Special enzyme preparation for walnut shells as well as preparation method and application of special enzyme preparation
By using an enzyme preparation composed of cellulase, xylanase, glutathione and fumaric acid, the problem of difficult utilization of walnut shells was solved, and the efficient extraction and quality improvement of walnut shell polysaccharides were achieved, with significant economic and ecological benefits.
Patent Information
- Application Number
- CN202510890945.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-30
- Publication Date
- 2025-09-26
AI Technical Summary
In the existing technology, walnut shells are difficult to be effectively utilized, resulting in resource waste and environmental pollution. There is a lack of research on walnut shell-specific enzyme preparations for efficient extraction of active ingredients.
An enzyme preparation composed of cellulase, xylanase, glutathione and fumaric acid is used to enzymatically hydrolyze walnut shells and synergistically extract walnut shell polysaccharides. The enzymatic hydrolysis function of cellulase and xylanase is maximized, and the enzyme activity is improved through the synergistic effect of glutathione and fumaric acid.
The efficient extraction of walnut shell polysaccharides is achieved, the anti-reduction ability of polysaccharides is significantly improved, and significant economic and ecological benefits are achieved.
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field of enzyme preparations, in particular to a special enzyme preparation for walnut shells, a preparation method and application thereof. Background Art
[0002] Walnuts, belonging to the Juglans genus of the Juglandaceae family, are known as the king of woody oilseeds and rank first among the world's four major dried fruits. As a major walnut producer and consumer, China produced 4.15 million tons of walnuts in 2017. Walnut shells are waste left after walnut processing or consumption. Assuming walnut shells account for 30% of the total walnut weight, walnut shell production in 2017 was approximately 1.245 million tons. Previously, walnuts were sold as dried fruit, making their shells difficult to recycle. However, many walnuts are now processed for further use, and the resulting large amounts of shells are often discarded or incinerated, resulting in significant resource waste and environmental pollution. Therefore, how to fully develop and utilize walnut shell resources and tap their potential value, thereby achieving an organic balance of economic, ecological, and social benefits, is an urgent issue that needs to be addressed.
[0003] Currently, walnut shells are primarily used for the preparation of activated carbon, extraction of brown pigments, filter media in filters, and the production of wood vinegar. Existing studies have also found that walnut shells contain a variety of active substances, such as phenolic acids, flavonoids, and glycosides, which have antioxidant, antibacterial, lipid-lowering, and anti-tumor effects. Enzyme preparations refer to biological products with catalytic functions that have been purified, processed, and supplemented with auxiliary ingredients. They are primarily used to catalyze various chemical reactions in the production process and have the characteristics of high catalytic efficiency, high specificity, mild reaction conditions, reduced energy consumption, and reduced chemical pollution. However, research on enzyme preparations specifically for walnut shells has not yet been reported. Therefore, there is an urgent need to provide an enzyme preparation product specifically for the efficient extraction of active ingredients from walnut shells. Summary of the Invention
[0004] The purpose of the present invention is to provide a walnut shell-specific enzyme preparation and a preparation method and application thereof, so as to solve the problems existing in the above-mentioned prior art. The walnut shell-specific enzyme preparation formula of the present invention maximizes the enzymatic hydrolysis function of cellulase and xylanase, and exerts the synergistic effect of glutathione and fumaric acid in protecting and improving enzyme activity, which has great economic significance, ecological benefits and broad application prospects.
[0005] To achieve the above object, the present invention provides the following solutions:
[0006] The present invention provides a special enzyme preparation for walnut shells, comprising enzymes and auxiliary materials;
[0007] The enzymes include cellulase and xylanase; and the auxiliary materials include glutathione and fumaric acid.
[0008] Preferably, the mass ratio of the enzyme to the auxiliary material is 2:1.
[0009] Preferably, the mass ratio of the cellulase to the xylanase is (3-1):(1-3).
[0010] Preferably, the mass ratio of the glutathione to the fumaric acid is (3-2):(2-3).
[0011] The present invention also provides a method for preparing the above-mentioned walnut shell-specific enzyme preparation, comprising the following steps:
[0012] The cellulase, the xylanase, the glutathione and the fumaric acid are weighed and mixed to obtain the walnut shell-specific enzyme preparation.
[0013] The present invention also provides an application of the above-mentioned enzyme preparation specifically for walnut shells in decomposing walnut shells.
[0014] The present invention also provides an application of the above-mentioned walnut shell special enzyme preparation in extracting walnut shell polysaccharide.
[0015] The present invention also provides a method for extracting walnut shell polysaccharide, comprising the step of using the above-mentioned walnut shell special enzyme preparation to enzymatically hydrolyze walnut shell powder.
[0016] Preferably, the method comprises the following steps:
[0017] The walnut shells were removed from the kernels, washed, dried, crushed, sieved, defatted, and filtered to obtain walnut shell powder after adding ethanol.
[0018] The walnut shell powder is mixed with distilled water, dissolved, and pH is adjusted, and the walnut shell special enzyme preparation is added for enzymolysis; the enzyme is inactivated, centrifuged, and the supernatant is collected, concentrated, and protein is removed to obtain a walnut shell polysaccharide extract.
[0019] Preferably, the enzymatic hydrolysis is performed at a pH of 4.8, a temperature of 40°C, and a time of 50 min;
[0020] The mass ratio of the walnut shell powder to the walnut shell-specific enzyme preparation is 100:(2-3).
[0021] The present invention discloses the following technical effects:
[0022] The raw materials of the walnut shell-specific enzyme preparation provided by the present invention include cellulase, xylanase, glutathione and fumaric acid. The present invention also provides the use of the walnut shell-specific enzyme preparation in decomposing walnut shells and extracting walnut shell polysaccharides. Experimental results show that the use of the walnut shell-specific enzyme preparation provided by the present invention can efficiently extract polysaccharides in walnut shells, and the anti-reduction ability of the obtained walnut shell polysaccharides is significantly better than that of the conventional walnut shell polysaccharide extraction method. The formula of the walnut shell-specific enzyme preparation of the present invention maximizes the enzymatic hydrolysis function of cellulase and xylanase, and plays a synergistic role of glutathione and fumaric acid in protecting and improving enzyme activity. The present invention provides a new strategy for the recycling of walnut shells, which has great economic significance, ecological benefits and broad application prospects. DETAILED DESCRIPTION
[0023] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.
[0024] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. The intermediate value within any stated value or stated range, and each smaller range between any other stated value or intermediate value within the stated range, is also encompassed within the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.
[0025] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.
[0026] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be exemplary only.
[0027] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.
[0028] The technical solution for extracting plant polysaccharides using enzyme preparations in the prior art includes: an enzyme preparation and method for extracting mulberry leaf polysaccharides, taking air-dried mulberry leaves, crushing them into 100 meshes, taking 9g of the powder, adding 180ml of distilled water, boiling and cooling for later use; taking a composite enzyme preparation: 0.02g of cellulase, 0.02g of pectinase, and 0.05g of papain, adding 0.9g of distilled water, and bathing in a 40°C water bath for 30min; mixing the solutions obtained in the first two steps evenly, adjusting the pH to 5.0, and performing enzymolysis at 50°C for 1h; decocting the enzymatic hydrolyzate, controlling the heating temperature, and keeping the solution in a slightly boiling state for 2h; centrifuging, taking the supernatant for filtration, and concentrating the filtrate to obtain mulberry leaf polysaccharides.
[0029] The technical solution for extracting plant polysaccharides using enzyme preparations in the prior art includes: an astragalus polysaccharide extraction process, pretreatment: the raw astragalus is picked, washed, moistened, and cut, and set aside; raw material enzymatic hydrolysis: the pretreated astragalus is put into 8-12 times the amount of distilled water, heated to 45-50°C, adjusted to pH = 5.5-7, added with enzyme preparation, and hydrolyzed for 2-3 hours; calcium hydroxide solution is added, the pH value is adjusted to 9-10, heated to 75°C, maintained for 60-100 minutes, naturally cooled to 50°C, filtered to remove residue, and filtrate I is obtained for set aside; residue enzymatic hydrolysis: the filtered residue is collected, put into 6-8 times the amount of distilled water, heated to 50-60°C, adjusted to pH = 5.5-7, added with enzyme preparation, and hydrolyzed for 2-3 hours. ; Add calcium hydroxide solution, adjust the pH value to 9-10, heat to 75°C, maintain for 40-60 minutes, cool naturally to 50°C, filter and remove the residue to obtain filtrate II-1; collect the residue, repeat the above operation to obtain filtrate II-2; combine filtrates I, II-1 and II-2; concentration: put the combined filtrate into a vacuum concentrator, concentrate under reduced pressure, and the concentrate is 70% of the original solution; alcohol precipitation: slowly add 95% ethanol to the concentrate until the ethanol concentration reaches 70%, let it stand and settle for 8-12 hours, the precipitate is astragalus polysaccharide, mechanically filter, and then wash with 95% ethanol 1-2 times; the washed astragalus polysaccharide precipitate is freeze-dried, then crushed, and passed through a 100-mesh sieve to obtain an astragalus polysaccharide extract.
[0030] Existing technical solutions for extracting polysaccharides using enzyme preparations include: a method for extracting polysaccharides from Morchella edulis. The method includes fermentation: dried Morchella edulis fruiting bodies are crushed, then expanded with 5-7 times the amount of water. A 1% (w / w) composite inoculum consisting of yeast and chitinase-secreting Bacillus subtilis (hereinafter referred to as "Bacillus") is added and fermented on a shaker. The weight ratio of yeast to Bacillus is 1:2, the fermentation temperature is 29°C, the pH is 4.5, and the fermentation time is 16 hours. Enzymolysis: After a period of fermentation, the fermentation broth is mixed and a composite enzyme preparation is added, comprising a 1:1 weight ratio of chitinase and acid protease. The addition amount of the complex enzyme preparation is 1.5%, the action temperature is 36°C, the pH value is 4.2, and the action time is 12 hours; the enzymatic hydrolyzate obtained after enzymatic hydrolysis is centrifuged, the supernatant is taken and concentrated, and then 4 times the volume of anhydrous ethanol is added to obtain an alcohol precipitate, and then the alcohol precipitate is spray-dried to obtain water-soluble morel polysaccharide.
[0031] The prior art technical solution for extracting biopolysaccharides using enzyme preparations includes: a method for extracting bamboo fungus polysaccharides, wherein dried bamboo fungus stalks are crushed through a 10-mesh sieve to obtain an undersize fraction, namely bamboo fungus powder; the bamboo fungus powder is placed in 40°C water at a solid-liquid ratio of 1g:20mL for extraction, and stirred at a stirring speed of 60r / min for 160min to obtain a reaction solution; the reaction solution temperature is controlled at 40°C, the pH is adjusted to 5-6, and a mixed enzyme preparation is added at 0.05% by weight of the bamboo fungus powder, and enzymatic hydrolysis is carried out for 15min to obtain an enzymatic hydrolyzate; The enzymatic hydrolyzate was heated to 75°C, and the extraction was continued with stirring at a speed of 60 r / min for 20 minutes to obtain an extract; after the extract was restored to room temperature, it was placed in a centrifuge at 7000 r / min, centrifuged for 5 minutes, and the supernatant was collected; anhydrous ethanol was added to the supernatant at a volume of 3 times that of the supernatant, and the supernatant was sealed and allowed to stand for 6 hours to obtain a mixture containing a precipitate, which was placed in a centrifuge at 7000 r / min, centrifuged for 5 minutes, and the solids were collected. The solids were placed in a cool, dark and ventilated place to air-dry for 20 hours to obtain bamboo fungus polysaccharide.
[0032] The cellulase (item number A002598), glutathione (item number A429652) and fumaric acid (item number A500472) of the present invention were purchased from Shanghai Shenggong Bioengineering Co., Ltd., and the xylanase (item number ml015177) was purchased from Shanghai ELISA Biotechnology Co., Ltd.
[0033] Example 1
[0034] 0.5 g of cellulase, 0.5 g of xylanase, 0.3 g of glutathione and 0.2 g of fumaric acid were mixed to obtain a special enzyme preparation for walnut shells.
[0035] The walnuts were shelled and washed, dried at 75°C for 2 h, crushed and passed through a 60-mesh sieve, and petroleum ether was added at a solid-liquid ratio of 1 g:30 mL. The mixture was refluxed at 65°C for 2 h in a constant temperature water bath for degreasing. 80% ethanol was then added at a solid-liquid ratio of 1 g:30 mL and refluxed for 2 h to remove monosaccharides and oligosaccharides. The mixture was then filtered and dried to obtain walnut shell powder.
[0036] 5 g of walnut shell powder was added to 150 mL of distilled water for dissolution, a citric acid buffer solution was added to adjust the pH of the system to 4.8, 0.15 g of a special enzyme preparation was added, the pH was 4.8, and enzymatic hydrolysis was carried out at 40° C. for 50 min; the enzyme was inactivated in a 95° C. boiling water bath for 15 min, the mixture was cooled and centrifuged at 3000 rpm for 20 min, the supernatant was collected, and the mixture was concentrated to 1 / 4 of the original volume, protein was removed by precipitation through a Sevage method, and the mixture was centrifuged at 3000 rpm for 20 min, and the supernatant was collected to obtain a walnut shell polysaccharide extract.
[0037] Example 2
[0038] The only difference from Example 1 is that the ingredients of the walnut shell enzyme preparation are: 0.75g cellulase, 0.25g xylanase, 0.3g glutathione and 0.2g fumaric acid.
[0039] Example 3
[0040] The only difference from Example 1 is that the ingredients of the walnut shell enzyme preparation are: 0.25g cellulase, 0.75g xylanase, 0.2g glutathione and 0.3g fumaric acid.
[0041] Example 4
[0042] 0.5 g of cellulase, 0.5 g of xylanase, 0.2 g of glutathione and 0.3 g of fumaric acid were mixed to obtain a special enzyme preparation for walnut shells.
[0043] The walnuts were shelled and washed, dried at 75°C for 2 h, crushed and passed through a 60-mesh sieve, and petroleum ether was added at a solid-liquid ratio of 1 g:30 mL. The mixture was refluxed at 65°C for 2 h in a constant temperature water bath for degreasing. 80% ethanol was then added at a solid-liquid ratio of 1 g:30 mL and refluxed for 2 h to remove monosaccharides and oligosaccharides. The mixture was then filtered and dried to obtain walnut shell powder.
[0044] 5 g of walnut shell powder was added to 150 mL of distilled water for dissolution, a citric acid buffer solution was added to adjust the pH of the system to 4.8, 0.1 g of a special enzyme preparation was added, the pH was 4.8, and enzymatic hydrolysis was carried out at 40° C. for 50 min; the enzyme was inactivated in a 95° C. boiling water bath for 15 min, the mixture was cooled and centrifuged at 3000 rpm for 20 min, the supernatant was collected, and the mixture was concentrated to 1 / 4 of the original volume, protein was removed by precipitation through a Sevage method, and the mixture was centrifuged at 3000 rpm for 20 min, the supernatant was collected, and a walnut shell polysaccharide extract was obtained.
[0045] Comparative Example 1
[0046] The only difference from Example 1 is that the ingredients of the walnut shell-specific enzyme preparation are: 1g cellulase, 0.2g glutathione and 0.3g fumaric acid.
[0047] Comparative Example 2
[0048] The only difference from Example 1 is that the ingredients of the walnut shell enzyme preparation are: 1g xylanase, 0.2g glutathione and 0.3g fumaric acid.
[0049] Comparative Example 3
[0050] The only difference from Example 1 is that the ingredients of the walnut shell enzyme preparation are: 0.5g cellulase, 0.5g xylanase and 0.5g glutathione.
[0051] Comparative Example 4
[0052] The only difference from Example 1 is that the ingredients of the walnut shell enzyme preparation are: 0.5g cellulase, 0.5g xylanase and 0.5g fumaric acid.
[0053] Comparative Example 5 Water Extraction Method of Walnut Shell Polysaccharide
[0054] Take 20g of dried walnut shells and crush them thoroughly in a grinder. Place the crushed walnut shells in a 250mL beaker, add 150mL of water, and soak at room temperature for 12 hours. After soaking, pour it into a 1000mL round-bottom flask, add 500mL of water, and extract at 90℃ for 4 hours. Collect the aqueous solution by suction filtration, and continue to extract the residue with 500mL of water for 4 hours, repeating twice. Combine the filtrate, concentrate it under reduced pressure to 200mL, add 80mL of chloroform and 20mL of n-butanol, stir at room temperature for 30min, and remove the protein from the polysaccharide. After the protein removal operation is completed, centrifuge and collect the supernatant to obtain the walnut shell polysaccharide extract.
[0055] Comparative Example 6 Microwave-assisted extraction method of walnut shell polysaccharide
[0056] The walnuts were shelled after being cleaned, dried naturally, and then ground and passed through a 20-mesh sieve. The resulting walnut shell powder was wrapped with filter paper and placed in a Soxhlet extractor. An appropriate amount of petroleum ether was added and cold-soaked for 24 hours. The extract was then placed in a water bath for Soxhlet extraction until the lipids and fat-soluble pigments in the sample were fully removed. The sample filter paper package was removed and air-dried until the petroleum ether was completely evaporated. The filter paper package was then dried in an oven at 60°C to obtain walnut shell dry powder with the pigment and oil removed.
[0057] Weigh 1 g of the walnut shell powder and place it in a microwave extraction tank. Add 40 mL of distilled water and microwave-assisted extraction at 70°C for 6 minutes. Centrifuge the extract (4°C, 4000 rpm for 5 minutes) and collect the supernatant to obtain the walnut shell polysaccharide extract.
[0058] Experimental Example 1 Calculation of walnut shell polysaccharide extraction rate
[0059] 1. Experimental Methods
[0060] 1.1 Standard curve drawing
[0061] Accurately weigh 100.0 mg of anhydrous glucose dried to constant weight, dissolve it in distilled water and transfer it to a 100 mL volumetric flask to make up the volume to obtain a glucose standard solution with a mass concentration of 1 mg / mL. Accurately pipette 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, and 0.8 mL of glucose standard solution into a 100 mL volumetric flask to make up the volume. Then pipette 2.0 mL of glucose solution into a test tube, add 1.0 mL of 5% phenol solution and 5.0 mL of concentrated sulfuric acid, and mix well. Heat in a constant temperature water bath at 70°C for 15 minutes, then cool with tap water for 15 minutes, and measure the absorbance at a wavelength of 490 nm. With glucose mass concentration as the horizontal axis and absorbance as the vertical axis, a glucose standard curve is drawn, and the regression equation is: y = 6.2783x-0.0054, R 2 =0.9993.
[0062] 1.2 Calculation of walnut shell polysaccharide extraction rate
[0063] Accurately pipette 2.0 mL of the walnut shell polysaccharide extract prepared in Examples 1-4 and Comparative Examples 1-6 into a stoppered test tube, add 1.0 mL of 5% phenol solution, shake well, then slowly add 5.0 mL of concentrated sulfuric acid, shake well, and heat in a 70°C water bath for 15 minutes. After taking out, cool with tap water for 15 minutes. Use distilled water as a blank control according to the same operation. Measure its absorbance at a wavelength of 490 nm using a visible spectrophotometer. Repeat the measurement three times and take the average value. Calculate the polysaccharide content of the sample using the standard curve regression equation. The polysaccharide extraction rate is calculated according to the following formula:
[0064] Walnut shell polysaccharide extraction rate (%) = C × V / M × 100;
[0065] In the above formula, C is the polysaccharide content of the walnut shell extract, V is the total volume of the walnut shell polysaccharide extract, and M is the mass of the walnut shell powder.
[0066] 2. Experimental Results
[0067] As shown in Table 1, the walnut shell polysaccharide extraction rates of Examples 1-4 of the present invention are significantly higher than those of Comparative Examples 1-2, indicating that the compounding of cellulase and xylanase within a certain range and the co-enzymatic hydrolysis can improve the enzymatic extraction efficiency of walnut shell polysaccharides; the walnut shell polysaccharide extraction rates of Examples 1-4 of the present invention are significantly higher than those of Comparative Examples 3-4, indicating that the combined use of glutathione and fumaric acid in the enzyme preparation exerts a synergistic effect of the two in protecting and enhancing enzyme activity. The walnut shell polysaccharide extraction rates of Examples 1-4 of the present invention are higher than those of Comparative Examples 5-6, indicating that the walnut shell-specific enzyme preparations prepared in Examples 1-4 can obtain a higher extraction rate in the enzymatic extraction of walnut shell polysaccharides than conventional extraction methods.
[0068] Table 1 Statistics of walnut shell polysaccharide extraction rate
[0069]
[0070]
[0071] Experimental Example 2 Determination of the reducing ability of walnut shell polysaccharide
[0072] 1. Experimental Methods
[0073] The concentration of walnut shell polysaccharide in the walnut shell polysaccharide extract prepared in Example 1-4 and Comparative Example 1-6 was adjusted to 0.05 mg / mL, and a 0.05 mg / mL vitamin C aqueous solution was prepared as a positive control. 0.4 mL of liquid was accurately pipetted, and 1 mL of 0.2 mol / L phosphate buffer solution and 1 mL of 0.4 mol / L potassium ferricyanide were added and mixed. The mixture was heated in a 55°C water bath for 20 min, followed by addition of 1 mL of 100 mg / mL trichloroacetic acid to the mixture, followed by centrifugation for 10 min. The supernatant was then added with 1 mL of filtered water and 0.2 mL of 1 mg / mL ferric chloride. After mixing, its absorbance was measured at 700 nm. The reducing power of the walnut shell polysaccharide was determined based on the absorbance at 700 nm. The greater the absorbance, the stronger the reducing power, indicating that its antioxidant activity was stronger.
[0074] 2. Experimental Results
[0075] The results are shown in Table 2. The absorbance values of the walnut shell polysaccharides of Examples 1-4 of the present invention after the reaction is completed are comparable to the absorbance values of vitamin C and significantly higher than those of Comparative Examples 1-6, indicating that the walnut shell polysaccharides extracted using the walnut shell-specific enzyme preparation provided by the present invention have significantly stronger reducing ability, indicating that the walnut shell-specific enzyme preparation of the present invention significantly improves the quality of walnut shell polysaccharides by combining cellulase and xylanase with auxiliary materials glutathione and fumaric acid.
[0076] Table 2 Statistics of reducing ability of walnut shell polysaccharides
[0077] Experimental groups Absorbance value Example 1 0.745 Example 2 0.727 Example 3 0.711 Example 4 0.722 Comparative Example 1 0.415 Comparative Example 2 0.492 Comparative Example 3 0.386 Comparative Example 4 0.437 Comparative Example 5 0.559 Comparative Example 6 0.512 Vitamin C 0.736
[0078] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art should fall within the scope of protection defined by the claims of the present invention.
Claims
1. A walnut shell enzyme preparation, characterized in that: Including enzymes and excipients; The enzymes include cellulases and xylanases; The auxiliary materials include glutathione and fumaric acid.
2. The walnut shell-specific enzyme preparation according to claim 1, wherein The mass ratio of the enzyme to the auxiliary material is 2:
1.
3. The walnut shell enzyme preparation according to claim 1, wherein The mass ratio of the cellulase to the xylanase is (3-1):(1-3).
4. The walnut shell-specific enzyme preparation according to claim 1, wherein The mass ratio of the glutathione to the fumaric acid is (3-2):(2-3).
5. A method for preparing the walnut shell enzyme preparation according to any one of claims 1 to 4, characterized in that: The following steps are involved: The cellulase, the xylanase, the glutathione and the fumaric acid are weighed and mixed to obtain the walnut shell-specific enzyme preparation.
6. Use of the walnut shell-specific enzyme preparation according to any one of claims 1 to 4 in decomposing walnut shells.
7. Use of the walnut shell-specific enzyme preparation according to any one of claims 1 to 4 in extracting walnut shell polysaccharides.
8. A method for extracting walnut shell polysaccharide, characterized in that: The method comprises the step of enzymatically hydrolyzing walnut shell powder by using the special enzyme preparation for walnut shell according to any one of claims 1 to 4.
9. The method according to claim 8, wherein The following steps are involved: The walnut shells were removed from the kernels, washed, dried, crushed, sieved, defatted, and filtered to obtain walnut shell powder after adding ethanol. The walnut shell powder is mixed with distilled water, dissolved, and pH is adjusted, and the walnut shell special enzyme preparation is added for enzymolysis; the enzyme is inactivated, centrifuged, and the supernatant is collected, concentrated, and protein is removed to obtain a walnut shell polysaccharide extract.
10. The method according to claim 9, wherein The enzymatic hydrolysis was performed at a pH of 4.8, a temperature of 40°C, and a time of 50 min; The mass ratio of the walnut shell powder to the walnut shell-specific enzyme preparation is 100:(2-3).