CN111218495B - Fish scale collagen, preparation method and application thereof, ice cream rich in fish scale collagen and preparation method thereof - Google Patents
Fish scale collagen, preparation method and application thereof, ice cream rich in fish scale collagen and preparation method thereof Download PDFInfo
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Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G9/00—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor
- A23G9/32—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds
- A23G9/38—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds containing peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G9/00—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor
- A23G9/32—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds
- A23G9/42—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds containing plants or parts thereof, e.g. fruits, seeds, extracts
-
- 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]
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Polymers & Plastics (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Inorganic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Food Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Toxicology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Cosmetics (AREA)
Abstract
The invention belongs to the technical field of food processing, and particularly relates to fish scale collagen, a preparation method and application thereof, ice cream rich in the fish scale collagen and a preparation method thereof. The preparation method of the fish scale collagen comprises the following steps: placing the fish scales in acid liquor for decalcification treatment to obtain decalcified fish scales; adding water into the decalcified fish scales, and extracting fish gelatin under heating to obtain fish gelatin extract; carrying out solid-liquid separation on the fish gelatin extracting solution to obtain fish gelatin; sequentially contacting fish gelatin with alkaline protease and trypsin for respective hydrolysis to obtain fish scale collagen. The fish scale collagen prepared by the invention has high protein hydrolysis degree and soluble protein content, and is used for making ice cream, and pomegranate and grape pulp are added, so that the prepared ice cream has good stability, has the original product characteristics, has higher nutritional value and fine mouthfeel, and improves the effects of oxidation resistance, beauty treatment, skin care and the like of the ice cream.
Description
Technical Field
The invention belongs to the technical field of food processing, and particularly relates to fish scale collagen, a preparation method and application thereof, ice cream rich in the fish scale collagen and a preparation method thereof.
Background
The traditional ice cream is loved and favored by people due to the fresh and bright sweet taste, the smooth and exquisite taste and the unique flavor. However, with the increasingly deep idea of healthy diet, people have new knowledge on ice cream. In addition to the emphasis on mouthfeel, the novel ice cream products begin to pay more attention to the nutritional and health-care effects of the products.
The fish scale is rich in protein, fat, vitamins, Fe, Zn, Ca, trace elements and colloid. The fish scale also contains various unsaturated fatty acids, and has effects of reducing accumulation of cholesterol on blood vessel wall, and preventing arteriosclerosis, hypertension and heart disease. The collagen in the fish scales has good effects of maintaining beauty, keeping young and resisting aging, and compared with other collagens, the collagen in the fish scales is easy to decompose, digest and absorb.
However, the existing fish scale collagen products have relatively low protein hydrolysis degree and soluble protein content, which are not beneficial to the decomposition, digestion and absorption of collagen, so that the nutritional value of the ice cream prepared by using the existing fish scale collagen is greatly reduced, and the prepared ice cream has poor stability and is not fine and smooth enough in taste.
Disclosure of Invention
The invention aims to overcome the defects of low protein hydrolysis degree and low soluble protein content in fish scale collagen in the prior art, and the requirements of nutritional value, taste and stability of ice cream prepared from the fish scale collagen are to be improved, and provides the fish scale collagen, the preparation method and the application of the fish scale collagen, the ice cream rich in the fish scale collagen and the preparation method of the ice cream.
In order to achieve the above object, one aspect of the present invention provides a method for preparing fish scale collagen, the method comprising:
(1) placing the fish scales in acid liquor for decalcification treatment to obtain decalcified fish scales;
(2) adding water into the decalcified fish scales, and extracting fish gelatin under heating to obtain fish gelatin extract;
(3) carrying out solid-liquid separation on the fish gelatin extracting solution to obtain fish gelatin;
(4) and sequentially contacting the fish gelatin with alkaline protease and trypsin to respectively carry out hydrolysis of the alkaline protease and hydrolysis of the trypsin, so as to obtain the fish scale collagen.
In a second aspect, the invention provides fish scale collagen prepared by the method.
The third aspect of the invention provides the application of the fish scale collagen in food, cosmetics, health products or medicines.
The invention provides an ice cream rich in scale collagen, which contains the scale collagen, a stabilizer, skimmed milk powder, monoglyceride, pomegranate-grape mixed fruit pulp, a sweetening agent, cream and vegetable oil;
wherein, the amount of the fish scale collagen is 1-3 wt% based on dry weight, the amount of the stabilizer is 0.2-0.8 wt%, the amount of the skimmed milk powder is 10-20 wt%, the amount of the monoglyceride is 0.1-0.3 wt%, the amount of the pomegranate-grape mixed fruit pulp is 10-30 wt%, the amount of the sweetener is 3-10 wt%, the amount of the cream is 1-3 wt%, and the amount of the vegetable oil is 1-3 wt% based on the total feed amount.
In a fifth aspect, the present invention provides a method for preparing an ice cream rich in fish scale collagen as described above, the method comprising: mixing fish scale collagen, stabilizer, skimmed milk powder, monoglyceride, pomegranate-grape mixed fruit pulp, sweetener, butter, vegetable oil and water; and then, homogenizing, sterilizing, carrying out solid-liquid separation, aging, freezing and hardening on the mixed material obtained by mixing in sequence to obtain the ice cream rich in the scale collagen.
Through the technical scheme, the invention can obtain the following beneficial effects:
1. according to the invention, the scale gelatin is treated by adopting the alkaline protease and the trypsin, so that the proteolysis degree and the soluble protein content of the prepared scale collagen can be effectively improved;
2. collagen polypeptide of fish scales treated by alkaline protease and trypsin can act together with the stabilizer of the invention, thus improving the stability of the prepared ice cream;
3. the fish collagen has effects of caring skin, resisting aging, and endowing the ice cream with high nutritive value;
4. the ice cream is added with pomegranate-grape pulp, is rich in mineral substances such as vitamin C, amino acid and trace elements, and has the effects of improving the oxidation resistance of a human body, softening blood vessels, reducing blood fat and the like;
5. the collagen polypeptide of fish scales treated by alkaline protease and trypsin can be used together with other raw materials to endow the prepared ice cream with fine taste.
Additional features and advantages of the invention will be set forth in the detailed description which follows.
Detailed Description
The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and such ranges or values should be understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.
In a first aspect, the present invention provides a method for preparing fish scale collagen, comprising:
(1) placing the fish scales in acid liquor for decalcification treatment to obtain decalcified fish scales;
(2) adding water into the decalcified fish scales, and extracting fish gelatin under heating to obtain fish gelatin extract;
(3) carrying out solid-liquid separation on the fish gelatin extracting solution to obtain fish gelatin;
(4) and sequentially contacting the fish gelatin with alkaline protease and trypsin to respectively carry out hydrolysis of the alkaline protease and hydrolysis of the trypsin, so as to obtain the fish scale collagen.
According to the invention, in step (1), the acid may be any acid capable of decalcifying the fish scales, and preferably, the acid is hydrochloric acid and/or citric acid.
The amount of the acid may vary within a wide range as long as it can completely impregnate the fish scales and effectively remove calcium therefrom, and preferably, the amount of the acid is 5 to 20 parts by volume, for example, 5 parts by volume, 7 parts by volume, 10 parts by volume, 12 parts by volume, 14 parts by volume, 15 parts by volume, 16 parts by volume, 18 parts by volume, 20 parts by volume, and preferably 5 to 10 parts by volume, relative to 1 part by volume of the fish scales. Wherein, the concentration of the acid solution can be 0.3-0.6 mol/L.
According to the invention, in order to improve the decalcification effect on the fish scales, stirring is carried out for 0.5-1.5min every 10-20min during the acid soaking decalcification.
According to the present invention, the conditions of the decalcification treatment are not particularly limited, and it is preferable that the temperature is 20 to 40 ℃ and the time is at least 1 hour, preferably 1 to 3 hours.
According to the invention, the fish scales can be obtained by conventional means, for example, from fish scales left over in the processing of fisheries. Preferably, before the fish scales are decalcified, removing impurities such as bone spurs and the like from the fish scales, intermittently beating the fish scales for 1 to 5 minutes by using a tissue mashing machine, and finally washing the fish scales for multiple times by using clear water.
The scale of the freshwater fish in the present invention is preferably a scale of freshwater fish, and the type of the freshwater fish is not particularly limited, and may be various kinds of freshwater fish that are conventionally commercially available, and according to a preferred embodiment of the present invention, the scale of the freshwater fish is mainly derived from tilapia, grass carp, and silver carp.
According to the present invention, it is preferable that the method further comprises washing the decalcified fish scales to neutrality with water before adding water to the decalcified fish scales and the medium for heating in step (2).
The amount of water added to the decalcified fish scales according to the invention may vary within wide limits, preferably between 3 and 10 parts by weight, preferably between 4 and 6 parts by weight, relative to 1 part by weight of the fish scales.
According to the present invention, the heating temperature is preferably 40-90 deg.C (for example, 40 deg.C, 42 deg.C, 44 deg.C, 46 deg.C, 48 deg.C, 50 deg.C, 52 deg.C, 54 deg.C, 56 deg.C, 58 deg.C, 60 deg.C, 62 deg.C, 64 deg.C, 66 deg.C, 68 deg.C, 70 deg.C, 72 deg.C, 74 deg.C, 76 deg.C, 78 deg.C.
According to the invention, the extraction time can vary within wide limits, preferably from 1 to 5 hours, preferably from 1 to 3 hours.
According to the present invention, in order to further improve the extraction effect of the fish gelatin, it is preferable to continuously perform stirring during the extraction, for example, using a mechanical stirrer.
The inventor of the invention finds that compared with the method of directly extracting fish gelatin by using a hot water method, ultrasonic treatment is introduced in the extraction process, the temperature of hot water in ultrasonic extraction can be reduced, so that energy consumption is saved, the degree of proteolysis and the content of soluble protein in the fish scale collagen prepared by the method are further improved, and the nutritional value, the taste and the stability of the ice cream prepared by the method are further improved. Therefore, preferably, the heating is carried out under ultrasonic conditions, and the heating temperature is 40-70 ℃ (for example, 40 ℃, 42 ℃, 44 ℃, 46 ℃, 48 ℃, 50 ℃, 52 ℃, 54 ℃, 56 ℃, 58 ℃, 60 ℃, 62 ℃, 64 ℃, 66 ℃, 68 ℃, 70 ℃).
According to the invention, the frequency of the overclocking can be changed in a wide range, preferably 180-220W, for example, 180W, 182W, 184W, 186W, 188W, 190W, 192W, 194W, 196W, 198W, 200W, 202W, 204W, 206W, 208W, 210W, 212W, 214W, 216W, 218W, 220W.
In the research process, the inventor of the invention further discovers that compared with the method of carrying out acid dipping decalcification by using hydrochloric acid and the like and then carrying out ultrasonic extraction on fish gelatin, the method preferably uses citric acid to decalcification and then carries out ultrasonic extraction on the fish gelatin, the temperature of hot water during ultrasonic extraction can be further reduced, so that the energy consumption is saved, the proteolysis degree and the soluble protein content in the fish scale collagen prepared by the method are further improved, and the nutritional value, the taste and the stability of the ice cream prepared by the method are further improved. Therefore, preferably, the acid is citric acid, and the heating is performed under ultrasonic conditions, the heating temperature is 40-55 ℃ (for example, 40 ℃, 42 ℃, 44 ℃, 46 ℃, 48 ℃, 50 ℃, 52 ℃, 54 ℃, 55 ℃).
According to the present invention, the solid-liquid separation method of the fish gelatin extract in step (3) is not particularly limited, and for example, centrifugation, filtration, or the like may be employed as long as solid residues such as fish scales are effectively removed, and a filtration method is preferable in the present invention.
According to the present invention, the liquid phase after the solid-liquid separation may be directly hydrolyzed with alkaline protease and trypsin, or may be dried and then hydrolyzed. According to a preferred mode of the invention, since the method of the invention can obtain the fish gelatin in the step (3), the step (3) further comprises drying the liquid phase after solid-liquid separation, namely, the fish gelatin solution, to obtain the fish gelatin dry powder (which is convenient for storage). The drying may be a drying method conventional in the art, and for example, the drying may be performed after the concentration.
According to the invention, the concentration is preferably carried out in vacuo, more preferably in a vacuum rotary evaporation at 50 to 70 ℃.
According to the invention, the drying is preferably a vacuum freeze drying, the drying time being 40-55 hours.
According to the present invention, in the step (4), when the fish gelatin is subjected to alkaline protease hydrolysis, the amount of the alkaline protease to be used may be in a wide range, and is preferably 1 to 5 parts by weight, for example, 1 part by weight, 2 parts by weight, 3 parts by weight, 4 parts by weight, or 5 parts by weight, based on 100 parts by weight of the fish gelatin on a dry weight basis.
According to a preferred embodiment of the invention, the hydrolysis of the fish gelatine by alkaline protease is carried out in an aqueous solution, wherein the amount of water is preferably such that the concentration of the alkaline protease is 0.1-0.5% by weight.
According to the present invention, the conditions for the hydrolysis of the alkaline protease may be the conventional conditions for the action of alkaline protease, preferably, the temperature is 45-55 deg.C, the time is 3-4 hours, and the pH is 7.5-8.5.
According to the present invention, it is preferred that the method further comprises inactivating the alkaline protease, for example, by heating, before subjecting the product after the alkaline protease hydrolysis to trypsin hydrolysis.
Further preferably, the method further comprises subjecting the product of the alkaline protease hydrolysis to solid-liquid separation (e.g., centrifugation), and drying the resulting liquid phase to obtain a powder. Wherein the drying is preferably freeze-drying.
According to the present invention, when the product after the alkaline protease hydrolysis (trypsin substrate) is subjected to trypsin hydrolysis, the amount of the trypsin may be in a wide range, and preferably is 1 to 3 parts by weight, for example, 1 part by weight, 1.5 parts by weight, 2 parts by weight, 2.5 parts by weight, or 3 parts by weight, based on 100 parts by weight of the trypsin substrate (i.e., the product after the alkaline protease hydrolysis), based on dry weight.
According to a preferred embodiment of the invention, the hydrolysis of trypsin is carried out in an aqueous solution, wherein the amount of water is preferably such that the concentration of the trypsin substrate is between 40 and 60 mg/ml.
According to the invention, the conditions for the hydrolysis of trypsin can be the conditions for the conventional trypsin action, preferably at a temperature of 45-55 ℃ for 3-4 hours and a pH of 7.5-8.5.
According to the present invention, preferably, the method further comprises inactivating the trypsin after completion of the hydrolysis of the trypsin, for example, by heating.
Further preferably, the method further comprises performing solid-liquid separation (centrifugation) on the product after the trypsin enzymolysis, and drying the obtained liquid phase into powder. Wherein the drying is preferably freeze-drying.
According to the invention, the fish scale collagen can be in a liquid form or a powder form after being freeze-dried.
According to a preferred embodiment of the present invention, the dry powder of the fish gelatin obtained in step (3) is added to an alkaline protease solution and subjected to enzymolysis at 45-55 ℃ for 3-4 hours, wherein the weight ratio of the dry powder of the fish gelatin: alkaline protease: the mass ratio of water is 10: 0.1-0.5: 100, heating to inactivate the alkaline protease after enzymolysis, centrifuging, and freeze-drying the obtained clear liquid into powder; then adding the obtained powder into a trypsin solution, and carrying out enzymolysis for 3-4 hours at the temperature of 45-55 ℃, wherein the substrate dry powder: trypsin: the ratio of water is 50: 1-3 g: 1ml, heating after enzymolysis to inactivate the trypsin, centrifuging, and freeze-drying the obtained clear liquid into powder to obtain the scale collagen powder. In this preferred case, the protein hydrolysis degree and the soluble protein content of the obtained scale collagen powder are further increased.
In a second aspect, the present invention provides fish scale collagen prepared by the method as described above.
In a third aspect, the invention provides an application of the fish scale collagen in food, cosmetics, health products or medicines.
In a fourth aspect, the invention provides an ice cream rich in scale collagen, which comprises the scale collagen, a stabilizer, skimmed milk powder, monoglyceride, pomegranate-grape mixed fruit pulp, a sweetener, cream and vegetable oil;
wherein, the amount of the fish scale collagen is 1-3 wt% based on dry weight, the amount of the stabilizer is 0.2-0.8 wt%, the amount of the skimmed milk powder is 10-20 wt%, the amount of the monoglyceride is 0.1-0.3 wt%, the amount of the pomegranate-grape mixed fruit pulp is 10-30 wt%, the amount of the sweetener is 3-10 wt%, the amount of the cream is 1-3 wt%, and the amount of the vegetable oil is 1-3 wt% based on the total feed amount.
According to the present invention, the mass ratio of the pomegranate fruit pulp to the grape fruit pulp in the pomegranate-grape mixed fruit pulp may be limited in a wide range, but in order to further improve the taste, flavor and color of the ice cream prepared, it is preferable that the mass ratio of the pomegranate fruit pulp to the grape fruit pulp in the pomegranate-grape mixed fruit pulp is 1: 0.5-1.5.
The preparation method of the pomegranate fruit pulp is not particularly limited, and the pomegranate fruit pulp can be prepared by the conventional method of the invention, such as pulping, solid-liquid separation and refining of pomegranate fruit grains in sequence. Preferably, no water is added during the preparation of the pomegranate pulp. The solid-liquid separation method may be a conventional technique in the art, and may be, for example, filtration. The refining can be carried out by refining the filtered filtrate in a colloid mill, so that the taste of the refining is more delicate.
The preparation method of the grape pulp is not particularly limited, and the grape pulp can be prepared by the conventional method of the invention, for example, grape fruit grains are peeled and then sequentially pulped, solid-liquid separated and refined. Preferably, no water is added during the preparation of the grape pulp. The solid-liquid separation method may be a conventional technique in the art, and may be, for example, filtration. The refining can be carried out by refining the filtered filtrate in a colloid mill, so that the taste of the refining is more delicate.
According to the present invention, the stabilizer is preferably a gel, and in order to further improve the mouthfeel and stability of the ice cream prepared, the gel is preferably selected from the group consisting of fish gelatin, xanthan gum, carrageenan and guar gum, and more preferably, the gel is a mixed gel of fish gelatin and at least one of xanthan gum, carrageenan and guar gum. Wherein, in the mixed gel, the ratio of fish gelatin: (at least one of xanthan gum, carrageenan and guar gum) is preferably 1:0.5 to 1.5 (mass ratio). In this preferable case, the fish scale collagen protein can exert the maximum synergistic effect, and the stability of the product can be improved.
According to the invention, the sweetener is preferably a low-sugar sweetener, wherein low sugar refers to a sweetener having less sweetness than sucrose at the same concentration, more preferably selected from the group consisting of isophytol, sorbitol and glucose, and even more preferably isophytol. Wherein, the sweetness determination method can be sweetness tester detection.
According to the present invention, the vegetable oil is preferably linseed oil and/or corn oil, more preferably linseed oil.
In a fifth aspect, the present invention provides a method for preparing an ice cream rich in fish scale collagen as described above, the method comprising: mixing fish scale collagen, stabilizer, skimmed milk powder, monoglyceride, pomegranate-grape mixed fruit pulp, sweetener, butter, vegetable oil and water; and then, homogenizing, sterilizing, carrying out solid-liquid separation, aging, freezing and hardening on the mixed material obtained by mixing in sequence to obtain the ice cream rich in the scale collagen.
According to the present invention, the conditions for homogenization may be selected from a wide range as long as the dispersion in the resulting mixture can be micronized and homogenized, and preferably include: homogenizing under 20-30MPa for 1-3 min.
According to the invention, the method of sterilization may be a conventional method, for example, pasteurization, the conditions of pasteurization comprising: sterilizing at 80-85 deg.C for 5-10 min.
According to the invention, the method of solid-liquid separation may be a conventional choice, for example, filtration may be carried out using 80-120 mesh filter cloth.
According to the invention, the aging method is preferably to allow the filtered filtrate to stand at a low temperature, for example, 2-6 ℃, for 8-12 h.
Wherein, in order to further improve the taste and stability of the ice cream, the filtrate after solid-liquid separation is preferably cooled at a rate of 0.3-0.5 ℃/s.
According to the present invention, the conditions of freezing may include: freezing and stirring the aged materials in a freezing machine, and discharging at the temperature of between 5 ℃ below zero and 3 ℃ below zero;
according to the present invention, the conditions for hardening may include: the hardening temperature is-35 deg.C to-25 deg.C, and the hardening time is 12-24 h.
The present invention will be described in detail below by way of examples. In the following examples and comparative examples:
alkaline protease was purchased from novacin biotechnology limited, enzyme activity: 6.12X 107U/g；
Trypsin was purchased from sulibao biotechnology ltd, enzyme activity: 4.07X 107U/g；
Pepsin was purchased from sulibao biotechnology ltd, enzyme activity: 4.92X 106U/g；
Papain was purchased from solibao biotechnology ltd, enzyme activity: 4.27X 106U/g；
The fish scale is from freshwater fish tilapia, grass carp and silver carp.
Examples 1 to 1
This example illustrates the preparation of collagen from fish scales
(1) Fish scale treatment: purchasing fish scales left in the processing process from aquatic products company, removing impurities, intermittently beating for 2min with a tissue mashing machine, and washing with clear water for multiple times. According to the fish scale: hydrochloric acid (concentration of 0.3mol/L) is 1:5, and the fish scales are placed in hydrochloric acid solution for decalcification at room temperature for 2.5h, and stirred for 1min every 15min during the decalcification. After the fish scale decalcification is finished, repeatedly washing and soaking the fish scale by using clear water until the pH value is washed to be neutral.
(2) Gelatin extraction: adding distilled water into the fish scales and the water in the ratio of 1:5 by volume, heating in a water bath at the temperature of 75 ℃ for 2 hours, and continuously stirring by using a mechanical stirrer in the period.
(3) Concentrating and drying: filtering to remove fish scale residue, concentrating the filtrate at 60 deg.C by rotary evaporation under vacuum, and lyophilizing in vacuum lyophilizer for 48 hr to obtain fish gelatin.
(4) Adding 10 parts by weight of the fish gelatin into 100 parts by weight of alkaline protease solution (with the concentration of 0.1 weight percent and the pH value of the solution of 8.0), carrying out enzymolysis for 4 hours at 50 ℃, heating to inactivate enzyme and centrifuging, freeze-drying supernatant, adding the supernatant into trypsin solution, and carrying out enzymolysis for 4 hours at 50 ℃, wherein the weight ratio of trypsin to substrate is 1: 50, the concentration of the substrate is 50mg/mL, and the pH value of the solution is 8.0. And after the enzymolysis is finished, heating the mixture for enzyme deactivation, centrifugally separating a solid phase and a liquid phase, and freeze-drying clear liquid into powder to obtain the freshwater fish scale collagen powder.
Examples 1 to 2
This example illustrates the preparation of collagen from fish scales
(1) Fish scale treatment: purchasing fish scales left in the processing process from aquatic products company, removing impurities, intermittently beating for 2min with a tissue mashing machine, and washing with clear water for multiple times. According to the fish scale: hydrochloric acid (pH value of 0.3mol/L) is 1:7, and the fish scales are placed in hydrochloric acid solution for decalcification at room temperature for 2h, and stirred for 1min every 15min during the decalcification. After the fish scale decalcification is finished, repeatedly washing and soaking the fish scale by using clear water until the pH value is washed to be neutral.
(2) Gelatin extraction: adding distilled water into the fish scales and the water in a volume ratio of 1:4, heating in a water bath at 85 ℃ for 2 hours, and continuously stirring by using a mechanical stirrer during the heating.
(3) Concentrating and drying: filtering to remove fish scale residue, concentrating the filtrate at 60 deg.C by rotary evaporation under vacuum, and lyophilizing in vacuum lyophilizer for 48 hr to obtain fish gelatin.
(4) Adding 10 parts by weight of the fish gelatin into 100 parts by weight of alkaline protease solution (with the concentration of 0.3 weight percent and the pH value of the solution of 8.0), carrying out enzymolysis for 3.5h at 55 ℃, heating to inactivate enzyme, centrifuging, freeze-drying supernatant, adding the supernatant into trypsin solution, and carrying out enzymolysis for 3.5h at 45 ℃, wherein the weight ratio of trypsin to substrate is 1.5: 50, the concentration of the substrate is 50mg/mL, and the pH value of the solution is 8.0. And after the enzymolysis is finished, heating the mixture for enzyme deactivation, centrifugally separating a solid phase and a liquid phase, and freeze-drying clear liquid into powder to obtain the freshwater fish scale collagen powder.
Examples 1 to 3
This example illustrates the preparation of collagen from fish scales
(1) Fish scale treatment: purchasing fish scales left in the processing process from aquatic products company, removing impurities, intermittently beating for 2min with a tissue mashing machine, and washing with clear water for multiple times. According to the fish scale: hydrochloric acid (pH value of 0.3mol/L) is 1:10, and the fish scales are placed in hydrochloric acid solution for decalcification at room temperature for 1.5h, and stirred for 1min every 15min during the decalcification. After the fish scale decalcification is finished, repeatedly washing and soaking the fish scale by using clear water until the pH value is washed to be neutral.
(2) Gelatin extraction: adding distilled water into the fish scales and the water in a volume ratio of 1:6, heating in a water bath at the temperature of 80 ℃ for 2 hours, and continuously stirring by using a mechanical stirrer during the heating.
(3) Concentrating and drying: filtering to remove fish scale residue, concentrating the filtrate at 60 deg.C by rotary evaporation under vacuum, and lyophilizing in vacuum lyophilizer for 48 hr to obtain fish gelatin.
(4) Adding 10 parts by weight of the fish gelatin into 100 parts by weight of alkaline protease solution (with the concentration of 0.5 weight percent and the pH value of the solution of 8.0), carrying out enzymolysis for 3 hours at 45 ℃, heating to inactivate enzyme and centrifuging, freeze-drying supernatant, adding the supernatant into trypsin solution, and carrying out enzymolysis for 3 hours at 55 ℃, wherein the weight ratio of trypsin to substrate is 0.5: 50, the concentration of the substrate is 50mg/mL, and the pH value of the solution is 8.0. And after the enzymolysis is finished, heating the mixture for enzyme deactivation, centrifugally separating a solid phase and a liquid phase, and freeze-drying clear liquid into powder to obtain the freshwater fish scale collagen powder.
Examples 1 to 4
This example illustrates the preparation of collagen from fish scales
The preparation of fish scale collagen was carried out in the same manner as in example 1-1, except that, in the step (2), the temperature of the water bath was 60 ℃.
Examples 1 to 5
This example illustrates the preparation of collagen from fish scales
The preparation of fish scale collagen was carried out in the same manner as in example 1-1, except that, in the step (2), the water bath heating was carried out under ultrasonic conditions of 200W frequency and the water bath temperature was 60 ℃.
Examples 1 to 6
This example illustrates the preparation of collagen from fish scales
The preparation of fish scale collagen was carried out in the same manner as in example 1-1, except that, in the step (1), hydrochloric acid was replaced with citric acid, and in the step (2), the heating in the water bath was carried out under ultrasonic conditions of 200W frequency, and the temperature of the water bath was 50 ℃.
Comparative examples 1 to 1
This comparative example illustrates the preparation of reference fish scale collagen
The preparation of fish scale collagen was performed according to the method of example 1-1, except that in step (4), trypsin was removed.
Comparative examples 1 to 2
This comparative example illustrates the preparation of reference fish scale collagen
The preparation of fish scale collagen was performed according to the method of example 1-1, except that in step (4), trypsin was replaced with an equal amount of pepsin.
Comparative examples 1 to 3
This comparative example illustrates the preparation of reference fish scale collagen
The preparation of fish scale collagen was performed according to the method of example 1-1, except that in step (4), trypsin was replaced with an equal amount of papain.
Test example 1
The hydrolysis degree and the content of soluble protein of the fresh water fish scale collagen peptide powder prepared in the examples 1 to 6 and the comparative examples 1 to 3 are respectively detected. The method for testing the hydrolysis degree refers to 'fractional enzymolysis preparation of fish scale collagen peptide and characteristic research thereof' 2011 of Master thesis of Huazhong university of agriculture. The method for testing the content of the soluble protein adopts a Coomassie brilliant blue method to test, firstly, a standard curve is made, then 10ml of enzymolysis liquid is taken in a 100ml volumetric flask, and distilled water is added to the scale to fix the volume. According to the steps of preparing a standard curve, absorbance at the wavelength of 595nm is respectively measured, and then the protein content of the sample is determined by using the standard curve. The results are shown in Table 1.
TABLE 1
| Degree of hydrolysis (%) | Soluble protein content (% by weight) | |
| Examples 1 to 1 | 25.11±0.03 | 43.05±0.58 |
| Examples 1 to 2 | 25.06±0.01 | 43.00±0.55 |
| Examples 1 to 3 | 25.22±0.04 | 43.89±0.75 |
| Examples 1 to 4 | 24.94±0.02 | 42.96±0.62 |
| Examples 1 to 5 | 27.24±0.03 | 44.57±0.91 |
| Examples 1 to 6 | 29.59±0.02 | 45.18±0.84 |
| Comparative examples 1 to 1 | 9.36±0.02 | 33.28±1.02 |
| Comparative examples 1 to 2 | 16.22±0.01 | 36.44±0.73 |
| Comparative examples 1 to 3 | 23.06±0.02 | 37.88±0.67 |
As shown in Table 1, the comparative examples and comparative examples show that the alkaline protease and the trypsin have the best effect of enzymolysis together, and the obtained soluble protein has the highest content, so that the application of the fresh water fish scale collagen in the ice cream is facilitated. Meanwhile, under the preferable ultrasonic condition, particularly under the condition of combining ultrasonic with citric acid, the enzymolysis effect and the content of soluble protein can be further improved, and the temperature of hot water extraction is reduced.
Example 2-1
This example illustrates the preparation of the ice cream according to the invention
(1) Preparing pomegranate-grape mixed pulp: selecting fresh pomegranate fruit grains and grapes which are not damaged by worms. Pulping the cleaned pomegranate fruit grains and grapes by a pulping machine respectively, filtering residues, and pulping the filtrate by a colloid mill to obtain pomegranate fruit pulp and grape fruit pulp. Mixing pomegranate fruit pulp and grape fruit pulp according to the weight ratio of 1:1 to obtain mixed slurry with mild color and moderate sweet and sour;
(2) mixing raw materials and auxiliary materials: accurately weighing each material according to the formula requirement, wherein 2 weight percent of the fish scale collagen powder prepared in the example 1, 0.4 weight percent of fish gelatin (purchased from 260-270 freezing power of ocean biology, Inc., Jiliding, Suzhou), 0.4 weight percent of guar gum, 12 weight percent of skim milk powder, 0.2 weight percent of monoglyceride, 20 weight percent of pomegranate-grape mixed fruit pulp, 5 weight percent of isophytol, 2 weight percent of animal cream (purchased from Nestle) and 2 weight percent of linseed oil are mixed, and then adding purified water to be fully stirred uniformly;
(3) high-pressure homogenization: homogenizing the uniformly mixed raw and auxiliary materials by a homogenizer with the pressure of 30 MPa;
(4) sterilizing and filtering: sterilizing the homogenized mixture at 85 deg.C for 5min, and filtering with 100 mesh nylon silk sieve to remove existing agglomeration;
(5) aging: rapidly cooling the filtered mixture to 2 ℃, and aging for 12 h;
(6) freezing: placing the mixture in a freezing machine for rapid (cooling rate of 0.3 ℃/s) freezing and stirring, and discharging at-4 ℃ to obtain soft ice cream;
(7) hardening: the soft ice cream was hardened at-30 ℃ for 24 h.
Examples 2-2 to 2-6
This example illustrates the preparation of the ice cream according to the invention
Ice cream was prepared according to the method of example 2-1, except that the scale collagen was replaced with the scale collagen of examples 1-2 to 1-6, respectively.
Comparative examples 2-1 to 2-3
This comparative example illustrates the preparation of an ice cream according to the invention with reference to
Ice cream was prepared according to the method of example 2-1, except that the scale collagen was replaced with the scale collagen of comparative examples 1-1 to 1-3, respectively.
Examples 2 to 7
This example illustrates the preparation of the ice cream according to the invention
Ice cream preparation was carried out according to the method of example 2-1, except that fish gelatin was replaced with an equal amount of carrageenan.
Examples 2 to 8
This example illustrates the preparation of the ice cream according to the invention
Ice cream was prepared according to the method of example 2-1, except that isophytol was replaced with an equal amount of sorbitol and linseed oil was replaced with an equal amount of corn oil.
Test example 2
The stability index of the ice cream produced was measured separately. The method for testing the stability index refers to Qilu industrial university Master's paper & study on soymilk ice cream processing and stability' 2016 & Chua sword. Values closer to 1 indicate better stability of the ice cream, and the results are shown in table 2.
Test example 3
Sensory scoring of ice cream smoothness: 30 test subjects were randomly selected, sensory scores of the 30 test subjects were recorded, and an average score was calculated as a final score of ice cream.
TABLE 2
As shown in Table 2, the stability and the smoothness of the texture of the ice cream prepared according to the present invention are improved as seen from the comparative examples and comparative examples. Meanwhile, the fish scale collagen prepared under the preferable ultrasonic condition, particularly under the condition of combining ultrasonic with citric acid, can further improve the stability and the mouthfeel fineness of the ice cream. And simultaneously, the preferable stabilizer combination, the sweetening agent and the vegetable oil are adopted, so that the stability and the mouthfeel fineness of the ice cream can be further improved.
The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including combinations of various technical features in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.
Claims (10)
1. A preparation method of fish scale collagen is characterized by comprising the following steps:
(1) placing the fish scales in citric acid solution for decalcification treatment to obtain decalcified fish scales; wherein, the dosage of the citric acid solution is 5-10 parts by volume relative to 1 part by volume of the fish scales; the decalcification conditions comprise: the temperature is 20-40 ℃, the time is at least 1h, and the mixture is stirred for 0.5-1.5min every 10-20min in the decalcification process;
(2) adding water into the decalcified fish scales, and extracting fish gelatin under heating condition to obtain fish gelatin extract, wherein the heating is carried out under ultrasonic condition, the heating temperature is 40-55 ℃, and the heating time is 1-5 hours; wherein, the amount of water is 3-10 parts by weight relative to 1 part by weight of the decalcified fish scales; the frequency of the ultrasonic wave is 180-220W;
(3) carrying out solid-liquid separation on the fish gelatin extracting solution to obtain fish gelatin;
(4) sequentially contacting the fish gelatin with alkaline protease and trypsin to respectively carry out hydrolysis of the alkaline protease and hydrolysis of the trypsin to obtain the fish scale collagen;
on a dry weight basis, the dosage of the alkaline protease is 1-5 parts by weight relative to 100 parts by weight of fish gelatin, and the hydrolysis conditions of the alkaline protease comprise that the temperature is 45-55 ℃, the time is 3-4 hours, and the pH value is 7.5-8.5;
the amount of trypsin is 1-3 parts by weight based on dry weight per 100 parts by weight of trypsin substrate, and the conditions for hydrolysis of the trypsin include a temperature of 45-55 ℃, a time of 3-4 hours, and a pH of 7.5-8.5.
2. Fish scale collagen produced by the process of claim 1.
3. The fish scale collagen of claim 2, for use in food, cosmetics, health products or medicine.
4. An ice cream rich in scale collagen, comprising the scale collagen of claim 2, a stabilizer, skim milk powder, monoglyceride, pomegranate-grape mixed fruit pulp, a sweetener, cream, and vegetable oil;
wherein, the amount of the fish scale collagen is 1-3 wt% based on dry weight, the amount of the stabilizer is 0.2-0.8 wt%, the amount of the skimmed milk powder is 10-20 wt%, the amount of the monoglyceride is 0.1-0.3 wt%, the amount of the pomegranate-grape mixed fruit pulp is 10-30 wt%, the amount of the sweetener is 3-10 wt%, the amount of the cream is 1-3 wt%, and the amount of the vegetable oil is 1-3 wt% based on the total feed amount.
5. The fish scale collagen-rich ice cream according to claim 4, wherein the mass ratio of pomegranate fruit pulp to grape fruit pulp in the pomegranate-grape mixed fruit pulp is 1: 0.5-1.5;
the pomegranate fruit pulp is prepared by sequentially pulping pomegranate fruit grains, carrying out solid-liquid separation and pulping;
the grape pulp is prepared by sequentially pulping grape granules, carrying out solid-liquid separation and pulping.
6. The ice cream of claim 4 or 5, wherein said stabilizer is selected from the group consisting of fish gelatin, xanthan gum, carrageenan and guar gum.
7. The ice cream of claim 4 or 5, wherein said sweetener is selected from the group consisting of isophytol, sorbitol and xylitol.
8. The ice cream of claim 4 or 5, wherein said vegetable oil is linseed oil and/or corn oil.
9. A process for the preparation of fish scale collagen-rich ice cream according to any one of claims 4 to 8, comprising: mixing fish scale collagen, stabilizer, skimmed milk powder, monoglyceride, pomegranate-grape mixed fruit pulp, sweetener, butter, vegetable oil and water; and then, homogenizing, sterilizing, carrying out solid-liquid separation, aging, freezing and hardening on the mixed material obtained by mixing in sequence to obtain the ice cream rich in the scale collagen.
10. The method of claim 9, wherein the conditions of homogenization comprise: homogenizing under 20-30MPa for 1-3 min; and/or
The sterilization conditions include: sterilizing at 80-85 deg.C for 5-10 min; and/or
The aging conditions include: cooling the liquid phase after solid-liquid separation to 2-6 ℃ at the speed of 0.3-0.5 ℃/s, and maintaining for 8-12 h; and/or
The freezing conditions include: freezing and stirring the aged materials in a freezing machine, and discharging at the temperature of between 5 ℃ below zero and 3 ℃ below zero;
the conditions for hardening include: the hardening temperature is-35 deg.C to-25 deg.C, and the hardening time is 12-24 h.
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