresh lentinus edodes, and carrying out blanching, crushing and pulping treatment to obtain lentinus edodes homogenate;
B. adding water into the lentinus edodes homogenate (lentinus edodes homogenate: water=1:20), stirring, performing dynamic high-pressure microjet treatment for three times, homogenizing under 130Mpa, centrifuging, filtering, and collecting supernatant;
C. alcohol precipitation is carried out according to the proportion of solution to absolute ethyl alcohol=1:1, and after standing for 12 hours at normal temperature, centrifugal filtration is carried out, and the sediment is crude polysaccharide;
D. purifying: re-dissolving the crude polysaccharide, regulating pH to 3.0 with citric acid, mixing, refrigerating in refrigerator for 24 hr, centrifuging at 8000r/min for 10min, filtering to obtain water phase, and freeze drying to obtain lentinan;
(2) Preparation of fish scale collagen peptide
A. Removing impurities: soaking fish scales in 0.2mol/LNaOH solution for 30min, and then washing with deionized water to remove most of pigment, foreign protein and other impurities;
B. decalcification: soaking fish scales in 0.25mol/L citric acid solution for decalcification treatment for 2h with solid-to-liquid ratio of 1:10, and stirring properly;
C. extracting: adding 10 times of deionized water in volume parts into the decalcified fish scales, adjusting the pH value of the solution to 4.0, and heating at 55 ℃ for 5 hours for extraction;
D. separating and concentrating: extracting fish gelatin, filtering, concentrating the supernatant, and freeze-drying to obtain fish gelatin;
E. ultrasonic-assisted enzymolysis: placing fish gelatin in an ultrasonic environment with the power of 250W for 30min, and respectively hydrolyzing with alkaline protease and flavourzyme after the completion of the process to obtain fish scale collagen peptide, wherein the enzymolysis conditions of the alkaline protease are as follows: pH7.5, 55 ℃ and 4 hours of enzymolysis time, wherein the enzymolysis conditions of the flavourzyme are as follows: pH6.5, temperature 55 ℃ and enzymolysis time 4h;
(3) Preparation of lentinan-fish scale collagen peptide conjugate
A. Compounding: the lentinan, the fish scale collagen peptide, the selenite and the quercetin=2000:1000:1:2 are compounded into a solution according to the mass portion;
B. ultrasonic pretreatment: the lentinan and the fish scale collagen peptide are compounded into a 1mg/ml solution, and the mixture is treated for 10min by adopting 200w ultrasonic power;
C. spray drying: spray drying the mixture in a spray tower after ultrasonic pretreatment, wherein the air inlet temperature is 175 ℃ and the air outlet temperature is 170 ℃;
D. strong magnetic field treatment: placing the dried mixture in a strong magnetic field environment, wherein the magnetic field strength is 2T;
E. superheated steam treatment: the solid powder mixture treated by the strong magnetic field is put under the condition of superheated steam for glycosylation at 170 ℃ for 3min;
example 2-1
(1) Weighing: the preparation method comprises weighing the following ingredients, based on the total amount of flour (common flour: low gluten flour=3:1), the butter addition amount is 40%, the erythritol addition amount is 10%, the baking soda addition amount is 0.8%, the lentinan-fish scale collagen peptide conjugate addition amount is 8%, the salt addition amount is 0.5%, the egg liquid addition amount is 30%,
(2) And (3) sending: heating butter, stirring with a whisk for 1-3min, adding erythritol and egg liquid in batches, and stirring for about 2min;
(3) Dough preparation: adding flour, lentinan-fish scale collagen peptide conjugate, baking soda and salt into the foaming mixture, stirring and mixing uniformly, and kneading dough to form;
(4) Baking: making biscuit into proper size and shape, and baking in oven at upper temperature of 185 deg.C and lower temperature of 85 deg.C for 12min.
Example 2-2
(1) Weighing: the preparation method comprises weighing the following ingredients, based on the total amount of flour (common flour: low gluten flour=2:1), the butter addition amount is 40%, the erythritol addition amount is 12%, the baking soda addition amount is 0.8%, the lentinan-fish scale collagen peptide conjugate addition amount is 10%, the salt addition amount is 0.5%, the egg liquid addition amount is 30%,
(2) And (3) sending: heating butter, stirring with a whisk for 1-3min, adding erythritol and egg liquid in batches, and stirring for about 2min;
(3) Dough preparation: adding flour, lentinan-fish scale collagen peptide conjugate, baking soda and salt into the foaming mixture, stirring and mixing uniformly, and kneading dough to form;
(4) Baking: making biscuit into proper size and shape, and baking in oven at upper temperature of 180deg.C and lower temperature of 80deg.C for 14min.
Examples 2 to 3
(1) Weighing: the preparation method comprises weighing the following ingredients, based on the total amount of flour (common flour: low gluten flour=1:1), the butter addition amount is 40%, the erythritol addition amount is 15%, the baking soda addition amount is 0.8%, the lentinan-fish scale collagen peptide conjugate addition amount is 12%, the salt addition amount is 0.5%, the egg liquid addition amount is 30%,
(2) And (3) sending: heating butter, stirring with a whisk for 1-3min, adding erythritol and egg liquid in batches, and stirring for about 2min;
(3) Dough preparation: adding flour, lentinan-fish scale collagen peptide conjugate, baking soda and salt into the foaming mixture, stirring and mixing uniformly, and kneading dough to form;
(4) Baking: making biscuit into proper size and shape, and baking in oven at upper temperature of 190 deg.C and lower temperature of 75 deg.C for 15min.
Comparative example 2-1
Biscuits were made according to the procedure of example 2-2, except that erythritol was added at 5% and the other operations were unchanged.
Comparative examples 2 to 2
Biscuits were made according to the procedure of example 2-2, except that the lentinan-fish scale collagen peptide conjugate was added at 5% and the other operations were unchanged.
Comparative examples 2 to 3
Biscuits were made according to the procedure of example 2-2, except that the lentinan-fish scale collagen peptide conjugate was added at 15% and the other operations were unchanged.
Comparative examples 2 to 4
Biscuits were made according to the procedure of example 2-2, except that the other operations were unchanged without adding low gluten flour to the flour.
Test example: and selecting 20 taste assessors for scoring, requiring the assessors to have no special taste preference and the same assessment environment, and before each assessment, carrying out mouth rinsing, scoring according to four parts of form, color, organization state, taste and taste, finally filling in a table 1, and recording the total score, wherein the result is shown in a table 2.
TABLE 1
TABLE 2
| Project | Sensory scoring |
| Example 2-1 | 80 |
| Example 2-2 | 85 |
| Examples 2 to 3 | 82 |
| Comparative example 2-1 | 73 |
| Comparative examples 2 to 2 | 75 |
| Comparative examples 2 to 3 | 77 |
| Comparative examples 2 to 4 | 73 |
The following modifications were made on the basis of examples 1-2: in comparative example 2-1, the erythritol addition amount was 5%; in comparative example 2-2, the added amount of the mushroom polysaccharide-fish scale collagen peptide conjugate was 5%; in comparative examples 2 to 3, the added amount of the mushroom polysaccharide-fish scale collagen peptide conjugate was 15%; in comparative examples 2 to 4, the flour used was all ordinary flour.
As can be seen from table 2, reducing the amount of erythritol added affects the taste score, in particular, the amount of erythritol affects the formation of gluten and the extent of caramelization reaction in addition to sweetness; the proper mushroom polysaccharide-fish scale collagen peptide conjugate can give a lot of flavors to the biscuits, but too much flavor is produced, too little flavor of the mushrooms is lost, and the addition amount is determined to be 8-12% by comparison; the addition of certain low gluten flour to the common flour can accelerate the formation of dough and make the biscuit more crisp.
The foregoing description of the preferred embodiments of the present invention has been presented only in terms of those specific and detailed descriptions, and is not, therefore, to be construed as limiting the scope of the invention. It should be noted that modifications, improvements and substitutions can be made by those skilled in the art without departing from the spirit of the invention, which are all within the scope of the invention. Accordingly, the scope of protection of the present invention is to be determined by the appended claims.
Claims (9)
1. A method for preparing a lentinan-fish scale collagen peptide conjugate, which is characterized by comprising the following steps:
(1) Extraction of lentinan: washing fresh mushrooms, and carrying out blanching, crushing and pulping treatment to obtain mushroom homogenate, wherein the dynamic high-pressure micro-jet treatment is carried out for three times, and the pressure of the dynamic high-pressure micro-jet is 110-130Mpa; extracting lentinan from the mushroom by water extraction and alcohol precipitation;
(2) Preparation of fish scale collagen peptide: removing impurities and decalcification from fish scales, adding deionized water for extraction, separating, concentrating, freeze-drying to obtain fish gelatin, and performing ultrasonic-assisted synergistic two-step enzymolysis to obtain fish scale collagen peptide;
the impurity removal is as follows: soaking fish scales in NaOH solution, and then washing with deionized water;
the extraction is as follows: adding deionized water into the decalcified fish scales, adjusting the pH value to 4.0, and then leaching by hot water;
the ultrasonic-assisted collaborative two-step enzymolysis method comprises the following steps: the fish gelatin is placed in an ultrasonic environment, and is subjected to hydrolysis by alkaline protease and flavourzyme respectively after the fish gelatin is finished, wherein the enzymolysis conditions of the alkaline protease are as follows: pH7.5-8.5, temperature 50-55deg.C, time 2-4h; the enzymolysis condition of the flavourzyme is that the pH is 6-6.5, the temperature is 45-55 ℃ and the time is 3-4h; ultrasonic power is 250W, ultrasonic time is 20-25min, and fish scale collagen peptide is obtained;
(3) Preparation of lentinan-fish scale collagen peptide conjugate: the lentinan and the fish scale collagen peptide prepared by the method are re-dissolved, selenite and quercetin are added to prepare a solution, the added lentinan, fish scale collagen peptide, selenite and quercetin=2000:1000:1-2:1-2 are finally prepared into a solution of 1mg/ml, the solution is pretreated in an ultrasonic environment, the mixture is treated for 10-20min by adopting 150-200w ultrasonic power, the mixture is treated by adopting 2-4T intensity in a strong magnetic field environment after spray drying, the spray drying condition is that the air inlet temperature is 175-180 ℃, the air outlet temperature is 170-180 ℃, and finally glycosylation is carried out under the condition of superheated steam, the temperature of the superheated steam is 170 ℃, and the treatment time is 1-3min.
2. The method for preparing a lentinan-fish scale collagen peptide conjugate according to claim 1, wherein the step (1) specifically comprises:
A. washing fresh lentinus edodes, and carrying out blanching, crushing and pulping treatment to obtain lentinus edodes homogenate;
B. adding water into the lentinus edodes homogenate, stirring, performing dynamic high-pressure microjet treatment for three times, wherein the lentinus edodes homogenate and water are 1:15-20, the dynamic high-pressure microjet pressure is 110-130Mpa, centrifuging, filtering, and collecting supernatant;
C. coarse extraction: alcohol precipitation is carried out according to the ratio of supernatant to absolute ethyl alcohol=1:1, standing is carried out for 12 hours at normal temperature, centrifugal filtration is carried out, and sediment is crude polysaccharide;
D. purifying: re-dissolving the crude polysaccharide, regulating pH to 3.0 with citric acid, mixing, refrigerating in refrigerator for 24 hr, centrifuging at 8000r/min for 10min, filtering to obtain water phase, and freeze drying to obtain lentinan.
3. The method for preparing a lentinan-fish scale collagen peptide conjugate according to claim 1, wherein in the step (2), specifically:
A. removing impurities: soaking fish scales in 0.2mol/L NaOH solution for 30min, and then washing with deionized water to remove most of pigments, foreign proteins and other impurities;
B. decalcification: soaking fish scales in citric acid solution for decalcification treatment, wherein the decalcification time is 1-2h, the solid-to-liquid ratio is 1:5-10, the concentration of the used citric acid is 0.25-0.5mol/L, and stirring is kept in the whole process;
C. extracting: adding 10 times of deionized water in volume parts into the decalcified fish scales, adjusting the pH value of the solution to 4.0, and heating at 55 ℃ for 3-5h for extraction;
D. separating and concentrating: extracting fish gelatin, filtering, concentrating the supernatant, and freeze-drying to obtain fish gelatin;
E. and (5) performing ultrasonic-assisted enzymolysis to obtain the fish scale collagen peptide.
4. A lentinan-fish scale collagen peptide conjugate prepared by the method of any one of claims 1-3.
5. Use of the lentinan-fish scale collagen peptide conjugate according to claim 4, characterized in that: the lentinan-fish scale collagen peptide conjugate is used in food.
6. The use of the lentinan-fish scale collagen peptide conjugate according to claim 5, wherein: the food comprises biscuits rich in the lentinan-fish scale collagen peptide conjugate, and the preparation method comprises the steps of mixing butter, erythritol and egg liquid, whipping, adding the lentinan-fish scale collagen peptide conjugate, baking soda, flour and salt, stirring to form dough, and baking after molding.
7. The use of the lentinan-fish scale collagen peptide conjugate according to claim 6, wherein: based on the total weight of flour, the addition amount of butter is 30-40%, the addition amount of erythritol is 10-15%, the addition amount of baking soda is 0.8%, the addition amount of lentinan-fish scale collagen peptide conjugate is 8-12%, the addition amount of salt is 0.5%, and the addition amount of egg liquid is 30%.
8. The use of the lentinan-fish scale collagen peptide conjugate according to claim 6, wherein: the flour uses mixed flour of low gluten flour and common flour, and the common flour is low gluten flour=1-3:1; heating and melting the butter before adding, and beating egg liquid and erythritol for multiple times; the baking condition is that the upper layer temperature is 180-200deg.C, the lower layer temperature is 60-100deg.C, the time is 12-15min, and the biscuit turns off-white.
9. A lentinan-fish scale collagen peptide conjugate-enriched biscuit prepared by the method of any one of claims 6 to 8.
Priority Applications (1)
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