Antioxidant ganoderma lucidum spore oil composition and preparation method thereof

Abstract

The invention discloses an antioxidant ganoderma lucidum spore oil composition and a preparation method thereof, and belongs to the technical field of edible and medicinal fungus extraction. The preparation method of the antioxidant ganoderma lucidum spore oil composition comprises the following steps: A1, swelling ganoderma lucidum spore powder with water, and performing ultrasonic extraction to obtain a first supernatant and a first material residue; a2, drying the first material residue, performing high-pressure supercritical carbon dioxide extraction, and separating to obtain ganoderma lucidum spore oil and second material residue; a3, separating out ganoderma lucidum spore powder protein from the first supernate; a4, carrying out hot water extraction on the second material residue to obtain a second supernatant and a third material residue; a5, mixing the third material residue with ganoderma lucidum spore powder protein, and performing enzymolysis extraction and centrifugation to obtain third supernate; a6, preparing an oil phase by taking ganoderma lucidum spore oil as a raw material; dissolving and homogenizing a wall material, adding xanthan gum, the second supernatant, the third supernatant and a lucid ganoderma extract, and continuously homogenizing to obtain a water phase; the water phase and the oil phase are mixed, homogenized, emulsified and then subjected to spray drying, and the antioxidant ganoderma lucidum spore oil composition is obtained.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of edible and medicinal fungus extraction, in particular to an antioxidant ganoderma lucidum spore oil composition and a preparation method thereof. BACKGROUND

[0002] Ganoderma lucidum has a long history of medicinal use and a wide range of biological activities as a traditional and valuable Chinese medicinal material. Ganoderma lucidum spores, which are reproductive cells of Ganoderma lucidum, concentrate the essence of Ganoderma lucidum and are rich in active substances such as Ganoderma lucidum polysaccharides, Ganoderma lucidum triterpenes, Ganoderma lucidum spore oil, and Ganoderma lucidum spore proteins. Among them, Ganoderma lucidum polysaccharides and Ganoderma lucidum spore proteins have significant immunomodulatory, anti-tumor, and anti-aging effects, while Ganoderma lucidum spore oil exhibits excellent antioxidant and liver-protecting effects due to its high content of unsaturated fatty acids and triterpenes. Therefore, developing high-value functional products from Ganoderma lucidum spores has broad market prospects.

[0003] Currently, the extraction of Ganoderma lucidum spore oil mainly uses supercritical carbon dioxide extraction technology, which is mild and solvent-free and can effectively extract fat-soluble components such as triterpenes and unsaturated fatty acids from Ganoderma lucidum spores. However, the Ganoderma lucidum spore oil obtained by this method has low content of Ganoderma lucidum polysaccharides, Ganoderma lucidum chitin, and Ganoderma lucidum cellulose, and does not have all Ganoderma lucidum components. In addition, Ganoderma lucidum spore oil itself is rich in unsaturated fatty acids, which are prone to oxidation and rancidity during extraction and storage, leading to a decrease in antioxidant activity and the generation of undesirable flavors, affecting the quality and efficacy of the product.

[0004] In summary, it is necessary to develop a preparation method for an antioxidant Ganoderma lucidum spore oil composition that can fully utilize the active components such as polysaccharides and spore proteins in Ganoderma lucidum spores and obtain high antioxidant activity. SUMMARY

[0005] The purpose of the present application is to provide an antioxidant Ganoderma lucidum spore oil composition and a preparation method thereof to solve the technical problems mentioned in the background.

[0006] The technical solution to achieve the purpose of the present application is as follows: In a first aspect, the present application provides a preparation method for an antioxidant Ganoderma lucidum spore oil composition, comprising the following preparation steps: A1. Swelling Ganoderma lucidum spores with water and extracting multiple times by ultrasonic extraction to obtain first supernatant and first residue; A2. Drying the first residue and then performing high-pressure supercritical carbon dioxide extraction and separation to obtain Ganoderma lucidum spore oil and second residue; A3. Precipitating and separating Ganoderma lucidum spore protein from the first supernatant; A4. Hot water extraction of the second residue to obtain second supernatant and third residue; A5. The third residue is mixed with Ganoderma lucidum spore powder protein and subjected to enzymatic extraction, and the obtained extraction liquid is centrifuged to obtain supernatant, thereby obtaining a third supernatant; A6. The Ganoderma lucidum spore oil is mixed with a first emulsifier and dissolved and homogenized to obtain an oil phase; a wall material, water, a second emulsifier are dissolved and homogenized, and then xanthan gum, the second supernatant, the third supernatant, Ganoderma lucidum extract are added, and homogenization is continued to obtain an aqueous phase of an emulsion; the aqueous phase and the oil phase are mixed and homogenized for several times, and then subjected to spray drying, thereby obtaining an antioxidant Ganoderma lucidum spore oil composition with a microcapsule structure.

[0007] Further, the specific steps of step A1 are as follows: the Ganoderma lucidum spore powder is fully swelled according to a material-to-water ratio of 1 g: 20-25 mL, and the pH is adjusted to 8.98-9.02, and then ultrasonic extraction is performed at 20-30℃ and a power of 350-450 W for 25-35 min, followed by water bath at 40-50℃ for 55-65 min, and then cooled to room temperature and centrifuged at 5000 r / min for 20 min to collect the supernatant, and the precipitate is subjected to secondary extraction by repeating the foregoing steps, and the supernatants are combined and subjected to suction filtration, thereby obtaining a first supernatant and a first residue.

[0008] Further, the high-pressure supercritical carbon dioxide extraction is performed at a pressure of 50-65 MPa, an extraction temperature of 38-42℃, and for a time of 30-40 min.

[0009] Further, the specific steps of step A3 are as follows: the pH of the first supernatant is adjusted to the isoelectric point 2.0, and then settled at 0-4℃ for 2.5-3.5 h, and then centrifuged to collect the precipitate and washed with water to neutral, and then dried in a vacuum freeze dryer to obtain Ganoderma lucidum spore powder protein, which is stored at -20℃ for standby use.

[0010] Further, the hot water extraction is performed at a temperature of 90-100℃ for a time of 90-150 min, and a material-to-liquid ratio of 1 g: 20-30 mL.

[0011] Further, the enzyme used for the enzymatic hydrolysis is neutral protease, the enzyme-to-substrate ratio is 1600-3000:1, the pH of the enzymatic hydrolysis is 4.6-6, the temperature of the enzymatic hydrolysis is 45-50℃, and the time of the enzymatic hydrolysis is 45-180 min.

[0012] Further, the mass ratio of the Ganoderma lucidum spore oil, the first emulsifier, the wall material, the water, the second emulsifier, the xanthan gum, the second supernatant, the third supernatant, and the Ganoderma lucidum extract is 40: 0.35-0.45: 120: 500-550: 0.11-0.13: 0.23-0.25: 40-60: 40-45; wherein the first emulsifier is monoglyceride, and the second emulsifier is sucrose ester.

[0013] Further, the wall material is composed of 90 parts by weight of malt dextrin, 14-16 parts by weight of egg yolk powder, and 14-16 parts by weight of vegetable protein; the vegetable protein includes any one or a combination of two of soybean protein isolate and ganoderma spore powder protein.

[0014] In a second aspect, the present application provides an antioxidant ganoderma spore oil composition prepared by the preparation method of the antioxidant ganoderma spore oil composition of the first aspect.

[0015] In a third aspect, the present application provides the use of the antioxidant ganoderma spore oil composition of the second aspect in the preparation of special dietary foods, health foods, and medicines.

[0016] The above technical solution has the following beneficial effects: (1) The preparation steps of the antioxidant ganoderma spore oil composition of the present application include: First, the ganoderma spore powder is swelled with water and extracted by ultrasonic multiple times to obtain first supernatant and first residue; the water molecules are fully penetrated into the spores, the cell structure is moderately expanded, the local structure of the cell wall is gently destroyed by the cavitation effect and mechanical disturbance of the ultrasonic waves, and the macromolecular substances such as ganoderma spore protein which are easily soluble in water and sensitive to heat are preferentially dissolved into the water phase, so as to pre-separate and protect the active protein from denaturation and inactivation in the process of high pressure and high temperature; Then, the first residue is dried and extracted by high-pressure supercritical carbon dioxide and separated to obtain ganoderma spore oil and second residue; in this process, the supercritical carbon dioxide has excellent permeability and solubility selectivity, and can efficiently extract the lipid-soluble active ingredients ganoderma spore oil in an oxygen-free and non-polar environment, while minimizing the oxidation risk of unsaturated fatty acids; at this time, the components such as ganoderma polysaccharide and chitin in the extracted ganoderma spore oil are low, and most of the components such as ganoderma polysaccharide are retained in the second residue after extraction; Subsequently, the ganoderma spore protein is separated from the first supernatant by the precipitation method; the protein is recovered in a targeted manner by adjusting the pH; then, the second residue is immersed in hot water to further release the ganoderma polysaccharide remaining in the solid phase, to obtain second supernatant containing ganoderma polysaccharide and third residue; the hot water immersion can effectively dissolve the water-soluble polysaccharides such as beta-glucan, thereby improving the yield of polysaccharides; then, the third residue and the separated ganoderma spore protein are mixed and subjected to enzymatic extraction, and the supernatant is obtained after centrifugation of the extraction liquid, to obtain third supernatant containing ganoderma polysaccharide and antioxidant peptides with antioxidant activity; the spore protein and the third residue are hydrolyzed by neutral protease to generate small-molecule antioxidant peptides, which are helpful to extract the residual polysaccharides; Finally, the ganoderma lucidum spore oil is mixed and dissolved with the first emulsifier and homogenized to form an oil phase; the wall material, water and the second emulsifier are dissolved and homogenized, then xanthan gum is added as a stabilizer, and the second supernatant rich in polysaccharide and the third supernatant containing polysaccharide and antioxidant peptide and ganoderma extract are added and continue to be homogenized to form a uniform and stable emulsion water phase; finally, the water phase and the oil phase are mixed and then subjected to multiple homogenization emulsification, and then subjected to spray drying to finally obtain an antioxidant ganoderma lucidum spore oil composition with a microcapsule structure; the antioxidant ganoderma lucidum spore oil composition is composed of polysaccharide, protein hydrolysate and wall material to form a compact shell, and the ganoderma lucidum spore oil is wrapped therein, which not only realizes efficient co-loading of fat-soluble and water-soluble active ingredients, but also effectively blocks the oxidation attack of oxygen, light and other factors on the oil through physical isolation, thereby significantly improving the chemical stability; meanwhile, the introduced ganoderma polysaccharide and antioxidant peptide can produce a free radical scavenging synergistic effect with triterpenoids in the spore oil, thereby effectively improving the overall antioxidant performance of the composition. The antioxidant ganoderma lucidum spore oil composition prepared in the application can fully utilize active components such as polysaccharide and spore protein in ganoderma lucidum spores, and has high antioxidant activity and good stability of the ganoderma lucidum spore oil.

[0017] (2) The wall material of the ganoderma lucidum spore oil composition of the application comprises, by weight fraction: 90 parts of malt dextrin, 14-16 parts of egg yolk powder and 15-16 parts of plant protein; wherein the plant protein is selected from any one of soybean protein isolate and ganoderma spore powder protein, or a combination of the two; wherein the malt dextrin serves as a main film-forming matrix and has good plasticity and carrier preparation performance, which helps to form a uniform microcapsule structure; the egg yolk powder is rich in lecithin and has natural emulsifying property and film strengthening function, which can effectively improve the stability of the emulsion system and enhance the compactness of the microcapsule wall; the plant protein not only provides good interfacial emulsification capacity, but also participates in the construction of the microcapsule wall to improve the embedding efficiency and physical strength; when the plant protein is ganoderma spore powder protein, not only the recycling of self-origin active components is realized, but also the microcapsule wall itself is endowed with antioxidant activity, further enhancing the overall efficacy; the raw materials used in the wall material system are safe and non-toxic, meet the relevant regulations and quality requirements of special diet food, health food and pharmaceutical preparations, and are suitable for the development of high-activity and high-stability functional products; furthermore, the microcapsule wall material formed by the above components has good solubility, with a solubility of at least 86%, and can be quickly dissolved in water during application to release active components. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to make the content of the application more easily understood, the application will be further described in detail below according to specific embodiments and in conjunction with the drawings, in which Figure 1 The preparation process of the antioxidant ganoderma lucidum spore oil composition of an embodiment of the application is shown in the figure. Detailed Implementation

[0019] To better understand the above technical solutions, the following will provide a detailed explanation of the technical solutions in conjunction with the accompanying drawings and specific implementation methods.

[0020] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The components of the embodiments of the present invention described and shown in the accompanying drawings can generally be arranged and designed in various different configurations.

[0021] Therefore, the following detailed description of the embodiments of the invention provided in the accompanying drawings is not intended to limit the scope of the claimed invention, but merely to illustrate selected embodiments of the invention. All other embodiments obtained by those skilled in the art based on the embodiments of the invention without inventive effort are within the scope of protection of the invention.

[0022] The present invention will be further described below with reference to the accompanying drawings. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and should not be used to limit the scope of protection of the present invention.

[0023] The Ganoderma lucidum spore powder is sourced from Anhui Hongzhi Technology Co., Ltd., batch number: KYYY24-01.

[0024] The broken-cell wall Ganoderma lucidum spore powder extract is from Anhui Hongzhi Technology Co., Ltd., batch number: KYYY24-02.

[0025] The Ganoderma lucidum extract is sourced from Anhui Hongzhi Technology Co., Ltd., batch number: KYYY24-03.

[0026] The neutral protease at 50 U / mg was obtained from Shanghai Maclean Biochemical Technology Co., Ltd.

[0027] Sucrose esters, soy protein isolate, maltodextrin, xanthan gum, and monoglycerides are all food-grade and sourced from Henan Wanbang Chemical Technology Co., Ltd.

[0028] The egg yolk powder is food grade and comes from Jiangsu Kangde Egg Industry Co., Ltd.

[0029] Example See Figure 1 A method for preparing an antioxidant Ganoderma lucidum spore oil composition includes the following preparation steps: A1. Swell the Ganoderma lucidum spore powder according to a material-water ratio of 1 g: 20-25 mL, adjust the pH to 8.98-9.02, and then perform ultrasonic extraction at 350-450 W power for 25-35 min at 20-30℃, followed by water bath at 40-50℃ for 55-65 min. After cooling to room temperature, centrifuge at 5000 r / min for 20 min to collect the supernatant. Repeat the above steps for the precipitate to perform secondary extraction. Collect the supernatant and then perform suction filtration to obtain the first supernatant and the first residue; A2. Dry the first residue and then perform high-pressure supercritical carbon dioxide extraction and separation to obtain Ganoderma lucidum spore oil and a second residue. The high-pressure supercritical carbon dioxide extraction is performed at a pressure of 50-65 MPa, an extraction temperature of 38-42℃, and for a time of 30-40 min. The separation is performed at a temperature of 50℃ and a pressure of 11 MPa. A3. Adjust the pH of the first supernatant to the isoelectric point 2.0, and then perform sedimentation at 0-4℃ for 2.5-3.5 h. Then, centrifuge to collect the precipitate and wash with water until neutral. Dry the precipitate in a vacuum freeze dryer to obtain Ganoderma lucidum spore powder protein, which is stored at -20℃ for standby use. A4. Perform hot water extraction on the second residue to obtain a second supernatant and a third residue. The hot water extraction is performed at a temperature of 90-100℃ for a time of 90-150 min, and the material-liquid ratio is 1 g: 20-30 mL. A5. Mix the third residue with the Ganoderma lucidum spore powder protein and perform enzymatic extraction. Inactivate the enzyme in a 100℃ water bath for 10 min. After cooling, centrifuge the obtained extraction liquid to obtain the third supernatant. The enzyme used for the enzymatic extraction is neutral protease. The enzyme substrate ratio U / g is 1600-3000:1. The enzymatic extraction is performed at a pH of 4.6-6, a temperature of 45-50℃, and a time of 45-180 min. A6. Mix and dissolve 40 parts by mass of Ganoderma lucidum spore oil and 0.35-0.45 parts by mass of the first emulsifier monoglyceride to obtain an oil phase. Dissolve and homogenize 90 parts by mass of malt dextrin, 14-16 parts by mass of egg yolk powder, 14-16 parts by mass of plant protein, 500-550 parts by mass of water, and 0.11-0.13 parts by mass of the second emulsifier sucrose ester to obtain an aqueous phase of an emulsion. Then, add 0.23-0.25 parts by mass of xanthan gum, 20-30 parts by mass of the second supernatant, 20-30 parts by mass of the third supernatant, and 40-45 parts by mass of Ganoderma lucidum extract, and continue to homogenize to obtain the aqueous phase of the emulsion. Mix the aqueous phase and the oil phase and homogenize several times, and then perform spray drying to obtain an antioxidant Ganoderma lucidum spore oil composition with a microcapsule structure.

[0030] The plant protein includes any one or a combination of two of soybean protein isolate and Ganoderma lucidum spore powder protein.

[0031] Example 1 A preparation method of an antioxidant ganoderma lucidum spore oil composition, comprising the following preparation steps: A1. Swell the ganoderma lucidum spore powder according to a material-water ratio of 1g:20mL and adjust the pH to 8.98, then extract at 20℃ for 25min under ultrasonic extraction with a power of 350W, and then extract in a water bath at 40℃ for 55min, and then centrifuge at 5000r / min for 20min after cooling to room temperature to collect the supernatant, and then repeat the above steps for secondary extraction of the precipitate, and then combine the supernatants and then perform suction filtration to obtain a first supernatant and a first residue; A2. Dry the first residue, and then perform high-pressure supercritical carbon dioxide extraction and separation to obtain ganoderma lucidum spore oil and a second residue; wherein the high-pressure supercritical carbon dioxide extraction is performed at a pressure of 50MPa, an extraction temperature of 38℃, for 30min, and a carbon dioxide flow rate of 26L / h, and the separation is performed at a separation temperature of 50℃ and a separation pressure of 11MPa; A3. Adjust the pH of the first supernatant to the isoelectric point 2.0, and then settle at 0℃ for 2.5h, and then collect the precipitate by centrifugation and then wash with water until neutral, and then dry in a vacuum freeze dryer to obtain ganoderma lucidum spore powder protein, and then store at -20℃ for standby; A4. Perform hot water extraction on the second residue to obtain a second supernatant and a third residue, wherein the hot water extraction is performed at a temperature of 90℃ for 90min, and a material-liquid ratio of 1g:20mL; A5. Mix the third residue with the ganoderma lucidum spore powder protein and then perform enzymatic extraction, and then inactivate the enzyme in a water bath at 100℃ for 10min, and then take the supernatant after centrifugation of the obtained extraction liquid to obtain a third supernatant, wherein the enzyme used for the enzymolysis is neutral protease, the enzyme substrate ratio U / g is 3000:1, the enzymolysis pH is 4.6, the enzymolysis temperature is 45℃, and the enzymolysis time is 45min; A6. Mix and dissolve 40 parts by mass of ganoderma lucidum spore oil with 0.35 parts by mass of the first emulsifier monoglyceride to obtain an oil phase; dissolve and homogenize 90 parts by mass of malt dextrin, 14 parts by mass of egg yolk powder, 16 parts by mass of plant protein, 500 parts by mass of water, 0.11 parts by mass of the second emulsifier sucrose ester, and then add 0.23 parts by mass of xanthan gum, 30 parts by mass of the second supernatant, 0.30 parts by mass of the third supernatant, and 40 parts by mass of ganoderma lucidum extract to continue homogenization to obtain an aqueous phase of an emulsion; mix and homogenize the aqueous phase and the oil phase several times, and then perform spray drying to obtain an antioxidant ganoderma lucidum spore oil composition with a microcapsule structure.

[0032] The plant protein is obtained by combining soybean protein isolate and ganoderma lucidum spore powder protein according to a mass ratio of 1:1.

[0033] Example 2 A preparation method of an antioxidant ganoderma lucidum spore oil composition, comprising the following preparation steps: A1. Swell the Ganoderma lucidum spore powder according to the material-water ratio of 1 g:20 mL and adjust the pH to 9, then extract at 400 W power for 30 min under ultrasonic at 25℃, and then water bath at 45℃ for 50 min. After cooling to room temperature, centrifuge at 5000 r / min for 20 min to collect the supernatant. Repeat the above steps for the second extraction of the precipitate, and then combine the supernatants and filter to obtain the first supernatant and the first residue; A2. Dry the first residue and then perform high-pressure supercritical carbon dioxide extraction and separation to obtain Ganoderma lucidum spore oil and a second residue. The high-pressure supercritical carbon dioxide extraction is performed at a pressure of 65 MPa, an extraction temperature of 40℃, a time of 35 min, a carbon dioxide flow rate of 26 L / h, a separation temperature of 50℃, and a separation pressure of 11 MPa; A3. Adjust the pH of the first supernatant to the isoelectric point of 2.0, and then centrifuge to collect the precipitate and wash with water until neutral. Dry the precipitate in a vacuum freeze dryer to obtain Ganoderma lucidum spore powder protein, and store it at -20℃ for standby; A4. Hot water leaching is performed on the second residue to obtain a second supernatant and a third residue. The hot water leaching is performed at a temperature of 95℃ for 120 min, and the material-liquid ratio is 1 g:20 mL; A5. Mix the third residue with the Ganoderma lucidum spore powder protein and perform enzymatic extraction. Inactivate the enzyme in a 100℃ water bath for 10 min. After cooling, centrifuge the obtained extract to obtain the third supernatant. The enzyme used for enzymolysis is neutral protease, the enzyme substrate ratio is 3000:1, the enzymolysis pH is 5, the enzymolysis temperature is 50℃, and the enzymolysis time is 120 min; A6. Mix and dissolve 40 parts by mass of Ganoderma lucidum spore oil and 0.4 parts by mass of the first emulsifier monoglyceride to obtain an oil phase. Dissolve and homogenize 90 parts by mass of malt dextrin, 15 parts by mass of egg yolk powder, 15 parts by mass of plant protein, 550 parts by mass of water, and 0.12 parts by mass of the second emulsifier sucrose ester, and then add 0.24 parts by mass of xanthan gum, 25 parts by mass of the second supernatant, 25 parts by mass of the third supernatant, and 40 parts by mass of Ganoderma lucidum extract. Continue to homogenize to obtain an aqueous phase of the emulsion. Mix the aqueous phase and the oil phase and homogenize several times, and then perform spray drying to obtain an antioxidant Ganoderma lucidum spore oil composition with a microcapsule structure.

[0034] The plant protein is obtained by combining soybean protein isolate and Ganoderma lucidum spore powder at a mass ratio of 1:1.

[0035] Example 3 A preparation method of an antioxidant Ganoderma lucidum spore oil composition, comprising the following preparation steps: A1. Ganoderma lucidum spore powder was fully swelled according to the material-water ratio of 1 g:23 mL and adjusted to pH 9.02, and then extracted at 30℃ for 35 min under ultrasonic extraction with a power of 450 W, followed by water bath at 50℃ for 65 min. After cooling to room temperature, the supernatant was collected by centrifugation at 5000 r / min for 20 min. The precipitate was subjected to the above-mentioned steps again for secondary extraction. The supernatants were combined, and then filtered to obtain the first supernatant and the first residue; A2. The first residue was dried and then subjected to high-pressure supercritical carbon dioxide extraction and separation to obtain Ganoderma lucidum spore oil and a second residue. The high-pressure supercritical carbon dioxide extraction was performed at a pressure of 65 MPa, an extraction temperature of 42℃, a time of 40 min, a carbon dioxide flow rate of 26 L / h, a separation temperature of 50℃, and a separation pressure of 11 MPa; A3. The pH of the first supernatant was adjusted to the isoelectric point of 2.0, and then the supernatant was subjected to sedimentation at 4℃ for 3.5 h. The precipitate was collected by centrifugation and then washed with water until neutral. The precipitate was dried in a vacuum freeze dryer to obtain Ganoderma lucidum spore powder protein, which was stored at -20℃ for later use; A4. The second residue was subjected to hot water extraction to obtain a second supernatant and a third residue. The hot water extraction was performed at a temperature of 100℃ for a time of 150 min, and the material-liquid ratio was 1 g:30 mL; A5. The third residue was mixed with the Ganoderma lucidum spore powder protein and then subjected to enzymatic extraction. The enzyme was inactivated by water bath at 100℃ for 10 min. After cooling, the supernatant was obtained by centrifugation of the obtained extraction solution to obtain a third supernatant. The enzyme used for the enzymolysis was neutral protease, the enzyme substrate ratio was 1600:1, the enzymolysis pH was 6, the enzymolysis temperature was 50℃, and the enzymolysis time was 180 min; A6. 40 parts by mass of Ganoderma lucidum spore oil was mixed and dissolved with 0.45 parts by mass of the first emulsifier monoglyceride to obtain an oil phase. 90 parts by mass of malt dextrin, 16 parts by mass of egg yolk powder, 14 parts by mass of plant protein, 550 parts by mass of water, and 0.13 parts by mass of the second emulsifier sucrose ester were dissolved and homogenized to obtain a water phase of the emulsion. Then, 0.25 parts by mass of xanthan gum, 20 parts by mass of the second supernatant, 20 parts by mass of the third supernatant, and 45 parts by mass of Ganoderma lucidum extract were added and continuously homogenized to obtain the water phase of the emulsion. The water phase and the oil phase were mixed and homogenized for several times, and then subjected to spray drying to obtain an antioxidant Ganoderma lucidum spore oil composition with a microcapsule structure.

[0036] The plant protein was obtained by combining soybean protein isolate and Ganoderma lucidum spore powder at a mass ratio of 1:1.

[0037] Example 4 A preparation method of an antioxidant Ganoderma lucidum spore oil composition, comprising the following preparation steps: A1. Ganoderma lucidum spore powder was swelled according to a material-water ratio of 1 g:20 mL and adjusted to pH 9, and then extracted at 400 W power for 30 min under ultrasonic extraction at 25℃, followed by water bath at 45℃ for 50 min. After cooling to room temperature, the supernatant was collected by centrifugation at 5000 r / min for 20 min. The precipitate was subjected to the above-mentioned steps again for secondary extraction. The supernatants were combined, filtered, and then the first supernatant and the first residue were obtained; A2. The first residue was dried and then subjected to high-pressure supercritical carbon dioxide extraction and separation to obtain Ganoderma lucidum spore oil and a second residue. The high-pressure supercritical carbon dioxide extraction was performed at a pressure of 65 MPa, an extraction temperature of 40℃, a time of 35 min, a carbon dioxide flow rate of 26 L / h, a separation temperature of 50℃, and a separation pressure of 11 MPa; A3. The pH of the first supernatant was adjusted to the isoelectric point 2.0, and then the precipitate was collected by centrifugation after being settled at 2℃ for 3 h. The precipitate was washed with water until neutral, and then dried in a vacuum freeze dryer to obtain Ganoderma lucidum spore powder protein, which was stored at -20℃ for standby use; A4. The second residue was subjected to hot water extraction to obtain a second supernatant and a third residue. The hot water extraction was performed at a temperature of 95℃ for 120 min, and a material-liquid ratio of 1 g:20 mL; A5. The third residue was mixed with the Ganoderma lucidum spore powder protein and subjected to enzymatic extraction. The enzyme was inactivated by water bath at 100℃ for 10 min. After cooling, the supernatant was obtained by centrifugation of the obtained extraction solution. The third supernatant was obtained. The enzyme used for enzymolysis was neutral protease, the enzyme substrate ratio was 3000:1, the enzymolysis pH was 5, the enzymolysis temperature was 50℃, and the enzymolysis time was 120 min; A6. 40 parts by mass of Ganoderma lucidum spore oil was mixed and dissolved with 0.4 parts by mass of the first emulsifier monoglyceride to obtain an oil phase. 90 parts by mass of malt dextrin, 15 parts by mass of egg yolk powder, 15 parts by mass of plant protein, 550 parts by mass of water, and 0.12 parts by mass of the second emulsifier sucrose ester were dissolved and homogenized to obtain a water phase of the emulsion. The water phase and the oil phase were mixed and homogenized several times, and then subjected to spray drying to obtain an antioxidant Ganoderma lucidum spore oil composition with a microcapsule structure.

[0038] The plant protein is soybean protein isolate.

[0039] Example 5 A preparation method of an antioxidant Ganoderma lucidum spore oil composition, comprising the following preparation steps: A1. Ganoderma lucidum spore powder was swelled according to a material-water ratio of 1 g:20 mL and adjusted to pH 9, and then extracted at 400 W power for 30 min under ultrasonic at 25℃, and then extracted in a water bath at 45℃ for 50 min. After cooling to room temperature, the supernatant was collected by centrifugation at 5000 r / min for 20 min. The precipitate was subjected to the above-mentioned steps again for secondary extraction. The supernatants were combined, and filtered to obtain a first supernatant and a first residue; A2. The first residue was dried and subjected to high-pressure supercritical carbon dioxide extraction and separation to obtain Ganoderma lucidum spore oil and a second residue. The high-pressure supercritical carbon dioxide extraction was performed at a pressure of 65 MPa, an extraction temperature of 40℃, and for 35 min. The carbon dioxide flow rate was 26 L / h. The separation was performed at a temperature of 50℃ and a pressure of 11 MPa; A3. The first supernatant was adjusted to an isoelectric point of 2.0, and then subjected to sedimentation at 2℃ for 3 h. The precipitate was collected by centrifugation and washed with water until neutral. The precipitate was dried in a vacuum freeze dryer to obtain Ganoderma lucidum spore powder protein, which was stored at -20℃ for later use; A4. The second residue was subjected to hot water extraction to obtain a second supernatant and a third residue. The hot water extraction was performed at a temperature of 95℃ for 120 min, and the material-liquid ratio was 1 g:20 mL; A5. The third residue was mixed with the Ganoderma lucidum spore powder protein and subjected to enzymatic extraction. The enzyme was inactivated by water bath at 100℃ for 10 min. After cooling, the supernatant was obtained by centrifugation of the obtained extract. The third supernatant was obtained. The enzyme used for enzymolysis was neutral protease. The enzyme substrate ratio was 3000:1. The enzymolysis pH was 5. The enzymolysis temperature was 50℃. The enzymolysis time was 120 min; A6. 40 parts by mass of Ganoderma lucidum spore oil was mixed and dissolved with 0.4 parts by mass of the first emulsifier monoglyceride to obtain an oil phase. 90 parts by mass of malt dextrin, 15 parts by mass of egg yolk powder, 15 parts by mass of plant protein, 550 parts by mass of water, and 0.12 parts by mass of the second emulsifier sucrose ester were dissolved and homogenized to obtain a water phase of the emulsion. Then, 0.24 parts by mass of xanthan gum, 25 parts by mass of the second supernatant, 25 parts by mass of the third supernatant, and 40 parts by mass of Ganoderma lucidum extract were added and homogenized to obtain the water phase of the emulsion. The water phase and the oil phase were mixed and homogenized several times, and then subjected to spray drying to obtain an antioxidant Ganoderma lucidum spore oil composition with a microcapsule structure.

[0040] The plant protein is Ganoderma lucidum spore powder protein.

[0041] Comparative Example 1 The preparation steps of the antioxidant Ganoderma lucidum spore oil composition of Comparative Example 1 are as follows: A1. Preparation of primary Ganoderma lucidum spore oil by supercritical extraction: Ganoderma lucidum spore powder was subjected to a double-roller freeze-drying process to break the cell walls, achieving a cell wall breakage rate of 99.99%. Granulation was then performed with 95% ethanol, followed by drying at 30°C for 3 hours. Finally, supercritical carbon dioxide extraction was conducted at a temperature of 58°C and a pressure of 33 MPa, with a carbon dioxide flow rate of 26 L / h. The extraction time was 4 hours to obtain primary spore oil for later use. A2. Prepare mixed granules: Ganoderma lucidum spore powder (100-mesh ultrafine powder), broken-cell wall Ganoderma lucidum spore powder extract (100-mesh ultrafine powder), and Ganoderma lucidum extract (100-mesh ultrafine powder) in a ratio of 1:1:1. Mix and granulate with 95% ethanol and dry at 30°C for 5 hours. The moisture content of the granules is controlled at 10-12%. A3. Preparation of an antioxidant Ganoderma lucidum spore oil composition: Take 100 parts of crude Ganoderma lucidum spore oil A and 100 parts of Ganoderma lucidum granules B, pour them into an extraction vessel, and extract with carbon dioxide fluid. The entrainer is 5% ethanol. After extraction, the products are passed through a primary separation vessel and a secondary separation vessel. The separated products are centrifuged. In the extraction vessel, the temperature is maintained at 58℃ and the pressure at 33MPa; the flow rate of the carbon dioxide fluid is 26L / h; and the extraction time is 4h. Discard the residue to obtain the antioxidant Ganoderma lucidum spore oil composition.

[0042] Example of effect Antioxidant efficacy test: a. Total reducing power test Take 1 mL of PBS aqueous solution with different material-to-liquid ratios (0.1, 0.5, 1.0, 1.5 and 2.0 mg / mL) of the antioxidant Ganoderma lucidum spore oil compositions of Examples 1-5 and Comparative Example 1, add 2 mL of PBS buffer solution (0.2 M, pH=6.6) and 2 mL of 1% potassium ferricyanide aqueous solution, mix well and react at 50°C for 20 min; then add 2 mL of trichloroacetic acid aqueous solution (10%, w / v) and centrifuge at 3000×g for 10 min; take 2.5 mL of supernatant and add an equal volume of distilled water and 1 mL of ferric chloride aqueous solution (0.1%, w / v), mix well and let stand at room temperature for 10 min; measure the absorbance value at 700 nm; Vc is used as a positive control; the reducing power of the sample is directly proportional to the absorbance value.

[0043] The experimental results are shown in Table 1. Table 1 shows the total extracellular reducing power data of the Ganoderma lucidum spore oil composition: Table 1

[0044] As shown in Table 1 above, the higher the concentration of divalent iron ions, the higher the ability of the ganoderma spore oil composition to reduce trivalent iron ions, and the stronger the reducing power of the ganoderma spore oil composition; within the range of 0.1-2 mg / mL of the material-to-liquid ratio of the ganoderma spore oil composition, the reducing power is positively correlated with the sample concentration; the maximum value is reached at a material-to-liquid ratio of 2.0 mg / mL; this shows that the ganoderma spore oil composition has good reducing power, and the ganoderma spore oil compositions prepared in Examples 1-5 have better reducing power.

[0045] b. DPPH free radical scavenging test The ganoderma spore oil composition was prepared into different material-to-liquid ratios (0.1, 0.2, 0.4 and 0.6 mg / mL) of PBS aqueous solution 1 mL, mixed with 1 mL of 0.2 mM DPPH anhydrous ethanol solution uniformly, reacted at room temperature for 30 min in the dark, and the absorbance value was measured at 517 nm.

[0046] The blank group used an equal volume of ethanol instead of the DPPH anhydrous ethanol solution, the control group used an equal volume of anhydrous ethanol instead of the ganoderma spore oil solution, and Vc was used as a positive control.

[0047] The scavenging rate calculation formula is: scavenging rate (%) = [(A0-Ai+Aj) / A0] x 100% In the formula, A0 is the absorbance value of the blank group; Aj is the absorbance value of the control group; and Ai is the absorbance value of the sample group.

[0048] The experimental results are shown in Table 2, which is the DPPH free radical scavenging test data of the ganoderma spore oil composition: Table 2

[0049] As shown in Table 2, within the range of 0.1-0.6 mg / mL of the material-to-liquid ratio of the ganoderma spore oil composition, the DPPH scavenging power is positively correlated with the sample concentration; this shows that the ganoderma spore oil composition has good DPPH scavenging capacity, and the ganoderma spore oil compositions prepared in Examples 1-5 have the best DPPH scavenging capacity.

[0050] 3. Hydroxyl radical scavenging test 2.5 mL of 4 mM aqueous salicylic acid solution, 2.5 mL of 4 mM aqueous ferrous sulfate solution, 2.5 mL of 4 mM aqueous hydrogen peroxide solution, and 2.5 mL of different concentrations of Ganoderma lucidum spore oil composition PBS aqueous solution (0.1, 0.5, 1.0, 1.5, and 2.0 mg / mL) were uniformly mixed, reacted at 37°C in the dark for 30 min, centrifuged at 3000 x g for 10 min, and the supernatant was taken to measure the absorbance value at 517 nm. The blank group was replaced with an equal volume of absolute ethanol instead of aqueous salicylic acid solution, and the control group was replaced with an equal volume of ultrapure water instead of Ganoderma lucidum spore oil solution, and Vc was used as a positive control.

[0051] The clearance calculation formula is: clearance (%) = [(A0-Ai+Aj) / A0] x 100%.

[0052] In the formula: A0 is the absorbance value of the blank group; Aj is the absorbance value of the control group; and Ai is the absorbance value of the sample group.

[0053] The experimental results are shown in Table 3, which is the data of the hydroxyl radical scavenging test of Ganoderma lucidum spore oil composition: Table 3

[0054] As shown in Table 3, within the range of 0.1-0.6 mg / mL of Ganoderma lucidum spore oil composition, the hydroxyl radical scavenging capacity is positively correlated with the sample concentration, indicating that Ganoderma lucidum spore oil composition has good hydroxyl radical scavenging capacity, and the Ganoderma lucidum spore oil composition prepared in Examples 1-5 has the best hydroxyl radical scavenging capacity.

[0055] c.ABTS + Radical scavenging test (ABTS+ reagent from Adamas Company) Prepare the stock solution: take 40 mg of ABTS+ and 6.88 mg of potassium persulfate, dissolve in 10 mL of distilled water, and store in the dark for 12 h (room temperature).

[0056] Prepare the working solution: add an appropriate amount of phosphate buffer (pH = 7.4) to the above solution until its absorbance at 734 nm is 0.70 ± 0.02.

[0057] Take Ganoderma lucidum spore oil composition as the sample PBS aqueous solution (0.1, 0.5, 1.0, 1.5, 2.0 mg / mL).

[0058] Mix 180 μL of working solution with 20 μL of sample solution to form the AS group; mix 180 μL of working solution with 20 μL of H2O to form the Ab group. Let stand for 5 min, measure the absorbance (734 nm), use VC as a positive control, and repeat the test three times.

[0059] The clearance rate calculation formula is: The clearance rate (%) = [1-(Ab-As) / Ab]x100% In the formula, Ab is the absorbance value of the control group, and As is the absorbance value of the sample group.

[0060] The experimental results are shown in Table 4, which is the ganoderma spore oil composition ABTS + Free radical scavenging test data: Table 4

[0061] As can be seen from Table 4, within the range of 0.1-0.6 mg / mL of the ganoderma spore oil composition, the ABTS + The size of the free radical scavenging capacity is positively correlated with the sample concentration, indicating that the ganoderma spore oil composition has good ABTS + Free radical scavenging capacity, wherein the ABTS + Free radical scavenging capacity of the ganoderma spore oil composition prepared in Examples 1-5 is the best.

[0062] In summary, the ganoderma spore oil composition prepared in Examples 1-5 has strong antioxidant capacity, and can achieve a relatively high clearance rate on ABTS + Free radicals, DPPH free radicals and hydroxyl radicals at a relatively low concentration, and also has a certain reducing power.

[0063] The above specific examples further illustrate the purpose, technical solutions and beneficial effects of the present application. It should be understood that the above description is only a specific embodiment of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A method of preparing an antioxidant Ganoderma spore oil composition, characterized by, The preparation steps include: A1. Swelling the ganoderma spore powder with water and extracting multiple times by ultrasonic to obtain a first supernatant and a first residue; A2. After drying the first residue, performing high-pressure supercritical carbon dioxide extraction and separation to obtain ganoderma spore oil and a second residue; A3. Precipitating and separating the ganoderma spore powder protein from the first supernatant; A4. Hot water leaching the second residue to obtain a second supernatant and a third residue; A5. Mixing the third residue with the ganoderma spore powder protein and performing enzymatic extraction, centrifuging the obtained extract to obtain a supernatant, and obtaining a third supernatant; A6. Mixing and dissolving the ganoderma spore oil with the first emulsifier to obtain an oil phase; dissolving and homogenizing the wall material, water and the second emulsifier, then adding xanthan gum, the second supernatant, the third supernatant and ganoderma extract, and continuing to homogenize to obtain an aqueous phase of emulsion; mixing the aqueous phase and the oil phase, homogenizing and emulsifying multiple times, and then performing spray drying to obtain an antioxidant ganoderma spore oil composition with a microcapsule structure.

2. The process for the preparation of the antioxidant Ganoderma lucidum spore oil composition as claimed in claim 1, wherein, The specific steps of step A1 are as follows: swelled the ganoderma spore powder according to the material water ratio of 1g:20-25mL, adjusted the pH to 8.98-9.02, ultrasonic extracted at 20-30℃ with a power of 350-450W for 25-35min, then water bathed at 40-50℃ for 55-65min, cooled to room temperature, centrifuged at 5000r / min for 20min to collect the supernatant, repeated the above steps twice for the precipitate, combined the supernatants, and obtained the first supernatant and the first residue by filtration.

3. The process for the preparation of the antioxidant Ganoderma lucidum spore oil composition as claimed in claim 1, wherein, The pressure of the high-pressure supercritical carbon dioxide extraction is 50-65MPa, the extraction temperature is 38-42℃, and the time is 30-40min.

4. The process for the preparation of the antioxidant Ganoderma lucidum spore oil composition as claimed in claim 1, wherein, The specific steps of step A3 are as follows: adjust the pH of the first supernatant to the isoelectric point 2.0, precipitate at 0-4℃ for 2.5-3.5h, then centrifuge to collect the precipitate and wash to neutral, dry in a vacuum freeze dryer to obtain ganoderma spore powder protein, and store at-20℃ for standby.

5. The process for the preparation of the antioxidant Ganoderma lucidum spore oil composition as claimed in claim 1, wherein, The hot water leaching temperature is 90-100℃, the time is 90-150min, and the solid-liquid ratio is 1g:20-30mL.

6. The process for the preparation of the antioxidant Ganoderma lucidum spore oil composition as claimed in claim 1, wherein, The enzyme used in the enzymolysis is neutral protease, the enzyme substrate ratio U / g is 1600-3000:1, the enzymolysis pH is 4.6-6, the enzymolysis temperature is 45-50℃, and the enzymolysis time is 45-180min.

7. The process for the preparation of the antioxidant Ganoderma lucidum spore oil composition as claimed in claim 1, wherein, The mass ratio of the ganoderma spore oil, the first emulsifier, the wall material, water, the second emulsifier, xanthan gum, the second supernatant, the third supernatant and the ganoderma extract is 40:0.35-0.45:120:500-550:0.11-0.13:0.23-0.25:40-60:40-45; wherein the first emulsifier is monoglyceride, and the second emulsifier is sucrose ester.

8. The process for the preparation of the antioxidant Ganoderma lucidum spore oil composition as claimed in claim 1, wherein, The wall material includes 90 parts by mass of malt dextrin, 14-16 parts by mass of egg yolk powder and 14-16 parts by mass of plant protein; the plant protein includes any one or a combination of two of soybean protein isolate and ganoderma spore powder protein.

9. An antioxidant Ganoderma lucidum spore oil composition, characterized by, The antioxidant Ganoderma lucidum spore oil composition is prepared by the method of any one of claims 1-8.

10. Use of the antioxidant Ganoderma lucidum spore oil composition according to claim 9, characterized in that, The antioxidant Ganoderma lucidum spore oil composition is applied to the preparation of special dietary food, health food and medicine.

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