Polygonatum cyrtonema flower polysaccharide with antioxidant activity and extraction method thereof
Through ultrasonic pretreatment and enzyme complex enzymatic extraction method, combined with the purification dialysis step, the problem of low extraction efficiency and purity of Polygonatum polysaccharide is solved, and efficient and high-purity polysaccharide extraction is achieved, and the extract is given antioxidant activity.
Patent Information
- Application Number
- CN202411876781.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-19
- Publication Date
- 2025-03-11
AI Technical Summary
There is little research on Polygonum multiflorum in the prior art, especially how to deeply explore its functional active ingredient polysaccharides, which leads to the failure to effectively utilize Polygonum multiflorum resources, and the extraction process is cumbersome and time-consuming, which affects the extraction efficiency and results.
Ultrasonic pretreatment and multiple enzyme complex enzymatic extraction methods, combined with purification and dialysis steps, the efficient and high-purity extraction of polysaccharides of polysaccharides was achieved.
The extraction efficiency and purity of polysaccharide polysaccharides have been significantly improved, making the extracted polysaccharides have antioxidant activity, and meeting the market's demand for high-purity polysaccharide products.
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field of polysaccharide extraction, and in particular to a polygonatum cyrtonema flower polysaccharide with antioxidant activity and an extraction method thereof. Background Art
[0002] Polygonatum is one of the original species of Chinese medicinal materials. It belongs to the genus Polygonatum of the Liliaceae family, and its rhizome is used as medicine. According to research, Polygonatum has a history of medicinal use of more than 2,000 years. Traditional medicine believes that Polygonatum has the effects of nourishing the kidney and essence, widening the stomach and replenishing qi, nourishing yin and moistening dryness, and promoting body fluid and tonifying the spleen. The chemical composition of Polygonatum is closely related to the clinical application of Polygonatum. Studies on the chemical composition of Polygonatum have shown that the main components of Polygonatum are saponins and polysaccharides. In addition, it also contains compounds such as anthrone compounds, alkaloids, cardiac glycosides, lignin, vitamins and amino acids. Among them, Polygonatum polysaccharide is the component with the highest content in Polygonatum, and its functionality is also the most significant. Modern pharmacological studies have shown that Polygonatum polysaccharide has multiple pharmacological effects such as anti-oxidation and anti-aging, anti-tumor, lowering blood sugar and blood lipids, and improving memory, indicating that Polygonatum and its polysaccharides have good application prospects in the development of functional foods and drugs. Polysaccharide is the core indicator for evaluating the quality and composition of Polygonatum sibiricum. Due to the different growth environments and species of Polygonatum sibiricum, the polysaccharide content varies. Based on anhydrous glucose, the polysaccharide content of Polygonatum sibiricum must be ≥7%. In the prior art, there are many studies on the rhizomes of Polygonatum sibiricum, but few studies on Polygonatum sibiricum flowers, especially how to deeply explore the value of Polygonatum sibiricum flowers.
[0003] The medicinal and edible part of polygonatum is mainly its rhizome, and the research on the extraction, processing and pharmacological effects of bioactive ingredients is mainly for the root and stem parts, while the non-medicinal part of polygonatum, the flower, is discarded as a by-product, resulting in failure to be effectively utilized and causing a large amount of resource waste. At present, there are many studies on the extraction process of polygonatum rhizome polysaccharides, including solvent extraction, ultrasonic assisted extraction, microwave assisted extraction, enzymatic hydrolysis, etc. However, in the process of extracting polygonatum polysaccharides, it is often extracted repeatedly in order to obtain natural ingredients with high purity. The whole process requires repeated extraction, filtration, purification and purification steps to ensure that the polysaccharides are fully obtained and made into high-quality polygonatum granules. However, the process is cumbersome and time-consuming, which affects the extraction efficiency and results. At present, there is no patent to study the functional active ingredient polysaccharides of multi-flowered polygonatum flower, so the present invention takes the non-medicinal part of multi-flowered polygonatum flower as the research object, and develops an extraction process for efficiently extracting multi-flowered polygonatum flower polysaccharides. Summary of the invention
[0004] The present invention aims at the deficiencies in the prior art and provides a polysaccharide from the flower of Polygonatum cyrtonema with antioxidant activity and an extraction method thereof. After ultrasonic pretreatment and multiple enzyme complex enzymatic extraction, the polysaccharide is subjected to a purification and dialysis step to achieve efficient and high-purity extraction of the polysaccharide from the flower of Polygonatum cyrtonema; the extraction method not only improves the extraction efficiency and purity of the polysaccharide from the flower of Polygonatum cyrtonema, but also the extracted polysaccharide from the flower of Polygonatum cyrtonema has antioxidant activity.
[0005] In order to solve the above technical problems, the first aspect of the present invention provides a method for extracting polysaccharides from Polygonatum cyrtonema with antioxidant activity, comprising the following steps:
[0006] S1. After defatting, the ultrafine powder of Polygonatum cyrtonema is mixed with pure water and placed in an ultrasonic water bath for ultrasonic treatment;
[0007] S2, enzymatic hydrolysis: add cellulase and xylanase, and bathe at 50-70℃ for 2-4h; then add thermostable α-amylase, and bathe at 70-100℃ for 1-2h; cool the water bath to 50-70℃, and add saccharifying enzyme for water bath treatment for 2-3h; after the extraction is completed, heat to 100℃ to inactivate the enzyme for 10min, centrifuge, and collect the supernatant;
[0008] S3. The supernatant is sequentially subjected to primary alcohol precipitation, decolorization and purification, secondary alcohol precipitation, dialysis, and freeze-drying treatment to obtain Polygonatum cyrtonema polysaccharide with antioxidant activity.
[0009] Further, in S1, the ratio of defatted Polygonatum cyrtonema flower superfine powder to pure water is 1: (30-60) g / mL;
[0010] Furthermore, in S1, the power of the ultrasonic treatment is 200-400 W, the temperature is 40-60° C., and the time is 60-120 min.
[0011] Furthermore, in S2, the added amounts of cellulase and xylanase are 800-1000U / g of the defatted Polygonatum cyrtonema flower superfine powder, respectively; the added amount of α-amylase is 1800-2000U / g of the defatted Polygonatum cyrtonema flower superfine powder; and the added amount of saccharifying enzyme is 1800-2000U / g of the defatted Polygonatum cyrtonema flower superfine powder.
[0012] Furthermore, in S2, the enzymatic hydrolysis process is stirred in a water bath at a speed of 180-250 r / min.
[0013] Furthermore, in S2, the centrifugal condition is a rotation speed of 8000-9000 r / min.
[0014] Furthermore, in S3, the decolorization and purification is specifically as follows: AB-8 macroporous resin is added to the polysaccharide solution after the first alcohol precipitation (the aqueous solution of polysaccharide precipitation), and the decolorization treatment is carried out at 30-37°C and 100-200r / min for 10-15h, and the amount of the macroporous resin added is 20%-30% of the mass of the polysaccharide solution after the alcohol precipitation.
[0015] Furthermore, in S3, the alcohol precipitation is specifically as follows: adding anhydrous ethanol to the polysaccharide solution obtained in the previous step until the ethanol mass concentration is 10%-20%, standing overnight, centrifuging for 10-20 minutes to remove the precipitate, adding anhydrous ethanol until the ethanol mass concentration is 80-95%, standing overnight, and centrifuging to collect the precipitate.
[0016] Furthermore, the supernatant is concentrated to 1 / 4-1 / 6 of the original volume by a rotary evaporator before the first alcohol precipitation.
[0017] Furthermore, in S3, the dialysis is specifically as follows: dissolving the polysaccharide after the secondary alcohol precipitation in water, adding it into a 3000-5000Da dialysis bag, dialyzing it with flowing water for 2-3 days, and then dialyzing it with pure water for 1-2 days.
[0018] Furthermore, in S1, the degreasing treatment is specifically: reflux with petroleum ether as solvent for 2-4 hours at 60-70°C. The ratio of the Polygonatum cyrtonema superfine powder to petroleum ether is 1: (30-60) g / mL.
[0019] Furthermore, the Polygonatum cyrtonema flowers are dried at 40-60° C., crushed through a 20-mesh sieve, and then ground by an ultrafine grinder to obtain micron-grade ultrafine powder.
[0020] The second aspect of the present invention provides Polygonatum cyrtonema flower polysaccharide with antioxidant activity prepared by the preparation method described in the first aspect.
[0021] The third aspect of the present invention provides the use of the Polygonatum cyrtonema flower polysaccharide described in the second aspect in an antioxidant product.
[0022] Beneficial effects of the present invention:
[0023] The invention extracts the polysaccharide from the flower of Polygonatum cyrtonema by ultrasound-assisted enzymatic hydrolysis, which can dissolve all the substances constituting the cell wall, make the substances in the cell more easily dissolved, effectively decompose the proteins in the cell, and the extraction process is more gentle, which significantly improves the yield and purity of the polysaccharide from the flower of Polygonatum cyrtonema.
[0024] The invention adopts two-stage alcohol precipitation, macroporous resin and dialysis combined purification to achieve high-purity extraction of polygonatum cyrtonema flower polysaccharide, and meets the market demand for high-purity polysaccharide products.
[0025] The extraction method of the invention maintains the biological activity of the polysaccharide, so that the extracted polysaccharide has significant antioxidant performance. BRIEF DESCRIPTION OF THE DRAWINGS
[0026] In order to more clearly illustrate the technical solution of the present invention, the drawings required for use in the embodiments are briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
[0027] Figure 1 To characterize the effect of ultrasonic temperature on polysaccharide yield in Example 1;
[0028] Figure 2 To characterize the effect of ultrasound time on polysaccharide yield in Example 1;
[0029] Figure 3 To characterize the effect of ultrasonic power on polysaccharide yield in Example 1;
[0030] Figure 4 To characterize the effect of the material-liquid ratio on the polysaccharide yield in Example 1;
[0031] Figure 5 To characterize the effect of xylanase and cellulase enzymatic hydrolysis time on polysaccharide yield in Example 1;
[0032] Figure 6 To characterize the effect of the enzymatic hydrolysis temperature of xylanase and cellulase on the polysaccharide yield in Example 1;
[0033] Figure 7 To characterize the effect of the enzymatic hydrolysis time of the thermostable α-amylase on the polysaccharide yield in Example 1;
[0034] Figure 8 To characterize the effect of the enzymatic hydrolysis temperature of the thermostable α-amylase on the polysaccharide yield in Example 1;
[0035] Fig. 9 To characterize the effect of saccharification enzyme hydrolysis time on polysaccharide yield in Example 1;
[0036] Fig.10 To characterize the effect of saccharification enzyme hydrolysis temperature on polysaccharide yield in Example 1;
[0037] Fig.11 The results of the macroporous resin decolorization rate and polysaccharide retention rate in Example 2;
[0038] Fig.12 The result of scavenging DPPH free radical by Polygonatum cyrtonema polysaccharide in Test Example 1;
[0039] Fig.13The result of the scavenging rate of ABTS free radicals by Polygonatum cyrtonema polysaccharide in Test Example 1;
[0040] Fig.14 This is the result of the iron ion reducing ability of Polygonatum cyrtonema flower polysaccharide in Test Example 1. DETAILED DESCRIPTION
[0041] The technical solution of the present invention will be described clearly and completely below in conjunction with the specific embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0042] Example 1
[0043] This embodiment relates to a method for extracting polysaccharides from Polygonatum cyrtonema with antioxidant activity, comprising the following steps:
[0044] (1) Select dried Polygonatum cyrtonema flowers, put them into an oven at 50° C. and dry them to constant weight, grind them with a tissue grinder and pass them through a 20-mesh sieve, and further grind the sieved Polygonatum cyrtonema pollen in an ultrafine grinder to obtain micron-level Polygonatum cyrtonema ultrafine powder.
[0045] (2) Defatting: The ultrafine powder of Polygonatum sibiricum flower was refluxed at 65°C with petroleum ether (boiling range 60-90°C) as solvent for 2 h, and the petroleum ether was recovered to obtain defatted Polygonatum sibiricum flower powder, wherein the ratio of the ultrafine powder of Polygonatum sibiricum flower to the petroleum ether was 1:30 g / mL.
[0046] (3) Defatted Polygonatum cyrtonema flower powder and pure water were mixed at a ratio of 1:40 g / mL and placed in a 400W ultrasonic water bath, and ultrasonicated at 40°C for 1 h. After the ultrasonication, 1000 U / g of defatted Polygonatum cyrtonema flower powder xylanase and cellulase were added respectively, and the mixture was incubated at 60°C for 3 h. 2000 U / g of defatted Polygonatum cyrtonema flower powder thermostable α-amylase was added, and the mixture was incubated at 80°C for 1 h. The water bath temperature was lowered to 60°C, and after the extract was cooled, 2000 U / g of defatted Polygonatum cyrtonema flower powder saccharifying enzyme was added, and the mixture was incubated at 100°C for 10 min to inactivate the enzyme. The supernatant was collected after centrifugation at 9000 r / min for 10 min.
[0047] (4) After the supernatant is concentrated to 1 / 4 of the original volume under reduced pressure, an alcohol precipitation is performed: anhydrous ethanol is added to an ethanol concentration of 20%, the mixture is allowed to stand at 4°C overnight, and the precipitate is removed by centrifugation for 10-20 min. Anhydrous ethanol is then added to an ethanol concentration of 95%, the mixture is allowed to stand at 4°C overnight, and the precipitate is collected by centrifugation and freeze-dried to obtain crude polysaccharide.
[0048] Taking polysaccharide yield as an indicator, the effects of ultrasonic power, ultrasonic time, ultrasonic temperature, enzymatic hydrolysis time and enzymatic hydrolysis temperature on the crude polysaccharide yield were investigated using a single factor approach.
[0049] 1.1 Ultrasonic temperature: The ultrasonic temperature of Example 1 was adjusted to 30°C, 50°C, 60°C, and 70°C, and other parameters were kept unchanged. The effect of ultrasonic temperature on the polysaccharide yield was obtained, as shown in FIG. Figure 1 As shown, it can be seen that the yield is highest at 40°C.
[0050] 1.2 Ultrasonic time: The ultrasonic time of Example 1 was adjusted to 20 min, 40 min, 80 min, and 100 min, and other parameters remained unchanged. The effect of ultrasonic time on the polysaccharide yield was obtained, as shown in FIG. Figure 2 As shown, it can be seen that the yield is highest at 60 min.
[0051] 1.3 Ultrasonic power: The ultrasonic power of Example 1 was adjusted to 100 W, 200 W, 300 W, and 500 W, and other parameters remained unchanged. The effect of ultrasonic power on the polysaccharide yield was obtained, as shown in FIG. Figure 3 As shown, it can be seen that the highest efficiency is achieved at 300W.
[0052] 1.4 Material-liquid ratio: The ratio of the defatted Polygonatum cyrtonema flower powder to pure water in step (3) of the above Example 1 was adjusted to 1:20 g / mL, 1:30 g / mL, 1:50 g / mL, and 1:60 g / mL, while other parameters remained unchanged. The effect of the material-liquid ratio on the polysaccharide yield was obtained, as shown in FIG. Figure 4 As shown, it can be seen that the yield is highest when the ratio of defatted Polygonatum cyrtonema flower powder to pure water is 1:50 g / mL.
[0053] 1.5 Xylanase and cellulase enzymolysis time: The xylanase and cellulase enzymolysis time of Example 1 was adjusted to 1 h, 2 h, 4 h, and 5 h, and other parameters remained unchanged. The effect of xylanase and cellulase enzymolysis time on polysaccharide yield was obtained, as shown in FIG. Figure 5 As shown, it can be seen that the yield is highest at 2h.
[0054] 1.6 Xylanase and cellulase enzymolysis temperature: The xylanase and cellulase enzymolysis temperatures of Example 1 were adjusted to 30°C, 40°C, 50°C, and 70°C, while other parameters remained unchanged. The effects of xylanase and cellulase enzymolysis temperatures on polysaccharide yields were obtained, as shown in FIG. Figure 6 As shown, it can be seen that the yield is highest at 60°C.
[0055] 1.7 α-amylase hydrolysis time: The α-amylase hydrolysis time of Example 1 was adjusted to 30 min, 40 min, 50 min, and 70 min, and other parameters remained unchanged. The effect of α-amylase hydrolysis time on polysaccharide yield was obtained, as shown in FIG. Figure 7As shown, it can be seen that the yield is highest at 60 min.
[0056] 1.8α-amylase hydrolysis temperature: The α-amylase hydrolysis temperature of Example 1 was adjusted to 60°C, 70°C, 90°C, and 100°C, and other parameters remained unchanged. The effect of α-amylase hydrolysis temperature on polysaccharide yield was obtained, as shown in FIG. Figure 8 As shown, it can be seen that the yield is highest at 90°C.
[0057] 1.9 Saccharification enzyme hydrolysis time: The saccharification enzyme hydrolysis time of the above Example 1 was adjusted to 2h, 3h, 4h, and 5h, and other parameters remained unchanged. The effect of saccharification enzyme hydrolysis time on polysaccharide yield was obtained, as shown in FIG. Fig. 9 As shown, it can be seen that the yield is highest at 1 h.
[0058] 1.10 Saccharification enzyme hydrolysis temperature: The saccharification enzyme hydrolysis temperature of Example 1 was adjusted to 30°C, 40°C, 50°C, and 70°C, and other parameters remained unchanged, and the effect of saccharification enzyme hydrolysis temperature on polysaccharide yield was obtained, as shown in FIG. Fig.10 As shown, it can be seen that the yield is highest at 60°C.
[0059] In summary, under the conditions of ultrasonic temperature of 40℃, ultrasonic time of 60min, ultrasonic power of 300W, solid-liquid ratio of 1:50g / mL, xylanase and cellulase hydrolysis time of 2h, xylanase and cellulase hydrolysis temperature of 60℃, high temperature resistant α-amylase hydrolysis time of 60min, high temperature resistant α-amylase hydrolysis temperature of 90℃, saccharifying enzyme hydrolysis time of 2h, and saccharifying enzyme hydrolysis temperature of 60℃, the yield of extracted polysaccharides is the highest, and this is used as the optimized extraction condition for subsequent purification treatment.
[0060] Example 2
[0061] This example relates to a method for purifying crude polysaccharides obtained under the optimized conditions of Example 1, comprising the following steps:
[0062] (1) Decolorization and purification of AB-8 macroporous resin: Before use, the macroporous resin was soaked in 95% ethanol aqueous solution for 24 hours and washed with distilled water until there was no alcohol smell. It was soaked in HCl (1 mol / L) for another 4 hours and washed with distilled water until the pH was neutral. Finally, it was soaked in sodium hydroxide (1 mol / L) for 4 hours and washed with distilled water until the pH was neutral. The concentrated aqueous solution of crude polysaccharides extracted under the optimized conditions of Example 1 was placed in a conical flask, and pretreated macroporous resins with a mass of 20%, 30%, 40%, 50%, and 60% of the concentrated aqueous solution of polysaccharides were added respectively. The solution was placed in a shaking incubator at 37°C, 200 r / min, decolorized for 10 hours, and filtered. The decolorization rate and polysaccharide retention rate were determined according to the following formula. The decolorization rate was determined at 450nm, and the optimal concentration was evaluated by a comprehensive score.
[0063] Polysaccharide retention rate (%) = content of polysaccharide after purification / mass of polysaccharide before purification * 100%
[0064] Decolorization rate (%) = (absorbance value before decolorization - absorbance value after decolorization) / absorbance value before decolorization * 100% Comprehensive score (%) = 0.5 * (polysaccharide retention rate + decolorization rate)
[0065] refer to Fig.11 It can be seen that the polysaccharide decolorization rate increases slowly with the increase of the content of macroporous resin AB-8, while the polysaccharide retention rate decreases significantly, indicating that AB-8 has a good ability to adsorb pigments and also has a strong adsorption capacity for polysaccharides. Therefore, when the AB-8 content is 20%, its effect is the best, the decolorization rate is 72.25%, and the polysaccharide retention rate is 75.41%.
[0066] Example 3
[0067] This embodiment relates to a further purification method of the decolorized and purified polysaccharide obtained under the optimized conditions of Example 2, comprising the following steps:
[0068] (1) The polysaccharide solution decolorized and purified in Example 2 with an AB-8 content of 20% was subjected to a secondary alcohol precipitation using the same parameters as in step (4) of Example 1.
[0069] (2) The precipitate from the second alcohol precipitation was redissolved in water, placed in a 3000Da dialysis bag and dialyzed against flowing water for 2 days, and then against pure water for 1 day.
[0070] (3) The dialyzed polysaccharide extract was concentrated to 1 / 3 of the original volume by rotary evaporation, poured into a culture dish, placed in a low-temperature refrigerator for freezing, and then placed in a freeze vacuum dryer for freeze drying, and finally a flocculent polysaccharide product was obtained, namely, Polygonatum cyrtonema polysaccharide. The purity of the purified sample was measured to be 90.04%.
[0071] Test Example 1
[0072] In this test example, DPPH free radical scavenging activity, ABTS free radical scavenging ability and iron ion reducing ability were used as indicators to evaluate the antioxidant properties of the purified Polygonatum cyrtonema flower polysaccharide in Example 3.
[0073] The purified Polygonatum cyrtonema polysaccharide from Example 3 was dissolved in water to prepare sample solutions of different concentrations; and vitamin C was dissolved in water to prepare sample solutions of different concentrations for comparison.
[0074] The DPPH free radical scavenging activity was determined as follows: 1 mL of sample solution was reacted with 0.1 mmol / L DPPH solution (3 mL) at room temperature for 30 min. The absorbance at 517 nm was measured. The DPPH free radical scavenging ability was calculated using the following formula:
[0075] Clearance / % = (A0-A1) / A0
[0076] Where: A0 is the absorbance of the solution in the control group, and A1 is the absorbance of the sample solution.
[0077] Determination of ABTS free radical scavenging ability: 7mmol / L ABTS aqueous solution was mixed with potassium persulfate (4.95mmol / L) and reacted in the dark for 12 hours to generate ABTS free radicals. The solution was diluted with distilled water and the dilution was stopped when the absorbance reached 0.7 (734nm). Finally, 1mL of sample solution was mixed with 3mL of ABTS free radical solution and reacted for 30 minutes, and the absorbance at 734nm was measured. The scavenging rate of ABTS free radicals was calculated using the following formula:
[0078] Clearance / % = (A0-A1) / A0
[0079] Wherein: A0 is the absorbance of the solution in the control group, and A1 is the absorbance of the sample solution. Determination of iron ion reducing power: add an equal volume of sodium phosphate buffer (0.2 mol / L, pH 6.6) to 2.5 mL of sample solution, then add 2.5 mL of potassium ferrocyanide solution (1%, w / v), then react in a water bath (50°C), cool to room temperature after 20 minutes, add 2.5 mL of 10% (w / v) trichloroacetic acid, vortex mix and centrifuge (6000g, 10 minutes), take 2.5 mL of supernatant and add an equal volume of distilled water and 0.5 mL of ferric chloride (0.1%, w / v), and let it react for 5 minutes. The iron ion reducing power is expressed by the absorbance value at 700 nm.
[0080] refer to Figure 12-14 It can be seen that the antioxidant property of the purified Polygonatum cyrtonema polysaccharide in Example 3 of the present invention is significantly better than that of vitamin C, and with the increase of polysaccharide concentration, the scavenging activity of DPPH and ABTS free radicals and the iron ion reducing power increase significantly, and show a certain dose dependence.
[0081] In summary, the present invention extracts polysaccharides from the flowers of Polygonatum cyrtonema through ultrasound-assisted enzymatic hydrolysis, which can dissolve all the substances constituting the cell wall, make the substances in the cells more easily dissolve, effectively decompose the proteins in the cells, and the extraction process is more gentle, which significantly improves the yield and purity of the polysaccharides from Polygonatum cyrtonema. The two-stage alcohol precipitation, macroporous resin and dialysis combined purification are used to achieve high-purity extraction of polysaccharides from Polygonatum cyrtonema, meeting the market demand for high-purity polysaccharide products. The extraction method maintains the biological activity of the polysaccharide, so that the extracted polysaccharide has significant antioxidant properties.
[0082] The present invention has been described in detail above in conjunction with specific implementations and exemplary examples, but these descriptions cannot be understood as limiting the present invention. Those skilled in the art understand that, without departing from the spirit and scope of the present invention, a variety of equivalent substitutions, modifications or improvements may be made to the technical solution of the present invention and its implementation methods, all of which fall within the scope of the present invention. The scope of protection of the present invention shall be subject to the attached claims.
Claims
1. A method for extracting polysaccharides from Polygonatum cyrtonema with antioxidant activity, characterized in that: The steps include: S1. After defatting, the ultrafine powder of Polygonatum cyrtonema is mixed with pure water and placed in an ultrasonic water bath for ultrasonic treatment; S2, enzymatic hydrolysis: add cellulase and xylanase, and bathe at 50-70℃ for 2-4h; then add thermostable α-amylase, and bathe at 70-100℃ for 1-2h; cool the water bath to 50-70℃, and add saccharifying enzyme for water bath treatment for 2-3h; after the extraction is completed, heat to 100℃ to inactivate the enzyme, centrifuge, and collect the supernatant; S3. The supernatant is sequentially subjected to primary alcohol precipitation, decolorization and purification, secondary alcohol precipitation, dialysis, and freeze-drying treatment to obtain Polygonatum cyrtonema polysaccharide with antioxidant activity.
2. The method for extracting the polysaccharide from Polygonatum cyrtonema with antioxidant activity according to claim 1, characterized in that: In S1, the power of the ultrasonic treatment is 200-400 W, the temperature is 40-60° C., and the time is 60-120 min.
3. The method for extracting the polysaccharide from Polygonatum cyrtonema with antioxidant activity according to claim 1, characterized in that: In S2, the enzymatic hydrolysis process is carried out in a water bath with stirring at a rotation speed of 180-250 r / min.
4. The method for extracting polysaccharides from Polygonatum cyrtonema with antioxidant activity according to claim 1, characterized in that: In S3, the decolorization and purification is specifically as follows: AB-8 macroporous resin is added to the polysaccharide solution after the first alcohol precipitation, and the decolorization treatment is carried out at 30-37°C and 100-200r / min for 10-15h, and the addition amount of the macroporous resin is 20%-30% of the mass of the polysaccharide solution after the first alcohol precipitation.
5. The method for extracting polysaccharides from Polygonatum cyrtonema with antioxidant activity according to claim 1, characterized in that: In S3, the alcohol precipitation is specifically as follows: adding anhydrous ethanol until the mass concentration of ethanol is 10%-20%, standing and centrifuging to remove the precipitate, adding anhydrous ethanol until the mass concentration of ethanol is 80-95%, standing and centrifuging to collect the precipitate.
6. The method for extracting polysaccharides from Polygonatum cyrtonema with antioxidant activity according to claim 1, characterized in that: In S3, the dialysis is specifically as follows: dissolving the polysaccharide after the secondary alcohol precipitation in water, adding it into a 3000-5000Da dialysis bag, dialyzing it with flowing water, and then dialyzing it with pure water.
7. The method for extracting polysaccharides from Polygonatum cyrtonema with antioxidant activity as claimed in claim 1, characterized in that: In S1, the degreasing treatment is specifically: reflux with petroleum ether as solvent at 60-70°C for 2-4 hours.
8. The method for extracting polysaccharides from Polygonatum cyrtonema with antioxidant activity as claimed in claim 1, characterized in that: In S1, the preparation method of the Polygonatum cyrtonema flower superfine powder is: drying the Polygonatum cyrtonema flower at 40-60° C., crushing and sieving, and then grinding the Polygonatum cyrtonema flower with an ultrafine pulverizer to obtain micron-grade superfine powder.
9. A Polygonatum cyrtonema flower polysaccharide with antioxidant activity obtained by the preparation method according to any one of claims 1 to 8.
10. Use of the Polygonatum cyrtonema flower polysaccharide as claimed in claim 9 in antioxidant products.