CN115736087A - Method for enhancing nutrition of soybean protein isolate based on solid state fermentation of edible fungi - Google Patents
Method for enhancing nutrition of soybean protein isolate based on solid state fermentation of edible fungi Download PDFInfo
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Abstract
The invention relates to the technical field of microbial fermented foods, in particular to a method for enhancing the nutrition of soybean protein isolate based on solid state fermentation of edible fungi. The preparation method comprises the following specific steps: (1) Activating the edible fungus strain, inoculating the activated edible fungus strain into a PDB liquid culture medium, and culturing to obtain an edible fungus seed liquid; (2) Sterilizing the isolated soy protein powder and distilled water in a fermentation container to obtain a soy protein isolate culture medium; (3) Adding the edible fungus seed liquid into a soy protein isolate culture medium, fermenting and culturing, drying and crushing to obtain the soy protein isolate nutrition enhancing powder. The isolated soybean protein powder after solid state fermentation has obviously enhanced nutrition and oxygen resistance. In addition, the method for enhancing the nutrition of the soybean protein isolate powder is suitable for large-scale production and wide application.
Description
Technical Field
The invention relates to the technical field of microbial fermented foods, in particular to a method for enhancing the nutrition of isolated soy protein based on solid state fermentation of edible fungi.
Background
The soybean protein isolate is widely applied to the food industry, is a plant protein separated from soybean products, has amino acid composition similar to that of protein in milk, contains rich trace elements such as calcium, iron and phosphorus, vitamins and isoflavone and the like, and has rich nutritive value. Wherein, the polyphenol substance represented by isoflavone provides stronger antioxidant activity for the soybean protein isolate, and can effectively resist aging, tumors, cardiovascular diseases and the like. However, in soy protein isolate, isoflavones are present in many forms of glycosides such as daidzin and genistin, which are not absorbed by the human body, and thus have low bioacessability.
The edible fungi refer to large fungi which can be used by human beings, have delicious taste and good edible and medicinal values. The mycelium is rich in high-quality protein, fatty acid, polysaccharide, vitamins, inorganic mineral substances and other functional components, and has the health-care functions of resisting tumors, reducing blood sugar and blood fat, preventing arteriosclerosis and the like. In addition, researches show that the edible fungi can convert the glucoside isoflavone into free aglycon isoflavone and improve the absorptivity of the isoflavone for human bodies.
Chinese patent with publication No. CN106036810A discloses a preparation method of bean dreg food raw material with the flavor of stropharia rugoso-annulata edible fungi. The bean dregs fermented by the stropharia rugoso-annulata edible fungi have improved components, the content of amino acid nitrogen, crude protein, fat and dietary fiber is reduced after fermentation, and the content of soluble total sugar is improved after fermentation. Therefore, the bean dregs fermented by the method have improved flavor and aroma to a certain extent after being prepared into food, and have higher nutritional and health-care values than the bean dregs.
The bean dregs are the main by-products of processing bean products such as soybean oil, soy sauce, bean curd and the like. The bean dregs contain rich nutrient components, and researches show that the content of crude protein in the dry bean dregs is about 25 to 28 percent, the content of crude fat is about 9 to 11 percent, and the content of dietary fiber is about 50 to 60 percent. The soybean protein isolate is a high molecular organic polymer formed by combining a series of amino acids through peptide bonds, and mainly comprises albumin and globulin, wherein the albumin accounts for about 5 percent, and the globulin accounts for about 90 percent. The two have different nutritional ingredients, and the soybean protein isolate has more abundant nutritional ingredients than the bean dregs, and the enhancement of the nutrition is not easy.
In addition, the existing research is mostly based on laboratory research, and the large-scale application of putting the laboratory process into a factory is not realized.
Therefore, based on the above, how to improve the nutrition of the isolated soy protein and put it into mass production still remains a problem to be solved urgently.
Disclosure of Invention
The invention provides a method for enhancing the nutrition of isolated soy protein based on solid-state fermentation of edible fungi, which aims to overcome the defects that the nutrition of isolated soy protein cannot be enhanced in the prior art and the conventional means cannot be used for large-scale production and application.
In order to achieve the above purpose, the invention provides the following technical scheme:
a method for enhancing the nutrition of isolated soy protein based on solid state fermentation of edible fungi comprises the following steps:
(1) Preparing edible fungus seed liquid: activating edible fungus strains, inoculating the activated edible fungus strains into a PDB liquid culture medium, and culturing to obtain an edible fungus seed liquid;
(2) Preparing a soy protein isolate culture medium: sterilizing the isolated soy protein powder and distilled water in a fermentation container to obtain a soy protein isolate culture medium;
(3) Solid state fermentation culture: adding the edible fungus seed liquid into a soy protein isolate culture medium, fermenting and culturing, drying and crushing to obtain the soy protein isolate nutrition enhancing powder.
The first purpose of the invention is to use edible fungi as strains and adopt a solid state fermentation mode to ferment the soybean protein isolate powder, and finally obtain the soybean protein isolate nutrition enhancing powder. The soybean protein powder fermented by edible fungi such as oyster mushroom, hericium erinaceus and needle mushroom has greatly improved components and increased amino acid content; in the two glucoside type soybean isoflavones, the content of daidzein is obviously reduced, and genistin is not detected, and in the two free aglycone type soybean isoflavones, the content of daidzein is increased by 100.68 to 226.02 percent, and the content of genistein is increased by 147.87 to 333.33 percent; the rising rate of the phenolic substances reaches 292.31 to 360.76 percent; the four oxidation resistance indexes of the soybean protein isolate nutrition enhancing powder with the same quality are obviously improved compared with the four oxidation resistance indexes of the soybean protein isolate powder. The data can prove that the bioacessability of the soybean protein isolate nutrition enhancing powder obtained after the fermentation treatment of the edible fungi is greatly improved, and the soybean protein isolate nutrition enhancing powder is beneficial to human body absorption; the effects of resisting tumor, inhibiting bacteria, reducing blood fat and the like are improved; the nutrition and the oxidation resistance are obviously enhanced.
Wherein, solid state fermentation is a process for culturing microorganisms by using insoluble solid substrates, and compared with liquid state fermentation widely applied in industry, the solid state fermentation has the advantages that: the water activity is low, which is beneficial to the growth of edible fungi; the aseptic requirement is lower than that of liquid fermentation, the contamination of mixed bacteria is not easy to occur or the pollution range is small, and the industrial production is facilitated; the space utilization rate is high, the post-treatment is simple, the pollutant emission is less, and the environment is more friendly. Liquid state fermentation has the defects of high energy consumption, high equipment requirement, large wastewater yield and the like, and meanwhile, the aseptic requirement is generally higher than that of solid state fermentation. In the existing research on the culture mode of liquid-solid dual-state fermentation, for example, chinese patent document CN110074403A, "an edible fungus powder and a preparation method thereof" discloses an edible fungus powder prepared by using coarse cereals and edible fungi and a preparation method thereof, which comprises the steps of inoculating a mycelium of edible fungi obtained by culturing a mother strain in a liquid medium together with the liquid medium into a solid medium for dual-state fermentation culture, and then drying and crushing to obtain the edible fungus powder. The method is not easy to control the water content in the solid-state fermentation process of the solid-state culture medium, is not favorable for the solid-state fermentation of non-hygrophilous strains, and is not suitable for wide application. Thus, the mode of solid state fermentation is preferred in the present invention.
In addition, in the solid state fermentation culture process, if areas on the soy protein culture medium cannot grow mycelia, the water distribution ratio and the water adding mode are adjusted according to specific conditions to ensure that the culture medium maintains proper humidity.
The second purpose of the invention is that the fermentation of the strain is not limited by the fermentation container, and the vessel with any shape and specification can be used as the fermentation container. Even tray-like containers that are not sealed may be used under certain aseptic conditions. Meanwhile, based on the selection of the scheme in the first invention object, the scheme designed by the invention has higher possibility of large-scale application compared with the general scheme.
When a general laboratory process is developed to actual production of a factory, the problem that the laboratory effect cannot be achieved in the factory production is easy to occur, and the problem is a common phenomenon after errors are amplified when a trace experiment is changed to a large process experiment. However, in consideration of such problems, the inventors of the present application tried to obtain the soybean protein isolate nutrition-enhancing powder by fermentation culture in a sterile room using a stainless steel tray at the pilot scale stage, and succeeded in the experiment. Meanwhile, in the test process, the inventor of the application finds that large-scale production is expected to be realized as long as fermentation is carried out in an aseptic environment and even large-scale fermentation tanks, fermentation cylinders and other vessels can be tried to be selected.
Preferably, the edible fungi in the step (1) are any one or more of flammulina velutipes, pleurotus ostreatus, hericium erinaceus, ganoderma lucidum, bolete and pleurotus eryngii.
The patent with publication number CN113068824A discloses a high-protein composite nutritional coarse cereal edible fungus powder food and a preparation method thereof, and the high-protein composite nutritional coarse cereal edible fungus powder is prepared by utilizing the advantages of edible fungi in solid state fermentation. In addition, the international enterprise Myco technology utilizes mushroom mycelium to ferment pea protein and rice protein mixture (65%: 35%), the protein fermentation product prepared by the method is easier to absorb by human body and lower in sensitization rate, and the protein fermentation product is applied to products such as plant meat, plant-based dairy products, yoghourt and ice cream. The use of Pleurotus Ostreatus mycelium for solid state fermentation of whole lentil powder increases the protein, resistant starch and polyphenol content, while the digestibility of the fermented powder is higher.
A great deal of research can prove the advantages of the edible fungi in fermentation, so the invention summarizes and develops the utility of various edible fungi in the pioneering of obtaining the soy protein isolate nutrition enhancing powder based on the research.
Preferably, in the step (1), the edible fungus strain is inoculated into a PDA solid culture medium for culture.
The edible fungus strain activation is usually carried out at 20 to 25 ℃, and the culture time is based on that the edible fungus mycelium grows full of PDA solid culture medium. The culture time is generally 5 to 7 days, and if the growth condition is not ideal, the culture is continued for 1 to 2 cycles on a PDA culture medium until the mycelium of the edible fungi grows to be full of the PDA solid culture medium.
Preferably, in the step (1), the inoculation operation is: after the mycelium of the edible fungus grows over the PDA solid culture medium, cutting a small block of the solid culture medium under the aseptic condition and inoculating the small block of the solid culture medium into the PDB liquid culture medium.
In order to ensure the normal development of subsequent experiments, the condition that the experiments are interrupted due to the non-ideal subsequent growth of the solid culture medium is prevented. In an experiment, 2 to 3 blocks of PDA solid culture medium full of edible fungus mycelium are usually cut, and all the cut small blocks of the PDA solid culture medium are transferred to a PDB liquid culture medium for continuous culture.
Preferably, in the step (1), the inoculated PDB liquid culture medium is cultured until the edible fungus seed solution is clear and transparent and contains white mycelium pellets.
And (3) after the PDA solid culture medium full of the edible fungus mycelia is inoculated into a PDB liquid culture medium, standing the PDB liquid culture medium for 24 h at the temperature of 20-25 ℃, and continuously carrying out shake culture for 2 days at the temperature to obtain the edible fungus seed liquid. The culture of the edible fungus seed liquid is greatly influenced by environmental factors, or the situation that the fermentation is not ideal in a general culture period (2 to 3 days) occurs. In order to ensure a higher biomass content during inoculation, the culture time can be appropriately prolonged (continued for 5 to 7 days) when the fermentation is not optimal. The well-fermented bacterial liquid is clear and transparent, and a large amount of white mycelium pellets can be seen to be suspended in the bacterial liquid.
Preferably, in the step (2), the mass-to-volume ratio of the isolated soy protein powder to the distilled water is 100 g: (50 to 100) mL.
In the addition of distilled water, in order to ensure the water content of the soybean protein powder, it should be noted that the water should be added as uniformly as possible. When necessary, the isolated soybean protein powder can be placed in distilled water, and the water content of the isolated soybean protein powder is ensured by filtering the redundant water after soaking for 30 to 60 min.
Preferably, in the step (2), the sterilization mode may be autoclaving or ultraviolet sterilization.
In practice, high temperature sterilization is often used, but functional components such as polyphenols and proteins are easily decomposed or inactivated under high temperature conditions. For example, publication No. CN113068824A discloses a high-protein composite nutritional coarse cereal edible fungus powder food and a preparation method thereof, wherein coarse cereals are steamed and boiled for 5 hours and then sterilized at high temperature, so that the energy consumption is high, and the loss of active ingredients is high. Therefore, the present invention is biased toward autoclaving or UV sterilization, thereby minimizing decomposition and inactivation of the beneficial ingredients.
Preferably, in the step (3), the drying manner may be any one of vacuum drying, microwave drying and freeze-drying.
In actual operation, various drying modes such as vacuum drying, microwave drying, freeze drying and the like can be selected for drying. The advantage of lyophilization over other drying methods is that the original flavor can be retained more completely, but its cost is higher.
Preferably, in the step (3), the mass-to-volume ratio of the soy protein isolate powder in the soy protein isolate culture medium to the edible fungus seed liquid is 100 g: (5 to 10) mL.
Distilled water is added into the soy protein isolate culture medium to ensure the water content of the soy protein isolate powder, and the subsequent fermentation is facilitated when the water content of the soy protein isolate powder is 60 to 70 percent in general. The addition amount of the edible fungus seed liquid can influence the fermentation period, a longer fermentation period is needed when the amount of the seed liquid is small, and the fermentation period can be shortened when the amount of the seed liquid is large.
Preferably, in the step (3), the fermentation culture time is 10-16 days.
In the fermentation culture process, if mycelium cannot grow in an area on the soybean protein culture medium, the water distribution ratio and the water adding mode are adjusted according to specific conditions to ensure that the culture medium maintains proper humidity.
Therefore, the invention has the following beneficial effects:
(1) The invention takes edible fungi as strains, and the nutrition and the oxygen resistance of the soybean protein isolate powder after solid state fermentation are obviously enhanced;
(2) The method for enhancing the nutrition of the soybean protein isolate powder is suitable for large-scale production and wide application;
(2) The invention utilizes edible fungus fermentation to convert glucoside isoflavone into free aglycone isoflavone, thus improving the biological accessibility;
(3) The soybean protein isolate nutrition enhancing powder processed by the scheme provided by the invention has more obvious flavor, relatively improved color and luster, favorable absorption and utilization by human bodies, health-care effects of reducing blood sugar and blood fat, resisting tumors and the like, and better market prospect.
Drawings
FIG. 1 is a process flow diagram of a method for enhancing the nutrition of soy protein isolate by solid state fermentation of edible fungi;
FIG. 2 is a graph showing the comparison of the content of phenolic substances in the soybean protein isolate powder, comparative example 1 and examples 1 to 3;
FIG. 3 is a graph comparing four oxidation resistance indexes of isolated soy protein powder, comparative example 1, and examples 1-3.
Detailed Description
The invention is further described with reference to specific examples. Those skilled in the art will be able to practice the invention based on these descriptions. Moreover, the embodiments of the present invention described in the following description are generally only some embodiments of the present invention, and not all embodiments. Therefore, all other embodiments obtained by a person of ordinary skill in the art based on the embodiments of the present invention without making creative efforts shall fall within the protection scope of the present invention.
[ PREPARATION PRE-TEST ]
Preparation method of PDA solid culture medium and PDB liquid culture medium
a. 200 g of peeled potatoes are weighed out and processed into small pieces of less than 5 mm, followed by addition of 600 mL of distilled water. Heating the distilled water with small pieces of potato to 100 deg.C, keeping boiling state for 30 min, and filtering with four layers of gauze to obtain potato juice.
b. And (B) weighing and adding 30 g of glucose, 3 g of monopotassium phosphate, 1.5 g of magnesium sulfate and 1 mg of vitamin B into the potato juice obtained in the step (a), using a 1000 mL volumetric flask for constant volume to obtain a PDB liquid culture medium, and continuously adding 20 g of agar to obtain a PDA solid culture medium.
c. Sterilizing the liquid/solid culture medium at 0.1 Mpa and 121 ℃ for 30 min, then placing the liquid/solid culture medium in a sterile environment for subpackaging, wherein the PDA solid culture medium is subpackaged in culture dishes with 15 mL of each part, and the PDB liquid culture medium is subpackaged in 500 mL conical flasks with 100 mL of each part.
[ examples ] A
Example 1
S1, preparing oyster mushroom seed liquid: inoculating Pleurotus Ostreatus strain into PDA solid culture medium at 25 deg.C, culturing and activating, subculturing once (at this time, pleurotus Ostreatus mycelium is overgrown with PDA solid culture medium), and cutting under aseptic condition to obtain a cut area of 17.5-22.5 mm 2 2-3 pieces of solid culture medium of (3) are inoculated into PDB liquid culture medium. And standing the inoculated PDB liquid culture medium for 24 hours at 25 ℃, keeping the temperature unchanged, transferring the PDB liquid culture medium into a shaking table for continuous culture for 48 hours, setting the rotating speed to be 150 rpm, and culturing to obtain the oyster mushroom seed liquid.
S2, preparing a soy isolate protein culture medium: 100 g of isolated soy protein powder is weighed into a 500 mL fermenter, 60 mL of distilled water is added, and the mixture is shaken gently to distribute the water evenly in the bottle. After the mixture is uniformly mixed, the fermentation tank cover is completely screwed and loosened for 1.5 rotation processes, the fermentation tank cover is heated to 121 ℃ for sterilization, the pressure is set to be 0.1 MPa, the sterilization time is 30 min, and the soy protein isolate culture medium is obtained after the sterilization is finished.
S3, fermentation culture: and (3) extracting 5 mL of oyster mushroom seed liquid prepared in the step (S1) (the biomass content of the oyster mushroom seed liquid is 245 mg/100 mL) in an aseptic environment, and adding the oyster mushroom seed liquid into the soy protein isolate culture medium prepared in the step (S2). And then, placing the fermentation tank fusing the oyster mushroom seed liquid and the soy protein isolate culture medium in a mushroom culture box, and culturing for 16 days at 25 ℃ until the oyster mushroom mycelia completely grow over the soy protein isolate culture medium without obvious change. Taking the oyster mushroom-soy protein isolate culture material compound out of the fermentation tank, vacuum drying at 60 ℃ for 48 h, crushing by using a pulverizer, and sieving by using a 40-mesh sieve to obtain the soy protein isolate nutrition enhancing powder.
Example 2
S1, preparing a hericium erinaceus seed solution: inoculating Hericium erinaceus strain into PDA solid culture medium at 25 deg.C, culturing and activating, subculturing once (at this time, mycelium of Hericium erinaceus grows over PDA solid culture medium), and cutting under aseptic condition to obtain Hericium erinaceus strain with area of 17.5-22.5 mm 2 2-3 pieces of solid culture medium are inoculated into PDB liquid culture medium. And standing the inoculated PDB liquid culture medium at 25 ℃ for 24 h, transferring the PDB liquid culture medium into a shaking table to continue culturing for 48 h while keeping the temperature unchanged, and culturing at the set rotating speed of 150 rpm to obtain the hericium erinaceus seed liquid.
S2, preparing a soy isolate protein culture medium: 100 g of isolated soy protein powder is weighed into a 500 mL fermenter, 70 mL of distilled water is added, and the mixture is shaken gently to distribute the water evenly in the bottle. After mixing uniformly, the cover of the fermentation tank is loosened by completely screwing for 1.5 rotation strokes, and the fermentation tank is irradiated by ultraviolet rays for 30 min, wherein the wavelength is set to be 220-290 nm. And (4) obtaining the soy protein isolate culture medium after sterilization.
S3, fermentation culture: and (3) extracting 5 mL of hericium erinaceus seed liquid prepared in the step (S1) (the biomass content of the hericium erinaceus seed liquid is 245 mg/100 mL) in a sterile environment, and adding the extracted liquid into the soy protein isolate culture medium prepared in the step (S2). And then, placing the fermentation tank fusing the hericium erinaceus seed liquid and the soy protein isolate culture medium in a fungus culture box, and culturing for 16 days at 24 ℃ until the hericium erinaceus mycelium completely grows over the soy protein isolate culture material and does not change obviously. Taking out the hericium erinaceus-soybean protein isolate culture material compound from the fermentation tank, freeze-drying at-80 ℃ for 48 hours, crushing by using a pulverizer, and sieving by using a 40-mesh sieve to obtain the soybean protein isolate nutrition enhancing powder.
Example 3
S1, preparing a needle mushroom seed solution: inoculating needle mushroom strain into PDA solid culture medium at 25 deg.C, culturing and activating, subculturing once (at this time, mycelium of needle mushroom grows over PDA solid culture medium), and cutting under aseptic condition to obtain a cut area of 17.5-22.5 mm 2 2-3 pieces of solid culture medium are inoculated into PDB liquid culture medium. And standing the inoculated PDB liquid culture medium at 25 ℃ for 24 h, transferring the PDB liquid culture medium into a shaking table to continue culturing for 48 h while keeping the temperature unchanged, and culturing at the set rotating speed of 150 rpm to obtain the flammulina velutipes seed liquid.
S2, preparing a soy isolate protein culture medium: 100 g of isolated soy protein powder is weighed into a 500 mL fermenter, 65 mL of distilled water is added, and the mixture is shaken gently to distribute the water evenly in the bottle. After the mixture is uniformly mixed, the fermentation tank cover is completely screwed and loosened for 1.5 rotation processes, the fermentation tank cover is heated to 121 ℃ for sterilization, the pressure is set to be 0.1 MPa, the sterilization time is 30 min, and the soy protein isolate culture medium is obtained after the sterilization is finished.
S3, fermentation culture: and (3) extracting 5 mL of needle mushroom seed liquid prepared in the step (S1) (the biomass content of the needle mushroom seed liquid is 245 mg/100 mL) in a sterile environment, and adding the extracted needle mushroom seed liquid into the soy protein isolate culture medium prepared in the step (S2). And then, putting the fermentation tank with the fused flammulina velutipes seed solution and the soy protein isolate culture medium into a fungus culture box, and culturing for 15 days at 25 ℃ until the flammulina velutipes mycelia are completely grown over the soy protein isolate culture material and no longer have obvious change. Taking the needle mushroom-soybean protein isolate culture material compound out of the fermentation tank, drying in a microwave oven at 3000 MHz for 15 min, pulverizing with a mill, and sieving with a 40 mesh sieve to obtain soybean protein isolate nutrition enhancing powder.
Example 4
S1, preparing oyster mushroom seed liquid: inoculating Pleurotus Ostreatus strain into PDA solid culture medium at 25 deg.C, culturing and activating, subculturing once (at this time, pleurotus Ostreatus mycelium is overgrown with PDA solid culture medium), and cutting under aseptic condition to obtain a cut area of 17.5-22.5 mm 2 2-solid medium3, inoculating into PDB liquid culture medium. And standing the inoculated PDB liquid culture medium at 25 ℃ for 24 h, transferring the PDB liquid culture medium into a shaking table to continue culturing for 48 h while keeping the temperature unchanged, and culturing at the set rotating speed of 150 rpm to obtain the oyster mushroom seed liquid.
S2, preparing a soy isolate protein culture medium: 100 g of soybean protein isolate powder is weighed in a stainless steel tray, 100 mL of distilled water is added, and the mixture is gently shaken to ensure that the water is uniformly distributed in a bottle. After the mixture is uniformly mixed, the fermentation tank cover is completely screwed and loosened for 1.5 rotation processes, the fermentation tank cover is heated to 121 ℃ for sterilization, the pressure is set to be 0.1 MPa, the sterilization time is 30 min, and the soy protein isolate culture medium is obtained after the sterilization is finished.
S3, fermentation culture: and (3) extracting 7 mL of oyster mushroom seed liquid prepared in the step (S1) (the biomass content of the oyster mushroom seed liquid is 245 mg/100 mL) in an aseptic environment, and adding the oyster mushroom seed liquid into the soy protein isolate culture medium prepared in the step (S2). And then, placing a stainless steel tray fusing the oyster mushroom seed liquid and the soy protein isolate culture medium in an aseptic platform, and culturing for 12 days at 25 ℃ until oyster mushroom mycelia completely grow over the soy protein isolate culture medium without obvious change. Taking the oyster mushroom-soybean protein isolate culture material compound out of the fermentation tank, vacuum drying at 60 ℃ for 48 hours, crushing by using a mill, and sieving by using a 40-mesh sieve to obtain the soybean protein isolate nutrition enhancing powder.
Example 5
S1, preparing oyster mushroom seed liquid: inoculating Pleurotus Ostreatus strain into PDA solid culture medium at 25 deg.C, culturing and activating, subculturing once (at this time, pleurotus Ostreatus mycelium grows over PDA solid culture medium), and cutting under aseptic condition to obtain pieces with area of 17.5-22.5 mm 2 2-3 pieces of solid culture medium of (3) are inoculated into PDB liquid culture medium. And standing the inoculated PDB liquid culture medium at 25 ℃ for 24 h, transferring the PDB liquid culture medium into a shaking table to continue culturing for 48 h while keeping the temperature unchanged, and culturing at the set rotating speed of 150 rpm to obtain the oyster mushroom seed liquid.
S2, preparing a soy isolate protein culture medium: 100 g of isolated soy protein powder is weighed into a cell culture flask, 60 mL of distilled water is added, and the mixture is gently shaken to ensure that the water is uniformly distributed in the flask. After the mixture is uniformly mixed, the fermentation tank cover is completely screwed and loosened for 1.5 rotation processes, the fermentation tank cover is heated to 121 ℃ for sterilization, the pressure is set to be 0.1 MPa, the sterilization time is 30 min, and the soy protein isolate culture medium is obtained after the sterilization is finished.
S3, fermentation culture: and (3) extracting 10 mL of oyster mushroom seed liquid prepared in the step (S1) (the biomass content of the oyster mushroom seed liquid is 245 mg/100 mL) in an aseptic environment, and adding the oyster mushroom seed liquid into the soy protein isolate culture medium prepared in the step (S2). And then, placing a cell culture bottle fusing the oyster mushroom seed liquid and the soy protein isolate culture medium in a fungus culture box, and culturing for 10 days at 25 ℃ until the oyster mushroom mycelia completely grow over the soy protein isolate culture medium without obvious change. Taking the oyster mushroom-soybean protein isolate culture material compound out of the fermentation tank, vacuum drying at 60 ℃ for 48 hours, crushing by using a mill, and sieving by using a 40-mesh sieve to obtain the soybean protein isolate nutrition enhancing powder.
Comparative example 1
S1, preparing a soy isolate protein culture medium: 100 g of isolated soy protein powder is weighed into a 500 mL fermenter, 60 mL of distilled water is added, and the mixture is shaken gently to distribute the water evenly in the bottle. After the mixture is uniformly mixed, the fermentation tank cover is completely screwed and loosened for 1.5 rotation processes, the fermentation tank cover is heated to 121 ℃ for sterilization, the pressure is set to be 0.1 MPa, the sterilization time is 30 min, and the soy protein isolate culture medium is obtained after the sterilization is finished.
S2, fermentation culture: the fermenter containing the soy protein isolate medium was placed in a bacterial incubator under sterile conditions and incubated at 25 ℃ for 12 days. And taking the soy protein isolate compost out of the fermentation tank, drying the compost in vacuum at 60 ℃ for 48 hours, crushing the compost by using a pulverizer, and sieving the compost by using a 40-mesh sieve to obtain the soy protein isolate nutrition enhancing powder.
[ Performance test ]
1. Color difference test
1) The measuring method comprises the following steps: measuring brightness (L), red-green value (a), and yellow-blue value (b) of the product with a color difference tester, randomly selecting 6 points of each sample, averaging, and calculating color difference (Δ E) according to formula (I).
The smaller the color difference value, the closer the color of the soybean protein isolate nutrition-enhancing powder is to that of the soybean protein isolate powder.
2) And (3) testing results: the results of the color difference performance test on the isolated soy protein nutrition-enhancing powders prepared in examples 1 to 3 and comparative example 1 are shown in Table 1.
TABLE 1 color difference of isolated soy protein powder before and after solid state fermentation
As can be seen from the observation of Table 1, the color of the isolated soy protein nutrition-enhancing powder obtained by the preparation process of the invention is deepened to a certain extent. According to the color difference test result, the color of the material is biased to red. Wherein, the color of the soybean protein isolate nutrition enhancing powder obtained by fermenting oyster mushroom serving as a strain is more yellow than that of needle mushroom and hericium erinaceus serving as strains.
2. Analysis of amino acid composition
1) The determination method comprises the following steps: accurately weighing 0.10 g of sample in an ampoule bottle, adding 10 mL of 6 mol/L hydrochloric acid into the ampoule bottle for wetting, blowing nitrogen for 15 min, heating by using an alcohol burner, and sealing the ampoule bottle. And (3) placing the completely sealed ampoule bottle at 110 ℃ for hydrolysis for 24-36 h, and transferring to a 50 mL volumetric flask for constant volume after the hydrolysis is completed. 2 mL of the solution was collected on a petri dish, evaporated to dryness at 100 ℃ to remove acid, 1 mL of buffer solution was added, mixed well, and then passed through a 0.45 μm filter for HPLC analysis.
2) And (3) testing results: the amino acid composition changes of the isolated soy protein powders obtained before and after the solid state fermentation in examples 1-3 and comparative example 1 are shown in Table 2.
TABLE 2 amino acid composition changes of soy protein isolate before and after solid state fermentation
As can be seen from the observation of Table 2, the autoclaving process results in a loss of some amino acids, such as lysine, arginine, etc. After the edible fungi are fermented, the amino acid content of each embodiment is increased, wherein the amino acid content of the soybean protein isolate nutrition enhancing powder obtained by fermenting oyster mushrooms serving as strains is increased most obviously.
3. Determination of isoflavone content in soybean
1) The determination method comprises the following steps: the content of soybean isoflavone is determined by referring to GB/T23788-200 high performance liquid chromatography.
2) And (3) testing results: the isoflavone contents of the isolated soy protein powders obtained before and after the solid state fermentation in examples 1-3 and comparative example 1 were varied as shown in Table 3.
TABLE 3 changes in Soy isoflavone content before and after solid fermentation
As can be seen from the observation of Table 3, the content of daidzin in the two glycoside type soybean isoflavones treated by the preparation process of the invention is remarkably reduced, and genistin is not detected. According to the existing research, isoflavone exists in the isolated soy protein mainly in the form of glycoside such as daidzin, genistin and the like, accounts for about 97 percent of the total content, is hardly absorbed and utilized by human bodies, and has low biological accessibility. Therefore, the soybean protein isolate nutrition enhancing powder obtained after solid state fermentation of edible fungi such as oyster mushrooms, needle mushrooms, hericium erinaceus and the like has greatly improved biological accessibility and is beneficial to absorption by human bodies.
Meanwhile, in the two free aglycone type soybean isoflavones, the content of daidzein is increased by 100.68 to 226.02 percent, and the content of genistein is increased by 147.87 to 333.33 percent. According to the existing research, the genistein has the strongest physiological activity, can inhibit the activity of the topoisomerase II and the tyrosine protein kinase PTK, and can induce the apoptosis of tumor cells. The content of genistein in the soybean protein isolate treated by the solid fermentation of the edible fungi is greatly improved, and the anti-tumor effect of the soybean protein isolate can be effectively enhanced.
In conclusion, the prepared soybean protein isolate nutrition enhancing powder has the effects of resisting tumor, inhibiting bacteria, reducing blood fat and the like, and is beneficial to the absorption of a human body.
4. Determination of phenolic substance content and antioxidant activity
1) The measuring method comprises the following steps: accurately weighing 0.10 g of sample, adding 10 mL of 80% ethanol, performing ultrasonic treatment for 2 h, centrifuging at 10000 rpm and 25 ℃ for 10 min, and obtaining a supernatant after centrifuging, wherein the supernatant can be used for measuring the total phenol content, the reducing power, the hydroxyl free radical scavenging capacity, the DPPH free radical scavenging capacity and the ABTS free radical scavenging capacity.
2) And (3) testing results: the content of phenolic substances and the antioxidant activity of the isolated soybean protein powder before and after solid fermentation are shown in figures 2 and 3, and the rise rate of the phenolic substances in the isolated soybean protein nutrition-enhancing powder in examples 1-3 can reach 292.31 to 360.76 percent; in addition, the soybean protein isolate nutrition enhancing powder with the same quality is remarkably improved compared with four antioxidant indexes of the soybean protein isolate powder in the comparative example 1. The isolated soybean protein powder treated by the solid fermentation of the edible fungi has obviously enhanced nutrition and oxidation resistance.
In addition, other edible fungi such as Ganoderma lucidum, bolete, and Pleurotus eryngii can exhibit the same effect as Pleurotus ostreatus, flammulina velutipes, and Hericium erinaceus, and are not repeated herein.
Claims (10)
1. A method for enhancing the nutrition of isolated soy protein based on solid state fermentation of edible fungi is characterized by comprising the following steps:
(1) Preparing edible fungus seed liquid: activating edible fungus strains, inoculating the activated edible fungus strains into a PDB liquid culture medium, and culturing to obtain an edible fungus seed liquid;
(2) Preparing a soy protein isolate culture medium: sterilizing the isolated soy protein powder and distilled water in a fermentation container to obtain a soy protein isolate culture medium;
(3) Solid state fermentation culture: adding the edible fungus seed liquid into a soy protein isolate culture medium, fermenting and culturing, drying and crushing to obtain the soy protein isolate nutrition enhancing powder.
2. The method for enhancing the nutrition of soybean protein isolate based on solid state fermentation of edible fungi according to claim 1,
the edible fungus in the step (1) is any one of needle mushroom, oyster mushroom, hericium erinaceus, lucid ganoderma, bolete and pleurotus eryngii.
3. The method for enhancing the nutrition of soybean protein isolate based on solid state fermentation of edible fungi according to claim 2,
in the step (1), the edible fungus strain is inoculated into a PDA solid culture medium for culture.
4. The method for enhancing the nutrition of soybean protein isolate based on solid state fermentation of edible fungi of claim 3,
in the step (1), the inoculation operation is as follows: after the mycelium of the edible fungus grows over the PDA solid culture medium, cutting a small block of the solid culture medium under the aseptic condition and inoculating the small block of the solid culture medium into the PDB liquid culture medium.
5. The method for enhancing the nutrition of soybean protein isolate based on solid state fermentation of edible fungi of claim 4,
in the step (1), the inoculated PDB liquid culture medium is cultured until the edible fungus seed liquid is clear and transparent and contains white mycelium pellets.
6. The method for enhancing the nutrition of soybean protein isolate based on the solid state fermentation of edible fungi of claim 1, 2 or 5,
in the step (2), the mass volume ratio of the soybean protein isolate powder to the distilled water is 100 g: (50 to 100) mL.
7. The method for enhancing the nutrition of soybean protein isolate based on the solid state fermentation of edible fungi according to claim 1, 2 or 5,
in the step (2), the sterilization mode can be high-pressure sterilization or ultraviolet sterilization.
8. The method for enhancing the nutrition of soybean protein isolate based on the solid state fermentation of edible fungi according to claim 1, 2 or 5,
in the step (3), the drying mode may be any one of vacuum drying, microwave drying and freeze drying.
9. The method for enhancing the nutrition of soybean protein isolate based on the solid state fermentation of edible fungi according to claim 1, 2 or 5,
in the step (3), the mass volume ratio of the soybean protein isolate powder to the edible fungus seed liquid in the soybean protein isolate culture medium is 100 g: (5 to 10) mL.
10. The method for enhancing the nutrition of soybean protein isolate based on the solid state fermentation of edible fungi according to claim 1, 2 or 5,
in the step (3), the fermentation culture time is 10-16 days.
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| CN117126906A (en) * | 2023-07-13 | 2023-11-28 | 中国农业大学 | Modification method of soybean protein isolate, modified soybean protein isolate and application thereof |
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