Ganoderma lucidum fermentation liquor composition with anti-oxidation effect as well as preparation method and application of ganoderma lucidum fermentation liquor composition
By mixing Ganoderma lucidum fermentation broth with Jiner fermentation broth in a specific volume ratio and using fermentation technology, a composition with better antioxidant ability was prepared, which solved the problem of rarely reported impact of compound antioxidant effects in the prior art, and achieved the goal of significantly improving the antioxidant effect.
Patent Information
- Application Number
- CN202411807476.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-10
- Publication Date
- 2025-03-28
AI Technical Summary
In the prior art, the impact of the antioxidant combination of Ganoderma lucidum and golden ear on antioxidant effects has been rarely reported, and it is difficult to effectively combine the antioxidant mechanisms of both.
By mixing Ganoderma lucidum fermentation broth with Jiner fermentation broth by volume ratio (0.5-2): 1, using the fermentation technology of Lactobacillus acidophilus and Lactobacillus plantarum, a composition with better antioxidant ability was prepared.
The optimal ratio of Ganoderma lucidum fermentation broth and Jiner fermentation broth is achieved, which significantly improves the superoxide anion radical scavenging rate and has better antioxidant effect.
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Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of traditional Chinese medicine fermentation, and particularly relates to a ganoderma lucidum fermentation liquid composition with antioxidant effect, and a preparation method and application thereof. Background Art
[0002] Ganoderma lucidum, known as the "immortal grass", has a long history in my country. Recent studies have shown that the active ingredients in Ganoderma lucidum, such as polysaccharides, triterpenoid compounds and amino acid peptides, have antioxidant, hypoglycemic and immunomodulatory effects. Ganoderma lucidum polysaccharides can terminate free radical chain reactions, improve cell vitality, reshape the activity of endogenous antioxidant enzymes such as superoxide dismutase (SOD) and catalase (CAT), thereby reducing the content of oxidation-related end products such as lactate dehydrogenase (LDH) and malondialdehyde (MDA), and play an antioxidant role. Ganoderma lucidum polysaccharides can regulate the metabolism of carbohydrates and lipids in the body, improve insulin sensitivity, reduce the level of pancreatic β-cell apoptosis, avoid insulin resistance, and control the body's blood sugar level in a stable state. Ganoderma triterpenoids E and ganoderic acid F protect cells from oxidative damage caused by H2O2 by scavenging 2,2-azino-bis(3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt (ABTS) and oxygen radical absorbance capacity (ORAC); ganoderic acid can lower blood sugar by inhibiting α-glucosidase activity.
[0003] Tremella fuciformis is an edible and medicinal fungus with high nutritional and medicinal value. It has multiple biological activities such as lowering blood sugar, lowering blood lipids, anti-oxidation, anti-inflammatory, anti-coagulation, and enhancing immunity. Studies have shown that Tremella fuciformis polysaccharides have strong antioxidant activity and can effectively reduce the negative impact of free radicals on cells. Tremella fuciformis fermentation broth polysaccharides have a certain reducing ability and strong activity in scavenging DPPH and hydroxyl free radicals. The polysaccharide TAP and the decomposition product H-TAP in Tremella fuciformis fruiting body can accelerate the metabolism of glucose in the blood; Tremella fuciformis fruiting body polysaccharides, mycelium polysaccharides, and extracellular polysaccharides all have the function of lowering blood sugar.
[0004] Both Ganoderma lucidum and Tremella fuciformis have antioxidant effects, but their mechanisms of action may be different. However, there are few reports on the effects of the combination of these two traditional Chinese medicines on antioxidant effects. Summary of the invention
[0005] To this end, the present invention provides a ganoderma lucidum fermented liquid composition with antioxidant efficacy, a preparation method and an application thereof. The composition uses ganoderma lucidum fermented liquid and Tremella fuciformis fermented liquid as raw materials. When the volume ratio of the ganoderma lucidum fermented liquid to the Tremella fuciformis fermented liquid is (0.5-2):1, the composition has a better antioxidant capacity, with the best superoxide anion radical scavenging rate.
[0006] The technical contents of the present invention are:
[0007] In one aspect, the present invention provides a ganoderma lucidum fermentation broth composition with antioxidant efficacy, comprising Tremella fuciformis fermentation broth and ganoderma lucidum fermentation broth.
[0008] Preferably, the volume ratio of the Tremella fuciformis fermentation liquid to the Ganoderma lucidum fermentation liquid is (0.5-2):1; further preferably, the volume ratio of the Tremella fuciformis fermentation liquid to the Ganoderma lucidum fermentation liquid is 1:1.
[0009] Preferably, the Ganoderma lucidum fermentation liquid is prepared by inoculating Lactobacillus acidophilus with the supernatant obtained by enzymatic hydrolysis, enzyme inactivation and solid-liquid separation of Ganoderma lucidum for fermentation, wherein the Lactobacillus acidophilus is the Lactobacillus acidophilus with the deposit number of CGMCC1.12735.
[0010] Preferably, the inoculation amount of Lactobacillus acidophilus is 1-3%; further preferably, the inoculation amount of Lactobacillus acidophilus is 2%.
[0011] More preferably, the viable count of Lactobacillus acidophilus is 1×10 8 CFU / mL and above.
[0012] Preferably, the enzyme preparation selected for enzymolysis includes papain, cellulase and pectinase.
[0013] Preferably, the added amount of the enzyme preparation is 0.5-1.0%; further preferably, the added amount of the enzymatic hydrolysis is 0.8%; further preferably, the weight ratio of papain, cellulase and pectinase is 1:2:2.
[0014] Preferably, the enzymatic activity of the cellulase is 3000-4000 U / g, the enzymatic activity of the pectinase is 10,000-30,000 U / g, and the enzymatic activity of the papain is 80,000-120,000 U / g; further preferably, the enzymatic activity of the cellulase is 3500 U / g, the enzymatic activity of the pectinase is 20,000 U / g; and the enzymatic activity of the papain is 100,000 U / g.
[0015] Preferably, the preparation method of the Ganoderma lucidum fermentation liquid is as follows: (1) adding an enzyme preparation to a mixture of Ganoderma lucidum fruiting bodies and water for enzymolysis and inactivating the enzyme to obtain a Ganoderma lucidum hydrolyzate; (2) separating the Ganoderma lucidum hydrolyzate from solid and liquid, inoculating Lactobacillus acidophilus into the obtained supernatant for fermentation, and separating the solid and liquid to obtain the Ganoderma lucidum fermentation liquid.
[0016] Further preferably, the solid-liquid ratio of the Ganoderma lucidum fruiting body to water is 1:(10-20); further preferably, the solid-liquid ratio of the Ganoderma lucidum fruiting body to water is 1:15.
[0017] Further preferably, the enzymatic hydrolysis conditions are: enzymatic hydrolysis temperature 40-60°C, enzymatic hydrolysis time 2-4h; further preferably, enzymatic hydrolysis temperature 50°C, enzymatic hydrolysis time 3h.
[0018] Further preferably, the enzyme inactivation conditions are: enzyme inactivation temperature 90-100°C, time 8-12 min; further preferably, the enzyme inactivation temperature 90°C, time 10 min.
[0019] More preferably, the fermentation temperature is 35-40°C, and the fermentation time is 48-72h; even more preferably, the fermentation temperature is 37°C, and the fermentation time is 60h.
[0020] Preferably, the Tremella fuciformis fermented liquid is prepared by extracting Tremella fuciformis and water and inoculating with Lactobacillus plantarum for fermentation, wherein the Lactobacillus plantarum is the Lactobacillus plantarum with a deposit number of CGMCC1.1856.
[0021] Preferably, the solid-liquid ratio of the golden ear to water is 1:(40-60), and further preferably, the solid-liquid ratio of the golden ear to water is 1:50.
[0022] Preferably, the inoculation amount of Lactobacillus plantarum is 1-3%, and further preferably, the inoculation amount of Lactobacillus plantarum is 2%; further preferably, the viable count of Lactobacillus plantarum is 1×10 8 CFU / mL and above.
[0023] Preferably, the preparation method of the golden ear fermentation liquid is as follows: (1) cutting fresh golden ear into pieces, mixing them evenly with water in a solid-liquid ratio of 1:(40-60), and extracting to obtain a golden ear water extract; (2) sterilizing the golden ear water extract, inoculating it with plant lactobacillus, and fermenting it. After the fermentation is completed, the solid and liquid are separated, and the supernatant is the golden ear fermentation liquid.
[0024] Preferably, the extraction temperature is 80-90°C and the extraction time is 40-50 min; further preferably, the extraction temperature is 85°C and the extraction time is 40 min.
[0025] Preferably, the sterilization temperature is 120-125°C and the time is 12-17 min; further preferably, the sterilization temperature is 121°C and the time is 15 min.
[0026] Preferably, the fermentation temperature is 35-40°C and the fermentation time is 48-72h; further preferably, the fermentation temperature is 37°C and the fermentation time is 72h.
[0027] In another aspect, the present invention provides a method for preparing the Ganoderma lucidum fermentation broth composition, which comprises mixing the Ganoderma lucidum fermentation broth and Tremella fuciformis fermentation broth.
[0028] In another aspect, the present invention provides use of the above composition in preparing antioxidant products.
[0029] In another aspect, the present invention provides an antioxidant product, which comprises the above-mentioned Ganoderma lucidum fermentation broth composition and food and pharmaceutically acceptable auxiliary materials.
[0030] Compared with the prior art, the present invention has the following advantages:
[0031] 1 The present invention provides a ganoderma lucidum fermentation liquid composition, comprising Tremella fuciformis fermentation liquid and Ganoderma lucidum fermentation liquid, and the applicant has found that when the volume ratio of Tremella fuciformis fermentation liquid to Ganoderma lucidum fermentation liquid is (0.5-2):1, it has a good superoxide anion free radical scavenging rate. When the volume of Tremella fuciformis fermentation liquid and Ganoderma lucidum fermentation liquid is 1:1, it has a better superoxide anion free radical scavenging rate, and can be applied to antioxidant products. DETAILED DESCRIPTION
[0032] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of the present invention.
[0033] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meanings as those generally understood by those of ordinary skill in the art to which the present invention belongs.
[0034] The present invention is further described below by way of specific examples. If no specific experimental steps or conditions are specified in the examples, the conventional steps or conditions described in the literature in the art can be used. All reagents without manufacturer specified can be obtained through conventional commercial channels.
[0035] In the following examples, the source information of some products is as follows:
[0036] Dendrobium officinale and Tremella fuciformis were from Zhejiang Fangge Pharmaceutical Co., Ltd. of Hu Qingyu Tang Group; Ganoderma lucidum was from Fujian Xianzhilou Biotechnology Group; cellulase was purchased from Xiasheng (Beijing) Biotechnology Development Co., Ltd. with an enzyme activity of 3500 U / g, pectinase was purchased from Henan Zhongchen Biotechnology Co., Ltd. with an enzyme activity of 20,000 U / g, and papain was purchased from Nanning Shanwan Biotechnology Co., Ltd. with an enzyme activity of 100,000 U / g.
[0037] Lactobacillus acidophilus was purchased from China General Microorganism Culture Collection Center with a collection number of CGMCC1.12735; Lactobacillus plantarum with a collection number of ACCC 11016 was purchased from China Agricultural Microorganism Culture Collection Management Center; Lactobacillus plantarum with a collection number of CGMCC1.1856 was purchased from China General Microorganism Culture Collection Center.
[0038] Preparation Example 1 Activation of Fermentation Bacteria
[0039] Take 52.2 g of MRS broth, add 1000 mL of distilled water, stir and heat to boil until completely dissolved, divide into portions, sterilize under high pressure at 121°C for 15 min, and set aside.
[0040] The lactic acid bacteria strains (Lactobacillus acidophilus with a preservation number of CGMCC1.12735, Lactobacillus plantarum with a preservation number of ACCC 11016, and Lactobacillus plantarum with a preservation number of CGMCC1.1856) stored in a -80°C refrigerator were taken out and inoculated into MRS medium at a 1% inoculum volume. The culture was cultured at 37°C for 24 h. The activation method was repeated three times until the viable cell count was 1.0×10 8 CFU / mL and above.
[0041] MRS broth was purchased from Hangzhou Best Biotechnology Co., Ltd., and its culture medium formula was 10.0 g / L peptone, 5.0 g / L beef extract powder, 4.0 g / L yeast extract powder, 20.0 g / L glucose, 2.0 g / L dipotassium hydrogen phosphate, 2 g / L triammonium hydrogen citrate, 5.0 g / L sodium acetate, 0.2 g / L magnesium sulfate, 0.05 g / L manganese sulfate, and 1 g / L Tween-80.
[0042] Preparation Example 2 Preparation of Ganoderma lucidum fermentation liquid
[0043] The fruiting bodies of Ganoderma lucidum were dried and crushed, and Ganoderma lucidum and water were mixed evenly at a solid-liquid ratio of 1:20 (g:mL) and then pulped to obtain Ganoderma lucidum slurry. A composite enzyme solution (papain: cellulase: pectinase = 1:2:2) was added to the Ganoderma lucidum slurry and mixed evenly. The enzymatic hydrolysis conditions were 0.8% composite enzyme dosage, enzyme solution pH 4.5, and enzymatic hydrolysis at 150r / min for 3h in a 50℃ constant temperature shaker to obtain Ganoderma lucidum enzymatic hydrolyzate. The enzyme was inactivated in a 90℃ water bath for 10 min, cooled and centrifuged at 5000×g for 10 min, and the supernatant was taken for fermentation. The activated Lactobacillus acidophilus with a preservation number of CGMCC1.12735 was inoculated into the supernatant at an inoculation rate of 2%, cultured at 37℃ for 60h, and the supernatant was centrifuged and pasteurized to obtain Ganoderma lucidum fermentation liquid.
[0044] Preparation Example 3 Preparation of Dendrobium officinale fermentation liquid
[0045] Take the stems of Dendrobium officinale, dry them, crush them, and pass them through a 60-mesh sieve. Mix them with pure water at a solid-liquid ratio of 1:15 (g:mL). The enzymatic hydrolysis conditions are 4% cellulase addition (based on the mass of Dendrobium officinale), enzymatic hydrolysis temperature of 50℃, and enzymatic hydrolysis for 4h. After the enzymatic hydrolysis is completed, inactivate them in a 90℃ water bath for 10 min, cool them, and obtain the enzymatic hydrolyzate of Dendrobium officinale.
[0046] The activated Lactobacillus plantarum culture liquid with the preservation number of ACCC 11016 was inoculated into the enzymatic hydrolyzed liquid of Dendrobium officinale at a 2% inoculation rate, and the fermentation was carried out at a constant temperature of 37°C for 48 hours and a fermentation speed of 150 r / min to obtain the fermented Dendrobium officinale juice, which was then pasteurized. Finally, the supernatant was taken as the fermented liquid of Dendrobium officinale at 4800 r / min for 10 minutes.
[0047] Preparation Example 4 Preparation of Tremella Fermentation Broth
[0048] Fresh golden ear was cut into small pieces and mixed with pure water at a solid-liquid ratio of 1:50 (g:mL), extracted at 85°C for 40 min, and cooled after extraction to obtain golden ear water extract.
[0049] The Tremella fuciformis water extract was sterilized at 121°C for 15 minutes, then inoculated with 2% activated Lactobacillus plantarum culture with a preservation number of CGMCC1.1856, and fermented at 37°C for 72 hours. After the fermentation, the Tremella fuciformis fermented liquid was obtained, and then pasteurized. Finally, centrifuged at 5000×g for 15 minutes, and the supernatant was taken, which was the Tremella fuciformis fermented liquid.
[0050] Example Antioxidant Composition Preparation
[0051] This embodiment provides a composition with antioxidant effect, which is obtained by mixing the Ganoderma lucidum fermentation liquid obtained in Preparation Example 2, the Tremella fuciformis fermentation liquid obtained in Preparation Example 4, or the Dendrobium officinale fermentation liquid obtained in Preparation Example 3 in a certain proportion. Specific raw materials and volume ratios are shown in Table 1.
[0052] Table 1 Composition and proportion of Dendrobium officinale fermentation liquid composition
[0053]
[0054] Experimental Example 1 Determination of superoxide anion free radical scavenging rate
[0055] The samples to be tested are: the Ganoderma lucidum fermentation liquid obtained in Preparation Example 2, the Tremella fuciformis fermentation liquid obtained in Preparation Example 4, the Dendrobium officinale fermentation liquid obtained in Preparation Example 3, and the combination obtained in Examples 1-9.
[0056] Pipette 1mL of the sample to be tested and 4.5mL of 0.05M Tris-HCL (pH=8.2) solution, mix well, and react in a water bath at 25℃ for 20min. Add 0.3mL of 0.05M pyrogallol, and react in a water bath at 25℃ for 4min. Add 1mL of 8mmol / L HCL to terminate the reaction, measure the absorbance at a wavelength of 325nm, and calculate the clearance rate according to the following formula:
[0057] R = (1 - )×100%
[0058] Where, R: superoxide anion free radical scavenging rate; A: absorbance of the experimental group; B: absorbance of the control group (pure water instead of pyrogallol); C: absorbance of the blank group (pure water instead of the sample)
[0059] See Table 2 for specific results.
[0060] Table 2 Superoxide anion radical scavenging rate
[0061]
[0062] Note: Different letters indicate significant differences among different groups.
[0063] As shown in Table 2, the superoxide anion radical scavenging rate of the Ganoderma lucidum fermented liquid obtained in Preparation Example 2 is the lowest, but the superoxide anion radical scavenging rate of the Tremella fuciformis fermented liquid and the Ganoderma lucidum fermented liquid when the ratio is (0.5-2):1 is higher than that of the single Ganoderma lucidum fermented liquid and Tremella fuciformis fermented liquid, and is significant; and when the volume ratio of Tremella fuciformis fermented liquid to Ganoderma lucidum fermented liquid is 1:1, it has the best superoxide anion radical scavenging rate.
[0064] The superoxide anion radical scavenging rate of the compound mixture of Tremella fuciformis fermentation liquid and Dendrobium candidum fermentation liquid or Ganoderma lucidum fermentation liquid and Dendrobium candidum fermentation liquid is lower than the superoxide anion radical scavenging rate of Tremella fuciformis fermentation liquid and Ganoderma lucidum fermentation liquid when the ratio is (0.5-2):1, and it is significant. Moreover, when the ratio of Ganoderma lucidum fermentation liquid and Dendrobium candidum fermentation liquid is (0.5-1):1, the superoxide anion radical scavenging rate is lower than the superoxide anion radical scavenging rate of Ganoderma lucidum fermentation liquid or Dendrobium candidum fermentation liquid alone, indicating that not all raw materials with antioxidant properties can enhance the antioxidant capacity of the composition, and the Tremella fuciformis fermentation liquid and Ganoderma lucidum fermentation liquid provided by the present invention are the best combination.
[0065] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.
Claims
1. A Ganoderma lucidum fermented liquid composition with antioxidant effect, characterized in that: The invention comprises Tremella fuciformis fermented liquid and Ganoderma lucidum fermented liquid, wherein the volume ratio of the Tremella fuciformis fermented liquid to the Ganoderma lucidum fermented liquid is (0.5-2):
1.
2. The composition according to claim 1, characterized in that The volume ratio of the Tremella fuciformis fermented liquid to the Ganoderma lucidum fermented liquid is 1:
1.
3. The composition according to claim 1, characterized in that The ganoderma lucidum fermentation liquid is obtained by inoculating the supernatant obtained by enzymolysis, sterilization and solid-liquid separation of ganoderma lucidum with lactobacillus acidophilus for fermentation, wherein the lactobacillus acidophilus is the lactobacillus acidophilus with the preservation number of CGMCC1.12735.
4. The composition according to claim 1, characterized in that The Tremella fuciformis fermented liquid is obtained by extracting Tremella fuciformis and water and inoculating Lactobacillus plantarum to ferment, wherein the Lactobacillus plantarum is the Lactobacillus plantarum with a preservation number of CGMCC1.1856.
5. The composition according to claim 3, characterized in that The enzyme preparations selected for enzymolysis of Ganoderma lucidum include cellulase, pectinase and papain.
6. The composition according to claim 5, characterized in that The added amount of the enzyme preparation is 0.5-1.0%, and the inoculation amount of Lactobacillus acidophilus is 1-3%.
7. The composition according to claim 3 or claim 4, characterized in that The fermentation temperature is 35-40°C and the fermentation time is 48-72h.
8. A method for preparing the composition according to any one of claims 1 to 7, characterized in that: Just mix the Tremella fuciformis fermented liquid and the Ganoderma lucidum fermented liquid.
9. Use of the composition according to any one of claims 1 to 7 in the preparation of antioxidant products.
10. An antioxidant product, characterized in that: The invention comprises the ganoderma lucidum fermentation liquid composition according to any one of claims 1 to 7 and pharmaceutically or food acceptable auxiliary materials.