CN110720641A - Protein freeze-dried block with fat-reducing and muscle-increasing effects and preparation method thereof - Google Patents

Protein freeze-dried block with fat-reducing and muscle-increasing effects and preparation method thereof Download PDF

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CN110720641A
CN110720641A CN201910995229.6A CN201910995229A CN110720641A CN 110720641 A CN110720641 A CN 110720641A CN 201910995229 A CN201910995229 A CN 201910995229A CN 110720641 A CN110720641 A CN 110720641A
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parts
chitosan
protein
solution
calcium carbonate
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胡章
卢思彤
孔松芝
李思东
张冬英
李程鹏
程瑜
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Guangdong Ocean University
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Guangdong Ocean University
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/18—Peptides; Protein hydrolysates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/16—Inorganic salts, minerals or trace elements
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/30—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation
    • A23L5/32—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation using phonon wave energy, e.g. sound or ultrasonic waves
    • 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

Abstract

The invention discloses a protein freeze-dried block with fat-reducing and muscle-increasing effects and a preparation method thereof. The invention firstly provides a composition with the effects of reducing fat and increasing muscle, which comprises the following components in parts by weight: 15-40 parts of chitosan, 35-65 parts of sodium carboxymethylcellulose, 1-4 parts of calcium carbonate, 5-30 parts of marine organism collagen peptide, 0.1-2 parts of cysteine methyl ester and 0.5-3.5 parts of sodium caproate. Based on the composition, the invention also provides a protein freeze-dried block with the effects of reducing fat and increasing muscle, which can quickly form hydrogel in the stomach, stay in the stomach for a long time, and can continuously release marine organism collagen peptide, so that the human body can feel satiety, the intake of carbohydrate is reduced, the absorption of lipid is prevented, and the protein freeze-dried block has remarkable effects of reducing fat and increasing muscle; in addition, the raw materials of the protein freeze-dried block are nontoxic and easy to degrade, and the preparation process is simple and has wide application prospect.

Description

Protein freeze-dried block with fat-reducing and muscle-increasing effects and preparation method thereof
Technical Field
The invention belongs to the technical field of food and food production. More particularly, relates to a protein freeze-dried block with the effects of reducing fat and increasing muscle functions and a preparation method thereof.
Background
With the gradual change of the aesthetic appearance of people, people pay more and more attention to figure management. However, most of the fitness personnel neglect the nutrition supplement after exercise for the purpose of weight reduction and life extension exercise, influence the recovery of physical strength, and have adverse reactions such as essence reduction and the like, or supplement a large amount of food after exercise, so that the weight reduction effect cannot be achieved, and even the weight can be increased; some body-building people do a lot of strength training to increase muscles, but do not supplement protein peptide in time, and can not repair damaged muscle fibers, so that the muscle-building effect can not be achieved.
Nowadays, most of the weight-reducing products on the market achieve the purpose of reducing the weight by reducing the water content and muscle content in the body. For example, the commercial low-calorie meal replacement food mostly takes dietary fiber as a main raw material, and has the properties of water absorption and expansion, so that the satiety is increased, and the excessive intake of energy is controlled; however, excessive intake of dietary fiber can affect the absorption of minerals by the human body; part of low-calorie meal replacement food does not contain protein or contains protein but is quickly decomposed in vivo, so that the muscle is gradually lost, the metabolism capability of the body is reduced, and the normal diet state is recovered in the future, and the rebound is very easy. In addition, the weight-reducing product is expensive, and brings certain economic burden to the weight-reducing person. Moreover, most weight-reducing products have dependence, the weight-reducing effect is obvious during eating, and once the weight-reducing products are stopped, the weight can rebound to the obesity state even if the weight is controlled. Therefore, the existing weight-reducing products have the defects of obvious side effect, strong dependence, easy relapse, high price and the like.
The weight-losing effect can be better achieved by increasing the muscle content of the organism, improving the basal metabolic rate and reducing the body fat rate. Therefore, the product which has good satiety, can continuously supply enough protein, increases the muscle content, assists in repairing damaged muscle fibers and makes the muscle fibers heal and become thick is provided, and has important significance for the weight loss of obese people.
Disclosure of Invention
The invention aims to solve the technical problems of obvious side effect, strong dependence, easy rebound and high price of the existing weight-reducing products and provides a protein freeze-drying block with the effects of reducing fat and increasing muscle and a preparation method thereof. The protein freeze-dried block with the effects of reducing fat and increasing muscle provided by the invention has good satiety, can continuously supply sufficient protein, increases the muscle content, assists in repairing damaged muscle fibers and makes the muscle fibers heal and become thick.
The invention aims to provide a composition with fat reducing and muscle building effects.
The invention also aims to provide the application of the composition in preparing products with the effects of reducing fat and increasing muscle.
The invention further aims to provide a protein freeze-dried block with the effects of reducing fat and increasing muscle.
The invention further aims to provide a preparation method of the protein freeze-dried block.
The above purpose of the invention is realized by the following technical scheme:
the invention firstly provides a composition with the effects of reducing fat and increasing muscle, which comprises the following components in parts by weight: 15-40 parts of chitosan, 35-65 parts of sodium carboxymethylcellulose, 1-4 parts of calcium carbonate, 5-30 parts of marine organism collagen peptide, 0.1-2 parts of cysteine methyl ester and 0.5-3.5 parts of sodium caproate.
Among them, chitosan is a linear polysaccharide obtained by deacetylation of chitin widely existing in the nature, is the only basic polysaccharide in nature so far, has the characteristics of diverse biological activities, excellent biocompatibility, biodegradability, nontoxicity and the like, and is a new biological functional material. The chitosan can well combine fat, bile acid and cholesterol through various ways such as electrostatic action, adsorption action, encapsulation action and the like.
The sodium carboxymethyl cellulose is a cellulose derivative with the glucose polymerization degree of 100-2000, is polyanion, straight chain and water-soluble cellulose ether, and can greatly change the viscosity of most common aqueous solution preparations. Carboxylate ions on the molecular chain can be rapidly subjected to ionic crosslinking with divalent (multivalent) cations or other polycations, particularly calcium ions, can form gel, and has the capability of rapidly absorbing water and swelling. Because the sodium carboxymethylcellulose is safe and reliable, the dosage of the sodium carboxymethylcellulose is not limited by the national food hygiene standard ADI, and the sodium carboxymethylcellulose has wide application prospect.
The protein peptide has good sports nutritive value, contains rich branched chain amino acids, and the branched chain amino acids have the functions of preventing muscle decomposition and promoting muscle synthesis.
Preferably, the composition with the effects of reducing fat and increasing muscle consists of the following components in parts by weight: 20-35 parts of chitosan, 40-60 parts of sodium carboxymethylcellulose, 2-3 parts of calcium carbonate, 10-25 parts of marine organism collagen peptide, 0.5-1.5 parts of cysteine methyl ester and 1-3 parts of sodium caproate.
More preferably, the composition with the effects of reducing fat and increasing muscle consists of the following components in parts by weight: 27 parts of chitosan, 50 parts of sodium carboxymethylcellulose, 2.5 parts of calcium carbonate, 17 parts of marine organism collagen peptide, 1 part of cysteine methyl ester and 2 parts of sodium caproate.
Preferably, the marine organism collagen peptide is any one or more of tilapia mossambica protein peptide, oyster protein peptide, cod protein peptide, salmon protein peptide or tuna protein peptide.
More preferably, the marine organism collagen peptide is tilapia protein peptide.
Preferably, the powder particle size of the calcium carbonate is 0.1-5 μm.
If the particle size of the calcium carbonate powder is too large (> 5 μm), the calcium carbonate will directly settle to the bottom of the solution in step S2, and the effect of uniform dispersion cannot be achieved; if the particle size of the calcium carbonate powder is too small (< 0.1 μm), the calcium carbonate is likely to be agglomerated in step S2 and is not likely to be uniformly dispersed throughout the solution system.
More preferably, the powder particle size of the calcium carbonate is 2.5 μm.
In addition, the application of the composition in preparing products with the effects of reducing fat and increasing muscle is also within the protection scope of the invention.
The invention also provides a protein freeze-dried block with the effects of reducing fat and increasing muscle, which is prepared from the composition.
The invention also provides a preparation method of the protein freeze-dried block, which comprises the following steps:
s1, adding chitosan in the acid solution according to the weight part to prepare a chitosan solution, adding cysteine methyl ester, uniformly stirring to obtain a sulfydryl functional chitosan solution, adjusting the pH value of the solution to be 5.5-6, adding calcium carbonate, performing ultrasonic treatment, freeze drying, and grinding to obtain chitosan-calcium carbonate coated particles;
s2, adding sodium caproate, marine organism collagen peptide and the chitosan-calcium carbonate coating particles prepared in the step S1 into the sodium carboxymethylcellulose solution, and uniformly stirring to obtain a paste solution;
and S3, injecting the paste solution prepared in the step S2 into a forming mold for pre-freezing, freeze-drying and sterilizing to obtain the protein freeze-dried block.
If the pH of the solution is too low (< 5.5), the acidity is too strong, so that the calcium carbonate added later is directly decomposed; if the pH of the solution is too high (> 6), chitosan will precipitate out of solution.
Preferably, the pH of the solution of step S1 is 5.8.
Preferably, the time of the ultrasonic treatment in the step S1 is 5-15 min.
If the ultrasonic treatment time is too short (< 5min), the calcium carbonate is not uniformly dispersed; if the ultrasonic treatment time is too long (more than 15min), the molecular structure of chitosan is affected, such as the breakage of molecular chains, the damage of intermolecular hydrogen bonds and the like, and the effect of the prepared chitosan-calcium carbonate coating particles is affected.
More preferably, the time of the ultrasonic treatment in step S1 is 10 min.
Preferably, the stirring temperature of step S1 is 24 to 26 ℃.
More preferably, the stirring temperature of step S1 is 25 ℃.
Preferably, the stirring time in the step S1 is 6-8 h.
If the stirring time is too short (< 6h), the reaction grafting rate is low; if the stirring time is too long (more than 8 hours), the reaction grafting rate is not obviously improved, and the efficiency is reduced.
More preferably, the stirring time of step S1 is 7 h.
Preferably, the pH of the chitosan solution in the step S1 is 2-2.5.
Under the condition, cysteine methyl ester is easy to react with amino on the chitosan chain, thereby achieving the modification effect. If the pH of the chitosan solution is too small (less than 2), the chitosan solution is too strong in acidity and can directly decompose cysteine methyl ester; if the pH of the chitosan solution is too high (> 2.5), the reaction effect is deteriorated.
More preferably, the chitosan solution of step S1 has a pH of 2.3.
Preferably, the mass concentration of the sodium carboxymethyl cellulose solution in step S2 is 2% to 3%.
More preferably, the mass concentration of the sodium carboxymethyl cellulose solution in step S2 is 2.5%.
The acidic solution used in step S1 is an acidic solution commonly used in the art, and any acidic solution that can dissolve chitosan can be used in the present invention.
Preferably, the mass concentration of the acidic solution in step S1 is 0.5% to 1.5%.
More preferably, the mass concentration of the acidic solution in the step S1 is 1%.
Preferably, the acidic solution in step S1 is an acetic acid solution.
The invention has the following beneficial effects:
(1) the invention provides a protein freeze-dried block with fat reducing and muscle increasing effects and a preparation method thereof. According to the invention, cysteine methyl ester is used for modifying chitosan to obtain sulfydryl functionalized chitosan, wherein sulfydryl of the sulfydryl functionalized chitosan forms a disulfide bond with a region rich in cysteine residues in mucin on the surface of epithelial cells of gastrointestinal mucosa to generate specific adhesion, so that the detention time of hydrogel in the gastrointestinal tract is effectively prolonged, marine organism collagen peptide is continuously released, the absorption of the marine organism collagen peptide is promoted, and muscle is boosted;
(2) the chitosan-calcium carbonate coating powder particles are uniformly dispersed in a sodium carboxymethylcellulose solution, when the chitosan-calcium carbonate coating powder particles enter the stomach of a human body, a gastric acid solution is used as an initiator, amino groups on a chitosan molecular chain are instantaneously initiated to rapidly generate protonation, and the protonation is combined with carboxymethyl cellulose through anions and cations to form a three-dimensional hydrogel structure to wrap marine organism collagen peptide; meanwhile, calcium carbonate is decomposed to release calcium ions, the hydrogel is solidified, the mechanical strength of the hydrogel is improved, marine organism collagen peptide is continuously released, a human body can feel full for a long time, and the intake of carbohydrate is reduced;
(3) the invention utilizes the electrostatic action generated by the sodium salt of caproic acid and the protonated chitosan to be attached to the chitosan chain, thus enhancing the binding capacity to fat and fatty acid, preventing the absorption of lipid and playing a role in reducing blood fat;
(4) the raw materials of the formula of the composition with the effects of reducing fat and enhancing muscle provided by the invention are nontoxic and easy to degrade, and other additives such as food preservatives and the like are not added in the formula, so that the composition is high in safety and has a certain health-care effect; in addition, the protein freeze-dried block with the fat reducing and muscle increasing effects provided by the invention is simple in preparation process, low in cost, easy for industrial large-scale production, and wide in popularization and application prospect in preparation of products with the fat reducing and muscle increasing effects.
Drawings
FIG. 1 is a graph of Fourier transform infrared spectrometry results of thiol-functionalized chitosan.
FIG. 2 is a scanning electron micrograph of calcium carbonate and chitosan-calcium carbonate coated particles; wherein, the picture (a) is a scanning electron micrograph of calcium carbonate; FIG. (b) is a scanning electron micrograph of chitosan-calcium carbonate coated particles
FIG. 3 is a scanning electron microscope image of a protein lyophilized block with effects of reducing fat and increasing muscle prepared by the present invention.
FIG. 4 is a graph showing the results of the body shape of mice after they were administered with the lyophilized protein block having the effect of reducing fat and increasing muscle prepared according to the present invention.
Fig. 5 is a graph showing the shape and color change of the lyophilized protein block with effects of reducing fat and increasing muscle, which is prepared by the present invention, after contacting artificial gastric juice and artificial intestinal juice.
FIG. 6 is a graph of the in vitro release assay result of the protein lyophilized block with fat reducing and muscle increasing effects on marine collagen peptide.
Detailed Description
The present invention is further illustrated by the following examples, which are not intended to limit the invention in any way. The reagents, methods and apparatus employed in the present invention are conventional in the art, unless otherwise specified.
Unless otherwise indicated, reagents and materials used in the following examples are commercially available.
Example 1 preparation of lyophilized cake of protein having effects of reducing fat and increasing muscle
A protein freeze-dried block with the effects of reducing fat and increasing muscle comprises the following components in parts by weight: 27 parts of chitosan, 50 parts of sodium carboxymethylcellulose, 2.5 parts of calcium carbonate, 17 parts of tilapia protein peptide, 1 part of cysteine methyl ester and 2 parts of sodium caproate.
The preparation method of the protein freeze-dried block comprises the following steps:
s1, adding chitosan in parts by weight into an acetic acid solution with the mass concentration of 1% to prepare a chitosan solution with the pH value of 2.3, adding cysteine methyl ester, stirring at 25 ℃ for 7 hours to obtain a sulfydryl functional chitosan solution, adjusting the pH value of the solution to 5.8, adding calcium carbonate with the particle size of 2.5 mu m into the solution, performing ultrasonic treatment for 10min, and performing freeze drying and grinding to obtain chitosan-calcium carbonate coated particles;
s2, adding sodium caproate, tilapia protein peptide and chitosan-calcium carbonate coating particles prepared in the step S1 into a sodium carboxymethylcellulose solution with the mass concentration of 2.5%, and uniformly stirring to obtain a paste solution;
and S3, injecting the paste solution prepared in the step S2 into a forming mold for pre-freezing, freeze-drying and sterilizing to obtain the protein freeze-dried block.
Example 2 preparation of lyophilized cake of protein having effects of reducing fat and increasing muscle
A protein freeze-dried block with the effects of reducing fat and increasing muscle comprises the following components in parts by weight: 20 parts of chitosan, 40 parts of sodium carboxymethylcellulose, 2 parts of calcium carbonate, 10 parts of oyster protein peptide, 0.5 part of cysteine methyl ester and 1 part of sodium caproate.
The preparation method of the protein freeze-dried block comprises the following steps:
s1, adding chitosan according to the weight part into an acetic acid solution with the mass concentration of 0.5% to prepare a chitosan solution with the pH value of 2, adding cysteine methyl ester, stirring for 6 hours at 24 ℃ to obtain a sulfydryl functional chitosan solution, adjusting the pH value of the solution to 5.5, adding calcium carbonate with the powder particle size of 0.1 mu m, carrying out ultrasonic treatment for 5min, carrying out freeze drying, and grinding to obtain chitosan-calcium carbonate coated particles;
s2, adding sodium caproate, oyster protein peptide and the chitosan-calcium carbonate coating powder particles prepared in the step S1 into a sodium carboxymethylcellulose solution with the mass concentration of 2%, and uniformly stirring to obtain a paste solution;
and S3, injecting the paste solution prepared in the step S2 into a forming mold for pre-freezing, freeze-drying and sterilizing to obtain the protein freeze-dried block.
Example 3 preparation of lyophilized cake of protein having effects of reducing fat and increasing muscle
A protein freeze-dried block with the effects of reducing fat and increasing muscle comprises the following components in parts by weight: 35 parts of chitosan, 60 parts of sodium carboxymethylcellulose, 3 parts of calcium carbonate, 25 parts of cod protein peptide, 1.5 parts of cysteine methyl ester and 3 parts of sodium caproate.
The preparation method of the protein freeze-dried block comprises the following steps:
s1, adding chitosan according to the weight part into an acetic acid solution with the mass concentration of 1.5% to prepare a chitosan solution with the pH value of 2.5, adding cysteine methyl ester, stirring at 26 ℃ for 8 hours to obtain a sulfydryl functional chitosan solution, adjusting the pH value of the solution to 6, adding calcium carbonate with the particle size of 5 mu m, carrying out ultrasonic treatment for 15min, carrying out freeze drying, and grinding to obtain chitosan-calcium carbonate coated particles;
s2, adding sodium adipate, cod protein peptide and the chitosan-calcium carbonate coating particles prepared in the step S1 into a sodium carboxymethylcellulose solution with the mass concentration of 3%, and uniformly stirring to obtain a paste solution;
and S3, injecting the paste solution prepared in the step S2 into a forming mold for pre-freezing, freeze-drying and sterilizing to obtain the protein freeze-dried block.
Example 4 preparation of lyophilized cake of protein having effects of reducing fat and increasing muscle
A protein freeze-dried block with the effects of reducing fat and increasing muscle comprises the following components in parts by weight: 15 parts of chitosan, 35 parts of sodium carboxymethylcellulose, 1 part of calcium carbonate, 5 parts of salmon protein peptide, 0.1 part of cysteine methyl ester and 0.5 part of sodium caproate.
The preparation method of the protein freeze-dried block comprises the following steps:
s1, adding chitosan in parts by weight into an acetic acid solution with the mass concentration of 0.8% to prepare a chitosan solution with the pH value of 2.2, adding cysteine methyl ester, stirring for 8 hours at 24 ℃ to obtain a sulfydryl functional chitosan solution, adjusting the pH value of the solution to 5.6, adding calcium carbonate with the powder particle size of 0.5 mu m, carrying out ultrasonic treatment for 8min, and carrying out freeze drying and grinding to obtain chitosan-calcium carbonate coated particles;
s2, adding sodium adipate, salmon protein peptide and the chitosan-calcium carbonate coating particles prepared in the step S1 into a sodium carboxymethylcellulose solution with the mass concentration of 2.2%, and uniformly stirring to obtain a paste solution;
and S3, injecting the paste solution prepared in the step S2 into a forming mold for pre-freezing, freeze-drying and sterilizing to obtain the protein freeze-dried block.
Example 5 preparation of lyophilized cake of protein having effects of reducing fat and increasing muscle
A protein freeze-dried block with the effects of reducing fat and increasing muscle comprises the following components in parts by weight: 40 parts of chitosan, 65 parts of sodium carboxymethylcellulose, 4 parts of calcium carbonate, 30 parts of tuna protein peptide, 2 parts of cysteine methyl ester and 3.5 parts of sodium caproate.
The preparation method of the protein freeze-dried block comprises the following steps:
s1, adding chitosan in parts by weight into an acetic acid solution with the mass concentration of 1.3% to prepare a chitosan solution with the pH value of 2.4, adding cysteine methyl ester, stirring at 26 ℃ for 6 hours to obtain a sulfydryl functional chitosan solution, adjusting the pH value of the solution to 5.9, adding calcium carbonate with the particle size of 3 mu m, carrying out ultrasonic treatment for 13min, carrying out freeze drying, and grinding to obtain chitosan-calcium carbonate coated particles;
s2, adding sodium adipate, tuna protein peptide and the chitosan-calcium carbonate coating particles prepared in the step S1 into a sodium carboxymethylcellulose solution with the mass concentration of 2.7%, and uniformly stirring to obtain a paste solution;
and S3, injecting the paste solution prepared in the step S2 into a forming mold for pre-freezing, freeze-drying and sterilizing to obtain the protein freeze-dried block.
Comparative example 1
A protein freeze-dried block comprises the following components in parts by weight: 27 parts of chitosan, 50 parts of sodium carboxymethyl cellulose, 2.5 parts of calcium carbonate, 17 parts of tilapia protein peptide and 2 parts of sodium caproate.
The preparation of the lyophilized cake of protein according to this comparative example was the same as in example 1, except that cysteine methyl ester was not added.
Comparative example 2
A protein freeze-dried block comprises the following components in parts by weight: 27 parts of chitosan, 50 parts of sodium carboxymethyl cellulose, 2.5 parts of calcium carbonate, 17 parts of tilapia protein peptide, 1 part of cysteine methyl ester and 2 parts of sodium caproate.
The preparation method of the lyophilized cake of protein according to this comparative example was the same as that of example 1 except that the particle size of calcium carbonate powder was 7 μm.
Comparative example 3
A protein freeze-dried block comprises the following components in parts by weight: 27 parts of chitosan, 50 parts of sodium alginate, 2.5 parts of calcium carbonate, 17 parts of tilapia protein peptide, 1 part of cysteine methyl ester and 2 parts of sodium caproate.
The preparation method of the lyophilized cake of protein described in this comparative example was the same as in example 1 except that sodium alginate was used instead of sodium carboxymethyl cellulose.
Comparative example 4
A protein freeze-dried block comprises the following components in parts by weight: 27 parts of chitosan, 50 parts of sodium carboxymethyl cellulose, 2.5 parts of calcium carbonate, 17 parts of tilapia protein peptide, 1 part of cysteine methyl ester and 2 parts of sodium caproate.
The preparation method of the protein freeze-dried block comprises the following steps:
s1, directly adding chitosan, cysteine methyl ester, calcium carbonate, sodium caproate and tilapia protein peptide into a sodium carboxymethylcellulose solution with the mass concentration of 2.5%, and uniformly stirring to obtain a paste solution;
s2, injecting the pasty solution prepared in the step S1 into a forming mold for pre-freezing, freeze-drying and sterilizing to obtain the protein freeze-dried block.
Comparative example 5
A protein freeze-dried block comprises the following components in parts by weight: 27 parts of chitosan, 50 parts of sodium carboxymethyl cellulose, 2.5 parts of calcium carbonate, 17 parts of tilapia protein peptide and 1 part of cysteine methyl ester.
The lyophilized cake of protein described in this comparative example was prepared in the same manner as in example 1, except that no sodium salt of caproic acid was added.
Application example 1 structural characterization of protein lyophilized cake with lipid-reducing and muscle-increasing effects
1. Experimental methods
The thiol-functionalized chitosan solution obtained in step S1 was purified by dialysis and characterized by PerkinElmer Spectrum 100 using Fourier transform infrared spectrometer. Then, the chitosan-calcium carbonate coating powder prepared in step S1 was gold-plated, and its microscopic morphology was observed by a scanning electron microscope. In addition, the lyophilized block of protein having effects of reducing fat and increasing muscle, which was prepared in example 1 of the present invention, was cut into a sheet having a thickness of 1mm, plated with gold, and observed for its microscopic morphology by a scanning electron microscope.
2. Results of the experiment
The Fourier transform infrared spectrometry result of the thiol-functionalized chitosan is shown in FIG. 1, and it can be seen that the thiol-functionalized chitosan is 3450cm-1The absorption peaks at (A) are the stretching vibration of N-H and O-H, 1650cm-1And 1526cm-1The absorption peak is the characteristic absorption peak of the amide I band and the amide II band, 1311cm-1The absorption peak is the absorption peak of amide III with amino; compared with the absorption peak of chitosan, the absorption peak is 1650cm-1And 1526cm-1The absorption peak is obviously enhanced, and at the same time, at 2100cm-1A weak sulfydryl frequency doubling absorption peak also appears. The above results illustrate that: cysteine methyl esters have been successfully grafted onto chitosan chains.
The scanning electron microscope images of the calcium carbonate and chitosan-calcium carbonate coated particles are shown in fig. 2, wherein, the image (a) is the scanning electron microscope image of calcium carbonate; FIG. (b) is a scanning electron microscope image of chitosan-calcium carbonate coated particles; therefore, the calcium carbonate without chitosan coating has an irregular shape and clear edges and corners; the surface morphology of chitosan-calcium carbonate coating particles is obviously changed, and a sulfydryl functional chitosan coating layer with a plurality of wrinkles appears on the surface.
The scanning electron microscope image of the protein freeze-dried block with the effects of reducing fat and increasing muscle is shown in fig. 3, and it can be seen that the protein freeze-dried block is composed of smooth porous structures which are connected with each other, the stable structure enables the protein freeze-dried block to have high specific surface area and high water content, and pores in the protein freeze-dried block are beneficial to the transportation of marine organism collagen peptide and the discharge of metabolic waste.
Application example 2 determination of fat-reducing efficacy of protein lyophilized cake having fat-reducing and muscle-increasing efficacy
Through establishing a nutritional obese mouse model, respectively administering the protein freeze-dried blocks with the fat reducing and muscle increasing effects prepared in the above examples 1 to 5 and the protein freeze-dried blocks prepared in the comparative examples 1 to 5 to a mouse, measuring the fat reducing effect, and observing the changes of the weight gain, the in vivo fat content and the daily average food intake of the mouse, the specific experimental method and the experimental result are as follows:
1. experimental methods
(1) Establishment of nutritional obesity mouse model
The male ICR mice are purchased from the medical animal center of Guangdong province, are weaning 1-week-old mice, and have the weight of 20 g +/-0.05, and are 132 mice in total; the temperature of the mouse breeding environment is 18-22 ℃ and the humidity is 50% -60% during the experiment, natural illumination is adopted, and the mouse can freely eat and drink water.
After 1 week of adaptive feeding, the mice were randomly divided into 2 groups, one group was Normal Mice (NM), the other group was obese model mice (Fat model mouse, FM), the Normal mice group was fed with maintenance diet, the obese model mice group was fed with high-Fat diet, and the mice were continuously fed for 30 days; the body weight is weighed once a week, and at the end of the model building, the body weight of the obese model mouse group exceeds 20 percent of that of the normal mouse group, so that the obese model is considered to be successful in model building.
(2) Experiment grouping
Model control group: feeding 12 mice with high-fat feed, and intragastrically feeding distilled water according to the weight of 2mL/kg of the mice;
examples 1 to 5 groups: feeding 12 high-fat feeds for each group, and respectively feeding the high-fat feeds into stomach according to the weight of 2mL/kg of mice to obtain the protein freeze-dried blocks with the effects of reducing fat and increasing muscle, which are prepared in the examples 1-5;
comparative examples 1 to 5 groups: and feeding 12 high-fat feeds in each group, and respectively feeding the high-fat feeds into the stomach according to the weight of 2mL/kg of the mice to obtain the protein freeze-dried blocks prepared in the comparative examples 1-5.
(3) Mouse treatment and index determination
The experiment lasts for 30 days, the gavage mouse is performed at 1 noon every day, the food intake, the food scattering amount and the food remaining amount of the mouse are recorded every week, and the weight is weighed for 1 time; at the end of the experiment, the mice were fasted for 12 hours without water deprivation, and the weights were weighed;
the method comprises the following steps of (1) taking eyeballs of a mouse to take blood, collecting the blood, removing cervical vertebrae to kill, centrifuging the blood at 3500r/min for 15min to obtain serum, immediately storing the serum at-60 ℃, and using the kit to detect indexes such as Total Cholesterol (TC), Triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) in the serum in the following step; after blood sampling, the mice were dissected to remove perirenal fat and testicular fat, weighed, and the fat content was calculated.
2. Results of the experiment
The body shape results of the mice taking the protein freeze-dried block with the effects of reducing fat and increasing muscle are shown in fig. 4, and it can be seen that the body shape of the mice taking the protein freeze-dried block is obviously changed, the effects of reducing fat and increasing muscle are achieved, and the hair of the mice is smooth, glossy and good in mental state.
The results of the changes of the weight gain, the in vivo fat content and the daily average food intake of the mice are shown in table 1, and it can be seen that the weight gain, the in vivo fat content and the daily average food intake of the mice in the groups of examples 1 to 5 are lower than those of the model control group and have very significant differences (p is less than 0.01) compared with the model control group in the groups of examples 1 to 5; compared with the model control group, the mice in the comparative example 1 group have obviously reduced weight gain and in-vivo fat content (p < 0.05), the mice in the comparative example 5 group have obviously reduced weight gain (p < 0.05) and very obviously reduced daily average food intake (p < 0.01), the mice in the comparative example 2 group, the mice in the comparative example 3 group and the mice in the comparative example 4 have no obvious difference in weight gain, in-vivo fat content and daily average food intake from the model control group, and the change of the formula composition of the protein freeze-dried block or the preparation method thereof can obviously influence the efficacy of the protein freeze-dried block in fat reduction and muscle increase.
The formulation of the protein lyophilized block in the group of comparative example 1 has no cysteine methyl ester (cannot improve chitosan functionalization), specific adhesion cannot be generated, but the amino group of chitosan is positively charged, namely, the chitosan presents cationic characteristics and acidic groups with negative electricity (anions) in mucosa realize adhesion through ionic interaction, the retention time of the protein lyophilized block in the stomach and intestine is increased to a certain extent, but the effect is not as good as that of thiol-functionalized chitosan obtained by adding cysteine methyl ester; the formula of the protein freeze-dried block in the comparative example 5 group contains no sodium caproate, and the data results in the table 1 show that the sodium caproate mainly influences the weight of fat in a mouse body, so that the sodium caproate plays a key role in the aspects of oil adsorption and the like.
In addition, compared with the group of the example 1, the weight gain, the in vivo fat content and the daily average food intake of the mice in the groups of the comparative examples 1 to 5 are all obviously improved, and the change of the formula composition of the protein freeze-dried block or the preparation method thereof is also proved to obviously influence the efficacy of the protein freeze-dried block in the aspects of fat reduction and muscle increase. The formula of the protein freeze-dried block in the group of the comparative example 1 has no cysteine methyl ester (cannot improve the chitosan functionalization), which shows that unmodified chitosan can still form a three-dimensional space structure, wraps marine organism collagen peptide and can achieve a better release effect, so that the marine organism collagen peptide can be combined with bile acid, the bile acid amount of liver and intestine circulation is reduced, the utilization of the bile acid is prevented, and the excretion is increased, so that the absorption of cholesterol and the deposition of cholesterol in vivo are inhibited.
The above results show that: the protein freeze-dried block prepared by the invention has obvious effects of reducing fat and increasing muscle.
TABLE 1 mouse weight gain, in vivo fat content and daily average food intake Change results
Figure BDA0002239524680000111
Figure BDA0002239524680000121
Note: "+" represents p < 0.05, "+" represents p < 0.01, compared to the model control group; in comparison with the group of example 1, "#" represents p < 0.05 and "##" represents p < 0.01.
The results of the changes of the concentrations of TC, TG, HDL-C and LDL-C in the mouse serum are shown in Table 2, and it can be seen that the concentrations of TC, TG and LDL-C in the mouse serum of the mice of the groups 1-5 in examples 1-5 are very significantly reduced (p < 0.01) and the concentration of HDL-C in the mouse serum of the groups 1-5 is significantly reduced (p < 0.05) compared with the model control group, which indicates that the protein freeze-dried block prepared by the invention has a significant inhibition effect on the increase of TC, TG and LDL-C in the mouse serum caused by high-fat diet; compared with the model control group, the concentrations of TC and TG in the mouse serum of the comparative example group 1 are remarkably reduced (p is less than 0.05), and the differences between the concentrations of TC, TG, HDL-C and LDL-C in the mouse serum of the other comparative example groups and the model control group are not remarkable, so that the change of the formula composition of the protein freeze-dried block or the preparation method thereof has remarkable influence on the lipid-lowering effect.
TABLE 2 results of changes in the concentrations of TC, TG, HDL-C and LDL-C in mouse serum (mmol/L)
Figure BDA0002239524680000122
Note: "+" represents p < 0.05 and "+" represents p < 0.01, compared to the model control group.
Application example 3 in vitro release assay of Marine collagen peptide from protein lyophilized Block with lipid reducing and muscle increasing effects
An ultraviolet spectrophotometer is adopted to measure the absorbance of the released marine organism collagen peptide (protein), the release content of the released marine organism collagen peptide (protein) is calculated by using the Lambert-Beer law, and then the in-vitro release of the protein freeze-dried block with the fat reducing and muscle increasing effects prepared in the examples 1 to 4 and the in-vitro release of the protein freeze-dried block prepared in the comparative examples 1 to 4 to the marine organism collagen peptide are measured, and the specific experimental method and the experimental result are as follows:
1. experimental methods
(1) Preparation of artificial gastric juice and artificial intestinal juice
Artificial gastric juice: taking 16.4mL of dilute hydrochloric acid, adding about 800mL of water, transferring the diluted hydrochloric acid to a 1000mL volumetric flask, adding water to a scale mark, and controlling the pH value to be 1-2;
artificial intestinal juice: 6.8g of monopotassium phosphate is taken, 500mL of water is added to dissolve the monopotassium phosphate, the pH value is adjusted to 6.8 by using 0.1mol/L sodium hydroxide solution, the mixture is transferred to a 1000mL volumetric flask, and water is added to the scale mark.
(2) Establishment of protein peptide standard curve
Accurately weighing 0.5g bovine serum albumin, dissolving in ultrapure water, and diluting to 100mL to obtain 5 × 10 concentration-3g/mL of marine active peptide solution; respectively diluted to 5 × 10-4g/mL、2.5×10-4g/mL、 1×10-4g/mL、5×10-5g/mL and 2.5X 10-5Measuring the absorbance of the g/mL solution at the maximum absorption wavelength of an ultraviolet-visible spectrophotometer by taking distilled water as a reference;
plotting the absorbance as ordinate and the concentration as abscissa to obtain standard protein peptide curve with fitting equation of 0.0185+17.206x, R2＝0.9996。
(3) In vitro release assay experiment of marine organism collagen peptide
Magnetic stirring is carried out at 37 ℃, 5mL of the protein freeze-dried block with the effects of reducing fat and increasing muscle prepared in examples 1-4 and the protein freeze-dried block prepared in comparative examples 1-4 are dissolved by warm water, and then are put into a dialysis bag, and are placed in 200mL of artificial gastric juice for 2 hours and then transferred into the artificial intestinal juice for 4 hours; taking out 1mL of sample solution at a proper time interval, measuring absorbance by using an ultraviolet-visible spectrophotometer, and calculating the in-vitro release amount of the marine organism collagen peptide; meanwhile, 1mL of fresh artificial gastric juice or artificial intestinal juice is used for replacing the buffer solution so as to keep the volume of the buffer solution the same; establishing an in-vitro release curve of the marine organism collagen peptide by taking the time as an abscissa and the release amount of the marine organism collagen peptide as an ordinate;
Figure BDA0002239524680000131
wherein R is0Adding amount of marine organism collagen peptide, RtThe release amount of the marine organism collagen peptide is shown.
2. Results of the experiment
The shape and color change results of the protein freeze-dried block with the effects of reducing fat and increasing muscle after contacting artificial gastric juice and artificial intestinal juice are shown in fig. 5, and it can be seen that the protein freeze-dried block is dissolved in warm water to form protein soup, and the protein soup can rapidly form hydrogel after contacting the artificial gastric juice (gastric acid); after dialysis in the artificial intestinal juice for a certain time, the hydrogel gradually decreases, and the color gradually changes from white to transparent; the protein freeze-dried block can gradually release marine organism collagen peptide in the gastrointestinal tract, and the protein freeze-dried block is gradually degraded.
The in vitro release measurement results of the protein freeze-dried block with the effects of reducing fat and increasing muscle prepared by the invention on the marine organism collagen peptide are shown in fig. 6, and it can be seen that the in vitro release rate of the marine organism collagen peptide in artificial gastric juice (pH is 1.2) is lower than that in artificial intestinal juice (pH is 7.4) in the groups of examples 1 to 4 and the groups of comparative examples 1 to 4, and the protein freeze-dried block can release the marine organism collagen peptide in the artificial intestinal juice for 4 hours; in addition, the in-vitro release rate of the marine organism collagen peptide in the artificial gastric juice in the groups of examples 1 to 4 is lower than that in the groups of comparative examples 1 to 4, while the in-vitro release rate of the marine organism collagen peptide in the artificial intestinal juice in the groups of examples 1 to 4 is higher than that in the groups of comparative examples 1 to 4, wherein the improvement effect of the group of example 1 is the most remarkable. The above results show that: the protein freeze-dried block with the effects of reducing fat and increasing muscle, which is prepared by the invention, can protect marine biological collagen peptide from being damaged by gastric juice to a certain extent, can be released in intestinal juice in a large amount, and is beneficial to the absorption of marine biological collagen peptide by human body.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, and simplifications are intended to be included in the scope of the present invention.

Claims (10)

1. The composition with the effects of reducing fat and increasing muscle is characterized by comprising the following components in parts by weight: 15-40 parts of chitosan, 35-65 parts of sodium carboxymethylcellulose, 1-4 parts of calcium carbonate, 5-30 parts of marine organism collagen peptide, 0.1-2 parts of cysteine methyl ester and 0.5-3.5 parts of sodium caproate.
2. The composition according to claim 1, characterized by consisting of, in parts by weight: 20-35 parts of chitosan, 40-60 parts of sodium carboxymethylcellulose, 2-3 parts of calcium carbonate, 10-25 parts of marine organism collagen peptide, 0.5-1.5 parts of cysteine methyl ester and 1-3 parts of sodium caproate.
3. The composition of claim 1, wherein the marine collagen peptide is one or more of tilapia, oyster, cod, salmon, or tuna protein peptides.
4. The composition according to claim 1, wherein the calcium carbonate has a powder particle size of 0.1 to 5 μm.
5. Use of the composition of any one of claims 1 to 4 for the preparation of a product having lipid-reducing and muscle-building effects.
6. A protein lyophilized block having effects of reducing fat and increasing muscle, which is prepared from the composition of any one of claims 1 to 4.
7. The method for preparing a lyophilized block of protein according to claim 6, comprising the steps of:
s1, adding chitosan according to any one of claims 1-2 in parts by weight into an acidic solution to prepare a chitosan solution, adding cysteine methyl ester, uniformly stirring to obtain a sulfydryl functional chitosan solution, adjusting the pH of the solution to 5.5-6, adding calcium carbonate, performing ultrasonic treatment, freeze drying, and grinding to obtain chitosan-calcium carbonate coated particles;
s2, adding sodium caproate, marine organism collagen peptide and the chitosan-calcium carbonate coating particles prepared in the step S1 into the sodium carboxymethylcellulose solution, and uniformly stirring to obtain a paste solution;
and S3, injecting the paste solution prepared in the step S2 into a forming mold for pre-freezing, freeze-drying and sterilizing to obtain the protein freeze-dried block.
8. The preparation method according to claim 7, wherein the time of the ultrasonic treatment in the step S1 is 5-15 min.
9. The method according to claim 7, wherein the stirring temperature of step S1 is 24-26 ℃; and S1, stirring for 6-8 h.
10. The method according to claim 7, wherein the chitosan solution of step S1 has a pH of 2-2.5.
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