CN111789205A - Preparation method of fermented agaric whey protein isolate composite beverage - Google Patents
Preparation method of fermented agaric whey protein isolate composite beverage Download PDFInfo
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- CN111789205A CN111789205A CN202010743290.4A CN202010743290A CN111789205A CN 111789205 A CN111789205 A CN 111789205A CN 202010743290 A CN202010743290 A CN 202010743290A CN 111789205 A CN111789205 A CN 111789205A
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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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
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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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
- A23L2/66—Proteins
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- A—HUMAN NECESSITIES
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- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
- A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
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- 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
- A23L33/19—Dairy proteins
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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Abstract
The invention discloses a preparation method of a fermented agaric whey protein isolate composite beverage, which takes dry agaric as a raw material and obtains a finished product beverage through powdering, leaching, enzymolysis, separation of leaching liquor, fermentation, blending with whey protein isolate, sterilization and filling. The extraction rate of 2.5% mg/mL of agaric polysaccharide is improved after enzymolysis, and the effect is obvious; on the other hand, the whey protein isolate is effectively combined with the agaric polysaccharide leaching liquor. In addition to replacing the separation and purification of the agaric polysaccharide with the agaric leaching liquor, lactobacillus plantarum is added into the agaric leaching liquor for fermentation to generate lactic acid, so that the agaric leaching liquor has the unique flavor of fermented food, the bitterness and the astringency of the beverage can be effectively reduced, and the wood fishy smell of the agaric is reduced; the agaric leaching liquor and the whey protein isolate are blended into the beverage, so that the beverage which is simple in preparation process, can be industrialized, is high in raw material utilization rate, is rich in nutrition and has excellent taste can be obtained.
Description
Technical Field
The invention belongs to the field of food processing, and particularly relates to a preparation method of a fermented agaric whey protein isolate compound beverage.
Background
Auricularia (Agaric) belongs to Basidiomycetes, Auriculariales, and Auriculariaceae. Dark brown color and soft texture. The black fungus fruiting body is colloid, has special flavor, and is an edible fungus with rich nutrition. Every 100g of dried black fungus contains 65.5g of carbohydrate, 10.6g of protein, 7.0g of cellulose and 0.2g of fat, and also contains various vitamins and mineral substances, and is known as "king in essence" and "black bird's nest" by nutriologists. The agaric polysaccharide has various biological activities as an important active ingredient.
In recent years, Whey Protein Isolate (WPI) has been used to develop various protein-rich beverages, such as meal replacement beverages and recovery sports beverages. During production, many whey protein isolate beverages are conditioned to an acidic environment (pH ≦ 3.5), where the astringency of the protein may cause sensory defects in the product. In order to solve this problem, an ideal whey protein isolate beverage should be formulated at a pH of 4-6. However, this pH range overlaps with the isoelectric point (pI) of whey protein. The pH of the solution changes around the isoelectric point, the whey protein can be precipitated, separated or gelatinized, which can have negative influence on the stability of the whey protein isolate beverage, and experiments prove that the auricularia auricula polysaccharide can be combined with the isolated whey protein around the isoelectric point of the whey protein, and the auricularia auricula polysaccharide and the isolated whey protein combination with certain concentration have good stability during heating and storage.
The university of Dalian industry applied for 'an edible fungus polysaccharide compounded high-concentration protein beverage and a preparation method thereof' in 2018, which proves that the edible fungus polysaccharide can be combined with whey protein isolate, but the extraction process of the polysaccharide is complex, and the process can be simplified for fast-food such as beverage, and the cost is reduced.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a preparation method of a fermented agaric whey protein compound beverage.
A preparation method of a fermented agaric whey protein isolate compound beverage comprises the following steps:
s1, raw material pretreatment: taking dry edible fungus, crushing into powder, and sieving by a 40-60-mesh sieve to obtain edible fungus powder; the dried black fungus meets the requirements of GB/T6192-2019, the front of the fungus is black brown, the fungus is glossy, the back of the fungus is dark gray, the fungus is complete, and the fungus has the peculiar smell of the black fungus and has no peculiar smell;
s2, leaching: uniformly stirring the agaric powder and the purified water in the mass ratio of 1: 30-50 in the step S1, heating at the temperature of 70-90 ℃ for 2-4 hours, and cooling to room temperature to obtain an agaric leaching mixture A;
s3, enzymolysis: adding cellulase into the agaric leaching mixture A obtained in the step S2 until the mass concentration of the cellulase is 0.6-0.8%, performing enzymolysis for 1-2 hours at the temperature of 35-55 ℃, and heating to inactivate enzyme to obtain an agaric leaching mixture B; the enzyme activity of the cellulase is 10000U/g;
s4, separation: centrifuging the agaric leaching mixture B obtained in the step S3 to obtain a supernatant to obtain an agaric leaching solution;
s5, fermentation of a leaching solution: adding cane sugar with the mass of 3% -6% of that of the agaric leaching solution in the step S4, stirring uniformly and then adding 10% of cane sugar6～107Adding activated lactobacillus plantarum into the CFU/mL inoculum size, and standing and fermenting for 18-24 hours at 35-38 ℃; after fermentation, carrying out sterilization treatment, and then rapidly cooling the fermented solution to room temperature to obtain fermentation liquor;
s6, blending: adding whey protein isolate accounting for 2-4% of the mass of the fermentation liquor in the step S5, uniformly stirring, and adjusting the pH value to 4.5-4.8 by using a citric acid buffer solution to obtain a beverage;
s7, sterilizing and canning: and (5) sterilizing the beverage obtained in the step S6, and then canning and cooling to obtain a finished product.
Preferably, the specific conditions for heating and inactivating the enzyme in step S3 are as follows: heating at 90-100 ℃ for 15-30 min.
Preferably, the sterilization process in step S5 specifically includes: heating at 85-95 ℃ for 15-30 min; the invention patent application with application number of 201910025420.8, namely 'a lactobacillus plantarum strain and application thereof in reducing biogenic amine content in fish tea', records a lactobacillus plantarum Yc-2 with the preservation number of CGMCC No.16614, and the lactobacillus plantarum related to the invention can be selected from the thalli.
Preferably, the sterilization conditions in step S7 are: heating at 85-95 ℃ for 15-30 min.
In a preferred mode, the preparation method of the fermented agaric whey protein isolate compound beverage comprises the following steps:
s1, raw material pretreatment: selecting high-quality glossy dry agaric without mildew, loose flowers, thick colloid and luster, crushing the agaric into powder by using a wall breaking machine, and sieving by using a 60-mesh sieve to obtain agaric powder;
s2, leaching: uniformly stirring the agaric powder and the purified water in the mass ratio of 1:40 in the step S1, heating the mixture in a water bath at the temperature of 90 ℃ for 2 hours, and cooling the mixture back to room temperature to obtain an agaric leaching mixture A;
s3, enzymolysis: adding cellulase into the agaric leaching mixture A in the step S2, wherein the mass concentration of the cellulase is 0.8%, performing enzymolysis for 1 hour at the temperature of 50 ℃, and then heating to 100 ℃ for 15 minutes to inactivate enzyme to obtain an agaric leaching mixture B;
s4, separation: centrifuging the agaric leaching mixture B obtained in the step S3 at 4 ℃ and 10000rpm for 10min, and taking supernate to obtain an agaric leaching solution;
s5, fermentation of a leaching solution: adding 3% of sucrose by mass into the agaric leaching solution obtained in the step S4, uniformly stirring, and adding 10% of sucrose by mass8CFU/mL lactobacillus plantarum Yc-2, adding 1mL bacterial liquid to every 100mL leaching liquor, and fermenting for 24 hours at 37 ℃; after the fermentation is finished, sterilizing for 15 minutes at the temperature of 85 ℃, and then quickly cooling the fermentation liquor back to room temperature to obtain fermentation liquor;
s6, blending: adding whey protein isolate with the mass of 2% into the fermentation liquor obtained in the step S5, uniformly stirring, and adjusting the pH to 4.7 by using a citric acid buffer solution to obtain a beverage; tests show that the agaric polysaccharide can be well combined with the isolated whey protein at the pH value of 4.7;
s7, sterilizing and canning: and (5) sterilizing the beverage obtained in the step (S6) for 15 minutes under the sterilization condition of 85 ℃, canning and cooling to obtain a finished product.
The room temperature is 20 to 25 ℃ unless otherwise specified.
The invention has the beneficial effects that:
the agaric beverage provided by the invention is simple to drink, rich in nutrition and suitable for people of all ages to drink. The agaric and the isolated whey protein are mixed according to different proportions, and the concentration of the isolated whey protein in the beverage is increased under the condition of not adding a stabilizing agent by utilizing the principle that agaric polysaccharide in the agaric can be combined with the isolated whey protein to prevent the isolated whey protein from aggregating and precipitating in the heat sterilization process. The utilization rate of the product of the edible fungus obtained by the process is high, the water-soluble vitamin and polysaccharide content of the prepared edible fungus beverage is high, the taste is soft and pure, the nutritional value is high, and the flavor is unique.
The invention provides a new technology, on one hand, the extraction rate of 2.5% mg/mL polysaccharide is improved after enzymolysis, and the effect is obvious; on the other hand, the whey protein isolate can be effectively combined with the agaric polysaccharide leaching liquor. In addition, the combination of the polysaccharide and the lactalbumin needs to adjust the pH value to be within the range of 4-5, the agaric leaching liquor is used for replacing the separated and purified agaric polysaccharide, and lactobacillus plantarum is added into the agaric leaching liquor for fermentation to generate lactic acid, so that the agaric leaching liquor has the unique flavor of fermented food, the bitter taste and the astringent taste of the beverage can be effectively reduced, and the wood smell of the agaric is reduced; the agaric leaching liquor and the whey protein isolate are blended into the beverage, and the beverage which is simple in preparation process, can be industrialized, is high in raw material utilization rate, is rich in nutrition and has excellent taste can be obtained.
Experiments prove that the agaric polysaccharide in the agaric leaching liquor can be combined with whey protein isolate under the condition of not removing protein, and the preparation method of the agaric leaching liquor is optimized.
Drawings
FIG. 1 shows the determination of the total sugar content in the leaching liquor after enzymolysis for 1 hour, wherein the mass concentrations of cellulase added during enzymolysis are respectively 0%, 0.4%, 0.8%, 1.6% and 3.2%. (Note: different lower case letters indicate that cellulase at different mass concentrations had a significant effect on total sugar content determination (p <0.05))
FIG. 2 is a graph showing the determination of the total sugar content in the leaching solution after the influence of cellulase is removed after enzymolysis is performed for 1 hour at the mass concentrations of 0%, 0.4%, 0.8%, 1.6% and 3.2% of cellulase added during enzymolysis in example 1 of the present invention. (Note: different lower case letters indicate that different mass concentrations of cellulase have a significant effect on the total sugar content determination.
(p<0.05))
FIG. 3 is a rheological characteristic curve of a leaching solution obtained after enzymolysis is performed for 1 hour by adding 0%, 0.4%, 0.8%, 1.6% and 3.2% of cellulase in enzymolysis according to example 1 of the invention.
FIG. 4 is a graph showing the determination of the total sugar content in the leaching solution after enzymolysis for 1h, 2h, 3h and 4h, respectively, with the mass concentration of 0.8% added cellulase in the enzymolysis in example 2 of the invention. (Note: different lower case letters indicate significant effects of different times of cellulase enzymatic hydrolysis on total sugar content determination. (p <0.05))
FIG. 5 is a graph showing the rheological characteristic of a leaching solution obtained in the embodiment 2 of the present invention after the cellulase is added in a mass concentration of 0.8% for enzymolysis for 1 hour, 2 hours, 3 hours, and 4 hours, respectively.
FIG. 6 is a graph of the rheology of a control solution of comparative example 1 of the invention containing 2%, 3%, 4% w/w whey protein isolate at pH 4.7.
FIG. 7 is a graph of the rheology of an unheated solution of comparative example 1 of the present invention formulated with 2%, 3%, 4% w/w whey protein isolate.
FIG. 8 is ZETA potentials of comparative example 1, control, unheated and heated solutions of the present invention. (Note: different capital letters indicate significant effects of different processing methods on the potential; different lower case letters indicate significant effects of different concentrations of whey protein isolate on the potential. (p <0.05))
Fig. 9 is a picture of a microstructure obtained at a magnification of 8000 × taken by a cold field scanning electron microscope of a whey protein isolate control solution of comparative example 1 to which no agaric extract was added.
FIG. 10 is a photograph of a microstructure of a whey protein isolate to which an Agaric extract was added, an unheated solution of comparative example 1 of the present invention, taken by a cold field scanning electron microscope at a magnification of 8000X.
FIG. 11 is a photograph of a microstructure of comparative example 1 of the present invention, obtained at 8000X magnification of a heated solution by cold field scanning electron microscopy, to which whey protein isolate from Auricularia leach solution was added.
Detailed Description
The present invention is further illustrated by the following specific examples.
A preparation method of a fermented agaric whey protein compound beverage comprises the following steps:
pretreatment of raw materials: selecting high-quality agaric with fleshy texture and no impurities. Crushing the agaric into powder by using a wall breaking machine, and then sieving the powder by using a 40-60-mesh sieve.
Hot water leaching: the agaric powder and the purified water are uniformly stirred according to the mass ratio of 1: 30-50, heated for 2-4 hours at the temperature of 70-90 ℃, and then cooled back to the room temperature.
Enzymolysis: and adding 0.6-0.8% of cellulase into the cooled agaric leaching mixture, performing enzymolysis for 1-2 hours at the temperature of 35-55 ℃, and then boiling for 5-15 minutes to inactivate enzyme.
Separation: and (4) centrifugally separating the enzyme-inactivated agaric leaching mixture, and storing a liquid part obtained after the centrifugation and separation into a container to be used as a raw material.
Fermentation of the leaching liquor: adding 3% -6% of cane sugar into the agaric leaching liquor, uniformly stirring, adding activated lactobacillus plantarum, and fermenting for 18-24 hours at 37 ℃. After the fermentation is finished, heating for 15-30 minutes at the temperature of 85-95 ℃, and then quickly cooling the mixture back to room temperature for sterilization treatment.
Blending: adding 2-4% of separated whey protein into the sterilized fermentation liquor, stirring uniformly, and adjusting the pH value to 4.5-4.8 by using a citric acid buffer solution.
Sterilizing and canning: and (3) sterilizing the blended beverage under the sterilization condition of 85-95 ℃ for 15-30 minutes, and then canning and cooling the finished product.
Unless otherwise specified, whey protein isolates used in the following examples: whey protein isolate manufactured by the NOW group of America, trade name 733739021748; cellulase used in each of the following examples, the enzyme activity of which was 10000U/g; the lactobacillus plantarum is lactobacillus plantarum Yc-2, and the preservation number of the China Committee for culture Collection of microorganisms (CGMCCNo.16614) in the common microorganism center is CGMCCNo..
Example 1:
a preparation method of a fermented agaric whey protein compound beverage comprises the following steps:
s1, raw material pretreatment: selecting high-quality glossy dry agaric without mildew, loose flowers, thick colloid and luster, crushing the agaric into powder by using a wall breaking machine, and sieving by using a 60-mesh sieve to obtain agaric powder;
s2, leaching: uniformly stirring the agaric powder and the purified water in the mass ratio of 1:40 in the step S1, heating the mixture in a water bath at the temperature of 90 ℃ for 2 hours, and cooling the mixture back to room temperature to obtain an agaric leaching mixture A; s3, enzymolysis: adding cellulase into the agaric leaching mixture A in the step S2, wherein the mass concentrations of the cellulase added are 0%, 0.4%, 0.8%, 1.6% and 3.2%, performing enzymolysis for 1 hour at 50 ℃, heating to 100 ℃, and inactivating enzyme for 15 minutes to obtain an agaric leaching mixture B; s4, separation: centrifuging the agaric leaching mixture B obtained in the step S3 at 4 ℃ and 10000rpm for 10min, and taking supernate to obtain an agaric leaching solution;
s5, fermentation of a leaching solution: adding 3% of sucrose by mass into the agaric leaching solution obtained in the step S4, uniformly stirring, and adding 10% of sucrose by mass8Adding 1mL of bacterial liquid into each 100mL of CFU/mL lactobacillus plantarum bacterial liquid, and fermenting at 37 ℃ for 24 hours; after the fermentation is finished, sterilizing for 15 minutes at the temperature of 85 ℃, and then quickly cooling the fermentation liquor back to room temperature to obtain fermentation liquor;
s6, blending: adding whey protein isolate with the mass of 2% into the fermentation liquor obtained in the step S5, uniformly stirring, and adjusting the pH to 4.7 by using a citric acid buffer solution to obtain a beverage; tests show that the agaric polysaccharide can be well combined with the isolated whey protein at the pH value of 4.7;
s7, sterilizing and canning: and (5) sterilizing the beverage obtained in the step (S6) for 15 minutes under the sterilization condition of 85 ℃, and then canning and cooling the finished product.
Fig. 1 shows the total sugar content in the agaric leach liquor obtained in step S4 after the enzymolysis is performed for 1 hour with the mass concentrations of 0%, 0.2%, 0.4%, 0.8%, 1.6%, and 3.2% of cellulase added during the enzymolysis in this embodiment, respectively;
since the addition of cellulase has an influence on the value of the total sugar content measured in the experiment, the total sugar content was determined for aqueous solutions having cellulase addition mass concentrations of 0%, 0.2%, 0.4%, 0.8%, 1.6%, and 3.2%, respectively. Fig. 2 is a total sugar content determination of the leaching solution without cellulase influence, which is obtained by calculating after the cellulase is removed, according to the embodiment, the mass concentrations of cellulase added during enzymolysis are respectively 0%, 0.2%, 0.4%, 0.8%, 1.6% and 3.2% for enzymolysis for 1 hour.
Since the cellulase also contains sugar, which has an influence on the total sugar content, the total sugar content in the leaching solution is the highest when the mass concentration of the added cellulase is 0.8% by combining with the graph shown in fig. 1 and 2, and the effect is obvious when the total sugar content is improved by 2.5mg/mL compared with the total sugar content without enzymolysis.
FIG. 3 is a graph showing the rheological property of the leaching liquor obtained after enzymolysis for 1 hour, wherein the mass concentrations of cellulase added during enzymolysis are 0%, 0.2%, 0.4%, 0.8%, 1.6% and 3.2%, respectively. The viscosity of the enzymolysis liquid with different cellulase addition amounts is about 10Pa.s at a low shear rate of 0.005-0.051/S, and the change is not obvious.
Therefore, the effect is better when cellulase is added into the agaric leaching mixture A in the step S2 until the mass concentration of the cellulase is 0.6-0.8%.
Example 2:
a preparation method of a fermented agaric whey protein compound beverage comprises the following steps:
s1, raw material pretreatment: selecting high-quality glossy dry agaric without mildew, loose flowers, thick colloid and luster, crushing the agaric into powder by using a wall breaking machine, and sieving by using a 60-mesh sieve to obtain agaric powder; s2, leaching: uniformly stirring the agaric powder and the purified water in the mass ratio of 1:40 in the step S1, heating the mixture in a water bath at the temperature of 90 ℃ for 2 hours, and cooling the mixture back to room temperature to obtain an agaric leaching mixture A;
s3, enzymolysis: adding cellulase into the agaric leaching mixture A in the step S2, wherein the mass concentration of the cellulase is 0.8%, performing enzymolysis for 1, 2, 3 and 4 hours respectively at the temperature of 50 ℃, heating to 100 ℃ and inactivating enzyme for 15 minutes to obtain an agaric leaching mixture B;
s4, separation: centrifuging the agaric leaching mixture B obtained in the step S3 at 4 ℃ and 10000rpm for 10min, taking supernatant and storing the supernatant in a container to obtain an agaric leaching solution;
s5, fermentation of a leaching solution: adding 3% of sucrose by mass into the agaric leaching solution obtained in the step S4, uniformly stirring, and adding 10% of sucrose8Adding 1mL of bacterial liquid into each 100mL of CFU/mL lactobacillus plantarum bacterial liquid, and fermenting at 37 ℃ for 24 hours; after the fermentation is finished, heating and sterilizing for 15 minutes at the temperature of 85 ℃, and then quickly cooling the fermentation liquor back to room temperature to obtain fermentation liquor;
s6, blending: adding whey protein isolate with the mass of 2% into the fermentation liquor obtained in the step S5, uniformly stirring, and adjusting the pH to 4.7 by using a citric acid buffer solution to obtain a beverage; tests show that the agaric polysaccharide can be well combined with the isolated whey protein at the pH value of 4.7;
s7, sterilizing and canning: sterilizing the blended beverage at 85 deg.C for 15min, and canning and cooling to obtain the final product.
FIG. 4 is a graph showing the determination of the total sugar content in the leaching solution after the cellulase is added in an amount of 0.8% for enzymolysis for 1h, 2h, 3h and 4h, respectively. There was no significant difference in total sugar content at different enzymatic hydrolysis times.
Fig. 5 is a graph of the rheological property of the leaching liquor obtained after the cellulase is added in an amount of 0.8% for enzymolysis for 1 hour, 2 hours, 3 hours and 4 hours respectively. The viscosity of the agaric leaching liquor in different enzymolysis time is about 7Pa.s at low shear rate in the rheological characteristic curve, and the change is small. Based on the time saving principle, the enzymolysis is properly carried out for 1 to 2 hours.
Comparative example 1:
the experimental sample preparation method for demonstrating that whey protein isolate can be combined with agaric leach liquor:
s1, raw material pretreatment: selecting high-quality glossy dry agaric without mildew, loose flowers, thick colloid and luster, crushing the agaric into powder by using a wall breaking machine, and sieving by using a 60-mesh sieve to obtain agaric powder;
s2, leaching: uniformly stirring the agaric powder and the purified water in the mass ratio of 1:40 in the step S1, heating the mixture in a water bath at the temperature of 90 ℃ for 2 hours, and cooling the mixture back to room temperature to obtain an agaric leaching mixture A;
s3, enzymolysis: adding cellulase into the agaric leaching mixture A in the step S2, wherein the mass concentration of the cellulase is 0.8%, performing enzymolysis for 1 hour at 50 ℃, heating to 100 ℃ and inactivating enzyme for 15 minutes to obtain an agaric leaching mixture B;
s4, separation: centrifuging the agaric leaching mixture B obtained in the step S3 at 4 ℃ and 10000rpm for 10min, and taking supernate to obtain an agaric leaching solution;
s5, preparing a whey protein isolate solution: dissolving whey protein isolate in water, stirring at 200rpm for 3 hours at room temperature, wherein the mass ratio of protein to water is 1:10, and fully hydrating the protein at 5 ℃ overnight to obtain whey protein isolate solution for later use.
S6, preparing a control solution, namely taking the whey protein isolate solution in the step S5, diluting with water to ensure that the mass concentration of the whey protein isolate is 2%, 3% and 4%, and adjusting the pH value of the solution from 7.0 to 4.7 to be used as the control solution.
S7, preparing a whey protein isolate-agaric leaching liquor composite solution: the total volume of the solution was 10mL, wherein 7mL of the Auricularia leach liquor (containing about 0.2% Auricularia polysaccharide) obtained in step S4 was added, the whey protein isolate solution obtained in step S5 was added, water was added to the solution so that the mass concentrations of the whey protein isolates were 2%, 3% and 4%, respectively, and the volume of the mixed solution was 10mL, and the pH of the solution was adjusted from 7.0 to 4.7 as an unheated solution.
S8, preparing a heating group solution: the solution of the non-heated group in step S7 is prepared, and is heated at 85 deg.C for 15min for sterilization to obtain the heated group solution.
FIG. 6 is a graph of the rheology of a control group at pH4.7 with 2%, 3%, 4% w/w whey protein isolate added (i.e., the control group described in step S6). Viscosity shear rate curve for control solution at Low shear Rate: (<0.1s-1) The following shows a clear fluctuation, indicating that the control solution has poor dispersibility at pH 4.7.
FIG. 7 is a graph of the rheology of an unheated solution of whey protein isolate at 2%, 3%, 4% w/w as added during formulation. Compare the viscosity shear rate curves of the solutions of FIG. 6 at low shear rates (<0.1s-1) There was no significant fluctuation in the following, indicating improved solution dispersibility.
Figure 8 is the ZETA potential of the control, unheated and heated solutions. When the mass concentration of the control group solution, namely Whey Protein Isolate (WPI), is increased from 2% to 4%, the ZETA potential is more than-15 mV, and after the leaching solution is added (namely the unheated group solution), the ZETA potential of the mixed solution is reduced to be below-25 mV. As the mass concentration of the whey protein isolate increases, the zeta potential of the mixed solution increases from-31.13 + -0.32 mV to-26.80 + -0.44. This is probably because after the negatively charged auricularia auricula polysaccharide is added, the positive charge on the whey protein isolate surface is combined with the electrostatic interaction thereof, thereby reducing the ZETA potential of the complex of the mixed solution and leading the solution to have excellent solubility; the ZETA potential did not change much after heat sterilization (i.e., heating the group solution) compared to the unheated, indicating that heating had no effect on the stability of the mixed solution.
Fig. 9 is a microstructure picture obtained at 8000 × magnification taken by a cold field scanning electron microscope of a whey protein isolate control solution to which no agaric extract was added. The freeze-dried whey protein isolate drops are irregular in interior and mainly have a spherical structure.
Fig. 10 and 11 are photographs of microstructures obtained at 8000 × magnification taken by a cold field scanning electron microscope of whey protein isolate to which the agaric extract was added, the solution of the non-heated group, and the solution of the heated group, respectively. The microstructure of freeze-dried liquid drops of the composite solution is of a sheet structure, internal spherical proteins are connected by filamentous auricularia auricula polysaccharides, a bridge structure is formed in a sheet layer, the sheet structure of a heating group is partially damaged probably due to heating, but the structure of connection of the filamentous auricularia auricula polysaccharides and the internal spherical proteins is not influenced, so that the stability of the whey protein isolate solution can be enhanced by the auricularia auricula polysaccharides, and the stability of the whey protein isolate solution can be still kept after heating treatment.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be able to cover the technical solutions and the inventive concepts of the present invention within the technical scope of the present invention.
Claims (6)
1. A preparation method of a fermented agaric whey protein isolate compound beverage is characterized by comprising the following steps:
s1, raw material pretreatment: taking dry edible fungus, smashing into powder, and sieving with a 40-60-mesh sieve to obtain edible fungus powder;
s2, leaching: uniformly stirring the agaric powder and the purified water in the mass ratio of 1: 30-50 in the step S1, heating at the temperature of 70-90 ℃ for 2-4 hours, and cooling to room temperature to obtain an agaric leaching mixture A;
s3, enzymolysis: adding cellulase into the agaric leaching mixture A obtained in the step S2 until the mass concentration of the cellulase is 0.6-0.8%, carrying out enzymolysis for 1-2 hours at the temperature of 35-55 ℃, and heating to inactivate the enzyme to obtain an agaric leaching mixture B; wherein the enzyme activity of the cellulase is 10000U/g;
s4, separation: centrifuging the agaric leaching mixture B obtained in the step S3 to obtain a supernatant to obtain an agaric leaching solution;
s5, fermentation of a leaching solution: adding cane sugar accounting for 3-6% of the mass of the agaric leaching liquor in the step S4, uniformly stirring the mixture according to the proportion of 106～107Adding lactobacillus plantarum into the strain with the inoculation amount of CFU/mL, and fermenting for 18-24 hours at 35-38 ℃; after fermentation, sterilizing, and cooling to room temperature to obtain fermentation liquor;
s6, blending: adding whey protein isolate accounting for 2-4% of the fermentation liquor in the step S5, uniformly stirring, and adjusting the pH value to 4.5-4.8 to obtain a beverage;
s7, sterilizing and canning: and (5) sterilizing the beverage obtained in the step (S6), canning and cooling to obtain a finished product.
2. The method for preparing the fermented black fungus whey protein isolate composite beverage according to claim 1, wherein the specific conditions for heating and enzyme deactivation in step S3 are as follows: heating at 90-100 ℃ for 15-30 min.
3. The method for preparing the fermented agaric whey protein isolate composite beverage according to claim 1, wherein the sterilization treatment in step S5 is specifically: heating at 85-95 ℃ for 15-30 min.
4. The method for preparing the fermented jelly fungus whey protein isolate composite beverage as claimed in claim 1, wherein the lactobacillus plantarum used in step S5 is lactobacillus plantarum Yc-2 with a preservation number of CGMCC No. 16614.
5. The method for preparing a fermented jelly fungus whey protein isolate composite beverage according to claim 1, wherein the sterilization conditions in step S7 are as follows: heating at 85-95 ℃ for 15-30 min.
6. The method for preparing the fermented jelly fungus whey protein isolate composite beverage according to claim 1, comprising the steps of:
s1, raw material pretreatment: crushing dry edible fungus into powder, and sieving with 60 mesh sieve to obtain edible fungus powder;
s2, leaching: uniformly stirring the agaric powder obtained in the step S1 and purified water according to the mass ratio of 1:40, heating in a water bath at the temperature of 90 ℃ for 2 hours, and then cooling to room temperature to obtain an agaric leaching mixture A;
s3, enzymolysis: adding cellulase into the agaric leaching mixture A in the step S2 until the mass concentration of the cellulase is 0.8%, carrying out enzymolysis for 1 hour at the temperature of 50 ℃, and heating for 15 minutes at the temperature of 100 ℃ to inactivate the enzyme, so as to obtain an agaric leaching mixture B; wherein the enzyme activity of the cellulase is 10000U/g;
s4, separation: centrifuging the agaric leaching mixture B obtained in the step S3 at 4 ℃ and 10000rpm for 10min, and taking supernate to obtain an agaric leaching solution;
s5, fermentation of a leaching solution: adding 3% of sucrose by mass into the agaric leaching solution obtained in the step S4, uniformly stirring, and adding 10% of sucrose by mass8Adding 1mL of the bacterial liquid of the CFU/mL lactobacillus plantarum Yc-2 bacterial liquid into every 100mL of leaching liquor, and fermenting for 24 hours at 37 ℃; after the fermentation is finished, heating and sterilizing for 15 minutes at the temperature of 85 ℃, and then cooling the fermentation liquor back to room temperature to obtain fermentation liquor; the lactobacillus plantarum Yc-2 has the preservation number of CGMCC No. 16614;
s6, blending: adding whey protein isolate with the mass of 2% into the fermentation liquor obtained in the step S5, uniformly stirring, and adjusting the pH to 4.7 by using a citric acid buffer solution to obtain a beverage;
s7, sterilizing and canning: and (5) heating and sterilizing the beverage obtained in the step S6 at 85 ℃ for 15 minutes, canning and cooling to obtain a finished product.
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