Monascus purpureus strain as well as culture method and application thereof
By using the squash red chorizo strain for solid fermentation, polysaccharide red chorizo powder is made, which solves the complex production process of the red chorizo product and food safety hazards, and achieves high content of crude polysaccharide and safety, which is suitable for a variety of food raw materials.
Patent Information
- Application Number
- CN202411438996.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-15
- Publication Date
- 2024-12-13
AI Technical Summary
The production process of red citrus products is complex and has low technical content, which leads to long production cycles and difficult to control hygiene conditions, which poses food safety risks; the existing standards do not set standards for the content of citrus secreted by Aspergillus Rhodobacteria, and the US FDA prohibits consumption Red citrus products containing trace statin drugs.
The polysaccharide red chorus powder was prepared by solid fermentation of Monascus pilosus Yang-Y1000. Through specific culture methods and medium formulations, including agarose and nanocellulose compounding, the crude polysaccharide content is significantly improved and the product is ensured. It does not contain harmful bacteria such as aflatoxin B1, citronellin and monaklin K.
It significantly improves the content of crude polysaccharides, ensures food safety, has certain functional ingredients, and is widely used in various food raw materials to meet food safety and efficacy needs.
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of food, and relates to a Monascus pubescens strain and a culture method and application thereof. Background Art
[0002] The red yeast rice industry is currently divided into two main categories of products: red yeast rice as a food additive and functional red yeast rice as a raw material for health food. However, the application and promotion of red yeast rice products and the development of the industry face the following three major challenges. First, traditional solid-state fermented red yeast rice is mainly used for food additives, such as colorants, saccharifiers, esterifiers, etc. Due to the production equipment and manual operation processes used in the red yeast rice industry, such as earth pits and cement pools, these processes are complex, low in technology, and labor-intensive, and the raw rice is not sterilized at high temperature and high pressure, resulting in a long production cycle, and it is difficult to control sanitary conditions and bacterial contamination. These factors have caused the traditional fermented red yeast rice products to seriously exceed the fungal toxin standard, poor product quality stability, and food safety risks.
[0003] Secondly, the national standard "GB 1886.19-2015 National Food Safety Standard Food Additive Red Yeast Rice" clearly stipulates that red yeast rice is limited to use as a food additive and limits the amount of use. However, this standard only limits the aflatoxin content secreted by Aspergillus flavus, and does not set a standard for the citrinin content secreted by Monascus.
[0004] Finally, the U.S. Food and Drug Administration (FDA) prohibits the consumption of red yeast rice products containing trace amounts of statins (Lovastatin, i.e., Monacolin K). At the same time, the Korean national standard for health functional foods stipulates that the content of Monacolin K in red yeast rice (powder) is greater than or equal to 0.5g / kg, and does not set a minimum content requirement for red yeast rice. Both countries limit the daily consumption of Monacolin K to between 4 and 8mg based on the standard, and clearly point out that children, pregnant women, breastfeeding women, patients with liver diseases, and those taking hyperlipidemia drugs are not suitable for consumption. Summary of the invention
[0005] The purpose of the present invention is to provide a strain of Monascus pubescens, a culture method and application thereof. Using Monascus pubescens as the strain, rice is solid-state fermented to prepare polysaccharide red yeast rice powder, which can significantly increase the content of crude polysaccharides. The polysaccharide red yeast rice powder does not contain harmful bacteria such as aflatoxin B1, citrinin and monacolin K, can ensure food safety and has certain effective ingredients.
[0006] The purpose of the present invention can be achieved by the following technical solutions:
[0007] A Monascus pilosus strain, wherein the Monascus pilosus strain is Monascus pilosus Yang-Y1000, the preservation number of the Monascus pilosus Yang-Y1000 is GDMCC No: 64056, the preservation unit is Guangdong Microbiological Culture Collection Center, and the preservation date is November 23, 2023; using Monascus pilosus as the strain, solid-state fermentation of rice is performed to prepare polysaccharide Monascus powder.
[0008] 2. A method for culturing the Monascus rubrum strain according to claim 1, characterized in that the culturing method comprises the following steps:
[0009] 1) Selecting bacterial strains: Monascus pilosus Yang-Y1000 was used as the bacterial strain;
[0010] 2) Strain culture: The Monascus rubrum obtained in step 1) is inoculated into a synthetic culture medium for culture, then inoculated into a natural culture medium for culture, and then inoculated into a liquid shake flask culture medium for culture.
[0011] As a preferred technical solution of the present invention, the culture condition of the synthetic culture medium is to culture at a temperature of 30-35°C for 6-8 days.
[0012] As a preferred technical solution of the present invention, the culture conditions of the natural culture medium are culture at a speed of 120-200 r / min and a temperature of 30-35° C. for 18-30 hours.
[0013] As a preferred technical solution of the present invention, the synthetic culture medium comprises 10-15% rice flour or soluble starch, 0.1-0.3% sodium nitrate, 0.1-0.3% yeast extract, 0.6-0.8% agarose, 0.8-1.0% nanocellulose, 2-3% agar and the balance is water; the natural culture medium comprises 8-20% malt extract, 2-3% agar and the balance is water.
[0014] As a preferred technical solution of the present invention, agarose and nanocellulose are compounded to reduce water loss from the culture medium and reduce the need for a curing agent required for the culture medium. Nanocellulose and agarose can also promote adhesion between elements of the culture medium by enhancing hydrophilicity, forming a more complex network structure, strengthening intermolecular interactions, and improving water absorption and swelling capacity and ion exchange capacity.
[0015] Specifically, agarose, as a natural polysaccharide derived from seaweed, has good gel-forming ability and stability. It can effectively fix cells, microorganisms or tissues, and provide a stable and suitable growth environment for the cultivation of bacterial species. Nanocellulose, as a new type of nanomaterial, has shown great potential in many fields with its unique nanosize effect, high specific surface area and excellent mechanical properties. Nanocellulose can not only significantly improve the mechanical strength of the culture medium, but also enhance the water retention capacity of the culture medium through its unique nanostructure, effectively extend the culture period, and reduce the inactivation of bacteria caused by water evaporation.
[0016] When agarose and nanocellulose are used in combination, the synergistic effect between the two significantly improves the moisture retention capacity of the culture medium. Nanocellulose can form a dense mesh structure, effectively locking the water in the culture medium and reducing evaporation. At the same time, due to the action of hydroxyl groups, agarose in water gradually changes from a flexible, irregularly curled state to a single helix and double helix conformation, and the interlacing between the double helix structure and the micelles constitutes a 3D lattice. This 3D lattice not only wraps the water molecules, but also connects to the grid through hydrogen bonds, and is compounded with nanocellulose to further stabilize the water molecules. It has a strong retention effect on water molecules, and its high specific surface area enables nanocellulose to fully adsorb and retain more water molecules, and can keep the culture medium moist even in a dry environment. This improvement not only prolongs the culture time, but also improves the stability and repeatability of the culture of the strain. In addition to improving the water retention capacity, the compound use of agarose and nanocellulose can also promote the adhesion between elements in the culture medium. The unique nanostructure of nanocellulose provides more attachment points for nutrients and growth factors in the culture medium, allowing these elements to be more tightly bound to the culture medium, improving their utilization and effectiveness. This effect of promoting element adhesion helps optimize the growth environment of the bacteria.
[0017] As a preferred technical solution of the present invention, the preparation of the liquid shake flask culture medium comprises selecting 8-15% rice flour, 0.1-0.3% sodium nitrate, and 0.1-0.5% yeast extract and mixing them with water, stirring them evenly, adjusting the pH value to 3.0-5.0 with lactic acid or acetic acid, boiling to form a liquid shake flask culture medium, and filling 40-70% by weight into a conical flask, sterilizing at a temperature of 121-125° C. and a pressure of 0.1-0.2Mpa for 30-40 minutes, cooling, and setting aside.
[0018] As a preferred technical solution of the present invention, the culture conditions of the liquid shake flask culture medium are a rotation speed of 120-160 r / min and a temperature of 32-35° C. for 18-28 hours.
[0019] As a preferred technical solution of the present invention, 5-10% of the seed liquid obtained by the culture method is inoculated into sterilized and cooled rice for fermentation culture for 5-8 days, and then dried at 60-80°C, crushed, sieved with a 100-200 mesh sieve, and packaged to obtain polysaccharide red yeast rice powder.
[0020] Beneficial effects of the present invention:
[0021] The present invention uses Monascus pubescens as the strain and solid-fermentes rice to prepare "polysaccharide red yeast rice powder". The polysaccharide red yeast rice powder is rich in crude polysaccharides reaching more than 200g / kg, which is equivalent to more than 15 times the crude polysaccharides of the first-level quality requirement of the ganoderma lucidum spore powder industry standard; and the polysaccharide red yeast rice powder does not contain harmful bacteria such as aflatoxin B1, citrinin and monacolin K, can ensure food safety and has certain effective ingredients.
[0022] Polysaccharide red yeast rice powder can be widely used as raw materials for various foods, including milk powder, solid beverages, convenience foods, biscuits, frozen drinks, candy products, cocoa and roasted coffee products, starch and starch products, cakes, soy products, health foods, special medical purpose formula foods, special dietary foods, etc. DETAILED DESCRIPTION
[0023] In order to further illustrate the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the specific implementation methods, structures, features and effects of the present invention are described in detail below in combination with the embodiments.
[0024] Wherein, the detection method adopted in the embodiment is as follows:
[0025] unit Detection limit Standard requirements Detection Methods Crude polysaccharides (dry basis) g / kg 0.0005 ≥20.0 NY / T1676-2008 Monacrine K (on dry basis) g / kg 0.01 ≤0.01 QB / T2847-2007 Citrinin (on dry basis) μg / kg 20 ≤50.0 QB / T2847-2007 Aflatoxin B1 (on dry basis) μg / kg 0.03 ≤5.0 GB5009.22-2016
[0026] Example 1
[0027] The Monascus pilosus strain is Monascus pilosus Yang-Y1000, the preservation number of the Monascus pilosus Yang-Y1000 is GDMCC No: 64056, the preservation unit is the Guangdong Provincial Microbial Culture Collection Center of the Institute of Microbiology of Guangdong Academy of Sciences (Guangdong Provincial Microbiological Analysis and Testing Center), and the preservation unit address is the 5th floor of Building 59, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province; using Monascus pilosus as the strain, solid-state fermentation of rice is carried out to prepare polysaccharide Monascus powder.
[0028] A method for culturing a Monascus rubrum strain, the method comprising the following steps:
[0029] 1) Selecting bacterial strains: Monascus pilosus Yang-Y1000 was used as the bacterial strain;
[0030] 2) Culture of strains: The Monascus rubrum obtained in step 1) is inoculated into a synthetic culture medium, wherein the synthetic culture medium comprises 10% rice flour, 0.1% sodium nitrate, 0.1% yeast extract, 0.6% agarose, 0.8% nanocellulose, 2% agar, and the balance is water; the natural culture medium comprises malt extract 8Bx, 2% agar, and the balance is water. The culture is carried out at a speed of 140 r / min and a temperature of 33° C. for 28 hours, and then the culture is inoculated into a liquid shake flask culture medium, and the culture is carried out at a speed of 160 r / min and a temperature of 32° C. for 18 hours.
[0031] 5% of the seed liquid is inoculated into sterilized and cooled rice, and after fermentation and cultivation at 33°C and pH 3.5 for 5 days, the polysaccharide red yeast rice powder is obtained after drying at 60°C, crushing, sieving through a 200-mesh sieve, and packaging.
[0032] Specifically, the preparation method of the red yeast rice fermentation medium comprises the following steps: 8% rice flour, 0.1% sodium nitrate, 0.1% yeast extract, 0.2% corn steep liquor and water are mixed and stirred evenly, pH value is adjusted to 3.5 by lactic acid, boiled, 40% by weight is filled into a conical flask, sterilized at 121° C. and 0.1 Mpa for 30 minutes, and cooled to obtain the product.
[0033] The crude polysaccharide content (on dry basis) was 202.4 g / kg, aflatoxin B1 was not detected (on dry basis, detection limit: 0.03 μg / kg), citrinin was not detected (on dry basis, detection limit: 20 μg / kg), and monacolin K was not detected (on dry basis, detection limit: 0.01 g / kg).
[0034] Example 2
[0035] The Monascus pilosus strain is Monascus pilosus Yang-Y1000, the preservation number of the Monascus pilosus Yang-Y1000 is GDMCC No: 64056, the preservation unit is the Guangdong Provincial Microbial Culture Collection Center of the Institute of Microbiology of Guangdong Academy of Sciences (Guangdong Provincial Microbiological Analysis and Testing Center), and the preservation unit address is the 5th floor of Building 59, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province; using Monascus pilosus as the strain, solid-state fermentation of rice is carried out to prepare polysaccharide Monascus powder.
[0036] A method for culturing a Monascus rubrum strain, the method comprising the following steps:
[0037] 1) Selecting bacterial strains: Monascus pilosus Yang-Y1000 was used as the bacterial strain;
[0038] 2) strain culture: the Monascus purpurogenum obtained in step 1) is inoculated into a synthetic culture medium, wherein the synthetic culture medium comprises 12% rice flour or soluble starch, 0.2% sodium nitrate, 0.2% yeast extract, 0.7% agarose, 0.9% nanocellulose, 2.5% agar and the balance is water; the natural culture medium comprises 10Bx malt extract, 2.5% agar and the balance is water, and the culture is carried out at a speed of 160 r / min and a temperature of 35° C. for 25 h, and then the culture is inoculated into a liquid shake flask culture medium, and the culture is carried out at a speed of 140 r / min and a temperature of 33° C. for 24 h.
[0039] 8% of the seed liquid is inoculated into sterilized and cooled rice, and after fermentation and cultivation at 32°C and pH 4.0 for 6 days, the polysaccharide red yeast rice powder is obtained after drying at 70°C, crushing, sieving through a 150-mesh sieve, and packaging.
[0040] Specifically, the preparation method of the fermentation medium comprises the following steps: 12% rice flour, 0.2% sodium nitrate, 0.3% yeast extract, 0.35% corn steep liquor and water are mixed and stirred evenly, pH value is adjusted to 4.0 by lactic acid, boiled, 60% by weight is filled into a conical flask, sterilized at 123° C. and 0.15 MPa for 35 minutes, and cooled to obtain the fermentation medium.
[0041] The crude polysaccharide content (on dry basis) was 204.3 g / kg, aflatoxin B1 was not detected (on dry basis, detection limit: 0.03 μg / kg), citrinin was not detected (on dry basis, detection limit: 20 μg / kg), and monacolin K was not detected (on dry basis, detection limit: 0.01 g / kg).
[0042] Example 3
[0043] The Monascus pilosus strain is Monascus pilosus Yang-Y1000, the preservation number of the Monascus pilosus Yang-Y1000 is GDMCC No: 64056, the preservation unit is the Guangdong Provincial Microbial Culture Collection Center of the Institute of Microbiology of Guangdong Academy of Sciences (Guangdong Provincial Microbiological Analysis and Testing Center), and the preservation unit address is the 5th floor of Building 59, No. 100 Xianlie Middle Road, Guangzhou City, Guangdong Province; using Monascus pilosus as the strain, solid-state fermentation of rice is carried out to prepare polysaccharide Monascus powder.
[0044] A method for culturing a Monascus rubrum strain, the method comprising the following steps:
[0045] 1) Selecting bacterial strains: Monascus pilosus Yang-Y1000 was used as the bacterial strain;
[0046] 2) Culture of strains: The Monascus rubrum obtained in step 1) is inoculated into a synthetic culture medium, wherein the synthetic culture medium comprises 15% rice flour or soluble starch, 0.3% sodium nitrate, 0.3% yeast extract, 0.8% agarose, 1.0% nanocellulose, 3% agar, and the balance is water; the natural culture medium comprises 18Bx malt extract, 3% agar, and the balance is water. The culture is carried out at a speed of 190 r / min and a temperature of 35° C. for 18 hours, and then the culture is inoculated into a liquid shake flask culture medium, and the culture is carried out at a speed of 120 r / min and a temperature of 35° C. for 28 hours.
[0047] 10% of the seed liquid is inoculated into sterilized and cooled rice, and after fermentation and culture at 35°C and pH 4.8 for 8 days, the polysaccharide red yeast rice powder is obtained after drying at 60°C, crushing, sieving through a 100-mesh sieve, and packaging.
[0048] Specifically, the preparation method of the fermentation medium comprises the following steps: 15% rice flour, 0.3% sodium nitrate, 0.3% yeast extract, 0.3% corn steep liquor and water are mixed and stirred evenly, the pH value is adjusted to 4.8 by lactic acid, boiled, 70% by weight is filled into a conical flask, sterilized at 125° C. and 0.17 Mpa for 30 minutes, and cooled to obtain the medium.
[0049] The crude polysaccharide content (on dry basis) was 207.8 g / kg, aflatoxin B1 was not detected (on dry basis, detection limit: 0.03 μg / kg), citrinin was not detected (on dry basis, detection limit: 20 μg / kg), and monacolin K was not detected (on dry basis, detection limit: 0.01 g / kg).
[0050] Comparative Example 1
[0051] Compared with Example 3, the difference is that nanocellulose is used instead of agarose, and the other components, preparation steps and parameters are the same.
[0052] The crude polysaccharide (on dry basis) was detected to be 192.8 g / kg, aflatoxin B 10.01 μg / kg, citrinin 22 μg / kg, and monacolin K 0.01 g / kg.
[0053] Comparative Example 2
[0054] Compared with Example 3, the difference is that agarose is used instead of nanocellulose, and the other components, preparation steps and parameters are the same.
[0055] The crude polysaccharide (on dry basis) was detected to be 187.4 g / kg, aflatoxin B 10.02 μg / kg, citrinin 23 μg / kg, and monacolin K 0.02 g / kg.
[0056] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technical personnel in this field can make some changes or modify the technical contents disclosed above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the content of the technical solution of the present invention are still within the scope of the technical solution of the present invention.
Claims
1. A Monascus rubrum strain, characterized in that: The Monascus pilosus strain is Monascus pilosus Yang-Y1000, the preservation number of the Monascus pilosus Yang-Y1000 is GDMCC No: 64056, and the preservation unit is Guangdong Microbiological Culture Collection Center; Monascus pilosus is used as the strain, and rice is solid-state fermented to prepare polysaccharide Monascus powder.
2. A method for culturing the Monascus rubrum strain according to claim 1, characterized in that: The culture method comprises the following steps: 1) Selecting bacterial strains: Monascus pilosus Yang-Y1000 was used as the bacterial strain; 2) Strain culture: The Monascus rubrum obtained in step 1) is inoculated into a synthetic culture medium for culture, then inoculated into a natural culture medium for culture, and then inoculated into a liquid shake flask culture medium for culture.
3. The method for cultivating a Monascus rubrum strain according to claim 2, characterized in that: The culture condition of the synthetic culture medium is to culture at a temperature of 30-35° C. for 6-8 days.
4. The method for cultivating a Monascus rubrum strain according to claim 2, characterized in that: The culture conditions of the natural culture medium are culturing at a speed of 120-200 r / min and a temperature of 30-35° C. for 18-30 hours.
5. The method for cultivating a Monascus rubrum strain according to claim 2, characterized in that: The synthetic culture medium comprises 10-15% rice flour or soluble starch, 0.1-0.3% sodium nitrate, 0.1-0.3% yeast extract, 0.6-0.8% agarose, 0.8-1.0% nanocellulose, 2-3% agar and the balance is water; the natural culture medium comprises 8-20% malt extract, 2-3% agar and the balance is water.
6. The method for cultivating a Monascus rubrum strain according to claim 2, characterized in that: The preparation of the liquid shaking flask culture medium comprises the following steps: selecting 8-15% rice flour, 0.1-0.3% sodium nitrate, and 0.1-0.5% yeast extract, mixing them with water, stirring them uniformly, adjusting the pH value to 3.0-5.0 with lactic acid or acetic acid, boiling to form a liquid shaking flask culture medium, and filling 40-70% of the weight of the liquid shaking flask culture medium into a triangular flask, sterilizing the culture medium at a temperature of 121-125° C. and a pressure of 0.1-0.2Mpa for 30-40 minutes, cooling the culture medium, and setting it aside.
7. The method for cultivating a Monascus rubrum strain according to claim 2, characterized in that: The culture conditions of the liquid shake flask culture medium are as follows: a rotation speed of 120-160 r / min and a temperature of 32-35° C. for 18-28 hours.
8. An application of the culture method according to any one of claims 2 to 7, characterized in that: 5-10% of the seed liquid obtained by the culture method is inoculated into sterilized and cooled rice for fermentation culture for 5-8 days, dried at 60-80°C, crushed and sieved through a 100-200 mesh sieve, and packaged to obtain polysaccharide red yeast rice powder.