Liquor added with Monascus purpureus and preparation method thereof

Abstract

The invention discloses a wine added with Monascus purpureus and a preparation method thereof. The Monascus purpureus added to the wine has a preservation number of CCTCC NO: M 20231512. The wine comprises amino acids, wherein the amino acids serve as a nitrogen source for the Monascus purpureus, and the total amount of the amino acids is greater than 0.3g / 100g. The Monascus purpureus in the wine has high acetaldehyde dehydrogenase activity and can tolerate 65% ethanol. After being added, the Monascus purpureus can utilize the amino acids in the wine to maintain good activity. The strain can accelerate the oxidative metabolism of ethanol into carbon dioxide and water in the body, thereby reducing the damage of ethanol in high-concentration wine to the body. At the same time, the strain also has a significant liver-protecting effect, and its liver-protecting effect is comparable to that of the liver-protecting drug bifendate, thereby significantly reducing the damage of high-concentration wine to the stomach and liver, and reducing the damage of drinking to the body.

Description

Technical Field

[0001] The invention belongs to the field of liquor, and more specifically relates to a liquor added with Monascus purpurogenus and a preparation method thereof. Background Art

[0002] Wine is closely related to Chinese culture. Wine is widely integrated into people's lives, and the wine culture close to "life" has been unprecedentedly enriched and developed.

[0003] However, it is well known that excessive drinking is harmful to the body, especially strong liquor. How to balance Chinese wine culture and physical health requires overcoming many difficulties. The present invention proposes a green and healthy wine, and develops a kind of wine that can turn losses into benefits and take into account the efficacy on the basis of traditional brewing, which is conducive to promoting Chinese wine culture. Summary of the Invention

[0004] In response to the above defects or improvement needs of the prior art, the present invention provides a liquor with added Monascus purpureus and a preparation method thereof, the purpose of which is to discover a strain of Monascus purpureus that has the effect of sobering up, and which can tolerate an alcohol concentration of more than 60%. The amino acids in the liquor can be used to improve its biological activity. The bacteria can accelerate the oxidation and decomposition of alcohol into carbon dioxide and water in the body, thereby reducing the damage of alcohol to the body, thereby solving the technical problem that existing high-alcohol liquor is easily harmful to the body after drinking.

[0005] To achieve the above-mentioned object, according to one aspect of the present invention, a wine with added Monascus purpureus is provided, wherein the Monascus purpureus has a preservation number of CCTCC NO: M 20231512, and the wine includes amino acids, wherein the amino acids serve as a nitrogen source for the Monascus purpureus, and the total amount of amino acids is above 0.3g / 100g.

[0006] Preferably, the total amount of amino acids in the wine added with Monascus purpureus is (0.3g-1g) / 100g, and the number of viable bacteria of Monascus purpureus is 1*10 3 CFU / mL or above.

[0007] Preferably, the wine added with Monascus purpurogenum has an alcohol content of 50% vol to 65% vol.

[0008] Preferably, the wine added with Monascus purpurogenum has an alcohol content of 60% vol to 65% vol.

[0009] Preferably, the amino acids in the wine added with Monascus purpurogenum include aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, isoleucine, tyrosine, phenylalanine, histidine, lysine, arginine and leucine.

[0010] Preferably, the contents of aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, isoleucine, phenylalanine, lysine, arginine and leucine in the wine added with Monascus purpurogenum are all above 0.01g / 100g.

[0011] Preferably, the contents of glutamic acid, proline and glycine in the liquor added with Monascus purpurogenum are all (0.03-0.06g) / 100g, and the contents of aspartic acid, threonine, serine, alanine, valine, isoleucine, phenylalanine, lysine, arginine and leucine are all (0.01-0.03g) / 100g.

[0012] According to another aspect of the present invention, there is also provided a method for preparing the wine added with Monascus purpurogenus as described in the present invention, which comprises the following steps:

[0013] Powder of Monascus purpureus with a preservation number of CCTCC NO: M20231512 is added to the wine containing amino acids according to a preset ratio to obtain wine containing Monascus purpureus.

[0014] Preferably, the method for preparing wine with added Monascus purpureus, wherein the preset ratio is such that the initial viable count of Monascus purpureus in the wine is 1*10 5 Set and add above CFU / mL.

[0015] Preferably, in the method for preparing the wine with added Monascus purpurogenum, the amino acids are derived from sturgeon protein peptides.

[0016] In general, the above technical solution conceived by the present invention can achieve the following beneficial effects compared with the prior art due to the addition of probiotics that can survive in high-concentration wine:

[0017] The liquor provided by the present invention comprises ingredients including Monascus purpureus and amino acids. Since the Monascus purpureus with a preservation number of CCTCC NO: M20231512 has high activity in producing acetaldehyde dehydrogenase and can tolerate 65% ethanol, and the strain can utilize the amino acids in the liquor to enhance its biological activity, it can accelerate the oxidative metabolism of ethanol in the body to decompose into carbon dioxide and water, and can reduce the damage of high-concentration alcohol to the body. At the same time, the strain also has obvious liver-protecting effects, and its liver-protecting effects are comparable to those of the liver-protecting drug bifendate. After drinking the liquor, not only can the oxidative metabolism of alcohol be accelerated, but the liquor also has a liver-protecting effect. Compared with existing brewed liquor, the liquor can significantly reduce the damage of high-concentration alcohol to the body without affecting the quality of the liquor, which is conducive to achieving the purpose of turning damage into benefit and creating green and healthy liquor. DETAILED DESCRIPTION

[0018] In order to make the objectives, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely for the purpose of explaining the present invention and are not intended to limit the present invention. In addition, the technical features involved in the various embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

[0019] The present invention is based on multiple strains isolated from koji in experiments, and a new strain of Monascus purpureus, recorded as Monascus purpureus CEWL18, was discovered. This strain can produce acetaldehyde dehydrogenase (ALDH) and the enzyme activity is the highest among the several isolated strains, which is beneficial to promoting the oxidative degradation of ethanol in the body and thus reducing the damage of alcohol to the body. It was verified by an animal liver injury model that this strain has a significant liver-protecting effect, and its liver-protecting effect is comparable to that of the liver-protecting drug bifendate. However, microorganisms are usually difficult to survive in high concentrations of alcohol. The present invention unexpectedly discovered that this strain can tolerate 65% ethanol and can maintain good activity in amino acid-containing wine. It can be added to amino acid-containing liquor to reduce the damage of high-concentration liquor to the body, thereby preparing health-care liquor containing probiotics.

[0020] The invention provides a liquor added with Monascus purpureus. The Monascus purpureus has a preservation number of CCTCC NO: M20231512 and is named Monascus purpureus CESWGF001. The liquor comprises amino acids, wherein the amino acids serve as a nitrogen source for the Monascus purpureus, and the total amount of the amino acids is greater than 0.3 g / 100 g.

[0021] In some embodiments, the total amount of amino acids in the wine is (0.3g-1g) / 100g, and the number of viable Monascus purpurogenum bacteria is 1*10 3 CFU / mL or above.

[0022] In some embodiments, the alcohol content of the liquor is 50% vol to 65% vol, and preferably, the alcohol content of the liquor is 60% vol to 65% vol.

[0023] The amino acids include aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, isoleucine, tyrosine, phenylalanine, histidine, lysine, arginine and leucine;

[0024] The contents of aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, isoleucine, phenylalanine, lysine, arginine and leucine in the wine are all above 0.01 g / 100 g;

[0025] Preferably, the contents of glutamic acid, proline and glycine are all (0.03g-0.06) / 100g, and the contents of aspartic acid, threonine, serine, alanine, valine, isoleucine, phenylalanine, lysine, arginine and leucine are all (0.01-0.03g) / 100g.

[0026] The total acid content (calculated as acetic acid) in the wine is ≥0.3 g / L. In some embodiments, the total acid content in the wine is 2.53 g / L.

[0027] In addition, the present invention also provides a method for preparing the wine containing Monascus purpurogenum according to the present invention, which is prepared according to the following method:

[0028] Adding powder of Monascus purpureus with a preservation number of CCTCC NO: M20231512 to the wine containing amino acids according to a preset ratio to prepare a wine containing Monascus purpureus;

[0029] The preset ratio is based on the initial viable count of Monascus purpurogenus in the wine at 1*10 5 CFU / mL, and after adding, the number of viable Monascus purpureus in the wine can be maintained at 1*10 3 CFU / mL or above.

[0030] In some embodiments, the amino acids in the wine are derived from sturgeon protein peptides, such as by adding sturgeon protein peptides to the raw materials for brewing wine and obtaining amino acids through enzymatic hydrolysis.

[0031] The following are examples:

[0032] Example 1 Strain Isolation

[0033] Sample: koji, the koji used by a winery to brew wine was used to isolate strains, indicating that the safety and genetic stability of the strains in the koji have been verified in practice, ensuring that the screened strains can be used directly.

[0034] Potato culture medium: 200g potatoes, 20g glucose, 1L purified water, natural pH.

[0035] Specific implementation steps:

[0036] a. Dilute the sample: Crush the koji in a mortar, weigh 2 g, add it to 10 ml of phosphate buffer solution, mix well, and dilute to obtain a dilution. Spread the dilution on a potato culture medium plate and incubate anaerobically at 30°C for 5 days.

[0037] b. Purification of single colonies: Single colonies were selected according to colony morphology, color, and size and three lines were drawn on potato culture medium plates. After multiple selections of hyphae from the edge of single colonies for purification, the resulting pure bacteria were recorded as CEWL-18. Colonies were scraped and subjected to 16s identification and seed preservation. The strain was identified as a new strain of Monascus purpurogenus.

[0038] The isolated new strain (CEWL-18) was sent to the China Type Culture Collection (Wuhan University Collection Center) on the campus of Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province on August 21, 2023 for preservation. Its preservation number is CCTCC NO: M 20231512, the preservation date is August 21, 2023, and it is named Monascus purpureus CESWGF001.

[0039] Example 2 Alcohol tolerance of different strains

[0040] (1) Survival rate of different strains in high concentration ethanol

[0041] Each strain isolated in Example 1 was taken out and placed in a liquid culture medium constant temperature incubator at 37°C for 24 hours at a 10% inoculum size to obtain a first-generation bacterial suspension. The first-generation bacterial suspension was re-inoculated into MRS broth at a 10% inoculum size and incubated at 37°C for 18 hours to obtain a second-generation bacterial suspension. The activated strain was inoculated at 5% into culture medium containing 65%, 50%, 42%, 24%, and 15% ethanol, mixed and allowed to stand, and anaerobically incubated at 37°C for 24 hours. MRS ethanol culture medium without bacterial solution was used as a control to detect CFU. The results are shown in Table 1.

[0042] Table 1 Survival ability of different strains in high concentration ethanol medium

[0043]

[0044] As shown in Table 1, among the above strains, Monascus purpureus CEWL-18 has a higher survival rate in 50% ethanol, and only Monascus purpureus CEWL-18 can survive in a culture medium containing 65% ethanol, indicating that Monascus purpureus CEWL-18 can tolerate high concentrations of alcohol. In addition, the strain has been verified by a mouse liver injury model to have a significant liver-protecting effect, and its effect is even better than that of the existing liver-protecting drug bifendate. The strain was deposited in the China Center for Type Culture Collection in Wuhan on August 21, 2023, with the deposit number CCTCC NO: M 20231512, named Monascus purpureus CESWGF001. It can be added to high-concentration wine for the preparation of wine products containing Monascus purpureus.

[0045] Furthermore, in order to study the biological activity of Monascus purpurogenus CEWL-18 in different types of wine, the survival of the strain in wine samples with an alcohol content of 65% vol and without amino acids and with amino acids was compared as follows:

[0046] Experimental group 1: 100 mL of wine sample with no amino acid residue and 65% alcohol content was added at a ratio of 1.5*10 8 CFU / mL Monascus purpurogenum CEWL-18 was added to the wine sample;

[0047] Experimental group 2: Using the wine sample in experimental group 1 as the matrix, amino acids were added to the wine to make the amino acid content in the wine 0.3g / 100g. 100mL of wine sample containing amino acids was taken and the addition ratio was 1.5*10 8 CFU / mL Monascus purpurogenum CEWL-18 was added to the wine sample;

[0048] The two groups were placed at room temperature and then sampled for testing. After 24 hours, the number of viable bacteria of Monascus purpurogenus CEWL-18 in the wine samples was tested. The results are as follows:

[0049] After 24 hours, the number of viable bacteria of Monascus purpureus CEWL-18 in experimental group 1 was 3.2*10 3 CFU / mL;

[0050] After 24 hours, the number of viable bacteria of Monascus purpureus CEWL-18 in experimental group 2 was 4.3*10 4 CFU / mL, indicating that the strain can utilize the amino acids in wine to grow, which is beneficial to improving its biological activity.

[0051] Example 3 Determination of ADH and ALDH activities of different strains

[0052] Each strain in Table 1 of Example 2 was activated, and the bacteria were separated by centrifugation at 4100 rpm for 10 minutes. The bacteria were detected using an alcohol dehydrogenase (ADH) detection kit and an acetaldehyde dehydrogenase (ALDH) detection kit. The results are shown in Table 2.

[0053] Table 2 Results of ADH and ALDH activity determination of different strains

[0054]

[0055]

[0056] First-pass metabolism (FPM) of ethanol refers to the phenomenon in which some ethanol is oxidized and metabolized in the stomach after drinking. The specific metabolic process of ethanol in the body is as follows: alcohol is oxidized to acetaldehyde by alcohol dehydrogenase (ADH) in the gastric mucosa. Acetaldehyde is then converted to acetic acid by acetaldehyde dehydrogenase (ALDH), and ultimately decomposed into carbon dioxide and water. Therefore, accelerating the oxidative metabolism of ethanol in the body helps reduce the body's bioavailability of ethanol and can reduce the toxic effects of alcohol on organs such as the liver and brain.

[0057] As shown in Table 2, among the aforementioned strains, the Monascus purpurogenum CEWL-18 strain exhibited the highest ALDH activity, reaching 4.18 U / mL. After ethanol is oxidized to acetaldehyde in the body, it is further converted to acetic acid under the action of ALDH and ultimately decomposed into carbon dioxide and water. Therefore, higher ALDH activity facilitates the rapid oxidative metabolism of ethanol in the body, thus reducing the damage caused by alcohol to the body. This indicates that this strain can promote the oxidative metabolism of ethanol in the body and has a significant effect in alleviating alcohol-induced alcohol damage.

[0058] Example 3

[0059] In this embodiment, 10% Monascus purpurogenus CEWL-18 powder (1*10 5 CFU / mL), and after 3 days of storage, the viable counts of Monascus purpurogenus CEWL-18 in the two samples were tested. The results are as follows:

[0060] The content of Monascus purpurogenum CEWL-18 in the finished wine 1 produced from the koji raw material is 3*10 3 CFU / mL, the content of Monascus purpurogenum CEWL-18 in the finished wine 2 produced from the koji raw material was 1*10 3 CFU / mL; This shows that the strain can survive in high-alcohol liquor with an alcohol content of 65% vol, and that Monascus purpurogenus CEWL-18 can utilize the amino acids in the liquor to maintain good activity. The bacterium can accelerate the oxidation and decomposition of alcohol into carbon dioxide and water in the body, which can reduce the damage of high-concentration alcohol to the body and has a good effect in sobering up and reducing damage.

[0061] The finished wine 2 with Monascus purpurogenus CEWL-18 added was sent to Titanium and Zhongpu Testing Technology (Jiangsu) Co., Ltd. for inspection, and the inspection results are shown in Table 3 below.

[0062] Table 3 Test results

[0063]

[0064]

[0065] The testing methods used for the above-mentioned testing items are all carried out in accordance with the currently valid national standard methods for the corresponding items.

[0066] It will be easily understood by those skilled in the art that the above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A wine containing Monascus purpurogenus, characterized in that: The Monascus purpurogenum has a preservation number of CCTCC NO: M20231512. The wine contains amino acids, wherein the amino acids serve as a nitrogen source for the Monascus purpurogenum, and the total amount of amino acids is above 0.3g / 100g. The alcohol content of the wine is 50%vol~65%vol.

2. The wine containing Monascus purpurogenum according to claim 1, characterized in that: The total amount of amino acids in the wine is (0.3g-1g) / 100g, and the number of viable Monascus purpurogenus bacteria is 1*10 3 CFU / mL or above.

3. The wine containing Monascus purpurogenum according to claim 2, characterized in that: The alcohol content of the liquor is 60% vol to 65% vol.

4. The wine containing Monascus purpurogenum according to claim 1, characterized in that: The amino acids include aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, isoleucine, tyrosine, phenylalanine, histidine, lysine, arginine and leucine.

5. The wine containing Monascus purpurogenum according to claim 4, characterized in that: The contents of aspartic acid, threonine, serine, glutamic acid, proline, glycine, alanine, valine, isoleucine, phenylalanine, lysine, arginine and leucine are all above 0.01 g / 100 g.

6. The wine containing Monascus purpurogenum according to claim 5, characterized in that: The contents of glutamic acid, proline and glycine are all within the range of (0.03-0.06g) / 100g; the contents of aspartic acid, threonine, serine, alanine, valine, isoleucine, phenylalanine, lysine, arginine and leucine are all within the range of (0.01-0.03g) / 100g.

7. The method for preparing liquor containing Monascus purpurogenum according to any one of claims 1 to 6, characterized in that: The following steps are involved: Powder of Monascus purpureus with a preservation number of CCTCC NO: M 20231512 is added to the wine containing amino acids according to a preset ratio to obtain wine containing Monascus purpureus.

8. The method for preparing liquor with Monascus purpurogenum added according to claim 7, characterized in that: The preset ratio is based on the initial viable count of Monascus purpurogenus in the wine at 1*10 5 Set and add above CFU / mL.

9. The method for preparing liquor with Monascus purpurogenum added according to claim 8, characterized in that: The amino acids are derived from sturgeon protein peptides.

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