Prevention and control method for post-turbidity of green plum fruit wine

Abstract

The invention discloses a post-turbidity prevention and control method for green plum fruit wine. The method comprises the step of adding polysaccharide or compound polysaccharide or a combination of the polysaccharide and the compound polysaccharide and bentonite into the green plum fruit wine for post-turbidity prevention and control of the fruit wine. Research finds that compared with carboxymethyl cellulose, guar gum and solek, Arabic gum, sodium alginate and glucan have obvious interference effects on four protein-polyphenol models when polysaccharide or compound polysaccharide or a combination of the polysaccharide and the compound polysaccharide and bentonite are added into the green plum fruit wine, the Arabic gum, the sodium alginate and the glucan have obvious interference effects on the four protein-polyphenol models, and by combining the Arabic gum, the sodium alginate and the glucan, the green plum fruit wine has the obvious interference effects on the four protein-polyphenol models. The interference effect is better than that of single polysaccharide. The polysaccharide or the compound polysaccharide is combined with the common bentonite for use, and the bentonite TOP + Arabic gum and the bentonite TOP + Arabic gum + sodium alginate have a better clarification effect compared with single bentonite TOP. Therefore, the polysaccharide or the compound polysaccharide or the combination of the polysaccharide and the bentonite is applied to turbidity prevention and control of the green plum fruit wine.

Description

Technical Field

[0001] The invention relates to a method for preventing and controlling the after-effect turbidity of fruit wine, and in particular to a method for preventing and controlling the after-effect turbidity of greengage fruit wine. Background Art

[0002] During the storage period, fruit wine will produce turbid precipitation. This has a negative impact on the appearance and sales of the wine. Protein is the main cause of the post-turbidity of fruit wine, mainly because protein can interact with polyphenols to produce turbidity. The occurrence of post-turbidity is also related to pH and microbial species. Studies have shown that when the pH value is acidic, the particle size of the precipitate is smaller. Beverages contaminated by microorganisms will produce turbidity, and the metabolites produced by microorganisms will also cause changes in the taste of the wine.

[0003] The post-turbidity phenomenon of wine has attracted the attention of scholars in the early days, but the research on fruit wine at home and abroad is mainly focused on wine, especially white wine. The clarity of white wine is one of the most intuitive and important sensory quality indicators. Since the current research is limited to wine, which restricts the development of the fruit wine industry, it is necessary to study other types of fruit wine.

[0004] Bentonite adsorption has been proven to be an effective method for removing unstable proteins from white wine. When bentonite combines with proteins, these complexes will be deposited at the bottom of the wine storage tank or fermentation tank in a relatively loose manner, occupying 3% to 10% of the total volume of the wine. However, the effectiveness of bentonite is not limited to removing proteins associated with precipitation, but may also cause loss of wine aroma and color. Therefore, it is particularly urgent and necessary to develop a new technology that is both economically feasible and can effectively maintain the quality of fruit wine.

[0005] Greengage wine is a kind of fruit wine that is made from greengage fruit as the main raw material and carefully brewed through a specific winemaking process. Greengage wine is bright in color, usually amber or light yellow, and the wine is clear and transparent. In terms of taste, greengage wine has both the sweet and sour taste of greengage and the soft taste of alcohol, and the overall flavor is mellow and refreshing. When drinking, the aroma of greengage wine is intoxicating and brings a pleasant sensory experience. Greengage wine is divided into fermented greengage wine and soaked greengage wine. Fermented greengage wine is made from greengage or greengage juice as the main raw material, and is fermented under the action of yeast to form a greengage wine with a certain alcohol content, usually 5-14% vol.

[0006] However, due to the turbidity caused during storage and transportation, the quality loss of fruit wine cannot be ignored. This problem has become a common technical problem with the continuous development of the global beverage industry. Moreover, different clarification methods have different effects on different fruit wines, so it is necessary to study the turbidity prevention and control methods for green plum wine. Summary of the invention

[0007] The purpose of the present invention is to provide a method for preventing and controlling the after-turbidity of greengage wine. In view of the fact that there is no method for preventing and controlling the after-turbidity of greengage wine at present, polysaccharides are applied to the prevention and control of the after-turbidity of greengage wine, so as to improve the clarity of greengage wine.

[0008] In order to achieve the above object, the present invention provides a method for preventing and controlling post-turbidity of greengage wine, which comprises: adding polysaccharides or composite polysaccharides or a combination of them and bentonite into greengage wine to prevent and control post-turbidity of the wine.

[0009] Preferably, the polysaccharide is selected from gum arabic, sodium alginate or dextran.

[0010] Preferably, the concentrations of gum arabic, sodium alginate and dextran are all 0.005 mg / mL.

[0011] Preferably, the complex polysaccharide is selected from gum arabic and sodium alginate, gum arabic and dextran.

[0012] Preferably, the complex polysaccharide comprises gum arabic and sodium alginate, gum arabic and dextran in equal concentrations and volumes.

[0013] Preferably, the complex polysaccharide and greengage wine are in equal volumes.

[0014] Preferably, the wine post-haze prevention and control is carried out at room temperature.

[0015] The method for preventing and controlling the post-turbidity of greengage wine of the present invention has the following advantages:

[0016] The present invention studies and finds that polysaccharides or complex polysaccharides or their combination with bentonite are added to greengold wine. Compared with carboxymethyl cellulose, guar gum, and β-glucan, gum arabic, sodium alginate, and dextran have obvious interference effects on four protein-polyphenol models, and gum arabic, sodium alginate, and dextran are combined. It is found that there is a certain interference effect on the protein-polyphenol model, and gum arabic-glucan> gum arabic-sodium alginate> dextran-sodium alginate, which is stronger than the interference effect of a single polysaccharide. The present invention uses polysaccharides or complex polysaccharides in combination with commonly used bentonite, and finds that bentonite TOP+gum arabic, bentonite TOP+gum arabic+sodium alginate have better clarification effects than single bentonite TOP. Therefore, polysaccharides or complex polysaccharides or their combination with bentonite are used in the prevention and control of turbidity after greengold wine.

[0017] The present invention adopts polysaccharides or complex polysaccharides or a combination of them and bentonite. The polysaccharides or complex polysaccharides are mainly used to remove turbid substances in fruit wine, and have little effect on aroma and color. The addition of polysaccharides will not significantly adsorb or remove aroma components. On the contrary, they may help maintain the stability of aroma and color, especially when used in combination with bentonite, which can reduce the adsorption of bentonite on aroma and pigments. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The results of applying different clarifiers to greengage wine in Experimental Example 2 of the present invention are shown.

[0019] Figure 2 The results are shown in Figure 2, showing the application effects of different clarifier combinations in greengage wine in Experimental Example 2 of the present invention. DETAILED DESCRIPTION

[0020] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. 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.

[0021] Note: The experimental methods in the following examples are conventional methods unless otherwise specified, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the following examples are all commercially available unless otherwise specified.

[0022] The present invention constructs a turbidity model with bovine serum albumin (BSA) and gelatin, epigallocatechin gallate (EGCG) and proanthocyanidins (PC), analyzes the effects of different clarification methods on the prevention and control of post-turbidity of greengage wine, and analyzes the clarification effects of different methods in greengage wine, so as to find a method suitable for the prevention and control of post-turbidity of greengage wine.

[0023] The reagent preparation in the following experimental examples is as follows:

[0024] 0.1mol / L citric acid-sodium citrate buffer (pH 4.2, hereinafter referred to as citric acid buffer): Pipette 1.6mL of solution A and 18.4mL of solution B, and mix well. Solution A (0.1mol / L citric acid): weigh 19.213g of citric acid, dissolve, and dilute to 1000mL. Solution B (1mol / L sodium citrate): weigh 25.813g of sodium citrate, dissolve, and dilute to 1000mL.

[0025] Gum arabic, sodium alginate, carboxymethyl cellulose, guar gum, Solubleco, dextran working solution: weigh 0.08 g of polysaccharide respectively and dissolve it in 0.02 M pH 4.2 citric acid buffer to obtain a polysaccharide stock solution with a concentration of 8.0 mg / mL, and dilute the working solution to the required concentration for the experiment and set aside.

[0026] EGCG solution: prepared by dissolving in pH 4.2 citric acid buffer.

[0027] PC solution: Prepared by dissolving in pH 4.2 citric acid buffer.

[0028] Imported bentonite TOPGRAM TM+ (Hereinafter referred to as imported bentonite TOP) solution: prepared with UP water.

[0029] Experimental Example 1 Study on polysaccharide prevention and control technology

[0030] (1) Selection of polysaccharide types

[0031] Under pH 4.2, 1 mg / mL EGCG or PC, 1 mg / mL bovine serum albumin or gelatin (prepared with pH 4.2 citric acid buffer) and different concentrations of polysaccharides were added to centrifuge tubes (phenolic substances 100 μL + protein 100 μL + polysaccharide 100 μL), and a vortex was used to mix them thoroughly. The tubes were placed in a 25°C constant temperature incubator for incubation for 5 min, and 200 μL of each tube was added to a 96-well plate for determination. The absorbance value A of the reactants was determined using an enzyme marker. 540At the same time, a group of polyphenol-protein with the same concentration without adding polysaccharides was set as a blank control, and the absorbance change value-polysaccharide concentration response curve was drawn. Each group of experiments was repeated three times (n=3). The six polysaccharides were: gum arabic, sodium alginate, carboxymethyl cellulose, guar gum, β-glucan, and dextran. The polysaccharide concentration gradient was set as 0, 0.5, 1, 2, 4, and 8 mg / mL.

[0032] The results showed that sodium alginate, dextran and gum arabic had significant interference effects on the four protein-polyphenol models.

[0033] (2) Selection of optimal polysaccharide concentration

[0034] The interference effect of sodium alginate, dextran and gum arabic concentration on protein-polyphenol turbidity was further studied as follows:

[0035] At pH 4.2, polyphenols, proteins and polysaccharides of different concentrations were added to a centrifuge tube (phenolic substances 100 μL + protein 100 μL + polysaccharide 100 μL), mixed after addition, placed in a constant temperature incubator at 25 ° C for 5 minutes, 200 μL was added to a 96-well plate, and the absorbance A of the system was measured using an enzyme marker. 540 At the same time, a group of polyphenol-protein with the same concentration but without polysaccharide was set as blank control B, and the absorbance change value-polysaccharide concentration response curve was drawn. Each group of experiments was repeated three times (n=3). The polyphenols were: 1mg / mL EGCG, 2mg / mL PC; the proteins were: 1mg / mL BSA, 2mg / mL gelatin; the polysaccharides were: gum arabic, dextran, sodium alginate. The concentrations of polysaccharides were set to 0.005, 0.05, 0.5, 1, 2mg / mL.

[0036] The results showed that when the turbidity of gum arabic was 0.005 mg / mL, the absorbance value of turbidity was the lowest, and the interference effect on these models was good; sodium alginate had a significant effect on the decrease of the overall turbidity of protein-polyphenol turbidity change. After adding sodium alginate, gelatin-EGCG and BSA-EGCG decreased significantly. It is speculated that in the EGCG system, the effect of sodium alginate is better; when the addition amount of dextran is 0.05 mg / mL, the turbidity of BSA-PC and gelatin-PC systems is the lowest at this time, and the polysaccharide has the best effect in inhibiting turbidity. In summary, within a certain concentration range, when the concentration of gum arabic is 0.005 mg / mL, the concentration of sodium alginate is 0.005 mg / mL, and the concentration of dextran is 0.05 mg / mL, the effect of interfering with protein and EGCG, PC turbidity is better.

[0037] (3) Effect of complex polysaccharides on protein-polyphenol turbidity

[0038] The combination of gum arabic-dextran, gum arabic-sodium alginate, and dextran-sodium alginate was used to study the interference effect of the composite polysaccharide on the four protein-polyphenol models, as follows:

[0039] At pH 4.2, polyphenols, proteins and polysaccharides of different concentrations were added to a centrifuge tube (phenolic substances 100 μL + protein 100 μL + complex polysaccharides 100 μL). After adding polyphenols, proteins and complex polysaccharides, they were mixed and the absorbance value A of the system was determined by an ELISA instrument. 540 At the same time, a group of polyphenol-protein models with the same concentration but without the addition of complex polysaccharides was set up as a blank control. Each experiment was repeated three times (n=3). The polyphenols were: 1mg / mL EGCG, 2mg / mL PC, and the proteins were 5mg / mL BSA, 2mg / mL gelatin. The polysaccharide combinations were: A, gum arabic: dextran (volume ratio 1:1); B, gum arabic: sodium alginate (volume ratio 1:1); C, dextran: sodium alginate (volume ratio 1:1). In each group, the concentration of gum arabic was 0.005mg / mL, the concentration of sodium alginate was 0.005mg / mL, and the concentration of dextran was 0.05mg / mL. Among them, the decline rate calculation formula is as follows:

[0040]

[0041] In formula (1), A1 is the absorbance value of polyphenol-protein at the same concentration without adding polysaccharide; A2 is the absorbance value of polyphenol-protein at the same concentration with adding polysaccharide.

[0042] The results showed that the addition of gum arabic-glucan, gum arabic-sodium alginate, and glucan-sodium alginate had a certain interference effect on the four protein-polyphenol models, especially the decline rate of the gelatin-PC model increased. The interference effect of the composite polysaccharide: gum arabic-glucan> gum arabic-sodium alginate> glucan-sodium alginate; the composite gum arabic-glucan had a stronger interference effect than the single use of gum arabic and glucan.

[0043] Table 1 shows the decrease rate of absorbance of the interaction between protein and polyphenols by complex polysaccharides

[0044]

[0045] Experimental Example 2 Application of different clarifiers in green plum wine

[0046] 1. Preparation of greengage wine

[0047] In order to further explore the late turbidity problem of fermented greengage wine, it is very important to maximize the effective substances in greengage wine. Therefore, the primary fermented greengage wine was prepared by turbid juice fermentation, and only a simple clarification treatment was performed. The main process route is as follows:

[0048] Pre-treatment: Select fresh fruits that are highly mature, plump, and not rotten, wash them repeatedly, and then put them into a wall-breaking machine to break them into green plum pulp;

[0049] Adjust sugar and acid: add sugar to make the soluble solid content reach 20°Brix;

[0050] Pasteurization: Place the green plum juice in a 60℃ constant temperature water bath for 30 minutes;

[0051] Enzymatic hydrolysis: adding 0.02% (w / w) compound enzyme (pectinase: cellulase mass ratio = 1:1.5);

[0052] Yeast activation: First, dissolve the yeast in warm water at a ratio of 1:20, and activate the bacteria at 35°C for 60 minutes. Under a sterile environment, transfer the green plum juice to a sterilized 1L triangular bottle, with the volume of each bottle of fermentation liquid not exceeding 70%, inoculate 0.5% SY Angel Yeast, and seal the bottle with a sealing film for later use.

[0053] Fermentation: Transfer the flask containing the fermentation liquid to a constant temperature shaker set to a certain temperature and ferment for 12 days.

[0054] 2. Application of different clarifiers

[0055] In order to study the inhibitory effect of arabinose and sodium alginate on the turbidity of green plum wine, they were applied to the green plum wine system. In addition, according to the inventor's discovery, alkaline protease has the deepest hydrolysis of green plum wine turbidity protein compared with papain and neutral protease. Therefore, alkaline protease, imported bentonite TOP, polysaccharide and their combination were used in the actual application of wine turbidity to find the optimal clarification method, as follows:

[0056] (1) In order to accelerate the formation of precipitation, four models were established using different methods: a 48-hour hot and cold cycle model, a tannin system model, an EGCG system model, and a group of fruit wine control system models. Alkaline protease (enzyme addition amount was 20U / g), imported bentonite TOPGRAM TM+ (0.2 mg / mL) (purchased from Epsler Group (Germany), referred to as bentonite TOP), gum arabic (0.005 mg / mL), sodium alginate (0.005 mg / mL) were used as experimental reagents. Add as follows:

[0057] Group 1: 1000 μL green plum wine;

[0058] Group 2: 500 μL alkaline protease + 500 μL green plum wine;

[0059] Group 3: 500 μL alkaline protease + 500 μL green plum wine;

[0060] Group 4: 500 μL bentonite + 500 μL green plum wine;

[0061] Group 5: 500 μL gum arabic + 500 μL green plum wine;

[0062] Group 6: 500μL sodium alginate + 500μL green plum wine.

[0063] 48-hour hot and cold cycle model: Define the temperature range of hot and cold cycles, freezing temperature -5°C and heating temperature 50°C, set the cycle period (cycle every 12 hours). Store the sample at the set freezing temperature for 12 hours, then transfer to the set heating temperature for 12 hours, for a total of 48 hours of cycles.

[0064] Tannin system model: prepare a 2 mg / mL tannin solution and dissolve the tannin with pH 4.2 citric acid buffer; divide the fruit wine samples into several groups, add 2 mg / mL tannin solution to each group, and fully mix the tannin solution and the fruit wine samples to ensure uniform distribution.

[0065] EGCG system model: prepare 2 mg / mL EGCG solution, the solvent is pH 4.2 citric acid buffer; divide the fruit wine samples into several groups, add 2 mg / mL EGCG solution to each group, and mix the EGCG solution and the fruit wine samples thoroughly.

[0066] All the above-mentioned clarifiers (alkaline protease, bentonite, gum arabic, sodium alginate) were dissolved in green plum wine. The reaction system was centrifuged at 10000rpm for 5min. After centrifugation, 200μL of the supernatant was taken and added to a 96-well plate. After sufficient reaction, the absorbance of the system was measured at 540nm using a Molecular Devices multifunctional microplate reader. A group of room temperature blank green plum wines without any added substances was set up as a control.

[0067] Depend on Figure 1 It can be seen that after adding tannins and EGCG and performing a 48-hour hot and cold cycle, green plum wine becomes turbid and the absorbance value increases. After adding clarifiers such as alkaline protease, bentonite and polysaccharides, there is a certain effect on reducing the absorbance in different systems of green plum wine. At the same time, bentonite has a better inhibitory effect on the generation of turbidity in green plum wine. Bentonite TOP has a better effect in the EGCG system, and both single polysaccharides and composite polysaccharides have an inhibitory effect on precipitation.

[0068] (2) Preparation of alkaline protease (enzyme addition amount is 20U / g) and imported bentonite TOPGRAMTM+ (concentration of 0.2 mg / mL), gum arabic (0.005 mg / mL), sodium alginate (concentration of 0.005 mg / mL), and gum arabic-sodium alginate composite polysaccharide (volume ratio of 1:1), alkaline protease, bentonite TOP, alkaline protease + gum arabic, alkaline protease + sodium alginate, chitosan + gum arabic, bentonite TOP + gum arabic, bentonite TOP + sodium alginate, bentonite TOP + composite polysaccharide and a group of blank controls were treated. The treated supernatant was reacted with EGCG (2 mg / mL) and tannin (2 mg / mL) respectively and placed at room temperature for 48 hours. After that, 200 μL of the supernatant was added to a 96-well plate, and the absorbance value A of the system was determined using a Molecular Devices multifunctional microplate reader. 540 .

[0069] Depend on Figure 2 It can be seen that alkaline protease + sodium alginate, alkaline protease + gum arabic, bentonite TOP + gum arabic, bentonite TOP + gum arabic + sodium alginate can improve the clarity of green plum wine. When green plum wine reacts with EGCG, compared with single alkaline protease, alkaline protease + sodium alginate and alkaline protease + gum arabic have little change in clarity, while bentonite TOP + gum arabic, bentonite TOP + gum arabic + sodium alginate have better clarification effect than single bentonite TOP. Therefore, polysaccharides or composite polysaccharides or their combination with bentonite are used in the prevention and control of post-turbidity of green plum wine.

[0070] Although the content of the present invention has been described in detail through the above preferred embodiments, it should be appreciated that the above description should not be considered as a limitation of the present invention. After reading the above content, it will be apparent to those skilled in the art that various modifications and substitutions of the present invention will occur. Therefore, the protection scope of the present invention should be limited by the appended claims.

Claims

1. A method for preventing and controlling turbidity of greengage wine, characterized in that: The method includes: The polysaccharide or the composite polysaccharide or the combination of the polysaccharide and bentonite is added into the greengage wine to prevent and control the post-harvest turbidity of the wine.

2. The method for preventing and controlling post-turbidity of greengage wine according to claim 1, characterized in that: The polysaccharide is selected from gum arabic, sodium alginate or dextran.

3. The method for preventing and controlling post-turbidity of greengage wine according to claim 2, characterized in that: The concentrations of the gum arabic, sodium alginate and dextran are all 0.005 mg / mL.

4. The method for preventing and controlling post-turbidity of greengage wine according to claim 1, characterized in that: The complex polysaccharide is selected from gum arabic and sodium alginate, gum arabic and dextran.

5. The method for preventing and controlling post-turbidity of greengage wine according to claim 4, characterized in that: The composite polysaccharide adopts gum arabic and sodium alginate, gum arabic and dextran with equal concentration and volume.

6. The method for preventing and controlling post-turbidity of greengage wine according to claim 1, characterized in that: The volumes of the complex polysaccharide and greengage wine are equal.

7. The method for preventing and controlling post-turbidity of greengage wine according to claim 1, characterized in that: The fruit wine post-haze prevention and control is carried out at room temperature.

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