Grape wine clarifying agent as well as preparation method and application thereof
A wine clarifying agent was prepared by extracting and enzymatically hydrolyzing potato protein using thermal flocculation, which solved the allergy risk and cost issues of animal-derived clarifying agents and achieved efficient clarification and antioxidant effects, making it suitable for wine clarification.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- LANZHOU INSTITUTE OF CHEMICAL PHYSICS CHINESE ACADEMY OF SCIENCES
- Filing Date
- 2026-04-08
- Publication Date
- 2026-05-12
AI Technical Summary
Existing animal-derived and chemical clarifying agents pose allergy risks and are costly in wine, making it difficult to find alternatives that are less allergenic, provide good clarification, and are inexpensive.
Potato protein was used as a clarifying agent. Potato protein was extracted by thermal flocculation, and then combined with pH adjustment and papain hydrolysis to prepare potato protein hydrolysate for wine clarification.
It achieves low allergenicity and good clarification, reduces the content of tannins and phenols, maintains the flavor of the wine, and increases the antioxidant capacity and drinking window of the wine.
Smart Images
Figure CN122012211A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of food manufacturing technology, specifically relating to a wine clarifying agent, its preparation method, and its application. Background Technology
[0002] Animal proteins are commonly used in winemaking to adjust the astringency of red wine. Astringency is primarily caused by the interaction between salivary proteins and polyphenols such as condensed tannins and ellagitannins. Wines are clarified before bottling to remove substances that cause turbidity and improve stability. Commonly used clarifying agents include bentonite, gelatin, fish glue, casein, egg white protein, and PVPP. Using bentonite as a clarifying agent affects the color of the wine and leaves residues, while using PVPP not only leaves residues but also increases winemaking costs. In existing technologies, casein, egg white, and gelatin are used as clarifying agents to reduce astringency in wine; however, due to their animal origin, they pose a potential risk of allergies and are detrimental to human health. Therefore, providing a plant-derived protein with low allergenicity, simple preparation, good clarifying effect, stable wine body, and low usage cost to replace animal proteins as a clarifying agent for red wine has become a problem urgently needing to be solved by those skilled in the art.
[0003] Potato protein is mainly composed of patatin (tuber storage protein), protease inhibitors, and other proteins. Protease inhibitors account for approximately 50% of the total protein, making them the most abundant protein, and possess excellent antioxidant, anticancer, and broad-spectrum antibacterial activities. From a nutritional perspective, they are considered an anti-nutritional factor with high application value in functional foods and biomedicine. Patatin protein is the main protein in potato tubers (39-45 kDa), a family of glycoproteins that accounts for more than 40% of the total soluble protein in potato tubers. Patatin protein has DPPH free radical scavenging ability and resistance to low-density lipoprotein peroxidation. This is due to the 12 soluble amino acids (methionine, tryptophan, tyrosine, phenylalanine, cysteine, and histidine) with free radical scavenging capabilities contained in patatin protein. These amino acids are exposed on the surface of the patatin protein molecule, determining its antioxidant properties. Potato protein has good solubility, foaming, and emulsifying properties, making it a suitable food additive. Potato protein is widely used in food processing, such as as a water-based binder in meat and sausages, a foaming agent in confectionery, baking, and dairy products, and an emulsifier in sauces, gravies, desserts, and condiments. Therefore, potato protein can replace animal protein as a clarifying agent for red wine, not only reducing allergic reactions but also increasing the amino acid content of the wine. Summary of the Invention
[0004] The purpose of this invention is to provide a method for preparing a wine clarifying agent to address the problem of allergic risks associated with animal-derived and chemical clarifying agents.
[0005] Another object of the present invention is to provide a wine clarifying agent.
[0006] A third objective of this invention is to provide an application of a wine clarifying agent.
[0007] The technical solution of this invention is: (a) A method for preparing a wine clarifying agent, comprising the following steps: S1. Extraction of potato protein: S1.1. Potato tubers, peeled and cut into small pieces (about 1-2 cm square), add deionized water, blend into a homogenate to break down cells and release protein; S1.2 Separate the homogenate by passing it through gauze (4-8 layers) or by centrifugation (3000-5000 rpm, 10-15 min), collect the supernatant, remove solid residues such as starch and cellulose, and obtain potato juice; if the juice is turbid, it can be filtered or centrifuged repeatedly to improve the clarity. S1.3. Heating potato juice causes protein denaturation and molecular aggregation to form visible flocculent particles. After the thermal flocculation in step S1.4 and S1.3 is completed, cool the mixture to room temperature and centrifuge at 4000-6000 rpm for 30 minutes to allow the flocculated potato protein to precipitate at the bottom. S2. Purification of potato protein: S2.1 Add an equal volume of deionized water to resuspend the potato protein, so that the potato protein is evenly dispersed. S2.2 Add NaOH to adjust the pH of the dispersion obtained in step S2.1 to 9-10, heat to 45-55℃, and maintain for 1-1.5h to dissolve the potato protein; S2.3 Centrifuge the mixture obtained in step S2.2 at 4000-6000 rpm for 30 min, discard the precipitate, and collect the supernatant; S2.4 Adjust the pH of the supernatant to 4.5-5.5 (near the isoelectric point of potato protein) with 1M HCl, stir well, centrifuge the mixture at 4000-6000 rpm, discard the supernatant, and collect the precipitate; S2.5 Add an equal volume of deionized water to resuspend the precipitate, centrifuge at 4000-6000 rpm for 30 min, and repeat the washing process 2-3 times to obtain purified potato protein. S3. Enzymatic hydrolysis of potato protein: S3.1 Resuspend the purified potato protein in deionized water, add papain, and perform enzymatic hydrolysis; S3.2 After the enzymatic hydrolysis is completed, heat the mixture to inactivate the enzyme, centrifuge at 4000-6000 rpm for 30 min, discard the precipitate, and collect the supernatant; S3.3 Adjust the pH of the supernatant to 4.5-5.5, stir well, centrifuge the mixture at 4000-6000 rpm, discard the supernatant, and collect the precipitate; S3.4 Add an equal volume of deionized water to resuspend the precipitate, centrifuge at 4000-6000 rpm for 30 min, and repeat the washing process 2-3 times to remove salt. S3.5. Resuspend the precipitate, homogenize it for 30 min using a homogenizer, adjust the pH to 7, and spray dry to obtain potato protein hydrolysate, which is the wine clarifying agent.
[0008] As a further improvement of the present invention, in step S1.1, the mass ratio of potato to deionized water is 1:1-3 (adjusted according to protein concentration requirements).
[0009] As a further improvement of the present invention, in step S1.3, the potato juice is heated to 90°C and kept for 30 minutes. The purpose is to break the weak bonds in the protein molecules, causing their spatial structure to unfold, exposing the hydrophobic groups that were originally hidden inside the molecules to the surface. The protein molecules bind with each other through hydrophobic interactions, van der Waals forces, etc., to form larger aggregates (flocculents) that precipitate from the solution.
[0010] As a further improvement of the present invention, in step S2.2, ultrasonic-assisted treatment is used to increase the solubility of potato protein.
[0011] As a further improvement of the present invention, in step S2.2, when using ultrasonic-assisted processing, the power is set to 200W, the temperature is set to 50℃, and the processing time is 30min.
[0012] As a further improvement of the present invention, in step S3.1, when enzymatically hydrolyzing potato protein, the papain activity is 200,000 U-300,000 U, the substrate mass concentration is 3%, the enzyme-to-substrate mass ratio is 6:100, the pH is maintained at 6.5, the temperature is 55°C, and the hydrolysis time is 3 hours.
[0013] As a further improvement of the present invention, in step S3.2, the mixture is heated to 85°C and held for 10 minutes.
[0014] (ii) A wine clarifying agent, prepared by the above-mentioned method for preparing a wine clarifying agent.
[0015] (III) The application of a wine clarifying agent in the wine clarification process, including the following steps: A. Weigh the wine clarifying agent and place it in a container; B. Dissolve the wine clarifying agent in a small amount of the wine to be treated, and stir until it becomes a paste; C. Add the paste-like wine clarifying agent to the wine tank containing the wine to be treated, stir for 0.5-2 hours / day, continue for 2-4 days, and let stand; D. After settling, filter the wine, determine its clarity, and bottle it.
[0016] Furthermore, in step A, the amount of wine clarifying agent used is 0.1-0.3 g / L.
[0017] This invention extracts potato protein using thermal flocculation, purifies it using pH adjustment, and then hydrolyzes it with papain to obtain potato protein hydrolysate, which has biological activity. Adding the potato protein hydrolysate to red wine in a certain proportion and allowing it to precipitate for a certain period of time can achieve the purpose of clarification and removal of astringency.
[0018] Potato enzyme hydrolysates interact with phenolic substances and tannins in wine to form large molecular flocs that precipitate, while also causing other turbid substances to settle. This not only achieves clarification but also reduces the content of tannins and phenolic substances, essentially preserving the wine's color and reducing bitterness.
[0019] Compared with the prior art, the present invention has the following advantages: 1. The potato protein hydrolysate prepared by the method of the present invention, when used as a wine clarifying agent, will not cause a change in the color of red wine after treatment; and it significantly reduces tannins that cause astringency in red wine, thus maintaining the original flavor of the red wine.
[0020] 2. The removal capabilities of different clarifying agents for wine polyphenols are as follows: The wine clarifying agent of this invention = gelatin > egg white protein > casein ( P >0.05). On the one hand, using potato protein hydrolysate as a wine clarifying agent increases the protein or polypeptide content in the wine; on the other hand, it enhances the antioxidant capacity and antibacterial properties of the wine, thereby increasing the wine's optimal drinking window.
[0021] 3. The potato protein used in this invention is extracted from the juice separated during potato starch processing using a thermal flocculation method. This represents waste recycling, reducing environmental pollution and turning waste into treasure. The potato protein extracted using the thermal flocculation method, after enzymatic hydrolysis, retains biological activity and normal protein functional characteristics. Therefore, potato protein hydrolysate, as a wine clarifying agent, not only achieves excellent clarification but also reduces allergic reactions, increases antioxidant properties, and does not alter the wine's flavor. Attached Figure Description
[0022] Figure 1This is a diagram showing the salivation index of the potato protein hydrolysate prepared in Example 1 of the present invention and the wine treated with gelatin, egg white protein and casein. Figure 2 This is a graph showing the tannin content of the potato protein hydrolysate prepared in Example 1 of the present invention and the wine treated with gelatin, egg white protein, and casein. Figure 3 This is a graph showing the total phenol content of the potato protein hydrolysate prepared in Example 1 of the present invention and the wine treated with gelatin, egg white protein, and casein. Figure 4 This is the total ion chromatogram of the potato protein hydrolysate in Example 2 of the present invention; Figure 5 This is a chromatogram (a) of the antioxidant peptides screened in Example 2 of the present invention; Figure 6 This is a chromatogram (II) of the antioxidant peptides screened in Example 2 of the present invention. Detailed Implementation
[0023] The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.
[0024] Example 1 A method for preparing a wine clarifying agent includes the following steps: S1. Extraction of potato protein: S1.1 Select fresh potato tubers, wash to remove surface mud and impurities, peel and cut into small pieces (about 1-2cm square), place in a tissue homogenizer or homogenizer, add deionized water, the mass ratio of potato to deionized water is 1:3, use the tissue homogenizer to homogenize into a paste, so that the cells are broken and the protein is released. S1.2 Separate the homogenate through gauze (4-8 layers), collect the supernatant, remove solid residues such as starch and cellulose, and obtain potato juice; S1.3. Slowly heat the potato juice to 90°C and maintain for 30 minutes to denature the protein and cause the molecules to aggregate and form visible flocculent particles. After the thermal flocculation in step S1.4 and S1.3 is completed, cool the mixture to room temperature and centrifuge at 5000 rpm for 30 minutes to allow the flocculated potato protein to precipitate at the bottom.
[0025] S2. Purification of potato protein: S2.1 Add an equal volume of deionized water to resuspend the potato protein, so that the potato protein is evenly dispersed. S2.2 Add NaOH to adjust the pH of the dispersion obtained in step S2.1 to 10, heat to 50℃ and maintain for 1 hour to dissolve the potato protein, and then use ultrasound-assisted treatment (power set to 200W, temperature set to 50℃, treatment for 30 minutes) to increase the solubility of the potato protein. S2.3 Centrifuge the mixture obtained in step S2.2 at 5000 rpm for 30 min, discard the precipitate, and collect the supernatant; S2.4 Adjust the pH of the supernatant to 4.5-5.5 with 1M HCl, stir well, centrifuge the mixture at 5000 rpm, discard the supernatant, and collect the precipitate; S2.5 Add an equal volume of deionized water to resuspend the precipitate, centrifuge at 5000 rpm for 30 min, and repeat the washing process 3 times to obtain purified potato protein.
[0026] S3. Enzymatic hydrolysis of potato protein: S3.1 Resuspend the purified potato protein in deionized water, add papain (activity 200,000 U-300,000 U) for enzymatic hydrolysis, substrate concentration 3%, enzyme-to-substrate ratio 6:100, maintain pH 6.5, temperature 55℃, and hydrolysis time 3h. S3.2 After the enzymatic hydrolysis is completed, heat the mixture to 85°C and keep it for 10 min to inactivate the enzyme. Centrifuge at 5000 rpm for 30 min, discard the precipitate, and collect the supernatant. S3.3 Adjust the pH of the supernatant to 4.5-5.5, stir well, centrifuge the mixture at 5000 rpm, discard the supernatant, and collect the precipitate; S3.4 Add an equal volume of deionized water to resuspend the precipitate, centrifuge at 5000 rpm for 30 min, and repeat this washing process 3 times to remove salt. S3.5. Resuspend the precipitate, homogenize it for 30 minutes using a homogenizer, adjust the pH to 7, and spray dry to obtain potato protein hydrolysate, which is the wine clarifying agent.
[0027] 100 L of Cabernet Sauvignon red wine that needs clarification was added to a dry, refrigerated tank. The wine clarifying agent prepared in this embodiment was used for clarification during the production of the modified red wine, as follows: A. Weigh out 10g, 20g, and 30g of wine clarifying agent respectively and place them in containers; B. Dissolve the wine clarifying agent in 200mL of the original wine (the red wine to be treated) and stir until it becomes a paste; C. Add the paste-like wine clarifying agent to the original wine, stir at low speed with a pump for 0.5 hours / day, continue stirring for 3 days, and then let it stand at 4℃ for 2 weeks to clarify. D. After the red wine has settled and clarified, filter it using a filter press, measure its clarity, and then bottle it.
[0028] Evaluation of Clarifying Agent Effect: Gelatin, egg white, casein, and potato protein hydrolysate (as described in this example) were used as clarifying agents, prepared at concentrations of 10, 20, and 30 g / hL, respectively. The clarifying effects of the four clarifying agents on wine were compared. 30 mL of the mixture of the original wine and the clarifying agent was taken, stirred thoroughly, and allowed to stand for clarification. After clarification, the supernatant was taken to determine the salivary sedimentation index (SPI) to evaluate the clarifying effect of potato protein hydrolysate as a wine clarifying agent. The results are as follows: Figure 1 As shown, there was no difference in SPI after treatment with gelatin, casein, and egg white protein. The lower the SPI value, the lower the content of wine polyphenols that react with salivary proteins. When the treatment concentration reached 30 g / hL, the SPI values were: potato protein hydrolysate > gelatin = egg white protein > casein. These results indicate that within the concentration range of 10-30 g / hL, the SPI value of wine gradually decreases with increasing concentration of potato protein hydrolysate. All the clarifying agents tested (potato protein hydrolysate, gelatin, egg white protein, and casein) significantly reduced the total phenol content of red wine at concentrations of 10, 20, and 30 g / hL, with the polyphenol scavenging ability in the following order: potato protein hydrolysate > gelatin = egg white protein > casein. Figure 2 As shown, at a concentration of 30 g / hL, the tannin removal effect was: potato protein hydrolysate = gelatin > egg white protein > casein. This indicates that the tannin removal effect of potato protein hydrolysate is comparable to that of traditional gelatin and superior to other animal proteins. Therefore, potato protein hydrolysate is more effective as a wine clarifying agent than gelatin, egg white protein, and casein, while maintaining minimal change in the wine's color.
[0029] Example 2 A method for preparing a wine clarifying agent includes the following steps: S1. Extraction of potato protein: S1.1 Select fresh potato tubers, wash to remove surface mud and impurities, peel and cut into small pieces (about 1-2cm square), place in a tissue homogenizer or homogenizer, add deionized water, the mass ratio of potato to deionized water is 1:1, homogenize into a homogenate to break down cells and release protein. S1.2 Separate the homogenate by centrifugation (5000 rpm, 10 min), collect the supernatant, remove solid residues such as starch and cellulose, and obtain potato juice; S1.3. Slowly heat the potato juice to 90°C and maintain for 30 minutes to denature the protein and cause the molecules to aggregate and form visible flocculent particles. After the thermal flocculation in step S1.4 and S1.3 is completed, cool the mixture to room temperature and centrifuge at 5000 rpm for 30 minutes to allow the flocculated potato protein to precipitate at the bottom.
[0030] S2. Purification of potato protein: S2.1 Add an equal volume of deionized water to resuspend the potato protein, so that the potato protein is evenly dispersed. S2.2 Add NaOH to adjust the pH of the dispersion obtained in step S2.1 to 9, heat to 50℃ and maintain for 1.5 h to dissolve the potato protein, and then use ultrasound-assisted treatment (power set to 200W, temperature set to 50℃, treatment for 30 min) to increase the solubility of the potato protein. S2.3 Centrifuge the mixture obtained in step S2.2 at 5000 rpm for 30 min, discard the precipitate, and collect the supernatant; S2.4 Adjust the pH of the supernatant to 4.5-5.5 with 1M HCl, stir well, centrifuge the mixture at 5000 rpm, discard the supernatant, and collect the precipitate; S2.5 Add an equal volume of deionized water to resuspend the precipitate, centrifuge at 5000 rpm for 30 min, and repeat the washing process 3 times to obtain purified potato protein.
[0031] S3. Enzymatic hydrolysis of potato protein: S3.1 Resuspend the purified potato protein in deionized water, add papain (activity 200,000 U-300,000 U) for enzymatic hydrolysis, substrate concentration 3%, enzyme-to-substrate ratio 6:100, maintain pH 6.5, temperature 55℃, and hydrolysis time 3h. S3.2 After the enzymatic hydrolysis is completed, heat the mixture to 85°C and keep it for 10 min to inactivate the enzyme. Centrifuge at 5000 rpm for 30 min, discard the precipitate, and collect the supernatant. S3.3 Adjust the pH of the supernatant to 4.5-5.5, stir well, centrifuge the mixture at 5000 rpm, discard the supernatant, and collect the precipitate; S3.4 Add an equal volume of deionized water to resuspend the precipitate, centrifuge at 5000 rpm for 30 min, and repeat this washing process 3 times to remove salt. S3.5. Resuspend the precipitate, homogenize it for 30 minutes using a homogenizer, adjust the pH to 7, and spray dry to obtain potato protein hydrolysate, which is the wine clarifying agent.
[0032] In this embodiment, the supernatant collected in step S3.2 was analyzed for polypeptide composition of the enzymatic hydrolysate using LC-MS / MS. The total ion current chromatograms (TICs) of potato protein after papain hydrolysis were analyzed, as shown below. Figure 4As shown, the baseline signal was low and relatively stable in the early and middle retention regions (0–35 minutes), with a limited number of detectable peaks; in the late elution region (50–65 minutes), the signal intensity increased sharply, dominated by a main peak at 59.1 minutes, with several additional high-intensity peaks at 55.9, 57.9, and 62.6 minutes. The peptide spectrum produced by papain digestion was narrower but more hydrophobic, mainly concentrated in the late retention time.
[0033] Table 1 presents the predicted bioactive peptide data from potato protein hydrolysates. In the papain hydrolysis system, 6710 peptides were predicted to possess ACE inhibitory activity, and 6987 peptides were predicted to possess antioxidant activity. The number of overlapping peptides exhibiting both activities was 1424, accounting for 21.2% of the total ACE inhibitory peptides and 20.4% of the total antioxidant peptides in this system. Among these overlapping peptides, 22 sequences matched previously reported bioactive peptide sequences in the BIOPEP-UWM database, indicating that their activities have reference value at the database level.
[0034]
[0035] Table 2 shows the identification and characterization information of peptide sequences containing Tyr and Trp. The presence of tyrosine (Tyr) or tryptophan (Trp) residues in peptide molecules endows them with antioxidant activity. In the papain digestion system, 32 peptides contained tyrosine residues, 68.75% of which were derived from Patain protein and 31.25% from protease inhibitors; 6 peptides contained tryptophan residues, 66.67% of which were derived from Patain protein and 33.33% from protease inhibitors. Among the target peptides identified through the above screening, only one sequence had a matching record in the BIOPEP database. The remaining peptides were all predicted novel peptides, previously unreported, and based on bioactivity prediction, they all possessed potential antioxidant activity. This also indicates that papain can release peptides containing aromatic amino acid residues from potato protein, thus providing a richer candidate library for subsequent screening of peptides with high antioxidant activity. Further screening yielded 10 antioxidant peptides that act as wine clarifying agents, of which 7 were derived from Patain protein (…). Figure 5 There are 3 protease inhibitors ( ). Figure 6 The list of identified antioxidant peptides is shown in Table 3.
[0036]
[0037]
[0038] The potato protein hydrolysate prepared by this invention possesses biological activities, such as antioxidant and ACE inhibitory activities. Compared with animal protein clarifying agents, the potato protein hydrolysate prepared by this invention, when added to wine as a clarifying agent, results in stable wine body, excellent clarification effect, preservation of the original color and flavor of red wine, reduction of tannin and polyphenol content that cause astringency, lower risk of allergies, and a more delicate and mellow taste.
Claims
1. A method for preparing a wine clarifying agent, characterized in that, Includes the following steps: S1. Extraction of potato protein: S1.
1. Potato tubers, peeled and cut into small pieces, add deionized water, blend into a homogenate to break down cells and release protein; S1.2 Separate the homogenate, collect the supernatant, remove solid residue, and obtain potato juice; S1.
3. Heating potato juice causes protein denaturation and molecular aggregation to form visible flocculent particles. After the thermal flocculation in step S1.4 and S1.3 is completed, cool the mixture to room temperature and centrifuge it to allow the flocculated potato protein to precipitate at the bottom. S2. Purification of potato protein: S2.1 Add deionized water to resuspend the potato protein to make the potato protein evenly dispersed; S2.2 Adjust the pH of the dispersion obtained in step S2.1 to 9-10, heat to 45-55℃, and maintain for 1-1.5h to dissolve the potato protein; S2.3 Centrifuge the mixture obtained in step S2.2, discard the precipitate, and collect the supernatant; S2.4 Adjust the pH of the supernatant to 4.5-5.5, stir well, centrifuge the mixture, discard the supernatant, and collect the precipitate; S2.5 Add deionized water to resuspend the precipitate, centrifuge, and repeat the washing process 2-3 times to obtain purified potato protein. S3. Enzymatic hydrolysis of potato protein: S3.1 Resuspend the purified potato protein in deionized water, add papain, and perform enzymatic hydrolysis; S3.2 After the enzymatic hydrolysis is completed, heat the mixture to inactivate the enzyme, centrifuge, discard the precipitate, and collect the supernatant; S3.3 Adjust the pH of the supernatant to 4.5-5.5, stir well, centrifuge the mixture, discard the supernatant, and collect the precipitate; S3.4 Add deionized water to resuspend the precipitate, centrifuge, and repeat this washing process 2-3 times to remove salt. S3.
5. Resuspend the precipitate, homogenize, adjust the pH to 7, and spray dry to obtain potato protein hydrolysate, which is the wine clarifying agent.
2. The method for preparing a wine clarifying agent according to claim 1, characterized in that: In step S1.1, the mass ratio of potatoes to deionized water is 1:1-3.
3. The method for preparing a wine clarifying agent according to claim 1, characterized in that: In step S1.3, the potato juice is heated to 90°C and kept at that temperature for 30 minutes.
4. The method for preparing a wine clarifying agent according to claim 1, characterized in that: In step S2.2, ultrasonic-assisted treatment is used to increase the solubility of potato protein.
5. The method for preparing a wine clarifying agent according to claim 4, characterized in that: In step S2.2, when using ultrasonic-assisted treatment, the power is set to 200W, the temperature is set to 50℃, and the treatment time is 30 minutes.
6. A method for preparing a wine clarifying agent according to claim 1, characterized in that: In step S3.1, when enzymatically hydrolyzing potato protein, the papain activity is 200,000 U-300,000 U, the substrate concentration is 3%, the enzyme-to-substrate mass ratio is 6:100, the pH is maintained at 6.5, the temperature is 55℃, and the hydrolysis time is 3h.
7. The method for preparing a wine clarifying agent according to claim 1, characterized in that: In step S3.2, the mixture is heated to 85°C and held for 10 minutes.
8. A wine clarifying agent, characterized in that: It is prepared by any one of the methods for preparing a wine clarifying agent according to claims 1-7.
9. The application of the wine clarifying agent according to claim 8 in the wine clarification process, characterized in that, Includes the following steps: A. Weigh the wine clarifying agent and place it in a container; B. Dissolve the wine clarifying agent in the wine to be treated and stir until it becomes a paste; C. Add the paste-like wine clarifying agent to the wine tank containing the wine to be treated, stir for 0.5-2 hours / day, continue for 2-4 days, and let stand; D. After settling, filter the wine, determine its clarity, and bottle it.
10. The application of the wine clarifying agent according to claim 9 in the wine clarification process, characterized in that: In step A, the amount of wine clarifying agent used is 0.1-0.3 g / L.