CN115058370A - Antioxidant metazoan compound fermentation liquid, preparation method and application - Google Patents
Antioxidant metazoan compound fermentation liquid, preparation method and application Download PDFInfo
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- CN115058370A CN115058370A CN202210888077.1A CN202210888077A CN115058370A CN 115058370 A CN115058370 A CN 115058370A CN 202210888077 A CN202210888077 A CN 202210888077A CN 115058370 A CN115058370 A CN 115058370A
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
The invention belongs to the technical field of biology, and particularly discloses an antioxidant metazoan compound fermentation liquid, a preparation method and application thereof, wherein the metazoan compound fermentation liquid is a compound fermentation product obtained by taking fungus-phellinus igniarius as a fermentation substrate and performing fermentation treatment on the fungus-phellinus igniarius by using lactic acid bacteria with high antioxidant activity; the post-growth composite fermentation broth not only contains functional components such as lactobacillus metabolites, lysate, phellinus igniarius polysaccharide, chitin (chitosan), beta-glucan and the like, but also contains new active components formed by decomposing and converting specific medicines in the original medicines by the aid of complicated reactions such as oxidation, isomerization, acetylation, esterification and vitization of lactobacillus, and the components synergistically enhance the antioxidant level of the composite fermentation broth; the preparation method is simple in preparation process and easy in industrial amplification, and has an important promotion effect on promoting the development of the metagenetic industry.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to an antioxidant metazoan compound fermentation broth, a preparation method and application.
Background
The post-growth refers to that the lactobacillus still retains the thallus components and metabolites with the same activity as the viable bacteria after being processed by heat treatment, physical treatment, high hydrostatic pressure treatment, freeze drying, ultrasonic oscillation and other processing modes, and has incomparable advantages compared with the lactobacillus. The metazoan has various biological activities, wherein the extracellular polysaccharide can inhibit lipid peroxidation and enhance the free radical scavenging capacity, so that the metazoan has relatively good antioxidant effect, but has certain effect difference compared with the existing antioxidant, so that the research and development of the metazoan are limited, and the popularization and the application of the metazoan preparation are not facilitated.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide an antioxidant metazoan compound fermentation broth, a preparation method and application.
In order to achieve the technical effects, the invention adopts the following technical scheme:
an antioxidant metancholia composite fermentation liquid comprises phellinus igniarius and lactic acid bacteria, and specifically comprises a product obtained by performing composite fermentation on phellinus igniarius serving as a single fermentation substrate and the lactic acid bacteria serving as fermentation strains.
Preferably, the thallus density in the composite fermentation liquid is more than or equal to 10 9 cfu/ mL。
Preferably, the lactic acid bacteria are selected from one of lactobacillus plantarum, lactobacillus acidophilus and lactobacillus rhamnosus.
Preferably, the lactobacillus plantarum is lactobacillus plantarum (b)Lactobacillus plantarum) BLCC2-0015 was deposited at the chinese collection center on 3/18 of 2015 at the following locations: china Wuhan university, the preservation number is CCTCC NO: m2015126; it is disclosed in the patent application "a Lactobacillus plantarum strain having high antioxidant activity and its use" (application No. 201510419415.7, filing date: 2015.07.16).
Lactobacillus acidophilus (C.acidophilus: (C.acidophilus)Lactobacillus acidophilus) PBIL2-003 was deposited at the China center for type culture Collection on 28 th 4 th 2018 at the following deposition addresses: the preservation number of the Wuhan university in Wuhan, China is CCTCC NO: m2018208; the ganoderma lucidum probiotic compound fermentation liquor, the preparation method and the application thereof are disclosed in the patent application (application number: 201911424255.X, application date: 2019.12.31).
Lactobacillus rhamnosus (A), (B), (C)Lactobacillus rhamnosus) PBIL3-003 was deposited at the China center for type culture Collection on 28 th 4 th 2018 at the following deposition addresses: china Wuhan university, the preservation number is CCTCC NO: m2018206; the ganoderma lucidum probiotic compound fermentation liquor, the preparation method and the application thereof are disclosed in the patent application (application number: 201911424255.X, application date: 2019.12.31).
The second objective of the present invention is to provide a preparation method of the antioxidant metazoan composite fermentation broth, which specifically comprises the following steps:
(1) preparation of phellinus igniarius fermentation liquor: activating and expanding culturing Phellinus Linteus strain to obtain Phellinus Linteus fermentation broth;
(2) preparation of the phellinus igniarius enzymolysis liquid: homogenizing the phellinus igniarius fermentation liquor to obtain phellinus igniarius homogenate liquid; adding cellulase into the phellinus igniarius homogenate liquid to obtain phellinus igniarius enzymatic hydrolysate;
(3) preparing a lactobacillus secondary seed solution: sequentially carrying out culture activation, primary seed culture and secondary seed culture on the lactobacillus strains to obtain a lactobacillus secondary seed solution;
(4) adding the secondary seed liquid of the lactobacillus into the phellinus igniarius enzymolysis liquid, and culturing to obtain the post-growth-element composite fermentation liquid.
Preferably, the specific preparation process of the phellinus linteus fermentation liquor in the step (1) comprises the following steps: sucking 0.3-0.4 mLPDA broth culture medium by using a sterile pipette, dripping the broth culture medium into a phellinus igniarius strain ampoule bottle, and slightly shaking to dissolve the freeze-dried strain into a suspension state; taking about 0.2mL of thallus suspension, transplanting the thallus suspension into a PDA agar culture medium, and culturing for 7-10 days at 25-28 ℃ until the surface of the culture medium is full of mycelia;
take 1cm 3 Inoculating the PDA agar culture medium full of mycelia into a PDA broth culture medium, and culturing at 25-28 deg.C for 5-7 days to form a large amount of dense mycelia balls to obtain Phellinus igniarius seed culture solution;
inoculating the phellinus igniarius seed culture solution into a PDA broth culture medium with the inoculation amount of 10%, and culturing for 3-5 days at 25-28 ℃ until a large number of dense mycelium pellets are formed and the culture solution becomes clear, so as to obtain the phellinus igniarius fermentation solution.
Preferably, the PDA broth medium comprises the following main components: 0.3 percent of potato powder, 0.2 percent of monopotassium phosphate and 2 percent of glucose, wherein the initial pH value is 6.0-7.0, and the sterilization is carried out at 121 ℃ for 20 min.
The PDA agar culture medium comprises the following main components: 0.3% of potato powder, 0.2% of potassium dihydrogen phosphate, 2% of glucose and 1.5% of agar, wherein the initial pH value is 6.0-7.0, the temperature is 121 ℃, and the sterilization is carried out for 20 min.
Preferably, the specific preparation method of the phellinus linteus enzymatic hydrolysate in the step (2) comprises the steps of adjusting the pH of phellinus linteus homogenate to =4.5, adding 0.1-5% of cellulase, carrying out enzymolysis in a water bath kettle at 45-50 ℃ for 48-60 h at constant temperature until no obvious precipitate is formed in the homogenate, and then sterilizing at 115 ℃ for 15min to obtain the phellinus linteus enzymatic hydrolysate.
Preferably, the inoculation amount of the primary lactobacillus seed culture solution in the step (3) is 1-5%; the thallus density in the secondary lactobacillus seed liquid after fermentation is more than or equal to 10 9 cfu/mL。
Preferably, the culture activation conditions of the lactic acid bacteria in step (3) are: culturing for 22-26 h at 25-40 ℃; the culture conditions of the first-stage seed liquid are as follows: carrying out static culture for 12-24 h at the temperature of 25-40 ℃;
the conditions of fermentation culture are as follows: carrying out intermittent shaking culture for 48-96 h at the temperature of 25-40 ℃, wherein the stirring speed is 100r/min and 5min/2h of intermittent stirring;
preferably, the culture activation medium for the lactic acid bacteria in step (3) is a modified M6 medium, and comprises the following components: 10g of casein peptone, 10g of beef extract powder, 5g of yeast powder, 20g of glucose, 0.5g of lactose, 801 g of tween-801 g, 2g of 1.5% agar, 1L of purified water, pH6.5, 121 ℃, and sterilizing for 20 min; the first seed culture medium and the fermentation culture medium are both agar-free modified M6 culture medium.
Preferably, the preparation of the metazoan fermentation liquor in the step (4): the phellinus igniarius enzymolysis liquid is used as a fermentation culture medium (100%), and the inoculation amount of the lactobacillus secondary seed liquid is 3% -5% (v/v).
Preferably, the culture conditions of the metazoan complex fermentation broth in the step (4) are as follows: standing and culturing for 24-36 h at 37-38 ℃ until the pH is =3.5 or the thallus density is more than or equal to 10 9 cfu/ mL。
The third purpose of the invention is to provide a composite preparation containing the antioxidant metazoan composite fermentation liquid.
Preferably, the formulation may be in the form of a liquid, solid or semi-solid.
The fourth purpose of the invention is to provide a preparation method of different forms of antioxidant metaplasia compound preparation, in particular,
the liquid form of the antioxidant prebiotics compound preparation is as follows: crushing thalli in the post-biotic compound fermentation liquor prepared in the step (4) to obtain the post-biotic compound fermentation liquor;
the solid state form of the antioxidant metazoan compound preparation is powder, and the antioxidant metazoan compound preparation is prepared by performing spray drying on the metazoan compound fermentation liquor prepared in the step (4);
the semi-solid state form of the antioxidant metazoan compound preparation is prepared by removing partial water from the metazoan compound fermentation liquor prepared in the step (4) in a vacuum concentration mode.
The fifth purpose of the invention is to provide the application of the antioxidant metazoan compound fermentation liquor and the antioxidant compound preparation in the aspect of antioxidation.
Compared with the prior art, the invention has the following beneficial effects:
the metazoan compound fermentation broth disclosed by the invention not only contains antioxidant functional components such as phellinus igniarius polysaccharide, chitin (chitosan), beta-glucan and the like, but also contains antioxidant active substances (enzymes, polysaccharides and the like) generated by metabolism of lactic acid bacteria, and the components are synergistic, so that the antioxidant level of the compound fermentation broth is remarkably improved, and the oxidative damage level is effectively reduced;
according to the post-growth factor compound fermentation liquid prepared by the invention, the biological treatment is carried out by utilizing the probiotic functions of fungi and lactic acid bacteria in a double compound fermentation mode, the functional components are fully enriched, the treatment mode is soft, the process operation is simple, and the large-scale production is easy to realize;
the raw materials used in the preparation process of the metazoan compound fermentation liquor are food-grade components, can be directly drunk, and are safe, healthy and free of side effects.
Drawings
FIG. 1 is a statistical graph of the clearance of DPPH for the different treatment groups described in example 2;
FIG. 2 is a statistical plot of hydroxyl radical clearance for various treatment groups as described in example 2;
FIG. 3 is a statistical plot of the effect of different treatment groups on GSH content as described in example 2;
FIG. 4 is a statistical graph of the effect of different treatment groups on SOD activity as described in example 2;
FIG. 5 is a statistical graph of the DPPH clearance of post-fermentation complex fermentation broths prepared from various control strains in comparative example 1;
FIG. 6 is a statistical chart of hydroxyl radical scavenging rate of post-growth composite fermentation broth prepared from various control strains in comparative example 1;
FIG. 7 is a statistical chart of the effect of post-growth composite fermentation broth prepared from various control strains on GSH content in comparative example 1;
FIG. 8 is a statistical chart showing the effect of post-growth factor complex fermentation broth prepared from various control strains on SOD activity in comparative example 1;
FIG. 9 is a statistical plot of the GSH content in the serum of mice treated by different treatment groups;
FIG. 10 is a statistical chart of SOD content in serum of mice treated by different treatment groups;
FIG. 11 is a statistical chart of the MDA content in the serum of mice treated by different treatment groups;
FIG. 12 is a statistical graph of the protein carbonyl content in the serum of mice treated by different treatment groups;
note: in fig. 1 to 4, P < 0.01 indicates that the difference is very significant; wherein, marked on the uppermost horizontal line represents that the difference of the oxidation resistance of the phellinus igniarius enzymolysis liquid of the sample group and the phellinus igniarius homogenate of the blank group is very obvious, and the difference of the oxidation resistance of the metagenetic composite fermentation liquid prepared by lactobacillus plantarum BLCC2-0015 of the sample group and the oxidation resistance of lactobacillus plantarum BLCC2-0015 of the blank group is very obvious; the antioxidant performance of the metazoan compound fermentation liquid prepared by the lactobacillus acidophilus PBIL2-0013 of the sample group is remarkably different from that of the lactobacillus acidophilus PBIL2-0013 of the blank group; the antioxidant performance of the post-growth compound fermentation liquor prepared by the lactobacillus rhamnosus PBIL3-0013 in the sample group is very different from that of the lactobacillus rhamnosus PBIL3-0013 in the blank group;
in fig. 9 to 12, P represents P < 0.05, i.e., significant difference, and P < 0.01, i.e., significant difference, as compared with the blank control group; # indicates that P < 0.01, compared with the model control group, the difference is extremely significant; the corresponding and the corresponding parts are compared with the positive group, and the corresponding parts are marked as P < 0.05, namely, the difference is obvious, and the corresponding parts are marked as P < 0.01, namely, the difference is extremely obvious; t.xxx represents that the difference is very significant when P is less than 0.01 compared with the phellinus igniarius enzymatic hydrolysate.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described with the following embodiments, but is by no means limited thereto. The following is a description of the preferred embodiments of the present invention, and should not be taken as limiting the invention, but rather as embodying the invention in its broadest form and as indicating any variations, equivalents and modifications within the spirit and scope of the present invention.
The media components used in the following examples are as follows:
the PDA broth culture medium comprises the following main components: 0.3 percent of potato powder, 0.2 percent of monopotassium phosphate and 2 percent of glucose, wherein the initial pH value is 6.0-7.0, and the sterilization is carried out at 121 ℃ for 20 min.
The PDA agar culture medium comprises the following main components: 0.3% of potato powder, 0.2% of monopotassium phosphate, 2% of glucose and 1.5% of agar, wherein the initial pH value is 6.0-7.0, the temperature is 121 ℃, and the sterilization is carried out for 20 min.
The culture activation medium of the lactobacillus is an improved M6 culture medium, and comprises the following components: 10g of casein peptone, 10g of beef extract powder, 5g of yeast powder, 20g of glucose, 0.5g of lactose, 801 g of tween-801 g, 2g of 1.5% agar, 1L of purified water, pH6.5, 121 ℃, and sterilizing for 20 min; the first seed culture medium and the fermentation culture medium are both agar-free modified M6 culture medium.
The culture activation conditions of the lactic acid bacteria are as follows: culturing for 22-26 h at 25-40 ℃;
the culture conditions of the first-stage seed liquid are as follows: carrying out static culture for 12-24 h at the temperature of 25-40 ℃;
the culture conditions of the secondary seed liquid are as follows: carrying out intermittent shaking culture for 48-96 h at the temperature of 25-40 ℃, wherein the stirring speed is 100r/min and 5min/2h of intermittent stirring;
example 1 preparation of antioxidant metazoan Compound fermentation broth
(1) Activating the phellinus igniarius strain: sucking a broth culture medium of 0.3-0.4 mLPDA by using a sterile pipette, dripping the broth culture medium into an ampoule bottle of phellinus igniarius strain, and slightly shaking to dissolve the freeze-dried strain into a suspension state; taking about 0.2mL of thallus suspension, transplanting the thallus suspension into a PDA agar culture medium, and culturing for 7-10 days at the temperature of 25-28 ℃ until hyphae are distributed on the surface of the culture medium;
(2) activating the phellinus igniarius seed liquid: take 1cm 3 Inoculating the PDA agar culture medium into a PDA broth culture medium, and culturing at 25-28 deg.C for 5-7 days to form a large amount of dense mycelium pellets, i.e. Phellinus igniarius seed culture solution;
(3) preparation of phellinus igniarius fermentation liquor: inoculating a phellinus igniarius seed culture solution into a PDA broth culture medium with an inoculation amount of 10%, performing shaking culture at the temperature of 25-28 ℃, stirring at 100r/min for 3-5 days until a large number of dense mycelium pellets are formed, and clarifying the culture solution to obtain a phellinus igniarius fermentation solution;
(4) preparation of the phellinus igniarius enzymolysis liquid: homogenizing the phellinus igniarius fermentation liquor by using a colloid mill to obtain a suspension, namely phellinus igniarius homogenate; adjusting the pH =4.5 of the phellinus igniarius homogenate, then adding 0.1-5% of cellulase into the phellinus igniarius homogenate, carrying out enzymolysis for 48-60 h in a water bath kettle at the temperature of 45-50 ℃ at constant temperature until no obvious precipitate exists in the homogenate, and sterilizing for 15min at the temperature of 115 ℃ to obtain phellinus igniarius enzymatic hydrolysate;
(5) preparing a lactobacillus secondary seed solution: culturing and activating each strain of lactobacillus, selecting 1-5% strain, performing primary seed culture to obtain primary seed solution, performing secondary fermentation culture to obtain secondary seed solution of lactobacillus, wherein the thallus density in the secondary seed solution is not less than 10 9 cfu/mL；
(6) Adding the secondary lactobacillus seed solution into the phellinus igniarius enzymolysis solution according to the inoculation amount of 3-5% (v/v); standing and culturing for 24-36 h at 37-38 ℃ until the pH is =3.5 or the thallus density is more than or equal to 10 9 cfu/mL to obtain the post-growth composite fermentation liquid.
Example 2 in vitro validation of antioxidant Activity of metazoan Compound fermentation broth
Lactobacillus plantarum BLCC2-0015, Lactobacillus acidophilus PBIL2-003 and Lactobacillus rhamnosus PBIL3-003 are respectively fermented to prepare fermentation liquid, the fermentation liquid and the phellinus igniarius homogenate liquid are counted as blank groups, and the lactobacillus plantarum BLCC2-0015, the Lactobacillus acidophilus PBIL2-003 and the Lactobacillus rhamnosus PBIL3-003 are respectively prepared into the metagen compound fermentation liquid according to the method described in the embodiment 1, and the metagen compound fermentation liquid and the phellinus igniarius hydrolysate are all recorded as sample groups. The following kits are respectively adopted to detect the antioxidant activity of each group, and the detection results are shown in figures 1-4;
DPPH clearance rate: a detection kit for DPPH free radical scavenging ability of Solebao biotechnology Limited company;
hydroxyl radical scavenging rate: shanghai enzyme-linked Biotechnology Ltd. -hydroxy radical scavenging rate test kit;
SOD content: shanghai enzyme-linked Biotechnology Limited-superoxide dismutase (SOD) test kit;
content of GSH: shanghai enzyme-linked Biotechnology Ltd. -reduced Glutathione (GSH) test kit;
as can be seen from fig. 1 to 4, the oxidation resistance of the phellinus linteus homogenate liquid is remarkably improved after enzymolysis; when the phellinus igniarius enzymolysis liquid is used as a culture medium and different strains are respectively used for fermentation, the DPPH clearance rate, the hydroxyl radical clearance rate, the GSH content and the SOD activity of the metazoan compound fermentation liquid prepared by fermenting lactobacillus plantarum BLCC2-0015, lactobacillus acidophilus PBIL2-003 or lactobacillus rhamnosus PBIL3-003 are obviously improved, and the differences are extremely obvious, so that the metazoan compound fermentation liquid prepared by the invention has obvious synergistic effect compared with the phellinus igniarius enzymolysis liquid and each lactobacillus fermentation liquid in the aspect of oxidation resistance.
Comparative example 1
Selecting lactobacillus plantarum L1-019, lactobacillus acidophilus L2-001 and lactobacillus rhamnosus L3-001 as control strains to respectively prepare postbiotic compound fermentation liquor according to the method in the embodiment 1, and detecting the antioxidant activity of the postbiotic compound fermentation liquor prepared by the control strains by using the kit in the embodiment 2, wherein the detection results are shown in figures 5-8;
as can be seen from the fig. 5-8, the antioxidant property of the metazoan compound fermentation liquid prepared by fermenting the lactobacillus plantarum L1-019, the lactobacillus acidophilus L2-001 and the lactobacillus rhamnosus L3-001 is similar to that of the phellinus igniarius enzymolysis liquid, the difference is not obvious, and no synergistic effect exists.
Example 3 Experimental validation of antioxidant ethanol model animal
Grouping tests: blank group, model group, Vc positive group (100 mg/kg BW), phellinus igniarius enzymolysis liquid, anagen 1, anagen 2 and anagen 3;
the metazoan 1, the metazoan 2 and the metazoan 3 are metazoan compound fermentation liquid prepared by taking phellinus igniarius enzymolysis liquid as a culture medium and fermenting the phellinus igniarius enzymolysis liquid respectively by lactobacillus plantarum BLCC2-0015, lactobacillus acidophilus PBIL2-003 and lactobacillus rhamnosus PBIL3-003 according to the method in the embodiment 1.
Taking 90 male mice with the weight of about 20g, and pre-feeding the mice for three days; then, starting from the fourth day at 10 am, feeding physiological saline to a blank group and a model group, feeding-Vc to a positive group by intragastric administration, feeding the other experimental group mice by using phellinus igniarius enzymatic hydrolysate, a later prebiotic 1, a later prebiotic 2 and a later prebiotic 3 respectively, feeding the mice with intragastric administration at a gastric administration dose of 10mL/kg BW, continuously feeding 30d, fasting for 16h without water prohibition, feeding 0.4mL of physiological saline to the blank group by intragastric administration, feeding 0.4mL of 56-degree Hongxing Erguotou to the other groups of mice by intragastric administration, and detecting the contents of GSH, SOD, MDA and protein carbonyl in the blood serum of the mice by using detection kits of Shanghai enzyme-linked biotechnology Limited company after 6h respectively, wherein the detection results are shown in fig. 9-12;
SOD and GSH play a crucial role in the oxidation and antioxidant balance of organisms and are important indexes for reflecting the antioxidant capacity of the organisms;
as can be seen from fig. 9 and 10, the mouse serum treated by the model group had the least contents of SOD and GSH, which indicates that the red star Erguotou can reduce the contents of SOD and GSH in the mouse serum and cause peroxidation damage to the mouse; the contents of SOD and GSH in the blood serum of the mice treated by other treatment groups except the group of metazoan 1, the group of metazoan 2 and the group of metazoan 3 are reduced to different degrees, but the reduction degrees are smaller than those of the model group; the contents of SOD and GSH in the blood serum of the mice treated by the post-shengyuan 1, the post-shengyuan 2 and the post-shengyuan 3 groups are basically unchanged compared with the blank groups, and the differences between the contents and other treatment groups are very obvious, which indicates that other treatment groups have certain antioxidant performance, and the antioxidant effects of the post-shengyuan 1, the post-shengyuan 2 and the post-shengyuan 3 are most obvious; the GSH content in the serum of mice of the postbiotic 1 group, the postbiotic 2 group and the postbiotic 3 group has no obvious difference, which shows that the three postbiotic groups have almost the same capability of improving the GSH content of the organism; the SOD content in the serum of the mice in the postbiotic 1 group has no obvious difference with that in the blank group, and the SOD content in the serum of the mice in the postbiotic 2 group and the postbiotic 3 group has a little difference with that in the blank group, which shows that the SOD level of the postbiotic 1 group is better than that of the postbiotic 2 group and the postbiotic 3 group, and the influence on the metabolic pathway of the organism is slightly different due to different strains and the difference of metabolic products.
MDA is one of the final products of lipid peroxidation, and the content of MDA can reflect the speed of lipid peroxidation of matrix and the level of oxygen free radical; protein carbonyl is an early marker of various amino acids in the oxidative modification process of protein, and the content of the protein carbonyl indicates the degree of oxidative damage of the protein, and the protein carbonyl is an important index for measuring the oxidative damage of the protein.
As can be seen from fig. 11 and 12, the contents of MDA and protein carbonyl in the serum of the mouse treated by the model group are significantly increased, which indicates that the content of MDA and protein carbonyl in the serum of the mouse is increased by the red star Erguotou, and the mouse is damaged by peroxidation; the MDA content and the protein carbonyl content in the blood serum of the mice treated by other treatment groups except the group of the metazoan 1, the metazoan 2 and the metazoan 3 are increased to different degrees compared with the blank group, but the increment is smaller than that of the model group, while the MDA content and the protein carbonyl content in the blood serum of the mice treated by the metazoan 1, the metazoan 2 and the metazoan 3 are basically unchanged compared with the blank group, which indicates that other treatment groups have certain antioxidant performance, and the antioxidant effect of the metazoan 1, the metazoan 2 and the metazoan 3 is most obvious.
Example 4
The post-growth composite fermentation liquor can be processed into composite preparations with different forms; specifically, the thalli in the post-prebiotics compound fermentation liquor prepared in the embodiment 1 are crushed to obtain a liquid post-prebiotics compound preparation; spray drying the post-prebiotics compound fermentation liquor prepared in the embodiment 1 to prepare powder, namely a solid anti-oxidation post-prebiotics compound preparation; and (3) removing partial water from the metazoan compound fermentation liquor prepared in the example 1 in a vacuum concentration mode to prepare the semi-solid antioxidant metazoan compound preparation.
The preparation can be directly drunk or taken, and can also be processed into compositions in the forms of food, health products, medicines and the like, and the compositions can be in the forms of common preparations such as tablets, powder, granules, capsules, suspending agents and the like; the composition can be directly taken or drunk to improve the oxidation resistance of organisms.
The above-mentioned embodiments only express the specific embodiments of the present application, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present application. It should be noted that, for those skilled in the art, without departing from the technical idea of the present application, several changes and modifications can be made, which are all within the protection scope of the present application.
Claims (10)
1. An antioxidant metabiosis compound fermentation liquid is characterized by comprising phellinus igniarius and lactic acid bacteria, wherein the phellinus igniarius is used as a single fermentation substrate, and the lactic acid bacteria are used as a product of compound fermentation of fermentation strains.
2. The antioxidant metazoan compound fermentation broth as claimed in claim 1, wherein the thallus density in the compound fermentation broth is not less than 10 9 cfu/ mL。
3. The antioxidant metazoan complex fermentation broth of claim 1, wherein the lactic acid bacteria is selected from lactobacillus plantarum (lactobacillus)Lactobacillus plantarum) BLCC2-0015, Lactobacillus acidophilus (L.) (Lactobacillus acidophilus) PBIL2-003 and Lactobacillus rhamnosus (L.) (Lactobacillus rhamnosus) One of PBIL 3-003.
4. The method for preparing the antioxidant metazoan composite fermentation broth as claimed in any one of claims 1 to 3, comprising the steps of:
(1) preparation of phellinus igniarius fermentation liquor: activating and expanding culturing Phellinus Linteus strain to obtain Phellinus Linteus fermentation broth;
(2) preparation of the phellinus igniarius enzymolysis liquid: homogenizing the phellinus igniarius fermentation liquor to obtain phellinus igniarius homogenate liquid; adding cellulase into the phellinus igniarius homogenate liquid to obtain phellinus igniarius enzymatic hydrolysate;
(3) preparing a lactobacillus secondary seed solution: sequentially carrying out culture activation, seed culture and secondary fermentation culture on lactobacillus strains to obtain a lactobacillus secondary seed solution;
(4) adding the secondary seed liquid of the lactobacillus into the phellinus igniarius enzymolysis liquid, and culturing to obtain the post-growth-element composite fermentation liquid.
5. The method for preparing the antioxidant metazoan compound fermentation broth according to claim 4, wherein the Phellinus linteus fermentation broth in the step (1) is prepared by the following steps:
a. sucking the PDA broth culture medium by using a sterile straw, dripping the PDA broth culture medium into a phellinus igniarius strain ampoule bottle, and slightly shaking to dissolve the freeze-dried strain into a suspension state;
b. taking the thallus suspension, transplanting the thallus suspension into a PDA agar culture medium, and culturing until the surface of the culture medium is full of hyphae;
c. inoculating the PDA agar culture medium full of mycelia into PDA broth culture medium, and culturing to form a large amount of dense mycelium pellets to obtain Phellinus Linteus seed culture solution;
d. inoculating the phellinus igniarius seed culture solution into a PDA broth culture medium, culturing until a large number of dense mycelium pellets are formed, and clearing the culture solution to obtain the phellinus igniarius fermentation broth.
6. The method for preparing the antioxidative metabiotic compound fermentation broth as claimed in claim 4, wherein the Phellinus linteus enzymatic hydrolysate in the step (2) is prepared by adjusting the pH of Phellinus linteus homogenate to =4.5, adding 0.1-5% of cellulase, performing enzymolysis in a water bath kettle at 45-50 ℃ for 48-60 h at constant temperature until no obvious precipitate is formed in the homogenate, and sterilizing at 115 ℃ for 15min to obtain Phellinus linteus enzymatic hydrolysate.
7. The method for preparing the antioxidant metabiotic compound fermentation liquid according to claim 4, wherein the inoculation amount in the step (3) for the primary lactic acid bacteria seed culture is 1% -5%, and the thallus density in the secondary lactic acid bacteria seed liquid after fermentation is more than or equal to 10 9 cfu/ mL。
8. The method for preparing the antioxidant metazoan compound fermentation broth as claimed in claim 4, wherein the preparation of the metazoan compound fermentation broth in step (4): the phellinus igniarius enzymolysis liquid is used as a fermentation culture medium, and the volume ratio of the lactobacillus secondary seed liquid to the phellinus igniarius enzymolysis liquid is 3-5%.
9. An antioxidant metazoan complex preparation comprising the antioxidant metazoan complex fermentation broth of claim 1.
10. Use of the antioxidant metazoan complex fermentation broth of claim 1 or the antioxidant metazoan complex formulation of claim 9 for antioxidation.
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