High-antioxidant yellow rice wine, preparation method thereof and bacteria used in high-antioxidant yellow rice wine

Abstract

The invention belongs to the field of food fermentation, and particularly relates to yellow rice wine with high oxidation resistance, a preparation method of the yellow rice wine and bacteria used in the yellow rice wine. The invention provides talaromyces funiculosuus 7-2-B3-1, and the preservation number of the talaromyces funiculosuus 7-2-B3-1 is CGMCC (China General Microbiological Culture Collection Center) No.41921. The talaromyces funiculosuus 7-2-B3-1 can be used The invention also provides a preparation method of the yellow rice wine with high oxidation resistance at the same time. In the saccharification process, a talaromyces funiculi 7-2-B3-1 culture solution is used. The invention also provides a preparation method of the yellow rice wine with high oxidation resistance. By adopting the method disclosed by the invention, the yellow wine with good oxidation resistance can be obtained; and the fermentation time is short.

Description

Technical Field

[0001] The invention belongs to the field of food fermentation, and particularly relates to yellow rice wine with high antioxidant capacity, a preparation method thereof and the bacteria used therein. Background Art

[0002] The Shaoxing rice wine industry has achieved considerable success in the selection and breeding of specialized glutinous rice varieties, the cultivation of high-quality rice, and the optimization of traditional brewing processes. However, insufficient research is underway in the development of functional rice wines, hindering further development. The current Shaoxing rice wine product portfolio is monotonous and fails to meet the needs of diverse consumer groups. Substances with antioxidant activity in rice wine include polysaccharides, phenolics, and Maillard reaction products. However, the antioxidant capacity of conventional rice wine is relatively limited. Exogenous additions of Codonopsis pilosula, Polygonatum sibiricum, Citrus aurantium, and Matsutake mushrooms are effective methods for enhancing the antioxidant activity of rice wine, but these substances can affect the wine's taste and increase production costs. Optimizing rice wine fermentation processes is another effective approach to enhancing its antioxidant activity. The isolation and purification of polysaccharides and peptides with various active properties in rice wine has been a research hotspot in recent years, but research on optimizing fermentation processes using suitable strains to produce rice wine containing polysaccharides with high antioxidant activity is limited.

[0003] The invention of 201911306635.3 "A strain producing Hubei Ophiopogon polysaccharide and its application" provides a funicularia ( Talaromyces funiculosus strain )EF048, used to prepare Hubei Ophiopogon japonicus polysaccharide, the polysaccharide produced by the bacteria is resistant to four free radicals (DPPH, O2 - ,OH,ABTS + ) have certain in vitro scavenging activity.

[0004] The invention of 202410245885.5 "A strain of cellulase-producing fungi and its application" provides a strain of cellulase-producing fungi (CGMCC NO. 41032), which has a good cellulose degradation effect on woody plants such as mulberry and caragana, and can be coordinated with brewer's yeast for fermentation for the production of woody protein feed. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a yellow rice wine with high antioxidant capacity, a preparation method thereof and the bacteria used therein.

[0006] In order to solve the above technical problems, the present invention provides a rope-shaped basket fungus ( Talaromyces funiculosus )7-2-B3-1, its deposit number is CGMCC No. 41921, deposited at the General Microbiology Center of China Culture Collection Administration, and deposited on April 22, 2025.

[0007] The present invention also provides a method for preparing high-antioxidant rice wine, wherein the funicularia tularensis ( Talaromyces funiculosu s)7-2-B3-1.

[0008] As an improvement of the preparation method of the high-antioxidant yellow rice wine of the present invention, the method comprises the following steps: 1) Raw material pretreatment: The glutinous rice as a raw material is weighed, washed, and then soaked in water (adding 1.5 to 2 times the mass of pure water as the glutinous rice and soaking at room temperature for 36 to 48 hours) as a pretreatment to obtain soaked glutinous rice; 2) Steaming and pouring rice: Steam the soaked glutinous rice obtained in step 1) (steam it in a conventional pot for 30±2 minutes), and then cool it (rinse the steamed rice with purified water and cool it to 37±2°C) to obtain cooled rice; 3) Inoculation and fermentation: Mix the cooled rice obtained in step 2) with koji and rope-shaped basket fungus ( Talaromyces funiculosu s) Canning the 7-2-B3-1 culture solution (into a ceramic pot); saccharifying at 20±2°C (with the pot open) for 5-7 days, then adding malt koji and pure water equal in weight to the glutinous rice. Fermentation is then sealed at room temperature for 60±1 days (with the pot sealed) to obtain the fermented mash. The koji is 0.3-0.4% of the weight of the glutinous rice; Glutinous rice: The culture medium of T. cordiformis 7-2-B3-1 is (110-140) g / 2 mL, preferably (125±5) g / 2 mL; The malt koji accounts for 10-20% of the weight of the glutinous rice, preferably 15%; 4) Slag and liquid separation: The fermented mash obtained in step 3) is subjected to slag-liquid separation (using a press to separate the slag-liquid), and the separated liquid is allowed to stand (for 2±0.5 days) to obtain a supernatant; 5) Decoction of wine: The supernatant obtained in step 4) is directly put into a pot and boiled (at 80-100°C for 10 minutes) to obtain high-antioxidant rice wine.

[0009] As a further improvement of the preparation method of the high antioxidant rice wine of the present invention: the preparation method of the culture solution of Talaromyces funiculosus 7-2-B3-1 is as follows: ① Wash the rice and soak it in water (add 1.5-2 times the mass of the rice in pure water and soak at room temperature for 12-24 hours). Then steam the soaked rice (steam it in a conventional pot for 30±5 minutes) to obtain cooked rice. Put the cooked rice into a container, add water 10 to 20 times the mass of the rice, grind it evenly into a slurry, and sterilize it (121°C, 0.103 MPa for 20 min) to obtain a rice culture medium; ②Inoculate the funicularia 7-2-B3-1 into the rice culture medium and culture it at a constant temperature of 30±1℃ until the concentration of funicularia 7-2-B3-1 reaches 10 6 pieces / mL.

[0010] The present invention also provides high-antioxidant rice wine prepared by any of the above methods.

[0011] The method of the present invention can obtain yellow rice wine with good antioxidant properties and a short fermentation time. In summary, the present invention will have important scientific and economic significance for promoting the innovation and development of the yellow rice wine industry. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The specific embodiments of the present invention are further described in detail below with reference to the accompanying drawings.

[0013] Figure 1 The morphological characteristics of strain 7-2-B3-1 are shown; Figure 1 middle: A: Front view of the colony morphology of strain 7-2-B3-1 on PDA medium; B: Reverse view of the colony morphology of strain 7-2-B3-1 on PDA medium; C: Colony diameter of strain 7-2-B3-1 after 5 days of culture.

[0014] Figure 2 This is a comparison chart of the effect of strain 7-2-B3-1 on improving the DPPH free radical scavenging rate of rice wine; Figure 3 This is a comparison chart showing how strain 7-2-B3-1 improves the iron reducing power (FRAP) of rice wine; Figure 4 Comparison chart of strain 7-2-B3-1 improving the scavenging rate of ABTS cationic radicals in rice wine; illustrate: Figures 2 to 4 middle: Taihu glutinous rice wine + 7-2-B3-1 represents the high antioxidant rice wine obtained in Example 3 (Taihu glutinous rice wine was used as the raw material, and 7-2-B3-1 bacteria was added during the fermentation process), and Taihu glutinous rice wine represents the Taihu glutinous rice wine blank control group; Jianhu Glutinous Rice Wine No. 1 + 7-2-B3-1 represents the high antioxidant rice wine obtained in Example 4 (Jianhu Glutinous Rice Wine No. 1 is used as the raw material, and 7-2-B3-1 bacteria is added during the fermentation process), and Jianhu Glutinous Rice Wine No. 1 represents the blank control group of Jianhu Glutinous Rice Wine No. 1; Shaonuo 9714+7-2-B3-1 represents the high antioxidant rice wine obtained in Example 5 (Shaonu 9714 is used as the raw material, and 7-2-B3-1 bacteria is added during the fermentation process), and Shaonuo 9714 represents the blank control group of Shaonuo 9714.

[0015] Figures 2 to 4 The a, b, and c in the figure represent statistically significant differences, that is, different letters (a, b, c) indicate that the results are significantly different at the P<0.05 level. DETAILED DESCRIPTION

[0016] The present invention is further described below with reference to specific embodiments, but the protection scope of the present invention is not limited thereto: Example 1. Acquisition of strains (1) Grind the koji sample from Shaoxing, Zhejiang Province, weigh 30 g, and place it in a conical flask containing 270 mL of sterile saline and a stirrer. Stir on a magnetic stirrer at 500 rpm for 30 min to prepare a sample solution (1:10 sample solution).

[0017] (2) Take 1 mL of the sample solution and add it to a test tube containing 9 mL of sterile saline. Mix it evenly to make a 1:100 sample solution. Follow the above steps to make a ten-fold series of dilutions of the sample solution until the dilution is completed 6 times.

[0018] (3) Take 100 μL of the sample solution from each dilution and apply it to a PDA plate. Incubate at 30°C for about 72 h.

[0019] (4) After the culture is completed, observe the growth of bacteria on the PDA plate, use an inoculation needle to inoculate the spores of the bacteria on a new PDA plate, and repeat this operation 3 to 5 times to obtain a pure culture of the bacteria.

[0020] (5) The resulting strain 7-2-B3-1 was cyan on the front, gradually becoming lighter in color from the inside to the edge. The core of the colony on the bottom was orange, slightly darker than the surrounding area. Seven days after the inoculation, the edges were irregular, jagged, and had small protrusions. The surface was irregular, with fuzzy mycelium in the center.

[0021] The NCBI comparison results showed that Talaromyces funiculosus, Talaromyces verruculosus, Talaromyces sp. All of them have high similarity. Comparison based on the characteristics of strain 7-2-B3-1 Talaromyces funiculosus, Talaromyces verruculosus, Talaromyces sp. The characteristic characteristics of Talaromyces funiculosus The characteristics are similar, so it is initially judged to be Talaromyces funiculosus .

[0022] The deposit information of the strain 7-2-B3-1 of the present invention is as follows: Deposit name: Basilicum cordiformis Talaromyces funiculosu s, deposited at the General Microbiology Center of China Culture Collection Administration, deposit address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, deposit number: CGMCC No. 41921, deposit date: April 22, 2025.

[0023] Example 2, Rope-shaped basket fungus ( Talaromyces funiculosu s) Preparation of 7-2-B3-1 culture medium: ① Wash the rice (ordinary rice is fine), add 2 times the weight of the rice in water (purified water), and soak at room temperature for 12 hours; then steam it in a conventional pot for 30 minutes; Next, the rice was placed in a culture container, water (pure water) was added in an amount 10 times the mass of the rice, the rice was pulverized into a slurry using a homogenizer, and sterilized (121°C, 0.103 MPa for 20 min) to obtain the rice culture medium; ②、The rope-shaped basket fungus ( Talaromyces funiculosu s) Inoculate 7-2-B3-1 into 250 mL of rice culture medium using a sterile inoculating loop, gently stir, and incubate at 30°C for 7 days to obtain the initial bacterial solution. The initial bacterial solution was inoculated into the rice culture medium at a volume of 10% (volume %) and cultured at 30°C for about 7 days to obtain a concentration of 10 6 / mL of funicularia tularensis ( Talaromyces funiculosu s) 7-2-B3-1 culture medium.

[0024] Example 3: A method for preparing a high-antioxidant rice wine (Shaoxing Yuanhong rice wine), comprising the following steps: 1. Raw material pretreatment Weigh and wash the glutinous rice, add 2 times the weight of pure water, and soak at room temperature for 48 hours; The glutinous rice can be, for example, Taihu glutinous rice.

[0025] 2. Steaming and pouring rice Steam the soaked glutinous rice in step 1 for 30 min, then rinse the steamed rice with purified water and cool it to about 37°C to obtain cooled rice.

[0026] 3. Inoculation and fermentation Set the following usage ratio relationship: Yeast: glutinous rice in step 1 = 0.3% by mass; For every 125g of glutinous rice in step 1, add 2mL of a colony count of 10 6 / mL of funicularia tularensis ( Talaromyces funiculosus) 7-2-B3-1 culture medium; that is, the ratio of glutinous rice to 7-2-B3-1 culture medium is 125g / 2 mL.

[0027] Wheat koji: glutinous rice in step 1 = 15% by mass; The cooled rice obtained in step 2 is mixed with koji and culture solution of T. cordiformis 7-2-B3-1, placed in a ceramic jar, and saccharified at 20°C for 7 days; Then, add wheat koji and pure water of the same weight as glutinous rice, seal and ferment at room temperature for 60 days to obtain fermented mash; The koji is, for example, yellow wine koji AN01 purchased from Angel Yeast Co., Ltd. The preparation method of malt koji can be a conventional preparation method in the field, for example, refer to the preparation method of yellow wine malt koji on pages 76-77 of "Yellow Wine Production Technology and Technology" published by Chemical Industry Press.

[0028] 4. Slag and liquid separation The fermented mash obtained in step 3 was passed through a press to separate the residue and liquid. The separated liquid was allowed to settle for 2 days to obtain a supernatant.

[0029] 5. Decoction of wine The supernatant from step 4 was directly placed in a pot and boiled (80-100°C for 10 minutes) to obtain high-antioxidant rice wine. Testing (GB / T 13662-2018, "Yellow Wine") revealed an alcohol content of 9.3% by volume.

[0030] Taihu glutinous rice blank control group: relative to Example 3, the rope-shaped basket fungus ( Talaromyces funiculosu s) 7-2-B3-1 was used (i.e., the use of the culture solution of T. funiculum 7-2-B3-1 was eliminated), and the rest was the same as in Example 3.

[0031] Example 4: Compared with Example 3, the following changes are made: The Taihu glutinous rice was replaced with Jianhu glutinous rice No. 1; the rest was the same as in Example 3. After testing, the alcohol content was 9.8% vol.

[0032] Jianhu Glutinous Rice No. 1 blank control group: Compared with Example 4, the rope-shaped basket fungus ( Talaromyces funiculosu s) 7-2-B3-1 was used, and the rest was the same as in Example 4.

[0033] Example 5: With respect to Example 3, the following changes are made: The Taihu glutinous rice was replaced with Shaonuo 9714, and the rest was the same as in Example 3. After testing, the alcohol content was 10.6% vol.

[0034] Shaonuo 9714 blank control group: Compared with Example 5, the rope-shaped basket fungus ( Talaromyces funiculosu s) 7-2-B3-1 was used, and the rest was the same as in Example 5.

[0035] Experiment 1: DPPH free radical scavenging rate of rice wine 1. The experimental process and calculation method are as follows: (1) Experimental process 1.0 mL of rice wine sample was placed in a 5.0 mL EP tube, and then 1.0 mL of 0.2 mmol / L DPPH solution was added. After mixing, the mixture was reacted at 37 °C in the dark for 30 min. The absorbance value A1 was measured at 517 nm. Anhydrous ethanol was used instead of DPPH solution as the control group, and the absorbance value As was measured. Distilled water was used instead of rice wine sample as the blank group, and the absorbance value A0 was measured.

[0036] (2) Calculation method The DPPH free radical scavenging rate of the sample to be tested is calculated as follows:

[0037] In the formula, A1 represents the absorbance value of the measuring tube, A s represents the absorbance value of the control tube, and A0 represents the absorbance value of the blank tube.

[0038] The results are as follows Figure 2 As shown: In Example 3, the DPPH radical scavenging rate of the yellow wine prepared with Taihu glutinous rice (added with 7-2-B3-1) was 49.9%, while the DPPH radical scavenging rate of the yellow wine corresponding to the Taihu glutinous rice blank control group (without 7-2-B3-1) was 37.1%; The DPPH radical scavenging rate of the rice wine prepared with Jianhu Glutinous Rice No. 1 (added with 7-2-B3-1) in Example 4 was 56.1%, while the DPPH radical scavenging rate of the rice wine corresponding to the blank control group of Jianhu Glutinous Rice No. 1 (not added with 7-2-B3-1) was 35.8%.

[0039] The DPPH radical scavenging rate of the rice wine prepared with Shaonuo 9714 (added with 7-2-B3-1) in Example 5 was 48.5%, while the DPPH radical scavenging rate of the rice wine corresponding to the Shaonuo 9714 blank control group (without 7-2-B3-1) was 36.7%.

[0040] That is, compared with the corresponding blank control group, the addition of bacteria 7-2-B3-1 significantly improved the DPPH free radical scavenging rate of rice wine.

[0041] Experiment 2: Determination of the Iron Reducing Power (FRAP) of Yellow Rice Wine 1. The experimental process and calculation method are as follows: (1) Experimental process Place 1 mL of rice wine solution in a test tube, add 3 mL of FRAP working solution, mix thoroughly, and incubate in a 37°C water bath in the dark for 10 minutes. Measure absorbance at 593 nm. A standard curve was constructed using FeSO₄ solutions at concentrations of 0, 0.1, 0.2, 0.4, 0.6, 0.8, and 1.0 mmol / L. The antioxidant capacity of each sample was expressed as the number of millimoles of FeSO₄ required to achieve the same absorbance. The reducing capacity was calculated based on the standard curve.

[0042] (2) Calculation method A standard curve was prepared with the FeSO4 solution concentration standard as the horizontal axis and the absorbance values ​​corresponding to different concentrations of FeSO4 solutions as the vertical axis. The curve formula was obtained using an EXCEL table. The curve formula was Y=1.9549X+0.0698(R 2 =0.997), where X is the concentration of FeSO4 (mmol / L) and Y is the absorbance value of the corresponding sample.

[0043] Substitute the absorbance value measured in the sample measurement hole into the calculation formula to obtain the result. Figure 3 : Table 1

[0044] Note: The data of iron ion reducing power in Table 1 are the average values ​​of 3 measurements.

[0045] Experiment 3: Determination of ABTS cation radical scavenging rate of rice wine 1. The experimental process and calculation method are as follows: (1) Experimental process 0.2 mL of rice wine sample was added to 4 mL of ABTS solution in sequence and mixed. The mixture was reacted at room temperature for 10 min. The absorbance measured at 734 nm was A1. When ABTS solution was replaced by distilled water, the absorbance measured at 734 nm was A2. When distilled water was replaced by rice wine sample, the absorbance measured at 734 nm was A0.

[0046] (2) Calculation method

[0047] The ABTS cation radical scavenging rates obtained from Example 3 (Taihu glutinous rice, with 7-2-B3-1 added), the blank group of Example 3 (without 7-2-B3-1 added), Example 4 (Jianhu glutinous rice No. 1, with 7-2-B3-1 added), the blank group of Example 4 (without 7-2-B3-1 added), Example 5 (Shaonuo glutinous rice 9714, with 7-2-B3-1 added), and the blank group of Example 5 (without 7-2-B3-1 added) are as follows: Figure 4 shown.

[0048] Comparative Example 1-1: With respect to Example 3, the following changes were made: The usage ratio of Taihu glutinous rice to 7-2-B3-1 culture medium was changed from "125 g / 2 mL" to "125 g / 1 mL", and the rest was the same as in Example 3.

[0049] The DPPH free radical scavenging rate was 43.8%, the iron ion reducing power was 0.692 mmol / L, and the ABTS cation free radical scavenging rate was 59.6%.

[0050] Comparative Example 1-2, relative to Example 3, makes the following changes: The usage ratio of Taihu glutinous rice to 7-2-B3-1 culture medium was changed from "125 g / 2 mL" to "125 g / 3 mL", and the rest was the same as in Example 3.

[0051] The DPPH free radical scavenging rate was 48.6%, the iron ion reducing power was 0.763 mmol / L, and the ABTS cation free radical scavenging rate was 62.6%.

[0052] Comparative Example 2, relative to Example 4 The usage ratio of Jianhu Nuo No. 1 and 7-2-B3-1 culture medium was changed from "125 g / 2 mL" to "125 g / 1 mL", and the rest was the same as in Example 4.

[0053] The scavenging rate of DPPH free radicals was 47.8%, and that of ABTS cation free radicals was 68.2%.

[0054] The present invention has also carried out the following experiment: When the strain EF048 and the funicular fungus in 202410245885.5 were substituted for the strain 7-2-B3-1 of the present invention, yellow rice wine was prepared according to Example 3, and the DPPH radical scavenging rate had no significant difference compared with the blank group.

[0055] Finally, it should be noted that the above examples are merely specific embodiments of the present invention. Obviously, the present invention is not limited to the above examples and is subject to numerous variations. All variations that can be directly derived or conceived by a person of ordinary skill in the art from the disclosure of the present invention are considered to be within the scope of protection of the present invention.

Claims

1. Rope-shaped basket fungus ( Talaromyces funiculosus ) 7-2-B3-1, characterized in that: The deposit number is CGMCC No. 41921, the deposit unit is General Microbiology Center of China Culture Collection Administration, and the deposit time is April 22, 2025.

2. A method for preparing high-antioxidant yellow rice wine, characterized in that: In the saccharification process, the fungus Basilicum cordifolium ( Talaromyces funiculosu s)7-2-B3-1.

3. The method for preparing the high antioxidant rice wine according to claim 2, characterized in that The steps include: 1) Pre-treating the glutinous rice as a raw material to obtain soaked glutinous rice; 2) Steaming and pouring the soaked glutinous rice obtained in step 1) to obtain cooled cooked rice; 3) Inoculation and fermentation: The cooled rice obtained in step 2) is mixed with koji and culture solution of T. cordiformis 7-2-B3-1 and canned; saccharified at 20±2°C for 5-7 days, then mixed with malt koji and purified water equal in weight to the glutinous rice, and then sealed and fermented at room temperature for 60±1 days to obtain fermented mash; The koji is 0.3-0.4% of the weight of the glutinous rice; Glutinous rice: Culture solution of T. cordiformis 7-2-B3-1 is (110-140) g / 2 mL; The malt koji is 10-20% of the weight of glutinous rice; 4) Separating the fermented mash obtained in step 3) from the residue to obtain a supernatant; 5) Decoction of wine: The supernatant obtained in step 4) is boiled to obtain high-antioxidant rice wine.

4. The method for preparing high-antioxidant yellow rice wine according to claim 3, characterized in that: The preparation method of the culture solution of T. cordiformis 7-2-B3-1 is as follows: ①, prepare cooked rice, add 10-20 times the mass of water to the cooked rice, evenly grind into a slurry, and sterilize to obtain a rice culture medium; ②Inoculate the funicularia 7-2-B3-1 into the rice culture medium and culture it at a constant temperature of 30±1℃ until the concentration of funicularia 7-2-B3-1 reaches 10 6 pieces / mL.

5. The high-antioxidant yellow rice wine prepared by the method according to any one of claims 2 to 4.

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