Monascus purpureus strain as well as culture method and application thereof
By using the purple Monascus strain Yang-Y3000 for optimized culture, the problem of low crude polysaccharide content and harmful ingredients in the existing polysaccharide Monascus powder has been solved. The prepared polysaccharide Monascus powder is safe, of high quality and suitable for food applications.
Patent Information
- Application Number
- CN202411395181.2
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-08
- Publication Date
- 2025-01-24
AI Technical Summary
The existing polysaccharide red chorus prepared by purple Aspergillus strains have low content of crude polysaccharides and contain harmful ingredients such as citronomycin and monaklin K, which affects the safety of the product and market application.
The purple red yeast strain Yang-Y3000 (Monascus purpureus Yang-Y3000) was used for culture. Through the sequential culture method of synthetic medium and wort agar medium, the culture conditions were optimized to improve the color stability and mycelium uniformity of the strain.
The prepared polysaccharide red yeast rice powder has a crude polysaccharide content of more than 200g/kg and does not contain citrinin and monacoline K, which ensures food safety and improves raw material utilization and purity after saccharification.
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Figure CN119351223A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of purple monascus strains and foods, and specifically relates to a purple monascus strain and a cultivation method and application thereof. Background Art
[0002] Red yeast rice is obtained by fermenting rice with red yeast rice, and the fermentation product is red yeast rice. With the continuous development of fermentation technology, modern fermentation technology can more accurately control the fermentation process, but in general, the red yeast rice obtained by fermenting red yeast rice on the market still has the problem of low crude polysaccharide content and the presence of harmful ingredients such as citrinin and monacolin K in red yeast rice products, which seriously affects the safety of the product and limits the export and market application scope of my country's red yeast rice products.
[0003] Therefore, providing a purple Monascus strain to solve the problem that the polysaccharide Monascus powder prepared by the purple Monascus strain on the market has a low crude polysaccharide content and contains harmful components such as citrinin and monacolin K is of great practical significance. Summary of the invention
[0004] The purpose of the present invention is to provide a purple red yeast strain and a culture method and application thereof, so as to solve the problems in the prior art that the polysaccharide red yeast powder prepared by using the purple red yeast strain has a low crude polysaccharide content and contains harmful components such as citrinin and monacolin K.
[0005] The purpose of the present invention can be achieved through the following technical solutions:
[0006] A red yeast rice strain, named Monascus purpureus Yang-Y3000, was deposited in the Guangdong Provincial Microbial Culture Collection Center, Institute of Microbiology, Guangdong Academy of Sciences (Guangdong Provincial Microbial Analysis and Testing Center) on November 23, 2023, and the biological deposit number is: GDMCC No: 64057.
[0007] A method for culturing a purple monascus strain specifically comprises the following steps:
[0008] S1. Select a strain as the initial culture strain;
[0009] S2. The initial culture strain selected in step S1 is cultured in a synthetic culture medium and a malt extract agar culture medium in sequence to obtain a purple Monascus strain Yang-Y3000 (Monascus purpureus Yang-Y3000).
[0010] As a preferred technical solution of the present invention, step S1 specifically comprises: selecting naturally fermented red yeast rice and picking strains with large colony diameter and dark color from its surface as initial culture strains.
[0011] As a preferred technical solution of the present invention, step S2 specifically includes the following steps:
[0012] S21, synthetic culture medium composition, calculated by mass percentage: rice flour 8-20%, sodium nitrate 0.1-0.3%, yeast extract 0.1-0.5%, agar 2-3%, mixed with the remainder of distilled water; adjust pH, boil, then put into a triangular flask, sterilize, cool, inoculate the initial culture strain of step S1 for slant culture until the mycelium covers the slant;
[0013] S22, malt agar medium: barley malt powder and ultrapure water are mixed evenly and kept warm in a temperature-controlled water bath, and then the temperature is controlled and kept warm to obtain a mixed solution A, α-amylase, xylanase and glucoamylase are added, stirred for saccharification, filtered, diluted, temperature-controlled enzyme inactivation, filtered, pH adjusted to obtain a mixed solution B, agar is added, temperature-controlled sterilization is performed, and then the mixture is filled into a triangular flask, and the mycelium of step S21 is inoculated for cultivation, and the mycelium obtained after the cultivation is separated after repeating this operation step 3-5 times, and the obtained purple red yeast rice strain is the purple red yeast rice strain Yang-Y3000 (Monascus purpureus Yang-Y3000).
[0014] Furthermore, the rice flour in step S21 can be replaced by soluble starch.
[0015] Furthermore, the composition of the synthetic culture medium in step S21 also includes 0.2-0.5% corn steep liquor.
[0016] Furthermore, in step S21, the pH is adjusted to 3.0-5.0 with lactic acid or acetic acid; the amount loaded into the conical flask is 250 mL; the sterilization conditions are: temperature 121-125°C, pressure 0.1-0.2 MPa, time 20-30 min; the cooling is cooling to below 35°C; the conditions for culturing the slant culture medium are: temperature 30-34°C; pH = 3.0-5.0; rotation speed 150-200 r / min; time 6-7d.
[0017] Furthermore, in step S22, the mass ratio of the barley malt powder to the ultrapure water is 1:3; the added α-amylase accounts for 0.2-0.25% of the mass of the mixed solution A, the added xylanase accounts for 0.1-0.15% of the mass of the mixed solution A, and the added glucoamylase accounts for 6-7% of the mass of the mixed solution A; and the amount of agar added is 2-3% of the mass fraction of the mixed solution B.
[0018] Furthermore, in step S22, the temperature of the temperature-controlled water bath is 45-50°C, and the insulation time is 2-2.5h; the temperature of the re-temperature-controlled insulation is 60-65°C, and the insulation time is 1-1.5h; the saccharification time is 2-3h; the dilution is to add ultrapure water to dilute to 8-20Bx; the temperature-controlled enzyme inactivation is to control the temperature at 110-115°C to inactivate the enzyme for 15-20min; the pH is adjusted to 4.0-4.5 with lactic acid; the temperature-controlled sterilization is to control the temperature at 121-125°C for sterilization for 20min; the amount of the conical flask is 250mL; and the culture time is 2-3d.
[0019] An application of the purple monascus strain obtained by the above-mentioned culture method, wherein the application is to use the purple monascus strain as a fermentation strain to prepare polysaccharide monascus powder.
[0020] As a preferred technical solution of the present invention, the preparation of polysaccharide red yeast rice powder specifically comprises the following steps:
[0021] A1, liquid seed culture medium, containing per liter: glycerol 90 g, rice flour 20 g, peptone 10 g, KH2PO4 2.5 g, NaNO35 g, MgSO4·7H2O 1 g, ZnSO4·7H2O 2 g, citric acid 1.0 g, nicotinamide 5.0 g;
[0022] Solid-state fermentation medium: soak 30g rice in deionized water for 3-5h, drain and steam, cool, transfer to a conical flask, seal with gauze and sterilize, cool to room temperature for later use;
[0023] A2, inoculate the purple red yeast rice strain on PDA medium for cultivation, wash the spores on the PDA medium with sterile water, shake and break up the spore pellets to obtain a spore suspension, inoculate the spore suspension into the liquid seed medium reserved in step A1 and culture it on a constant temperature shaker, inoculate the cultured seed liquid into the solid fermentation medium at an inoculum amount of 10wt%, shake and spread it after static culture, culture it on a constant temperature shaker, and dry it to obtain polysaccharide red yeast rice powder.
[0024] As a preferred technical solution of the present invention, the amount of deionized water added to the solid-state fermentation medium in step A1 is just enough to immerse the rice.
[0025] As a preferred technical solution of the present invention, the steaming in step A1 is steaming at 0.1 MPa for 50-60 min; the cooling is cooling to 30-35°C, and the number of gauze sealing layers is 6-8 layers; the sterilization conditions are: temperature 121-125°C, pressure 0.08-0.18 MPa, and sterilization for 30 min.
[0026] As a preferred technical solution of the present invention, the conditions for culturing on PDA culture medium in step A2 are: temperature 28-30°C, culturing for 7 days; the conditions for culturing on a constant temperature shaker are: temperature 30-32°C, rotation speed 200r / min, and culturing time 2 days; the temperature of the static culture is 28-30°C, and the time is 48h; the conditions for culturing on a constant temperature shaker are: temperature 25-28°C, rotation speed 180-200r / min, and culturing time 15 days.
[0027] Beneficial effects of the present invention:
[0028] (1) The polysaccharide red yeast rice powder obtained by the cultivation method of the present invention can stably contain more than 200 g / kg of crude polysaccharides (on a dry basis), and the polysaccharide red yeast rice powder does not contain harmful components, including citrinin and monacolin K, which greatly ensures food safety and has certain effective components.
[0029] (2) The present invention uses a synthetic culture medium and a malt agar culture medium to culture the purple Monascus strain in sequence, innovatively designs the components of the synthetic culture medium, and optimizes the components of the malt agar culture medium. The α-amylase, xylanase and glucoamylase in the malt agar culture medium are compounded for saccharification, which effectively improves the raw material utilization rate and the purity after saccharification, and has an excellent pre-effect on culturing a purple Monascus strain with stable color and uniform and regular mycelium.
[0030] (3) In the process of preparing polysaccharide red yeast rice powder with purple red yeast rice strain, citric acid and nicotinamide are added to the liquid seed culture medium for compounding, which can effectively inhibit the production of citrinin and monacolin K in the process of preparing polysaccharide red yeast rice powder, thereby ensuring food safety. BRIEF DESCRIPTION OF THE DRAWINGS
[0031] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for describing the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative work.
[0032] Figure 1 Schematic diagram of the purple Monascus strain obtained in Example 3 of the present invention. DETAILED DESCRIPTION
[0033] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. 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.
[0034] Example 1
[0035] A method for culturing a purple monascus strain, the culturing method specifically comprising the following steps:
[0036] S1. Select strains as initial culture strains:
[0037] Select naturally fermented red yeast rice and pick strains with large colony diameter and dark color from its surface as initial culture strains;
[0038] S2, culturing the initial culture strain selected in step S1 in a synthetic culture medium and a malt extract agar culture medium in turn:
[0039] S21, synthetic culture medium composition, calculated by mass percentage: rice flour 8%, sodium nitrate 0.3%, yeast extract 0.1%, agar 2%, mixed with the remainder of distilled water; pH value is adjusted to 3.0 with lactic acid, boiled, and then filled into a conical flask, the filling volume is 250mL, sterilized, cooled to below 35°C, inoculated with the initial culture strain of step S1 for slant culture until the mycelium covers the slant;
[0040] The sterilization conditions are: temperature 123°C, pressure 0.1MPa, time 30min; the slant culture medium culture conditions are: temperature 30°C, pH=5.0, rotation speed 150r / min, time 7d;
[0041] S22, malt agar medium: barley malt powder and ultrapure water were mixed in a mass ratio of 1:3 and kept warm in a water bath at 48°C for 2.5 hours, and then kept warm at 65°C for 1 hour to obtain a mixed solution A, α-amylase, xylanase and glucoamylase were added, stirred and saccharified for 2 hours, filtered, ultrapure water was added to dilute to 20Bx, the enzyme was inactivated at 113°C for 15 minutes, filtered, the pH was adjusted to 4.5 with lactic acid, a mixed solution B was obtained, agar was added, the temperature was controlled at 121°C for sterilization for 20 minutes, and then the mixture was put into a triangular flask, the mycelium of step S21 was inoculated for cultivation, the mycelium obtained after cultivation was separated after repeating this operation step 3 times, and the purple red yeast rice strain obtained was the purple red yeast rice strain Yang-Y3000 (Monascus purpureus Yang-Y3000);
[0042] The added α-amylase accounts for 0.23% of the mass of the mixed solution A, the added xylanase accounts for 0.15% of the mass of the mixed solution A, and the added glucoamylase accounts for 7% of the mass of the mixed solution A; and the added amount of agar is 2.5% of the mass fraction of the mixed solution B.
[0043] It was observed that the purple Monascus strain prepared in this example had stable color and uniform mycelium.
[0044] Example 2
[0045] A method for culturing a purple monascus strain, the culturing method specifically comprising the following steps:
[0046] S1. Select strains as initial culture strains:
[0047] Select naturally fermented red yeast rice and pick strains with large colony diameter and dark color from its surface as initial culture strains;
[0048] S2, culturing the initial culture strain selected in step S1 in a synthetic culture medium and a malt extract agar culture medium in turn:
[0049] S21, synthetic culture medium composition, calculated by mass percentage: soluble starch 20%, sodium nitrate 0.1%, yeast extract 0.5%, agar 2.5%, corn steep liquor 0.2%, and the rest distilled water mixed; adjust the pH value to 5.0 with acetic acid, boil, and then put into a conical flask, the filling volume is 250mL, sterilize, cool to below 35°C, inoculate the initial culture strain of step S1 for slant culture until the mycelium covers the slant;
[0050] The sterilization conditions are: temperature 125°C, pressure 0.2MPa, time 20min; the slant culture medium culture conditions are: temperature 34°C, pH=3.0, rotation speed 175r / min, time 6d;
[0051] S22, malt agar medium: barley malt powder and ultrapure water were mixed in a mass ratio of 1:3 and kept warm in a water bath at 45°C for 2 hours, and then kept warm at 60°C for 1 hour to obtain a mixed solution A, α-amylase, xylanase and glucoamylase were added, stirred and saccharified for 2.5 hours, filtered, ultrapure water was added to dilute to 8Bx, the enzyme was inactivated at 110°C for 18 minutes, filtered, the pH was adjusted to 4.0 with lactic acid, a mixed solution B was obtained, agar was added, the temperature was controlled at 125°C for sterilization for 20 minutes, and then the mixture was put into a triangular flask, the mycelium of step S21 was inoculated for cultivation, the mycelium obtained after cultivation was separated after repeating this operation step 5 times, and the purple red yeast rice strain obtained was the purple red yeast rice strain Yang-Y3000 (Monascus purpureuss Yang-Y3000);
[0052] The added α-amylase accounts for 0.25% of the mass of the mixed solution A, the added xylanase accounts for 0.1% of the mass of the mixed solution A, and the added glucoamylase accounts for 6.4% of the mass of the mixed solution A; and the added amount of agar is 3% of the mass fraction of the mixed solution B.
[0053] It was observed that the purple Monascus strain prepared in this example had stable color and uniform mycelium.
[0054] Example 3
[0055] A method for culturing a purple monascus strain, the culturing method specifically comprising the following steps:
[0056] S1. Select strains as initial culture strains:
[0057] Select naturally fermented red yeast rice and pick strains with large colony diameter and dark color from its surface as initial culture strains;
[0058] S2, culturing the initial culture strain selected in step S1 in a synthetic culture medium and a malt extract agar culture medium in turn:
[0059] S21, synthetic culture medium composition, calculated by mass percentage: rice flour 14%, sodium nitrate 0.2%, yeast extract 0.3%, agar 3%, corn steep liquor 0.35%, mixed with the remainder distilled water; pH value is adjusted to 4.0 with lactic acid, boiled, then filled into a triangular flask, the filling volume is 250mL, sterilized, cooled to below 35°C, inoculated with the initial culture strain of step S1 and cultured on the slant medium until the mycelium covers the slant;
[0060] The sterilization conditions are: temperature 121°C, pressure 0.15MPa, time 25min; the slant culture medium culture conditions are: temperature 31°C, pH=4.0, rotation speed 200r / min, time 7d;
[0061] S22, malt agar medium: barley malt powder and ultrapure water were mixed in a mass ratio of 1:3 and kept warm in a water bath at 50°C for 2 hours, and then kept warm at 63°C for 1.5 hours to obtain a mixed solution A, α-amylase, xylanase and glucoamylase were added, stirred and saccharified for 3 hours, filtered, ultrapure water was added to dilute to 14Bx, the enzyme was inactivated at 115°C for 20 minutes, filtered, the pH was adjusted to 4.3 with lactic acid, a mixed solution B was obtained, agar was added, the temperature was controlled at 123°C for sterilization for 20 minutes, and then the mixture was put into a triangular flask, the mycelium of step S21 was inoculated for cultivation, the mycelium obtained after cultivation was separated after repeating this operation step 4 times, and the purple red yeast rice strain obtained was the purple red yeast rice strain Yang-Y3000 (Monascus purpureus Yang-Y3000);
[0062] The added α-amylase accounts for 0.2% of the mass of the mixed solution A, the added xylanase accounts for 0.13% of the mass of the mixed solution A, and the added glucoamylase accounts for 6% of the mass of the mixed solution A; and the added amount of agar is 2% of the mass fraction of the mixed solution B.
[0063] It was observed that the purple Monascus strain prepared in this example had stable color and uniform mycelium.
[0064] Example 4
[0065] An application of a purple Monascus strain, wherein the application is to use the purple Monascus strain obtained by the cultivation method of Example 1 as a fermentation strain to prepare polysaccharide Monascus powder, wherein the preparation of polysaccharide Monascus powder specifically comprises the following steps:
[0066] A1, liquid seed culture medium, containing per liter: glycerol 90 g, rice flour 20 g, peptone 10 g, KH2PO4 2.5 g, NaNO35 g, MgSO4·7H2O 1 g, ZnSO4·7H2O 2 g, citric acid 1.0 g, nicotinamide 5.0 g;
[0067] Solid-state fermentation medium: soak 30g rice in deionized water for 3h, drain and cook at 0.1MPa for 50min, cool to 33℃, transfer to a conical flask, seal with 6 layers of gauze, sterilize, and cool to room temperature for use;
[0068] The sterilization conditions are: temperature 125°C, pressure 0.13MPa, sterilization 30min;
[0069] The amount of deionized water added is just enough to submerge the rice;
[0070] A2, inoculating the purple red yeast rice strain on a PDA medium and culturing at 30° C. for 7 days, washing the spores on the PDA medium with sterile water, shaking and breaking up the spore pellets to obtain a spore suspension, inoculating the spore suspension into the liquid seed medium reserved in step A1 and culturing on a constant temperature shaker, inoculating the cultured seed liquid into a solid fermentation medium at an inoculum amount of 10wt%, shaking and flattening after static cultivation, culturing on a constant temperature shaker, and drying to obtain polysaccharide red yeast rice powder;
[0071] The conditions for the constant temperature shaker culture are: temperature 31°C, rotation speed 200r / min, and culture time 2d; the temperature of the static culture is 30°C, and the time is 48h; the conditions for the re-constant temperature shaker culture are: temperature 25°C, rotation speed 200r / min, and culture time 15d.
[0072] It was observed that the polysaccharide red yeast rice powder prepared in this embodiment:
[0073] Color: purple;
[0074] Properties: Powder, no visible foreign impurities;
[0075] Smell and taste: It has the inherent aroma and taste of rice fermented by Monascus koji.
[0076] Detection:
[0077] (1) According to NY / T1676-2008, the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder prepared in this embodiment was tested, and the result was: the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder in this embodiment = 201.62 g / kg;
[0078] (2) According to QB / T 2847-2007, citrinin and monacolin K in the polysaccharide red yeast rice powder prepared in this example were detected. The results were: no citrinin was detected, and no monacolin K was detected.
[0079] Example 5
[0080] An application of a purple Monascus strain, wherein the application is to use the purple Monascus strain obtained by the cultivation method of Example 2 as a fermentation strain to prepare polysaccharide Monascus powder, wherein the preparation of polysaccharide Monascus powder specifically comprises the following steps:
[0081] A1, liquid seed culture medium, containing per liter: glycerol 90 g, rice flour 20 g, peptone 10 g, KH2PO4 2.5 g, NaNO35 g, MgSO4·7H2O 1 g, ZnSO4·7H2O 2 g, citric acid 1.0 g, nicotinamide 5.0 g;
[0082] Solid-state fermentation medium: soak 30g rice in deionized water for 5h, drain and cook at 0.1MPa for 60min, cool to 35℃, transfer to a conical flask, seal with 7 layers of gauze, sterilize, and cool to room temperature for use;
[0083] The sterilization conditions are: temperature 123°C, pressure 0.18MPa, sterilization 30min;
[0084] The amount of deionized water added is just enough to submerge the rice;
[0085] A2, inoculate the purple red yeast rice strain on PDA medium and culture at 29 ° C for 7 days, wash the spores on the PDA medium with sterile water, shake and break up the spore pellets to obtain a spore suspension, inoculate the spore suspension into the liquid seed medium reserved in step A1 and culture it on a constant temperature shaker, inoculate the cultured seed liquid into the solid fermentation medium at an inoculum amount of 10wt%, shake and spread it after static culture, culture it on a constant temperature shaker, and dry it to obtain polysaccharide red yeast rice powder;
[0086] The conditions for the constant temperature shaker culture are: temperature 30°C, rotation speed 200r / min, and culture time 2d; the temperature of the static culture is 28°C, and the time is 48h; the conditions for the re-constant temperature shaker culture are: temperature 26.5°C, rotation speed 180r / min, and culture time 15d.
[0087] It was observed that the polysaccharide red yeast rice powder prepared in this embodiment:
[0088] Color: purple;
[0089] Properties: Powder, no visible foreign impurities;
[0090] Smell and taste: It has the inherent aroma and taste of rice fermented by Monascus koji.
[0091] Detection:
[0092] (1) According to NY / T1676-2008, the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder prepared in this embodiment was tested, and the result was: the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder in this embodiment = 200.73 g / kg;
[0093] (2) According to QB / T 2847-2007, citrinin and monacolin K in the polysaccharide red yeast rice powder prepared in this example were detected. The results were: no citrinin was detected, and no monacolin K was detected.
[0094] Example 6
[0095] An application of a purple Monascus strain, wherein the application is to use the purple Monascus strain obtained by the cultivation method of Example 3 as a fermentation strain to prepare polysaccharide Monascus powder, wherein the preparation of polysaccharide Monascus powder specifically comprises the following steps:
[0096] A1, liquid seed culture medium, containing per liter: glycerol 90 g, rice flour 20 g, peptone 10 g, KH2PO4 2.5 g, NaNO35 g, MgSO4·7H2O 1 g, ZnSO4·7H2O 2 g, citric acid 1.0 g, nicotinamide 5.0 g;
[0097] Solid-state fermentation medium: soak 30g rice in deionized water for 4h, drain and cook at 0.1MPa for 55min, cool to 30℃, transfer to a conical flask, seal with 8 layers of gauze, sterilize, and cool to room temperature for use;
[0098] The sterilization conditions are: temperature 121°C, pressure 0.08MPa, sterilization 30min;
[0099] The amount of deionized water added is just enough to submerge the rice;
[0100] A2, inoculating the purple red yeast rice strain on a PDA medium and culturing at 28° C. for 7 days, washing the spores on the PDA medium with sterile water, shaking and breaking up the spore pellets to obtain a spore suspension, inoculating the spore suspension into the liquid seed medium reserved in step A1 and culturing on a constant temperature shaker, inoculating the cultured seed liquid into a solid fermentation medium at an inoculum amount of 10wt%, shaking and flattening after static cultivation, culturing on a constant temperature shaker, and drying to obtain polysaccharide red yeast rice powder;
[0101] The conditions for the constant temperature shaker culture are: temperature 32°C, rotation speed 200r / min, and culture time 2d; the temperature of the static culture is 29°C, and the time is 48h; the conditions for the re-constant temperature shaker culture are: temperature 28°C, rotation speed 190r / min, and culture time 15d.
[0102] It was observed that the polysaccharide red yeast rice powder prepared in this embodiment:
[0103] Color: purple;
[0104] Properties: Powder, no visible foreign impurities;
[0105] Smell and taste: It has the inherent aroma and taste of rice fermented by Monascus koji.
[0106] Detection:
[0107] (1) According to NY / T1676-2008, the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder prepared in this embodiment was tested, and the result was: the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder in this embodiment = 202.40 g / kg;
[0108] (2) According to QB / T 2847-2007, citrinin and monacolin K in the polysaccharide red yeast rice powder prepared in this example were detected. The results were: no citrinin was detected, and no monacolin K was detected.
[0109] Control group 1
[0110] Compared with Example 3, the difference is that in step S22 of control group 1, no α-amylase is added, but the added xylanase accounts for 0.23% of the mass of the mixed solution A, and the added glucoamylase accounts for 6.1% of the mass of the mixed solution A; the other operating steps and parameters remain unchanged.
[0111] It was observed that the purple Monascus strain prepared in this example had a small amount of variegated colors and a small amount of spots, and the mycelium was relatively uniform and regular.
[0112] Control group 2
[0113] Compared with Example 3, the difference is that in step S22 of control group 2, no xylanase is added, but the added α-amylase accounts for 0.27% of the mass of the mixed solution A, and the added glucoamylase accounts for 6.06% of the mass of the mixed solution A; the other operating steps and parameters remain unchanged.
[0114] It was observed that the purple Monascus strain prepared in this example had a small amount of variegated color, no spots, and the mycelium was relatively uniform and regular.
[0115] Control group 3
[0116] Compared with Example 3, the difference is that in step S22 of control group 3, no glucoamylase is added, but the added α-amylase accounts for 3.2% of the mass of the mixed solution A, and the added xylanase accounts for 3.13% of the mass of the mixed solution A; the other operating steps and parameters remain unchanged.
[0117] It was observed that the purple Monascus strain prepared in this example had a small amount of variegated colors and a small amount of spots, and the mycelium was relatively uniform and regular.
[0118] Control group 4
[0119] Compared with Example 6, the difference is that in the control group 4, the purple Monascus strain obtained by the cultivation method of Example 3 is replaced by the purple Monascus strain obtained by the cultivation method of the control group 1; the other operating steps and parameters remain unchanged.
[0120] It was observed that the polysaccharide red yeast rice powder prepared in this embodiment:
[0121] Color: light purple;
[0122] Properties: Powder, with a small amount of visible foreign impurities;
[0123] Smell and taste: It has a lighter aroma and taste inherent to rice fermented with Monascus koji.
[0124] Detection:
[0125] (1) According to NY / T1676-2008, the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder prepared in this embodiment was tested, and the result was: the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder in this embodiment = 165.36 g / kg;
[0126] (2) According to QB / T 2847-2007, citrinin and monacolin K in the polysaccharide red yeast rice powder prepared in this example were detected. The results showed that 21 μg / kg of citrinin and 0.02 g / kg of monacolin K were detected.
[0127] Control group 5
[0128] Compared with Example 6, the difference is that in the control group 5, the purple Monascus strain cultured by the culture method of Example 3 is replaced by the purple Monascus strain cultured by the culture method of the control group 2; the other operating steps and parameters remain unchanged.
[0129] It was observed that the polysaccharide red yeast rice powder prepared in this embodiment:
[0130] Color: deep purple;
[0131] Properties: Powder, with a small amount of visible foreign impurities;
[0132] Smell and taste: It has a lighter aroma and taste inherent to rice fermented with Monascus koji.
[0133] Detection:
[0134] (1) According to NY / T1676-2008, the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder prepared in this embodiment was tested, and the result was: the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder in this embodiment = 170.82 g / kg;
[0135] (2) According to QB / T 2847-2007, citrinin and monacolin K in the polysaccharide red yeast rice powder prepared in this example were detected. The results showed that 20 μg / kg of citrinin and 0.01 g / kg of monacolin K were detected.
[0136] Control group 6
[0137] Compared with Example 6, the difference is that in the control group 6, the purple Monascus strain cultured by the culture method of Example 3 is replaced by the purple Monascus strain cultured by the culture method of the control group 3; the other operating steps and parameters remain unchanged.
[0138] It was observed that the polysaccharide red yeast rice powder prepared in this embodiment:
[0139] Color: light purple;
[0140] Properties: Powder, with a small amount of visible foreign impurities;
[0141] Smell and taste: It has a lighter aroma and taste inherent to rice fermented with Monascus koji.
[0142] Detection:
[0143] (1) According to NY / T1676-2008, the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder prepared in this embodiment was tested, and the result was: the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder in this embodiment = 162.04 g / kg;
[0144] (2) According to QB / T 2847-2007, citrinin and monacolin K in the polysaccharide red yeast rice powder prepared in this example were detected. The results showed that 22 μg / kg of citrinin and 0.02 g / kg of monacolin K were detected.
[0145] Control group 7
[0146] Compared with Example 6, the difference is that nicotinamide is not added to the liquid seed culture medium in step A1 of control group 7, that is, the liquid seed culture medium contains: 90 g glycerol, 20 g rice flour, 10 g peptone, 2.5 g KH2PO4, 35 g NaNO, 1 g MgSO4·7H2O, 2 g ZnSO4·7H2O, and 6.0 g citric acid per liter; the other operating steps and parameters remain unchanged.
[0147] It was observed that the polysaccharide red yeast rice powder prepared in this embodiment:
[0148] Color: light purple;
[0149] Properties: Powder, with a small amount of visible foreign impurities;
[0150] Smell and taste: It has a lighter aroma and taste inherent to rice fermented with Monascus koji.
[0151] Detection:
[0152] (1) According to NY / T1676-2008, the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder prepared in this embodiment was tested, and the result was: the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder in this embodiment = 153.90 g / kg;
[0153] (2) According to QB / T 2847-2007, citrinin and monacolin K in the polysaccharide red yeast rice powder prepared in this example were detected. The results showed that 23 μg / kg of citrinin and 0.03 g / kg of monacolin K were detected.
[0154] Control group 7
[0155] Compared with Example 6, the difference is that, in step A1 of control group 7, citric acid is not added to the liquid seed culture medium, that is, the liquid seed culture medium contains, per liter: 90 g glycerol, 20 g rice flour, 10 g peptone, 2.5 g KH2PO4, 35 g NaNO, 1 g MgSO4·7H2O, 2 g ZnSO4·7H2O, and 6.0 g nicotinamide; the other operating steps and parameters remain unchanged.
[0156] It was observed that the polysaccharide red yeast rice powder prepared in this embodiment:
[0157] Color: light purple;
[0158] Properties: Powder, with a small amount of visible foreign impurities;
[0159] Smell and taste: It has a lighter aroma and taste inherent to rice fermented with Monascus koji.
[0160] Detection:
[0161] (1) According to NY / T1676-2008, the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder prepared in this embodiment was tested, and the result was: the crude polysaccharide (on a dry basis) contained in the polysaccharide red yeast rice powder in this embodiment = 148.77 g / kg;
[0162] (2) According to QB / T 2847-2007, citrinin and monacolin K in the polysaccharide red yeast rice powder prepared in this example were detected. The results showed that 24 μg / kg of citrinin and 0.03 g / kg of monacolin K were detected.
[0163] In the description of the specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.
[0164] The above contents are merely examples and explanations of the concept of the present invention. The technicians in this technical field may make various modifications or additions to the specific embodiments described or replace them in a similar manner. As long as they do not deviate from the concept of the invention or exceed the scope defined by the claims, they should all fall within the protection scope of the present invention.
Claims
1. A purple Monascus strain, named Purple Monascus Yang-Y3000 ( Monascus purpureus Yang-Y3000), which was deposited in the Guangdong Microbial Culture Collection Center, Institute of Microbiology, Guangdong Academy of Sciences on November 23, 2023, and the biological deposit number is: GDMCC No: 64057.
2. A method for cultivating the purple Monascus strain according to claim 1, characterized in that: The specific steps include: S1. Select a strain as the initial culture strain; S2, the initial culture strain selected in step S1 was cultured in a synthetic medium and a malt extract agar medium in turn to obtain a purple Monascus strain Yang-Y3000 ( Monascus purpureus Yang-Y3000).
3. The method for cultivating the purple Monascus strain according to claim 2, wherein: Step S2 specifically includes the following steps: S21, synthetic culture medium composition, calculated by mass percentage: rice flour 8-20%, sodium nitrate 0.1-0.3%, yeast extract 0.1-0.5%, agar 2-3%, mixed with the remainder of distilled water; adjust pH, boil, then put into a triangular flask, sterilize, cool, inoculate the initial culture strain of step S1 for slant culture until the mycelium covers the slant; S22, malt agar medium: barley malt powder and ultrapure water are mixed evenly and kept warm in a temperature-controlled water bath, and then kept warm to obtain a mixed solution A, α-amylase, xylanase and glucoamylase are added, stirred for saccharification, filtered, diluted, temperature-controlled enzyme inactivation, filtered, pH adjusted to obtain a mixed solution B, agar is added, temperature-controlled sterilization is performed, and then the mixture is filled into a triangular flask, and the mycelium obtained in step S21 is inoculated for cultivation, and the mycelium obtained after the cultivation is separated after repeating this operation step 3-5 times. The purple Monascus strain obtained is the purple Monascus strain Yang-Y3000 ( Monascus purpureus Yang-Y3000).
4. The method for cultivating the purple Monascus strain according to claim 3, characterized in that: In step S21, the rice flour is replaced with soluble starch.
5. The method for cultivating the purple Monascus strain according to claim 3, characterized in that: The synthetic culture medium composition also includes 0.2-0.5% corn steep liquor.
6. The method for cultivating the purple Monascus strain according to claim 3, characterized in that: The sterilization conditions in step S21 are: temperature 121-125°C, pressure 0.1-0.2MPa, time 20-30min; the conditions for culturing the slant culture medium are: temperature 30-34°C; pH=3.0-5.0; rotation speed 150-200r / min; time 6-7d; the saccharification time in step S22 is 2-3h; the dilution is to add ultrapure water to dilute to 8-20Bx; the temperature-controlled enzyme inactivation is to control the temperature at 110-115°C to inactivate the enzyme for 15-20min; the temperature-controlled sterilization is to control the temperature at 121-125°C for sterilization for 20min; the culturing time is 2-3d.
7. The method for cultivating the purple Monascus strain according to claim 3, characterized in that: In step S22, the mass ratio of the barley malt powder to the ultrapure water is 1:3; the added α-amylase accounts for 0.2-0.25% of the mass of the mixed solution A, the added xylanase accounts for 0.1-0.15% of the mass of the mixed solution A, and the added glucoamylase accounts for 6-7% of the mass of the mixed solution A; the amount of agar added is 2-3% of the mass fraction of the mixed solution B.
8. An application of the purple Monascus strain obtained by culturing the purple Monascus strain according to any one of claims 2 to 7, characterized in that: The application is to use the purple red yeast rice strain as a fermentation strain to prepare polysaccharide red yeast rice powder.
9. The use according to claim 8, characterized in that: The specific steps include: A1, liquid seed culture medium, containing per liter: glycerol 90 g, rice flour 20 g, peptone 10 g, KH2PO4 2.5 g, NaNO35 g, MgSO4·7H2O 1 g, ZnSO4·7H2O 2 g, citric acid 1.0 g, nicotinamide 5.0 g; Solid-state fermentation medium: soak rice and deionized water for 3-5 hours, drain and steam, cool, transfer to a conical flask, seal with gauze and sterilize, cool to room temperature for use; A2, the purple red yeast strain was inoculated on PDA medium for cultivation, the spores on the PDA medium were washed off with sterile water, the spore pellets were shaken to obtain a spore suspension, the spore suspension was inoculated into the liquid seed medium reserved in step A1 and cultured on a constant temperature shaker, the cultured seed liquid was inoculated into a solid fermentation medium at an inoculum amount of 10wt%, the mixture was shaken and flattened after standing and cultured, the mixture was cultured on a constant temperature shaker, and the mixture was dried to obtain polysaccharide red yeast powder.
10. The use according to claim 9, characterized in that: In step A1, the amount of deionized water added to the solid-state fermentation medium is just enough to submerge the rice.