CN116574777A - Preparation method of crayfish shell polypeptide and application of crayfish shell polypeptide in puffed food - Google Patents
Preparation method of crayfish shell polypeptide and application of crayfish shell polypeptide in puffed food Download PDFInfo
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- CN116574777A CN116574777A CN202310569981.0A CN202310569981A CN116574777A CN 116574777 A CN116574777 A CN 116574777A CN 202310569981 A CN202310569981 A CN 202310569981A CN 116574777 A CN116574777 A CN 116574777A
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- China
- Prior art keywords
- microwave
- crayfish
- polypeptide
- crayfish shell
- puffed food
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The invention belongs to the technical field of food additives, and particularly relates to a preparation method of crayfish shell polypeptides and application of crayfish shell polypeptides in puffed foods. The preparation method of the crayfish shell polypeptide comprises the following steps: s1, crushing cleaned shrimp shells to prepare shrimp shell powder; s2, sequentially carrying out high hydrostatic pressure, super-energy electron beam irradiation and ultrasonic treatment on shrimp shell powder, preparing a solution, adding compound protease, and hydrolyzing at constant temperature to obtain protein hydrolysate, wherein the compound protease comprises the following components in percentage by mass of 0.3-0.8:1 and trypsin; and S3, filtering the protein hydrolysate, and freeze-drying the filtrate to obtain the crayfish shell polypeptide. The invention combines super-energy electron beam irradiation and ultrasonic treatment, adopts composite treatment of alkaline protease and trypsin, can effectively improve the content of active polypeptide, and the obtained shrimp shell polypeptide has excellent antioxidant activity, and is suitable for preparing puffed food with high added value.
Description
Technical Field
The invention relates to the technical field of food additives, in particular to a preparation method of crayfish shell polypeptides and application of the crayfish shell polypeptides in puffed foods.
Background
The puffed food is a snack food which takes grains and the like as main raw materials, is made into a structure with obviously increased volume and a certain puffing degree by adopting a puffing process, is crisp and loose and generates a series of aromatic odors, is popular with consumers due to the advantages of portability and flavor, and takes an important role in the food market. The puffing technology can improve the quality of food, improve the functional characteristics of the food such as digestibility and the like, relieve the production problem of food safety and provide driving force for the steering development of the food industry. A series of chemical changes can be generated in the frying process of the traditional puffed food, a large amount of harmful substances such as benzopyrene, acrylamide and the like are generated, and a series of health problems such as obesity, hyperlipidemia and the like can be caused. Non-traditional puffing technologies comprise microwave puffing, airflow puffing, extrusion puffing and the like, the equipment of the technologies is simple, the operation is convenient, the obtained product is healthy, and the product is widely loved by consumers and has high market acceptance. In recent years, research into the preparation of puffed foods by non-conventional puffing techniques has been increasingly focused.
For puffed food, the raw material composition may be one of the main factors affecting its puffing characteristics, and thus, grasping the basic ingredients of raw materials such as proteins and the like is very important for puffed products. The protein is one of seven elements of a human body, has remarkable meaning to the human body, and the protein is added into food in the form of an additive, so that the nutritional value of the food can be enhanced, the functional characteristics of the protein can also influence the product to a certain extent such as puffing characteristics and functional characteristics, the special appearance, physical and chemical indexes and the like, and the proper amount of protein can influence the swelling power and gelatinization performance of the product. At present, most of research on puffed foods is focused on technological parameters, and the cognition on the influence of proteins on the puffed foods is relatively backward, so that high-protein nutritional puffed foods cannot be developed.
Shrimp is one of important aquatic products in China, a large amount of shrimp shells and other byproducts are generated in the processing and eating processes of various shrimps such as crayfish, and most of the shrimp shells are not fully utilized, so that waste is caused, the environment is polluted, and the comprehensive utilization of the shrimp byproducts is urgent. Researches show that the shrimp shell is rich in various nutritional components, the chitin content is 10% -20%, the protein content is 20% -40%, and the crayfish shell protein is essentially a protein with a certain biological activity, so that the crayfish shell protein is extracted or hydrolyzed to obtain active polypeptide by adopting a proper method, and the active polypeptide is added into a raw material matrix as a food additive to improve the food quality or endow the food with a new function, thereby providing a new way for recycling shrimp byproducts. However, the related process for preparing the puffed food by taking the crayfish shell protein as the food additive is less in research, and the problems of poor puffing effect, insufficient flavor and texture, shortened shelf life and the like of the snack food caused by low protein content and less active ingredients extracted by the prior art are solved, so the crayfish shell protein extraction process more suitable for the microwave puffed food additive is to be further developed and explored.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a preparation method of the crayfish shell polypeptide, wherein the protein extraction rate can be up to more than 90%, the extracted crayfish shell polypeptide has high content of active ingredients, is rich in various essential amino acids and hydrophobic amino acids, and shows good antioxidant activity.
In order to achieve the above purpose, the present invention is specifically realized by the following technical scheme:
the first aspect of the invention provides a preparation method of crayfish shell polypeptide, which comprises the following steps:
s1, crushing cleaned shrimp shells to prepare shrimp shell powder;
s2, sequentially carrying out high hydrostatic pressure, super-energy electron beam irradiation and ultrasonic treatment on the shrimp shell powder to obtain pretreated shrimp shell powder, preparing the pretreated shrimp shell powder into a solution, adding compound protease to hydrolyze at a constant temperature to obtain protein hydrolysate, wherein the compound protease comprises the following components in percentage by mass of 0.3-0.8:1 and trypsin;
And S3, filtering the protein hydrolysate in the step S2, and freeze-drying the filtrate to obtain the crayfish shell polypeptide.
Further, in step S1, the preparation of the shrimp shell powder specifically includes the following steps: washing fresh shrimp shell with tap water to remove soil, residue and residue on the surface, boiling for sterilizing for 30min, draining water, oven drying at 80deg.C to constant weight, pulverizing, and micronizing to obtain shrimp shell powder with particle diameter of 10-25 μm.
Further, in the step S2, the pressure of the high hydrostatic pressure is 100-400MPa, and the dwell time is 10-30min; the accelerating voltage of the electron beam irradiation is 8-12kV, and the irradiation time is 10-60min; the ultrasonic power is 60-100W, the ultrasonic temperature is 15-55 ℃, and the ultrasonic time is 20-40min; the enzyme activity of the compound protease is 8000-12000U, the hydrolysis temperature is 45-55 ℃, and the hydrolysis time is 2-4h.
Further, in step S2, the accelerating voltage of the electron beam irradiation is 10kV, the irradiation time is 20min, the ultrasonic power is 90W, the ultrasonic temperature is 25 ℃, the ultrasonic time is 30min, the enzyme activity of the composite protease is 10200U, and the mass ratio of the alkaline protease to the trypsin is 0.65:1, hydrolysis time was 2.8h.
The second aspect of the invention provides an application of the crayfish shell polypeptide prepared by the preparation method of the crayfish shell polypeptide in puffed food.
The third aspect of the invention provides a crayfish shell polypeptide microwave puffed food, which comprises matrix flour, 1-5% of crayfish shell polypeptide, 2-4% of yeast, 0.5-1.5% of baking soda and 20-33% of seasoning auxiliary materials, wherein the mass ratio of the matrix flour is 1-5%.
Further, the compound feed comprises matrix flour, 2 to 4 percent of the crayfish shell polypeptide, 2 to 4 percent of yeast, 0.5 to 1.5 percent of baking soda and 20 to 33 percent of seasoning auxiliary materials, wherein the mass ratio of the crayfish shell polypeptide to the matrix flour is 2 to 4 percent. Still further, comprises matrix flour and 3% of the crayfish shell polypeptide, 3% of yeast, 1% of baking soda and 26% of flavoring agent.
Further, the matrix flour comprises wheat flour and corn starch, wherein the mass ratio of the wheat to the corn starch is 7:1-5. Further, the mass ratio of the wheat to the corn starch is 7:3.
further, the seasoning comprises: the mass ratio of the raw materials to the matrix flour is as follows: 1-3% of salt, 4-6% of white sugar, 13-18% of butter and 2-6% of black tea. Still further, the seasoning comprises: 2% of table salt, 5% of white sugar, 15% of butter and 4% of black tea water.
The fourth aspect of the invention provides a preparation method of a crayfish shell polypeptide microwave puffed food, which comprises the following steps:
t1, mixing and uniformly stirring the raw materials according to the proportion, stirring into dough in a dough mixer, and performing primary fermentation;
t2, making the dough subjected to primary fermentation into dough sheets, then cutting into biscuit dough, and carrying out secondary fermentation;
and T3, baking the biscuit dough subjected to secondary fermentation in a microwave oven, and naturally cooling to obtain the crayfish shell polypeptide microwave puffed food.
Further, in the step T1 and the step T2, the primary fermentation and the secondary fermentation are both carried out at 37 ℃ for 1 hour.
Further, in the step T2, the moisture content of the dough sheet is 30% -50%. Further, the water content was 35%.
Further, in the step T3, when the material is baked in a microwave oven, the microwave power is 600W-1400W, and the microwave time is 1.5min-5.5min. Further, the microwave power is 1200W, and the microwave time is 2.5min.
The invention has the advantages and positive effects that:
1. the invention combines super-energy electron beam irradiation and ultrasonic treatment, can fully release the content while keeping the active ingredients of the shrimp shell to the maximum extent, then adopts compound enzyme treatment of alkaline protease and trypsin, can effectively improve the content of active polypeptide, the protein extraction rate is up to more than 90%, the total amino acid content is more than 3mg/mL, the obtained polypeptide is rich in a plurality of essential amino acids and hydrophobic amino acids such as Glu, arg, asp, glu, arg, asp is rich in amino and carboxyl groups and can be used as proton donors, oxidation resistance is enhanced, and the hydrophobic amino acids provide a hydrophobic environment for oxidation resistance, so that the shrimp shell polypeptide prepared by the invention has excellent oxidation resistance, is suitable for being used as a high-activity additive to prepare puffed food with high added value, and remarkably improves the puffing rate, flavor substance formation, oxidation resistance, antibacterial property and other shelf life properties of the puffed food.
2. The invention adopts the microwave puffing technology, takes the prepared crayfish shell polypeptide as an additive, can obviously destroy the ordered crystal structure of starch in matrix flour, makes the network structure loose, greatly improves the puffing rate, and has rich amino acid content and nutrient content, thereby being beneficial to improving the volatile component content in the microwave puffed food and being beneficial to the formation of the flavor of the product. In addition, the excellent oxidation resistance of the puffed food is beneficial to prolonging the storage period of the puffed food, can delay the change of physical properties of the puffed food such as water solubility index, water absorbability index, oil absorbability index, chromaticity and the like, can improve the total phenol content and TBA inhibition rate of the puffed food, reduce the total colony count and the volatile basic nitrogen content, further inhibit the putrefaction acidification of the puffed food and the growth of microorganisms and degrade protein, and remarkably prolong the storage period.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for the description of the embodiments will be briefly described below, and it is apparent that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a graph showing the effect of the crayfish shell polypeptides of example 1 on DPPH radical scavenging rate;
FIG. 2 is a graph showing the effect of crayfish shell polypeptides prepared by various methods of example 1 of the present invention on ABTS radical scavenging rate;
FIG. 3 is a graph showing the effect of various factors of example 2 of the present invention on the sensory evaluation score, puffing rate and TBA inhibition rate of a microwave puffed food; wherein, graphs (A) - (D) respectively show CSPH addition, microwave power, microwave time and dough moisture content;
FIG. 4 is a graph showing the crystallinity of starch in the microwave puffed food prepared with the different additives of example 3 of the present invention;
FIG. 5 is a microstructure of the microwave puffed food prepared by different additives in example 3 of the present invention, wherein A1-A3 are electron microscope images 1000 times, 100 times and 10 times of the Control group, B1-B3 are electron microscope images 1000 times, 100 times and 10 times of the Whey protein group, and C1-C3 are electron microscope images 1000 times, 100 times and 10 times of the CSP group; D1-D3 are electron microscope images of 1000 times, 100 times and 10 times of CSPH group;
FIG. 6 is a graph showing the total number of colonies of the microwave puffed food prepared by different additives of example 3 of the present invention as a function of shelf life;
FIG. 7 is a graph showing the variation of the total phenol content of the microwave puffed food prepared with different additives of example 3 over shelf life of the food;
FIG. 8 is a graph showing the variation of TBA inhibition rate of microwave puffed food prepared with different additives of example 3 over shelf life;
fig. 9 is a graph showing the change of the TVB-N content of the microwave puffed food prepared with different additives of example 3 of the present invention during shelf life.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the following examples. The examples described herein are intended to illustrate the invention only and are not intended to limit the invention.
Various modifications to the precise description of the invention will be readily apparent to those skilled in the art from the information contained herein without departing from the spirit or scope of the appended claims. It is to be understood that the scope of the invention is not limited to the defined processes, properties or components, as these embodiments, as well as other descriptions, are merely illustrative of specific aspects of the invention. Indeed, various modifications of the embodiments of the invention which are obvious to those skilled in the art or related fields are intended to be within the scope of the following claims.
For a better understanding of the present invention, and not to limit its scope, all numbers expressing quantities, percentages, and other values used in the present invention are to be understood as being modified in all instances by the term "about". Accordingly, unless indicated otherwise, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. Each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. In addition, the terms "comprising," "including," "containing," "having," and the like are intended to be non-limiting, as other steps and other ingredients may be added that do not affect the result.
In order that the above-recited objects, features and advantages of the present invention will become more readily apparent, a more particular description of the invention briefly described above will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings.
The embodiment of the invention provides a preparation method of crayfish shell polypeptide, which comprises the following steps:
s1, crushing cleaned shrimp shells to prepare shrimp shell powder;
s2, sequentially carrying out high hydrostatic pressure, super-energy electron beam irradiation and ultrasonic treatment on the shrimp shell powder to obtain pretreated shrimp shell powder, preparing the pretreated shrimp shell powder into a solution, adding compound protease to hydrolyze at a constant temperature to obtain protein hydrolysate, wherein the compound protease comprises the following components in percentage by mass of 0.3-0.8:1 and trypsin;
and S3, filtering the protein hydrolysate in the step S2, and freeze-drying the filtrate to obtain the crayfish shell polypeptide.
According to the invention, the shrimp shell is crushed, then washed and sterilized by high hydrostatic pressure, the cell tissue structure of the shrimp shell is primarily destroyed by high pressure, super-energy electron beam irradiation and ultrasonic treatment are further combined, the super-energy electron beam energy is high, after the shrimp shell is irradiated, a porous micro-nano network structure and a larger specific surface area are formed on the shrimp shell, more active protease can be contacted and fixed in a network structure system of the shrimp shell, so that the contact site between the enzyme and a substrate is increased, the enzymolysis efficiency is improved, the enzymolysis is more thorough, the protein yield is improved, and more active polypeptide components are obtained. Moreover, the structural form of the natural shrimp shell powder can be changed by combining the super-energy electron beam irradiation and the ultrasonic technology, so that cell membranes and the like are destroyed while the active ingredients of the shrimp shell are reserved to the maximum extent, and further, the release of the content is facilitated, and the full enzymolysis is facilitated and the content of active polypeptide in the subsequent protein hydrolysate is obviously improved. The complex enzyme treatment of alkaline protease and trypsin can lead the enzyme cleavage site of protease to be cracked at arginine and lysine preferentially, ensure that the obtained polypeptide is rich in various essential amino acids (such as Glu, arg, asp) and hydrophobic amino acids, glu, arg, asp is rich in amino and carboxyl, can be used as a proton donor, has positive significance for enhancing oxidation resistance, and the hydrophobic amino acids provide a hydrophobic environment for oxidation resistance, are also helpful for enhancing the oxidation resistance of the extracted crayfish shell polypeptide, so that the obtained crayfish shell polypeptide shows good oxidation resistance, and has good extraction effect of crayfish shell protein, protein extraction rate is more than 90%, total amino acid content is more than 3mg/mL, active ingredient yield is greatly improved, and added value of crayfish shells is improved based on the process flow. The crayfish shell polypeptide disclosed by the invention is applied to puffed foods, and has remarkable improvement on puffing rate, flavor substance formation, oxidation resistance, antibacterial property and other shelf life properties of the puffed foods.
In step S1, the preparation of the shrimp shell powder specifically includes the following steps: washing fresh shrimp shell with tap water to remove soil, residue and residue on the surface, boiling for sterilizing for 30min, draining, oven drying at 80deg.C to constant weight, pulverizing, and micronizing to obtain shrimp shell powder with particle diameter of 10-25 μm.
In the step S2, the pressure of the high hydrostatic pressure is 100-400MPa, and the dwell time is 10-30min.
In the step S2, the accelerating voltage of the electron beam irradiation is 8-12kV, and the irradiation time is 10-60min.
In the step S2, the ultrasonic power is 60-100W, the ultrasonic temperature is 15-55 ℃, and the ultrasonic time is 20-40min.
In the step S2, the enzyme activity of the compound protease is 8000-12000U, the hydrolysis temperature is 45-55 ℃, and the hydrolysis time is 2-4h.
According to the single factor experiment and the response surface optimization result, preferably, in step S2, the accelerating voltage of the electron beam irradiation is 10kV, the irradiation time is 20min, the ultrasonic power is 90W, the ultrasonic temperature is 25 ℃, the ultrasonic time is 30min, the enzyme activity of the compound protease is 10200U, and the mass ratio of alkaline protease to trypsin is 0.65:1, the hydrolysis time is 2.8h, under the condition, the optimized protein extraction rate can reach 90.30 percent, the total amino acid content is 3.583mg/mL, the hydrolysis time has obvious antioxidant activity, the clearance rate of 10mg/mL of crayfish shell polypeptide to DPPH free radical and the clearance rate of ABTS free radical are (54.17 +/-0.37)%, and (97.85+/-0.80)%, respectively, and the EC of DPPH and ABTS are cleared 50 4.38mg/mL and 3.66mg/mL, respectively.
The invention further provides an application of the crayfish shell polypeptide prepared by the preparation method in puffed food.
The advantages of the crayfish shell polypeptide prepared by the preparation method in puffed food are the same as those of the crayfish shell polypeptide prepared by the preparation method in the prior art, and are not repeated here.
Based on the same inventive concept, a further embodiment of the invention provides a crayfish shell polypeptide microwave puffed food, which comprises matrix flour, 1% -5% of crayfish shell polypeptide, 2% -4% of yeast, 0.5% -1.5% of baking soda and 20% -33% of seasoning auxiliary materials, wherein the weight ratio of the matrix flour is 1% -5% of the crayfish shell polypeptide.
The ratio of each raw material is based on the total amount of the matrix flour, and represents the percentage of the raw material in the total amount of the matrix flour, for example, 1% -5% of the crayfish shell polypeptide represents that the mass of the crayfish shell polypeptide is 1% -5% of the total amount of the matrix flour, and when the matrix flour is 100g, the adding amount of the crayfish shell polypeptide is 1g-5g, specifically, 1g, 2g, 3g, 4g or 5g.
According to the invention, a microwave puffing technology is adopted, the crayfish shell polypeptide is used as an additive, the additive is added into matrix flour, the influence of the flavor texture and the physicochemical property of the microwave puffed food is studied, and the fact that the addition of the crayfish shell polypeptide can obviously destroy the ordered crystal structure of starch in the matrix flour, so that the net structure of the starch becomes loose, the surface of the obtained product becomes no longer smooth and flat, the puffing rate is greatly improved, and the abundant amino acid content and nutrient content of the crayfish shell polypeptide are beneficial to improving the volatile component content in the microwave puffed food, and the proportion of pyrazine substances is improved most obviously, so that the flavor of the product is facilitated. In addition, the superior oxidation resistance of the crayfish shell polypeptide is beneficial to prolonging the storage life of the puffed food; during the storage period, the addition of the crayfish shell polypeptide delays the change of physical properties of the puffed food such as water solubility index, water absorbability index, oil absorbability index, chromaticity and the like, and is beneficial to improving the appearance and the texture of the product; the puffed food has high total phenol content and TBA inhibition rate, and low colony count and volatile basic nitrogen content, which shows that the crayfish shell polypeptide can inhibit food spoilage, microbial growth and protein degradation, and has positive effect on the prolongation of storage period. The implementation of the invention provides a theoretical basis for the application of the crayfish shell polypeptide in food, and improves the added value of the crayfish shell polypeptide and the microwave puffed food to a certain extent.
It is understood that the oxidation resistance of the food tends to increase gradually with increasing addition of the shrimp shell polypeptide due to increasing oxidation resistance active substances therein, but the oxidation resistance decreases gradually with further increasing content, which may be that the molecular structure of the related substances having oxidation resistance activity is decomposed or changed under the high temperature environment of microwaves. Therefore, preferably, the addition amount of the crayfish shell polypeptide is 2% -4%, more preferably 3%.
Preferably, the crayfish shell polypeptide microwave puffed food comprises matrix flour, 2% -4% of crayfish shell polypeptide, 2% -4% of yeast, 0.5% -1.5% of baking soda and 20% -33% of seasoning auxiliary materials, wherein the mass ratio of the crayfish shell polypeptide to the matrix flour is 2% -4%. More preferably, the addition amount of each raw material accounting for the matrix flour is as follows: 3% of crayfish shell polypeptide, 3% of yeast, 1% of baking soda and 26% of seasoning auxiliary materials.
Optionally, the flavoring auxiliary material comprises: the mass ratio of the raw materials to the matrix flour is as follows: 1-3% of salt, 4-6% of white sugar, 13-18% of butter and 2-6% of black tea. In some preferred embodiments, the flavoring aid comprises: 2% of table salt, 5% of white sugar, 15% of butter and 4% of black tea water.
Optionally, the matrix flour comprises wheat flour and corn starch, wherein the mass ratio of the wheat to the corn starch is 7:1-5. In some preferred embodiments, the mass ratio of wheat to corn starch is 7:3.
the invention also provides a preparation method of the crayfish shell polypeptide microwave puffed food, which comprises the following steps:
t1, mixing and uniformly stirring the raw materials according to the proportion, stirring into dough in a dough mixer, and performing primary fermentation;
t2, making the dough subjected to primary fermentation into dough sheets, then cutting into biscuit dough, and carrying out secondary fermentation;
and T3, baking the biscuit dough subjected to secondary fermentation in a microwave oven, and naturally cooling to obtain the crayfish shell polypeptide microwave puffed food.
Alternatively, in step T1 and step T2, both the primary fermentation and the secondary fermentation are performed at 37℃for 1 hour.
Generally, the thickness of the dough sheet and the size of the biscuit dough are prepared according to actual requirements, such as the size of the packaging bag, the eating object, etc. The present invention is not particularly limited thereto. Illustratively, the dough is pressed in a sheeter to a thickness of 2mm and then cut into 1cm 3cm biscuit dough.
The power and time of microwave baking have a great influence on the content of antioxidant substances in the crayfish shell polypeptide, phenolic substances which are obviously related to antioxidant activity can be oxidized and degraded under the thermal condition, and at high temperature, phenolic compounds with low molecular weight are easy to volatilize and polymerize to generate insoluble compounds such as melanin and polycyclic aromatic hydrocarbon, so that the antioxidant activity of food is negatively influenced, therefore, preferably, the microwave power is 600-1400W, the microwave time is 1.5-5.5 min, more preferably 1200W, and the microwave time is 2.5min when the crayfish shell polypeptide is baked in a microwave oven.
The moisture content of the dough sheet and the loss of dough moisture before and after puffing are important for the puffing effect produced by microwave puffing, and preferably the moisture content of the dough sheet is 30% -50%, more preferably 35%.
The invention will be further illustrated with reference to specific examples. The experimental methods, which do not address specific conditions in the following examples, are generally in accordance with the conditions recommended by the manufacturer.
EXAMPLE 1 preparation of crayfish Shell polypeptide and Property study thereof
1. Preparation of crayfish shell polypeptides
The preparation method comprises the following steps:
s1, washing fresh shrimp shells with tap water to remove soil, residues and residues on the surfaces, boiling for disinfection for 30min, draining water, drying in an oven at 80 ℃ to constant weight, and carrying out superfine grinding after preliminary grinding to obtain shrimp shell powder with the particle size of 10-25 mu m;
s2, treating shrimp shell powder by high hydrostatic pressure of 200MPa for 30min, then irradiating for 20min by using super-energy electron beams with accelerating voltage of 10KV, and then performing ultrasonic treatment for 30min to obtain pretreated shrimp shell powder; preparing pretreated shrimp shell powder into a solution, adding compound protease, hydrolyzing for 2.8 hours at the constant temperature of 50 ℃, wherein the enzyme activity of the compound protease (prepared by alkaline protease and trypsin according to the mass ratio of 0.65:1) is 10200U, so as to obtain protein hydrolysate;
S3, filtering the protein hydrolysate, and freeze-drying the filtrate to obtain the crayfish shell polypeptide, and storing at the temperature of minus 20 ℃ for later use.
2. Performance study of crayfish shell polypeptides
The shrimp shell powder subjected to (1) electron beam irradiation ultrasonic treatment is subjected to alkaline protease, (2) the shrimp shell powder subjected to electron beam irradiation ultrasonic treatment is subjected to trypsin, (3) the shrimp shell powder not subjected to electron beam irradiation ultrasonic treatment is subjected to alkaline protease and trypsin complex enzyme, and (4) the proteolytic liquid obtained by the shrimp shell powder subjected to electron beam irradiation ultrasonic treatment is subjected to alkaline protease and trypsin complex treatment is freeze-dried, so that four crayfish shell polypeptides are obtained, and are respectively marked as UCSP-A, UCSP-T, CSP-AT and UCSP-AT and are configured into protein liquids with different concentrations (2, 4, 6, 8 and 10 mg/mL), and the performances of oxidation resistance, amino acid composition and the like of the crayfish shell polypeptides are studied.
2.1 measurement of antioxidant Capacity of shrimp Shell Polypeptides
Antioxidant Capacity determination reference "Zhao Jing. Investigation of enzymatic hydrolysis of shrimp processing byproducts for the preparation of antioxidant peptides [ D ]]DPPH and ABTS radical scavengers (DSA, ASA) were calculated as follows, respectively, with slight modifications, in the method of China institute of metrology, 2012), where absorbance was measured at 517nm, A 1 ，A 1 '，A 1j Respectively representing the absorbance of the sample, the blank control and the absorption correction; the absorbance in formula (2) was measured at 420nm, A 2 ，A 2 '，A 2j Represented as sample, blank, absorbance corrected for absorption, respectively:
DPPH and ABTS free radicals are widely used as relatively stable free radicals to detect substances in the form of free radical traps or hydrogen donors, thereby assessing their antioxidant capacity, and as positive controls, UCSP-A, UCSP-T, CSP-AT, UCSP-AT scavenging capacity for DPPH and ABTS is shown in FIGS. 1-2, with an initial DPPH concentration of 0.0394mg/mL and an initial ABTS concentration of 3.843mg/mL.
As can be seen from fig. 1, the DPPH clearance of each sample increases with increasing concentration, and UCSP-AT shows stronger DPPH clearance than the other three samples AT the same sample concentration, and thus has stronger antioxidant activity. The DPPH clearance rate of UCSP-AT is obviously increased along with the concentration AT the concentration of 2-6mg/mL, and reaches the highest value AT the concentration of 6mg/mL, and the DPPH clearance rate is as follows(54.17 +/-0.37)%, and then tend to be stable. Calculated EC of UCSP-AT 50 The value was 4.38mg/mL.
As can be seen from FIG. 2, the ABTS clearance of each sample increases with increasing concentration, and UCSP-AT exhibits higher ABTS clearance than other three samples AT the same sample concentration, and the ABTS clearance of UCSP-AT increases with increasing concentration of 2-6mg/mL, and becomes stable AT a concentration of 6-10mg/mL, and the highest ABTS clearance is (97.85+ -0.80)% AT a concentration of 10 mg/mL. The clearance rate of the CSP-AT and the UCSP-AT to the ABTS is (72.59 +/-1.94054) percent and (80.29 +/-1.48624) percent AT 2mg/mL respectively, which are obviously higher than the clearance rate of the UCSP-A and the UCSP-T (55.69+/-1.30849) percent and (56.79 +/-2.25874) percent, and the EC of the UCSP-AT is higher than that of the UCSP-A and the UCSP-T 50 The value of 3.66mg/mL was lower than 4.51mg/mL of CSP-AT.
The results show that at low concentration, the sample treated by the compound protease has stronger scavenging capability to free radicals than the sample treated by the single enzyme, and the ultrasonic sample has higher scavenging rate to free radicals than the sample without ultrasonic, which also shows that the crayfish shell polypeptide obtained by combining the electron beam irradiation and ultrasonic treatment with the enzymolysis of the compound protease contains more active peptide components related to oxidation resistance, which is important for improving the shelf life of the puffed food.
2.2 analysis of amino acid composition of shrimp Shell polypeptide
Amino acid composition analysis reference "Chen Jianlan. Amino acid automatic analysis method for determining taste components of farmed and wild grass carp [ J ]. Modern food 2020,10:162-163 ]. The amino acid groups of UCSP-A, UCSP-T, CSP-AT and UCSP-AT are shown in Table 1, respectively.
All four groups of samples detected 16 amino acid components and all other several essential amino acids except tryptophan. The total amino acid amount and each amino acid component detected by UCSP-AT are obviously higher than those of other three samples, which shows that the combination of electron beam irradiation and ultrasonic treatment and composite protease enzymolysis can effectively improve the amino acid content of the shrimp shell polypeptide and the nutritional value and the content of active ingredients. From the aspect of amino acid composition, the content of essential amino acid in UCSP-AT accounts for 43.06 percent of the total amino acid, and the nutrition is relatively rich; the highest content of amino acids is Glu, arg, asp, which respectively account for 13.23%, 10.30% and 9.24% of the total amino acids, and the amino acids are rich in amino groups and carboxyl groups, have the capability of chelating metal ions, can be used as proton donors, and are beneficial to enhancing the antioxidation of the sample. In addition, the UCSP-AT detected hydrophobic amino acid accounts for 33.52 percent of the total amino acid, and the hydrophobic amino acid comprises Phe, val, leu, ile, ala, met and the like, so that a hydrophobic environment can be provided for the antioxidant peptide, the repair of physiological processes such as DNA oxidative damage in cells and the like is facilitated, and the method is also a reason for the strong antioxidant property of the shrimp shell polypeptide.
TABLE 1 amino acid composition of shrimp shell polypeptides (unit: mg/mL)
Further separating the polypeptide by Gel Permeation Chromatography (GPC) and ultrafiltration membrane system, and finding molecular weight of 3000Da < M W Has highest antioxidant activity of less than 5000Da, and has molecular weight of 3000Da < M by gel permeation chromatography (Sephadex G-25 gel column), reversed phase liquid chromatography (RP-HPLC) and liquid chromatography-mass spectrometry (LC-MS/MS) W The polypeptide component with the weight less than 5000Da is purified and identified to obtain a polypeptide sequence IPVAPPLPPH with high antioxidant activity, which also proves that the preparation method of the invention extracts active polypeptide, which is the key of high antioxidant capacity.
Example 2 preparation of crayfish Shell polypeptide microwave puffed food and optimization of Process parameters thereof
1. Preparation of crayfish shell polypeptide microwave puffed food
A crayfish shell polypeptide microwave puffed food comprises wheat flour and corn starch according to a mass ratio of 7:3, 1% -5% of mixed matrix flour, 3% of yeast, 1% of baking soda, 2% of salt, 5% of white sugar, 15% of butter and 4% of black tea water, wherein the weight ratio of the raw materials is the weight ratio of the matrix flour. The preparation method comprises the following steps:
t1, mixing and uniformly stirring the raw materials according to the proportion, stirring into dough in a dough mixer, and primarily fermenting for 1h at 37 ℃;
T2, pressing the dough subjected to primary fermentation into dough sheets with the thickness of 2mm in a dough pressing machine, controlling the water content of the dough sheets to be 30% -50%, cutting into biscuit dough blanks with the thickness of 1cm multiplied by 3cm, and carrying out secondary fermentation for 1h at 37 ℃;
and T3, baking the biscuit dough subjected to secondary fermentation in a microwave oven with the microwave power of 600W-1400W and the microwave time of 1.5min-5.5min, and naturally cooling to obtain the crayfish shell polypeptide microwave puffed food, and storing in a dryer for later use.
2. Technological parameter optimization of preparation method of crayfish shell polypeptide microwave puffed food
The sensory evaluation and the puffing rate are important physical parameters for evaluating the puffed food, and the quality of the product with high sensory evaluation score and high puffing rate is higher. Lipid peroxidation is a major cause of food deterioration during food storage, and the lipid peroxidation inhibition activity of the microwave puffed food can be measured by measuring the TBA inhibition rate, thereby evaluating the antioxidant function of the product. Taking the evaluation index as a standard, carrying out single factor experiments on the addition amount (1%, 2%, 3%, 4%, 5%) of the crayfish shell polypeptide (hereinafter referred to as CPSH), the microwave power (600W, 800W, 1000W, 1200W, 1400W), the microwave time (1.5 min, 2.5min, 3.5min, 4.5min, 5.5 min) and the dough piece moisture (30%, 35%, 40%, 45%, 50%) in the biscuit preparation process and the microwave puffing process, and determining the optimal technological scheme of the microwave puffed food.
The organoleptic evaluation of the biscuits was carried out by the method of the reference "Samakradhamrongthai RS, jannu T, renaldi G.Physics chemical properties and sensory evaluation of high energy cereal bar and its consumer acceptability [ J ]. Heliyon,2021,7 (08): 07776" and modified slightly. The appearance, color, texture and taste of the biscuits were evaluated, respectively, and the sensory evaluation criteria are shown in table 2. The sensory evaluation group consisted of 10 professionally trained students, and after the evaluation was completed, the highest score and lowest score were removed when the final score was calculated and the average was taken.
TABLE 2 organoleptic criteria for microwave puffed foods
The puffing rate was measured by the millet-in-row volumetric method. Before puffing, placing the sample into a 100mL measuring cylinder, pouring millet into 100mL graduation line, recording the graduation value at the moment, taking out and recording the value again, and recording the difference value as V 1 The method comprises the steps of carrying out a first treatment on the surface of the The same method is used for measuring the puffed sample, and the numerical difference is recorded as V 2 The expansion rate is calculated as follows:
thiobarbituric acid (Thiobarbituric acid, TBA) inhibition ratio reference "Ahmad M, gani A, hassan I, et al production and characterization of starch nanoparticles by mild alkali hydrolysis and ultra-sonication process [ J ] ]Scientific Reports,2020,10:3533", wherein the concentration of TBA is 1% (w/v). TBA inhibition was calculated by the following formula A sample And A control Is the absorbance at 535nm of the sample and control:
the effect of CSPH addition (A), microwave power (B), microwave time C) and dough moisture content (D) on the sensory evaluation score, puffing rate and TBA inhibition of the microwave puffed food is shown in FIG. 3. When CSPH is added into the microwave puffed food, if the addition amount is not proper, the organoleptic properties of the product are affected, and the functional properties of the product are also affected to a certain extent. As shown in the graph (A), the sensory evaluation score, the puffing rate and the TBA inhibition rate of the microwave puffed food reach the highest value when the CSPH addition amount is 3%, and the index values are (79.88 +/-5.94), (68.05+/-0.48) percent and (44.58 +/-0.75) percent respectively. With increasing addition of CSPH, TBA inhibition is first gradually increased as expected due to the increase of antioxidant active substances therein, but then gradually decreased, which may be the decomposition or change of the molecular structure of the related substances having antioxidant activity under the high temperature environment of microwaves. As shown in the graph (B), the sensory evaluation score and the puffing rate of the product were the highest at 1200W, respectively (84.63.+ -. 5.66) and (83.97.+ -. 1.53), and the TBA inhibition rate was the highest at 1400W, respectively (45.84.+ -. 1.28), but the difference from the product at 1200W (45.61.+ -. 0.52) was not large. As shown in the graph (C), the puffed food obtained by microwave baking for 2.5min has the best quality, and the sensory evaluation score, puffing rate and TBA inhibition rate are (77.50 + -3.70), (84.44 + -1.20)% and (54.57 + -1.08)%, respectively. The power and time of microwave baking may have a negative effect on the oxidation resistance of the crayfish shell polypeptide, the total phenol content significantly associated with the oxidation resistance activity may also undergo oxidative degradation under thermal conditions, and low molecular weight phenolic compounds are easily volatilized and polymerized to form insoluble compounds such as melanin and polycyclic aromatic hydrocarbons at high temperature, thereby negatively affecting the oxidation resistance, so that 1200W of microwave power and 2.5min of microwave time are selected as the optimal process conditions. As shown in the graph (D), the TBA inhibition rate did not change much with the change of the moisture of the dough, but the relatively low moisture could maintain a high phenol content and improve the antioxidant activity, so the optimum moisture content was 35%, and at this time, the respective indexes were (72.00.+ -. 3.93), (83.97.+ -. 1.53) and (58.16.+ -. 1.13)%, respectively.
Based on a single factor experiment, a 4-factor 3 level experiment is designed by using software Design-Expert 13, 29 groups are added, and the Design scheme and the result of the response surface are shown in table 3. Wherein factor A is CSPH addition (%), level 1, 0, -1 represents 2%, 3% and 4%, factor B is microwave power (W), level 1, 0, -1 represents 1000W, 1200W and 1400W, factor C is microwave time (min), level 1, 0, -1 represents 1.5min, 2.5min and 3.5min, factor D is dough moisture (%), level 1, 0, -1 represents 30%, 35% and 40%, respectively.
TABLE 3 response surface design and results
Sensory scores (Y) obtained by multiple regression fitting with Design-Expert 13 1 ) Expansion ratio (Y) 2 ) And TBA inhibition (Y) 3 ) Regression equations for the CSPH addition (a), microwave power (B), microwave time (C), and slice moisture (D) were respectively:
Y 1 ＝77.5+0.32708A+1.1875B-0.17083C-0.302083D-0.4999AB+0.51875AC-1.0000AD-1.4375BC-0.8125BD+0.78125CD-3.646875A 2 -5.23125B 2 -7.1C 2 -5.121875D 2 ；
Y 2 ＝76.058-1.00916A-1.49583B-0.48C+2.15833D-0.47749AB+1.0725AC-1.1525AD+2.1025BC-3.3625BD-0.395CD-7.75233A 2 -6.2673B 2 -6.511083C 2 -8.421083D 2 ；
Y 3 ＝73.652+0.41083A-1.815B-2.14416C+0.36D+1.2175AB-5.2AC-1.815AD+3.3725BC-2.595BD-0.1800CD-8.61475A 2 -12.651B 2 -10.03975C2-10.6635D 2 。
the absolute value of each coefficient of the model represents the influence degree of a single factor on the protein extraction rate, and positive and negative represent the influence direction.
P values of the three models<0.0001, extremely obvious, F values of 0.3215, 0.5909 and 4.24 in the mismatch terms of the three models respectively, P values of 0.9338, 0.7724 and 0.0883 > 0.05 are not obvious, which indicates that the three models are well fitted, and variation in response can be well explained. Model R 2 0.9261, 0.9023 and 0.9494 respectively, which show that the fitting degree of the equation to the model is good and the error is small.
In the related model of sensory evaluation, the influence of the primary term B on the sensory evaluation score was remarkable (P<0.05 The interaction term CD has a highly pronounced effect on the sensory evaluation score (P<0.01 Secondary term a) 2 、B 2 、C 2 、D 2 The effect on the sensory evaluation score was very pronounced (P<0.001 The order of the impact of the four factors on the sensory evaluation is: microwave power > CSPH addition > slice moisture > microwave time.
In the related model of the expansion rate, the influence of the primary term B and the interactive term BD on the expansion rate is remarkable (P<0.05 Secondary term a) 2 、B 2 、C 2 、D 2 Has very remarkable effect on the puffing rate (P<0.001 The magnitude order of the impact of the four factors on the expansion rate is as follows: slice moisture > microwave power > CSPH addition > microwave time.
In the correlation model of TBA inhibition, the influence of the primary term B, C and the interactive term BC on TBA inhibition was remarkable (P<0.05 The interaction term AC has a highly pronounced effect on TBA inhibition (P<0.01 Secondary term a) 2 、B 2 、C 2 、D 2 Has very remarkable effect on TBA inhibition rate (P<0.001 The order of the effect of the four factors on TBA inhibition is: microwave time > microwave power > CSPH addition > slice moisture.
In the preparation method of the crayfish shell polypeptide microwave puffed food, the preferred adding amount of the crayfish shell polypeptide is 3%, the moisture content of the dough sheet is 35%, the microwave power is 1200W, and the microwave time is 2.5min.
EXAMPLE 3 Performance study of crayfish Shell polypeptide microwave puffed foods
The effect of the crayfish shell polypeptide on the physicochemical properties and shelf life of the microwave puffed food is explored by preparing the crayfish shell polypeptide microwave puffed food (CSPH group) under the optimal technological parameters, and no CSPH (Control group), whey protein (Whey protein group) and CSP (CSP group) are added as controls. Each experiment was set up in 3 groups in parallel and the experimental data was analyzed using Origin2019, SPSS Statistics 26 and Design-Expert 13 software.
1. Influence of additives on the properties of microwave puffed foods
1.1, X-ray diffraction analysis (X-ray Diffraction Analysis, XRD)
In order to study the filling characteristics and crystallinity of the material, bruker D8 advanced 25 is used, under 40kv and 40mA conditions, the scanning angle is 5-45 degrees, the scanning speed is 2 degrees/min, the influence of different additives on the diffraction intensity of the microwave puffed food is recorded every 5 degrees, and the results are analyzed by Origin2019 b and jade software.
The effect of each additive on the crystallinity of the microwave puffed food is shown in fig. 4, and diffraction peaks are not obvious in the scanning range of 5-45 degrees, which is probably that the ordered crystalline structure of the starch is destroyed by the microwaves. The matrix of the microwave puffed food is composed of wheat flour and corn starch, the wheat flour and the corn starch belong to A-type crystal structures, the XRD pattern of the microwave puffed food prepared by adding the additives has peaks at diffraction peaks of 15 DEG, 17 DEG, 18 DEG and 23 DEG, and is also a typical A-type crystal structure, which shows that the crystal form of the raw material matrix is not changed by adding the additives and microwave treatment, but the crystallinity of the matrix starch can be reduced by adding CSPH, and compared with other three samples, the ordered crystal structure of the mixed starch matrix can be damaged more remarkably, so that the molecular chains tend to be disordered, and the method has positive effects on improving the puffing rate.
1.2 scanning Electron microscope analysis (Scanning electron microscopy analysis, SEM)
Directly adhering trace amount to conductive adhesive, and spraying metal for 45s with a Quorum SC7620 sputtering film plating instrument, wherein the metal spraying is 10mA; and then, using a ZEISS Gemini SEM 300 scanning electron microscope to shoot the morphology, and respectively obtaining images with the magnification of 0, 100 and 1000 times at the accelerating voltage of 5kV so as to study the surface morphology of the microwave puffed food.
The change in microstructure is closely related to the characteristics of the product, and for flour products, the change in the network of gluten often reflects the change in the interactions between the dough components. The effect of several additives on the microstructure of the microwave puffed food is shown in fig. 5, and the surface of the microwave puffed food product becomes no longer smooth and flat under the action of microwave baking, which indicates that the microwave action breaks the covalent bonds and intermolecular forces that maintain the gluten structure. Compared with a Control group, the other three additives can loosen the network structure, the number of pores on the surface of the gluten protein is increased, and the pore diameter is increased, because microwaves can break intermolecular acting forces such as hydrogen bonds, van der Waals forces and the like for connecting the secondary structure of the protein, so that covalent bonds (-S-) among the protein molecules are broken, the arrangement of the protein molecular structure becomes disordered, and the gluten network structure is loose and porous. The CSP has no obvious effect compared with other two groups of additives, the effect of the Whey protein group and the effect of the CSPH are relatively similar, which is probably the reason of the proteins existing in the additives, and the protein content in the CSP is relatively less compared with the other two groups, so that the net structure is relatively compact, thereby improving the processing performance of dough, improving the quality characteristics of products and increasing the puffing rate.
1.3 flavour analysis
Reference "Yang Zhao, liang Ruijin, yao Yujing, et al. Effect of oyster enzymatic hydrolysate on biscuit quality research [ J ]. Food science, 2020,45 (01): 198-203" the method was modified slightly. And detecting the volatile flavor components of the biscuits by using a headspace solid-phase microextraction gas chromatography-mass spectrometry. 0.5g of ground sample is weighed, 1mL of NaCl solution with mass concentration of 20% is added, the mixture is placed in a 20mL headspace bottle, the mixture is capped and tested, and 3 parallel samples are prepared for each group of samples. The extraction head is inserted into a headspace bottle, and is adsorbed for 30min after being balanced for 10min at 60 ℃. DB-WAX type chromatographic column (30 m 0.25 mm); the inlet temperature is 240 ℃ and the pressure is 100kPa; high purity helium is the carrier gas. The initial temperature is kept at 60 ℃ for 4min, gradually rises to 240 ℃ and is kept for 2min, sample injection is not split, the temperature of a sample inlet is 240 ℃, the desorption temperature is 240 ℃, and the desorption time is 5min. The electrons bombard the EI ion source at 200℃and the interface temperature at 220℃with a solvent delay time of min. The compounds and amounts of flavor substances were retrieved by NIST 11 standard library, with compounds having a similarity of greater than 80%. The relative content of the flavor substances is obtained by integrating a peak area normalization method.
The analysis of the ingredients of the microwave puffed food prepared with each additive is shown in table 4. The volatile substances in the four groups of samples are mainly alkanes and aldehydes, and 26 types of the flavor substances of the samples added with CSPH are detected, which are higher than 12 types of CSP, 10 types of whey proteins and 6 types of blank samples. The volatile compounds of several foods mainly consist of alkane, aldehyde, ester, pyrazine, alkene, phenol, ketone, furan and other substances. The aroma threshold of the alkane is smaller and the contribution to the overall flavor is smaller. The carbon chains of the aldehydes detected in the samples are short, which are possibly generated by oxidative degradation of fat, and the volatile components of the samples contain benzaldehyde through analysis, so that the aldehydes have pleasant almond fragrance, fruit fragrance and nut fragrance, and are generated after amino acid reaction. The ratio of aldehydes in the sample added with CSP and CSPH is 31.25% and 18.52%, and the linear aldehydes such as nonanal and hexanal are detected in the two samples, and the linear aldehydes have a certain fishy smell, and the nonanal is a main smell-developing substance of the fishy smell. 4 and 6 pyrazines were detected in the microwave puffed food with CSP and CSPH added, respectively, which were higher than 1 for the blank and 2 for whey protein. The pyrazine substance has low aromatic threshold, has nut flavor, and plays an important role in the flavor formation of baked foods; this shows that CSP and CSPH can effectively improve pyrazine substances in microwave puffed food, and has positive effect on the overall flavor of the product. Compared with other three groups, the CSPH group samples additionally detect several substances including sulfur-containing compounds, ketone, alkene, furan and pyridine, so that the richness of the flavor components of the product is improved. It has been demonstrated that ketones play an important role in the product, whereas the presence of meat flavor is closely related to sulfur-containing compounds. In general, CSP and CSPH added products can provide flavor-related volatile materials, CSPH can increase the volatile component types in the products, wherein the types of pyrazine substances are obviously increased, and the formation of baking flavor of the products is facilitated.
Table 4 analysis of ingredients of microwave puffed foods prepared with additives
2. Influence of additives on shelf life of microwave puffed food
2.1 Water Absorption Index (WAI), water Solubility Index (WSI) and Oil Absorption Index (OAI)
Weigh m 0 Placing a sample with the absolute weight of m1 of 1.00g into a centrifuge tube with the absolute weight of m1, adding 20mL of distilled water, carrying out water bath at 30 ℃ for 30min, shaking for 30s after every 10min, centrifuging in a centrifuge with the absolute weight of m 3000r/min for 15min, and pouring the supernatant into a centrifuge tube with the absolute weight of m 2 In the evaporation pan, the evaporation pan is put into a drying oven at 105 ℃ to be dried to constant weight, and the weight of the evaporation pan after drying is m 3 Weigh the centrifuge tube weight m containing sediment after discarding the supernatant 4 Each sample was repeated three times and WAI and WSI were calculated separately according to the following formulas. Another 1g sample of the powder was placed in a centrifuge tube and mixed with 30mL sunflower oil, then the sample was allowed to stand for 30min and then centrifuged at 1500r/min for 30min. Pouring the upper layer of oil in the centrifuge tube into a measuring cylinder after centrifugation and measuring the volume, and calculating the OAI according to the following formula, wherein m 0 Is the mass of the sample; m is m 1 The centrifuge tube is the net weight; m is m 2 The net weight of the evaporating dish; m is m 3 The total mass of the dried substances and the evaporating dish; m is m 4 To discard the supernatant, the pellet and the total mass of the centrifuge tube. The results are shown in Table 5.
Table 5 changes in WSI, WAI and OAI of microwave puffed foods prepared with various additives over shelf life
The Water Solubility Index (WSI) is an important indicator for measuring the degree of starch degradation in foods. The WSI of puffed food with added CSPH showed a trend of decreasing and increasing during shelf life, and was significantly higher than the other three groups (P < 0.05), indicating that the addition of CSPH increased the water-soluble substances in the product. The decrease in WSI is a slower rate of decomposition of macromolecular substances such as starch and proteins in CSPH group foods during shelf life, resulting in a decrease in water-soluble substances, and the subsequent increase in WSI may be due to an increase in water content of the product and an increase in starch solubility with an increase in shelf life, and in addition, the loose structure of the product is also conducive to dissolution of macromolecular substances.
The Water Absorption Index (WAI) reflects the degree of starch gelatinization in the food, and the WAI of the food added with the additive is larger than that of a blank sample, and is consistent with the porosity of the tissue in the SEM, which indicates that the additive can loosen the structural tissue of the food, thereby positively playing a role in increasing the WAI. As shelf life increases, the moisture content increases gradually, and the WAI of each product increases and decreases, which is contrary to the trend of WSI, probably because the decomposition rate of macromolecular substances such as starch, protein and the like becomes slow, so that water-soluble substances decrease and water-absorbing substances become more.
The Oil Absorption Index (OAI) may reflect the degree of binding of the matrix to the oil in the food product, and is to some extent related to the hydrophobicity of the food product. The change trend of the OAI is basically consistent with that of WAI, the OAI of several samples is not obviously different in the earlier shelf life, and the increase speed of CSPH is slower, so that the combination speed of the CSPH product and oil is slower in the shelf life, and the combination capacity of the product and oil is weakened along with the dissolution of water-soluble substances such as starch, protein and the like, thereby being beneficial to the storage of the product.
2.2 chromaticity
The color of the microwave puffed food was measured by a colorimeter CR-20 (Konica Minolta, INC. Japan). The colorimeter is corrected by a white standard plate before the test, and each sample is measured three times at the same time and place by selecting a random part, and the average value is obtained.
With the change of the storage time, the color of CSP in each sample is the darkest, and the color of whey protein is the lightest, and the result is probably that, besides the color of the additive, the sample undergoes Maillard reactions with different degrees during the storage process, so that melanoidin and pigment with different contents are generated during the browning process, and the brightness of the sample is reduced. Too dark a color may affect consumer acceptance of the product, with each product becoming progressively darker in color during storage, while the color of the csch is the lightest and more acceptable to consumers.
2.3 determination of colony count (Total Plate Count, TPC)
TPC was measured for each sample according to the method of GB 4789.2-2016 "food safety national Standard food microbiology test colony count determination", and if colonies were aseptically grown on plates at all dilutions of the sample, the colony count for that sample was recorded as lg (CFU/g).
The TPC of biscuits added with different additives during storage is shown in FIG. 6, and according to NY/T1511-2015 green food puffed food, if TPC is lower than 500 (CFU/g), the product is considered to meet the regulations in food standards for microorganisms. From the graph, it can be seen that several samples are safe and stable in storage period, and TPC of the samples all shows rising trend along with the extension of storage time, but TPC of the microwave puffed food added with CSPH is lowest, TPC on 90 days is 2.27CFU/g, TPC of the microwave puffed food added with CSP is highest (TPC on 90 days is 2.48 CFU/g), which is probably that nutritional components in shrimp shells are complex and rich, growth of microorganisms is facilitated, and compared with other three groups, active components in CSPH show antibacterial effect. Within 90 days, the TPC of the CSPH-added products was lower than that of the control group, indicating that the substances can prolong the shelf life of the puffed food.
2.4 Total Phenol (TP) content
The total phenol content was determined by Folin-Ciocalteu colorimetry. After 0.5g of the sample was stored in 10mL of 80% methanol for 15 hours, it was centrifuged and homogenized. Will be 0.After 5mL of the extract, 7.5mL of distilled water and 1mL of Folin-Ciocalteu reagent were mixed and left for 5min, 1mL of saturated sodium carbonate (Na 2 CO 3 ) A solution. After 1h, a spectrophotometer reading was taken at 760 nm. TP is expressed as gallic acid equivalent per gram of dry matter (mg GAE/100 g) and is calculated according to the following formula.
The change in total phenol content during storage is shown in figure 7. The TP of each product fluctuates somewhat during the storage period, but tends to decrease overall. Most of the time, the TP of the CSPH added product was significantly higher than the other three groups (P < 0.05). The concentration of dissolved oxygen can be reduced through ultrasonic treatment, so that the decomposition of bioactive compounds is inhibited, and the ultrasonic treatment can also form pores in the raw material structure, so that the extraction of polyphenol in shrimp shells is improved; furthermore, extraction of polyphenols depends on starch-polyphenol complex interactions, and gelatinized starch granules can release more polyphenols, consistent with XRD-demonstrated results. The higher phenolic content is beneficial to improving the oxidation resistance of the food, relieving the acidification spoilage of the food and further prolonging the shelf life.
2.5 TBA inhibition Rate
The measurement method was the same as in example 2.
Food peroxidation is an important factor causing spoilage during food preservation, and the TBA inhibition rate can reflect the peroxidation degree of food to some extent, and in fig. 8, the food added with CSPH performs best, the TBA inhibition rate is significantly higher than that of other groups (P < 0.05), and TBA inhibition rates at 0d and 90d are (68.97 ±0.46)% and (32.70±1.42)%, respectively, and then whey protein, CSP and blank, respectively. The TBA inhibition rate gradually decreases in the whole shelf life, which is consistent with the trend of TP content, which shows that the TP content gradually decreases with the extension of the storage time, so that the effect of inhibiting lipid oxidation step is weakened, and the grease in the product is gradually oxidized, thus causing rancidity of food. Both the TP content and TBA inhibition rate of the CSPH group were significantly higher over time than the other groups, indicating that good oxidation resistance of CSPH had a positive impact on the oxidation stability of the product.
2.6 volatile basic Nitrogen (Total Volatile Base Nitrogen, TVB-N) content
The TVB-N content was determined using an automatic Kjeldahl nitrogen determination apparatus according to the method of GB 5009.228-2016, "determination of volatile basic Nitrogen in food safety national Standard food", and TVB-N-level was expressed as mg N/100g sample.
TVB-N is mainly associated with proteolytic degradation by microbial growth. As shown in FIG. 9, within 15 days, the TVB-N values of the three products are not greatly different except the blank sample, the TVB-N value of the three products is rapidly increased after 60 days, when the storage time is 90 days, the TVB-N value of the product added with CSPH is (2.56+/-0.07) mg/100g, which is obviously lower than (5.58+/-0.16) mg/100g (P < 0.05) of the blank sample, and the CSPH inhibits the growth of bacteria, so that the progress of decomposing protein of bacteria is slowed down, the contents of nitrogen, amine and amine substances generated by the growth and metabolism of microorganisms are reduced, and the TVB-N value of the product can be effectively reduced, thereby being beneficial to prolonging the shelf life of the product. The TVB-N value of the added CSPH is less than 30mg/100g specified by the national standard during the whole shelf life.
The results show that the crayfish shell polypeptide prepared by the method can obviously destroy the ordered crystal structure of the mixed starch matrix by taking wheat flour and corn starch as matrixes, so that the network structure becomes loose, the surface of the product becomes no longer smooth and flat, and the content of volatile components in the microwave puffed food can be improved by the crayfish shell polypeptide, wherein the proportion of pyrazine substances is most obvious, and the flavor of the puffed food is facilitated. The crayfish shell polypeptide can delay the change of physical properties (WAI, WSI, OAI, chromaticity) of the product during storage, and has positive influence on the appearance, texture and the like of the product. The TP content and TBA inhibition rate of the puffed food added with the crayfish shell polypeptide are greatly improved, the growth of microorganisms and the protein decomposition can be inhibited, and the puffed food has a remarkable effect of accumulating for the extension of the storage period. The invention provides a theoretical basis for the application of the crayfish shell polypeptide in food, and improves the added value of crayfish byproducts to a certain extent.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, and alternatives falling within the spirit and principles of the invention.
Claims (10)
1. A method for preparing crayfish shell polypeptides, comprising the steps of:
s1, crushing cleaned shrimp shells to prepare shrimp shell powder;
s2, sequentially carrying out high hydrostatic pressure, super-energy electron beam irradiation and ultrasonic treatment on the shrimp shell powder to obtain pretreated shrimp shell powder, preparing the pretreated shrimp shell powder into a solution, adding compound protease to hydrolyze at a constant temperature to obtain protein hydrolysate, wherein the compound protease comprises the following components in percentage by mass of 0.3-0.8:1 and trypsin;
and S3, filtering the protein hydrolysate in the step S2, and freeze-drying the filtrate to obtain the crayfish shell polypeptide.
2. The method for producing a crayfish shell polypeptide of claim 1, wherein in step S2, the high hydrostatic pressure is 100-400MPa and the dwell time is 10-30min;
the accelerating voltage of the electron beam irradiation is 8-12kV, and the irradiation time is 10-60min;
The ultrasonic power is 60-100W, the ultrasonic temperature is 15-55 ℃, and the ultrasonic time is 20-40min;
the enzyme activity of the compound protease is 8000-12000U, the hydrolysis temperature is 45-55 ℃, and the hydrolysis time is 2-4h.
3. The method for preparing the crayfish shell polypeptide of claim 2, wherein in step S2, the accelerating voltage of the electron beam irradiation is 10kV, and the irradiation time is 20min;
the ultrasonic power is 90W, the ultrasonic temperature is 25 ℃, and the ultrasonic time is 30min;
the enzyme activity of the composite protease is 10200U, and the mass ratio of the alkaline protease to the trypsin is 0.65:1, hydrolysis time was 2.8h.
4. The microwave puffed food of the crayfish shell polypeptide is characterized by comprising 1-5% of matrix flour, 2-4% of yeast, 0.5-1.5% of baking soda and 20-33% of seasoning auxiliary materials, wherein the mass ratio of the matrix flour to the crayfish shell polypeptide is 1-3.
5. The microwave puffed food of the crayfish shell polypeptide of claim 4, comprising matrix flour and 3% of crayfish shell polypeptide prepared by the preparation method of the crayfish shell polypeptide of any one of claims 1-3, 3% of yeast, 1% of baking soda and 26% of seasoning auxiliary materials in a mass ratio of the matrix flour.
6. The crayfish shell polypeptide microwave puffed food of claim 4 wherein the matrix flour comprises wheat flour and corn starch in a mass ratio of 7:1-5;
the seasoning auxiliary materials comprise: the mass ratio of the raw materials to the matrix flour is as follows: 1-3% of salt, 4-6% of white sugar, 13-18% of butter and 2-6% of black tea.
7. The crayfish shell polypeptide microwave puffed food of claim 6 wherein the mass ratio of the wheat to the corn starch is 7:3, a step of;
the seasoning auxiliary materials comprise: 2% of table salt, 5% of white sugar, 15% of butter and 4% of black tea water.
8. A method for preparing a microwave puffed food of a crayfish shell polypeptide, which is used for preparing the microwave puffed food of the crayfish shell polypeptide according to any one of claims 4 to 7, and comprises the following steps:
t1, mixing and uniformly stirring the raw materials according to the proportion, stirring into dough in a dough mixer, and performing primary fermentation;
t2, making the dough subjected to primary fermentation into dough sheets, then cutting into biscuit dough, and carrying out secondary fermentation;
and T3, baking the biscuit dough subjected to secondary fermentation in a microwave oven, and naturally cooling to obtain the crayfish shell polypeptide microwave puffed food.
9. The method for preparing the microwave puffed food of crayfish shell polypeptides according to claim 7, wherein in step T1 and step T2, the primary fermentation and the secondary fermentation are each performed at 37 ℃ for 1 hour.
In the step T2, the water content of the dough sheet is 30% -50%;
in the step T3, when the material is placed in a microwave oven for baking, the microwave power is 600W-1400W, and the microwave time is 1.5min-5.5min.
10. The method for preparing the crayfish shell polypeptide microwave puffed food of claim 9, wherein the method comprises the steps of,
in the step T2, the moisture content of the dough sheet is 350%;
in the step T3, the microwave power is 1200W, and the microwave time is 2.5min.
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