CN115058370B - Antioxidant metancholia composite fermentation liquid, preparation method and application - Google Patents
Antioxidant metancholia composite fermentation liquid, preparation method and application Download PDFInfo
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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 metagenesis industry.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to an antioxidant metaplasia 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 realize the technical effects, the invention adopts the following technical scheme:
an antioxidant metazoan compound fermentation liquid comprises Phellinus igniarius and lactobacillus, specifically, phellinus igniarius is used as a single fermentation substrate, lactobacillus is used as a fermentation strain, and a product is obtained by compound fermentation.
Preferably, the thallus density in the compound 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 has been deposited at the chinese collection center on 3 months and 18 days 2015 at the deposition site: china Wuhan university, the preservation number is CCTCC NO: m2015126; a strain of Lactobacillus plantarum having high antioxidant activity and its use has been disclosed in the patent application No.: 201510419415.7, filed: 2015.07.16) In (1).
Lactobacillus acidophilus (C.acidophilus: (C.acidophilus)Lactobacillus acidophilus) PBIL2-003 has been deposited at the China center for type culture Collection at 28/4.2018 with 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)Lactobacillus rhamnosus) PBIL3-003 has been deposited at the China center for type culture Collection at 28/4.2018 with 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 metagenesis composite fermentation liquid, which specifically comprises the following steps:
(1) Preparing a phellinus igniarius fermentation liquid: activating Phellinus linteus strain, and performing amplification culture 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; 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 broth in the step (1) is as follows: sucking a 0.3 to 0.4mLPDA broth culture medium by using a sterile suction pipe, dripping the broth culture medium into an ampoule bottle of phellinus igniarius strain, and slightly oscillating the broth culture medium to dissolve the freeze-dried strain into a suspension shape; taking about 0.2mL of the thallus suspension, and transplanting the thallus suspension into a PDA agar culture medium to culture for 7 to 10 days at 25 to 28 ℃ until the surface of the culture medium is full of hyphae;
take 1cm 3 The above P full of hyphaeInoculating DA agar culture medium into PDA broth culture medium, and culturing at 25-28 deg.C for 5-7 d to form a large amount of dense mycelium pellets to obtain Phellinus Linteus 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 to 5 days at the temperature of 25 to 28 ℃ until a large number of dense mycelium pellets are formed, and clearing the culture solution to obtain the phellinus igniarius fermentation liquid.
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 to 7.0, and the potato is sterilized at the temperature of 121 ℃ for 20 min.
The PDA agar culture medium comprises the following main components: 0.3 percent of potato flour, 0.2 percent of monopotassium phosphate, 2 percent of glucose and 1.5 percent of agar, wherein the initial pH value is 6.0 to 7.0, the temperature is 121 ℃, and the sterilization is carried out for 20 min.
Preferably, the specific preparation method of the phellinus igniarius enzymatic hydrolysate in the step (2) comprises the steps of adjusting the pH of the phellinus igniarius homogenate to be =4.5, adding 0.1-5% of cellulase, carrying out constant-temperature enzymolysis for 48-60h in a water bath kettle at 45-50 ℃ until no obvious precipitate exists in the homogenate, and then sterilizing at 115 ℃ for 15min to obtain the phellinus igniarius enzymatic hydrolysate.
Preferably, the inoculation amount of the lactobacillus primary seed culture solution in the step (3) is 1 to 5 percent; 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 at 25 to 40 ℃ for 22 to 26h; the culture conditions of the first-stage seed liquid are as follows: standing and culturing at 25 to 40 ℃ for 12 to 24h;
the conditions of fermentation culture are as follows: carrying out intermittent shaking culture for 48 to 96h under the condition of 25 to 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, 1g of tween-80, 2g of 1.5% agar, 1L of purified water, pH6.5, 121 ℃, and sterilizing for 20 min; the first-order seed culture medium and the fermentation culture medium are both modified M6 culture media without agar.
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 culturing conditions of the metazoan complex fermentation broth in the step (4) are as follows: standing and culturing for 24 to 36h under the condition of 37 to 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 metaplasia compound preparation is powder, and the antioxidant metaplasia compound preparation is prepared by spray drying the metaplasia compound fermentation liquor prepared in the step (4);
the semi-solid state form of the antioxidant metaplasia compound preparation is prepared by removing partial water from the metaplasia 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 factor 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 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 protein carbonyl content in 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 post-growth-element compound fermentation liquor prepared by the lactobacillus acidophilus PBIL2-0013 in the sample group is very different from that of the lactobacillus acidophilus PBIL2-0013 in the blank group; the antioxidant performance of the post-growth factor compound fermentation liquor prepared by the lactobacillus rhamnosus PBIL3-0013 of the sample group is very different from that of the lactobacillus rhamnosus PBIL3-0013 of 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; the # indicates that the P is less than 0.01 when compared with the model control group, namely, the difference is extremely significant; the green color and the green color are compared with the positive color and have the respective color indications of P < 0.05, namely, the difference is remarkable, and the green color indicate the P < 0.01, namely, the difference is remarkable; xxx represents that the difference is very significant when compared with phellinus igniarius enzymatic hydrolysate, wherein P is less than 0.01.
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 to 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 percent of potato flour, 0.2 percent of monopotassium phosphate, 2 percent of glucose and 1.5 percent of agar, wherein the initial pH value is 6.0 to 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 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, 1g of tween-80, 2g of 1.5% agar, 1L of purified water, pH6.5, 121 ℃, and sterilizing for 20 min; the first-order seed culture medium and the fermentation culture medium are both modified M6 culture media without agar.
The culture activation conditions of the lactic acid bacteria are as follows: culturing at 25-40 ℃ for 22-26h;
the culture conditions of the first-stage seed liquid are as follows: standing and culturing for 12 to 24h under the condition of 25 to 40 ℃;
the culture conditions of the secondary seed liquid are as follows: carrying out intermittent shaking culture for 48 to 96h under the condition of 25 to 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 0.3 to 0.4mLPDA broth culture medium by using a sterile suction pipe, dripping the broth culture medium into an ampoule bottle of phellinus igniarius strain, and slightly oscillating the culture medium to dissolve the freeze-dried strain into a suspension shape; taking about 0.2mL of the thallus suspension, transplanting the thallus suspension into a PDA agar culture medium, and culturing for 7 to 10 days under the conditions of 25 to 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 d to form a large amount of dense mycelium pellets, i.e. Phellinus Linteus seed culture solution;
(3) Preparation of phellinus igniarius fermentation liquor: inoculating a phellinus igniarius seed culture solution into a PDA broth culture medium with the inoculation amount of 10%, carrying out shake culture at the temperature of 25-28 ℃, wherein the stirring speed is 100r/min, and culturing for 3-5 d until a large number of dense mycelium pellets are formed and the culture solution becomes clear, so as 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 suspension, namely phellinus igniarius homogenate; regulating the pH of the phellinus igniarius homogenate to be =4.5, then adding 0.1-5% of cellulase into the phellinus igniarius homogenate, carrying out constant-temperature enzymolysis in a water bath kettle at 45-50 ℃ for 48-60h until no obvious precipitate exists in the homogenate, and carrying out sterilization at 115 ℃ for 15min to obtain a phellinus igniarius enzymatic hydrolysate;
(5) Preparing a lactobacillus secondary seed solution: culturing and activating each strain of lactobacillus, selecting 1-5% of strains, performing primary seed culture to obtain primary seed liquid, performing secondary fermentation culture to obtain milkAcid bacteria secondary seed liquid with thallus density 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 at 37-38 ℃ for 24-36h 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 element 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 a fermentation liquid, the fermentation liquid and the phellinus igniarius homogenate are counted as blank groups, and the metaplastic composite fermentation liquid prepared by the Lactobacillus plantarum BLCC2-0015, the Lactobacillus acidophilus PBIL2-003 and the Lactobacillus rhamnosus PBIL3-003 according to the method of the embodiment 1 and the mixture and the phellinus igniarius enzymolysis liquid are respectively recorded as a sample group. 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 clearance rate: shanghai enzyme-linked Biotechnology Limited-hydroxy radical scavenging test kit;
SOD content: shanghai enzyme-linked Biotechnology Limited-superoxide dismutase (SOD) test kit;
content of GSH: shanghai enzyme-linked Biotechnology Limited-reduced Glutathione (GSH) test kit;
as can be seen from fig. 1 to 4, the oxidation resistance of the phellinus linteus homogenate 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 metagenesis 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 metagenesis compound fermentation liquid prepared by the invention has obvious synergistic effect on the oxidation resistance compared with the phellinus igniarius enzymolysis liquid and each lactobacillus fermentation liquid.
Comparative example 1
Selecting lactobacillus plantarum L1-019, lactobacillus acidophilus L2-001 and lactobacillus rhamnosus L3-001 as control strains to prepare post-growth composite fermentation liquor respectively according to the method in the embodiment 1, and detecting the antioxidant activity of the post-growth composite 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 graphs in FIGS. 5 to 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, postnatal 1, postnatal 2 and postnatal 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 every day, physiological saline is given to a blank group and a model group, the positive group is subjected to intragastric administration-Vc, the mice of the other experimental groups are subjected to intragastric administration by utilizing phellinus igniarius enzymatic hydrolysate, prebiotic 1, prebiotic 2 and prebiotic 3 respectively, the intragastric administration dosage is 10mL/kg BW, after continuous administration for 30d, fasting is not prohibited for 16h, 0.4mL physiological saline is given to the blank group, 0.4mL of 56-degree Hongxing Erguotou is given to the intragastric administration of the other groups of mice, and after 6h, the content of GSH, SOD, MDA and protein carbonyl in the blood serum of the mice is detected by adopting detection kits of Shanghai enzyme-linked biology Limited company respectively, and the detection results are shown in figures 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 fig. 10, the contents of SOD and GSH in the serum of the mice treated by the model group are the least, which indicates that the content of SOD and GSH in the serum of the mice can be reduced by the red star Erguotou, and the mice are damaged by peroxidation; the contents of SOD and GSH in the blood serum of the mice treated by other treatment groups except the group of postbiotic 1, postbiotic 2 and postbiotic 3 are reduced to different degrees, but the reduction degrees are all 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 blood serum of the mice in the postnatal 1 group has no obvious difference with the blank group, and the SOD content in the blood serum of the mice in the postnatal 2 group and the postnatal 3 group has slight difference with the blank group, which shows that the SOD level of the postnatal 1 group is better than that of the postnatal 2 group and the postnatal 3 group in the aspect of improving the SOD level of the organism, 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 taken directly or drunk to improve the oxidation resistance of organism.
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 all belong to the protection scope of the present application.
Claims (9)
1. The antioxidant metazoan compound fermentation liquid is characterized by comprising phellinus igniarius and lactic acid bacteria, wherein the phellinus igniarius is used as a single fermentation substrate, the lactic acid bacteria are used as a product of compound fermentation of a fermentation strain, and the lactic acid bacteria are selected from lactobacillus which has a preservation number of CCTCC NO: lactobacillus plantarum of M2015126 (Lactobacillus plantarum) BLCC2-0015, preservation number CCTCC NO: m2018208 Lactobacillus acidophilus (Lactobacillus acidophilus) PBIL2-003 and the preservation number is CCTCC NO: m2018206 Lactobacillus rhamnosus (L.), (Lactobacillus rhamnosus) PBIL 3-003.
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 method for preparing the antioxidant metazoan composite fermentation broth as claimed in claim 1 or 2, which is characterized by comprising 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; 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 fermentation culture on the lactobacillus strain to obtain a secondary lactobacillus 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.
4. The method for preparing the antioxidant metazoan compound fermentation broth according to claim 3, wherein the specific preparation process of the phellinus linteus fermentation broth in the step (1) is as follows:
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, and transplanting the thallus suspension into a PDA agar culture medium for 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.
5. The method for preparing the antioxidant metabiotic compound fermentation liquid according to claim 3, wherein the Phellinus linteus enzymolysis liquid in the step (2) is specifically prepared by adjusting the pH of Phellinus linteus homogenate to =4.5, adding 0.1 to 5% of cellulase, carrying out enzymolysis in a water bath kettle at a constant temperature of 45 to 50 ℃ for 48 to 60h until no obvious precipitate is formed in the homogenate, and then carrying out sterilization at 115 ℃ for 15min to obtain the Phellinus linteus enzymolysis liquid.
6. The preparation method of the antioxidant metaplasia compound fermentation liquid according to claim 3, wherein the inoculation amount of the primary seed culture of the lactic acid bacteria in the step (3) is 1% -5%, and the thallus density in the secondary seed liquid of the lactic acid bacteria after the fermentation is finished is more than or equal to 10 9 cfu/mL。
7. The method for preparing the antioxidant metaplasia compound fermentation broth according to claim 3, wherein the preparation of the metaplasia compound fermentation broth in the step (4) comprises the following steps: 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%.
8. An antioxidant metazoan complex preparation comprising the antioxidant metazoan complex fermentation broth of claim 1.
9. The antioxidant metagenin compound fermentation broth of claim 1 or the antioxidant metagenin compound preparation of claim 8 is used for antioxidation.
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