A culture method of oat monascus

Abstract

A method for cultivating oat-based red yeast rice relates to the field of Monascus purpureus purification and cultivation technology. The method includes the following steps: Step 1, cleaning the red yeast rice raw material; Step 2, activating the Monascus purpureus; Step 3, purifying the Monascus purpureus; Step 4, propagating and culturing the Monascus purpureus; and Step 5, producing oat-based red yeast rice using alkali-treated oats as raw material. This method uses commercially available red yeast rice raw material to isolate and purify the strain. Through cultivation using oats as raw material, it has shown good adaptability, providing a feasible solution for the development of more functional oat products. Simultaneously, it effectively solves the problems of nutrient loss, raw material waste, and complex processes caused by the current traditional process of grinding or processing oats into flakes. Alkali treatment alters the permeability of the oat outer skin, while retaining the beneficial active ingredients in oats, increasing the absorption efficiency of oat nutrients, effectively promoting the healthy growth of Monascus purpureus, and simplifying the production process.

Description

Technical Field

[0001] This invention belongs to the field of Monascus purpureus purification and culture technology, specifically relating to a method for culturing oat-based Monascus purpureus. Background Technology

[0002] Oats, as a nutrient-rich grain, possess numerous health benefits. The soluble fiber beta-glucan and arnthramide in oats can effectively lower low-density lipoprotein cholesterol (LDL-C), promoting cardiovascular health and improving vascular elasticity. With a glycemic index (GI) of approximately 55, it is classified as a low-GI food, and its high fiber content can slow glucose absorption, helping type 2 diabetes patients stabilize post-meal blood sugar. Furthermore, oat fiber can promote the proliferation of beneficial gut bacteria (such as Bifidobacteria), inhibit the growth of harmful bacteria, and alleviate constipation.

[0003] As a filamentous fungus, Monascus purpureus is often accompanied by harmful fungi and bacteria such as Aspergillus flavus, Penicillium, and Burkholderia cocovenenans during the storage, transportation, and production of raw materials and red yeast rice. The presence of these harmful bacteria not only affects the quality and safety of Monascus purpureus products but may also pose a potential threat to human health. Therefore, selecting low-contamination raw materials and appropriate culture media to purify and cultivate a pure Monascus purpureus, effectively avoiding the problem of harmful bacteria co-occurrence, is of great practical significance.

[0004] Current oat-based red yeast rice production processes involve crushing or processing oats into oat flakes, which destroys the integrity of the oats. When cultivating red yeast rice using crushed or oat flakes, it is crucial to pay attention to the moisture content of the raw materials, otherwise saccharification may occur, prolonging the fermentation time. The dried product also has a high degree of fragmentation, requiring re-pressing and other processes. At the same time, the fermentation process requires continuous oxygen supply and moisture control, making the production process complex and costly. How to produce red yeast rice without destroying the integrity of the oats and reduce the number of processing steps is an urgent problem to be solved.

[0005] Red yeast rice fermentation products are the main ingredient in Xuezhikang capsules. Xuezhikang capsules have the effects of regulating blood lipids, protecting the cardiovascular system, and relieving symptoms of hyperlipidemia such as dizziness, chest tightness, abdominal distension, and limb numbness. They also have the functions of strengthening the spleen and promoting digestion, and invigorating blood circulation. With increasing public awareness of health, the quality and safety requirements for Monascus purpureus products are becoming increasingly stringent. Obtaining pure and highly effective Monascus purpureus strains has become a key research focus. Red yeast rice products produced using oats as a culture medium do not destroy the functional components such as β-glucan contained in oats, and can produce a synergistic effect of "1+1 greater than 2" in regulating human functions. Developing red yeast rice products based on oats, a specialty agricultural product of Qinghai, can effectively promote the improvement of the quality and efficiency of Qinghai's specialty agriculture and increase farmers' income. Summary of the Invention

[0006] Purpose of the invention

[0007] The purpose of this invention is to provide a method for cultivating oat red yeast rice, in order to solve the problem of the growth of harmful fungi and bacteria such as Aspergillus flavus, Penicillium, and Burkholderia cocovenenans during the storage, transportation, and production of raw materials and red yeast rice. This method selects low-pollution raw materials, preserves the complete oat structure, simplifies the production process and steps, and produces safe, efficient red yeast rice products with specific functions.

[0008] Technical solution

[0009] The present invention also provides a method for isolating and purifying Monascus strains, characterized by comprising the following steps:

[0010] Step 1: Cleaning the red yeast rice raw materials: Using commercially available red yeast rice raw materials as the source of bacteria, clean the red yeast rice raw materials several times with 75% alcohol and sterile water in sequence, and then drain the water.

[0011] Step 2: Activation of Monascus purpureus: Add the cleaned Monascus purpureus raw material to the prepared seed activation solution for activation culture for 12 hours;

[0012] Step 3: Purification of Monascus purpureus: The activated Monascus raw material is inoculated into a solid culture medium and cultured to obtain a single Monascus purpureus colony;

[0013] Step 4: Propagation and culture of Monascus purpureus: Inoculate the single Monascus purpureus colony described in Step 3 into the seed culture medium for culture. When the color of the seed culture medium gradually changes from clear to orange-yellow and a large number of mycelial clumps grow, the propagation and culture of Monascus purpureus is completed.

[0014] Step 5: Production of oat red yeast rice using alkali-treated oats as raw material: Seed culture containing Monascus purpureus mycelium is inoculated into a pre-prepared oat solid culture medium and cultured for 240-360 hours, controlling the temperature at 25-40℃ and humidity at 70%, to obtain oat red yeast rice fermentation product. The oat red yeast rice fermentation product is then dried until the humidity is below 25% to obtain oat red yeast rice. The oat solid culture medium is prepared by alkalizing oats to adjust the pH.

[0015] The specific process and methods are as follows:

[0016] Step 1: Cleaning the red yeast rice raw materials.

[0017] Using commercially available red yeast rice as the strain, weigh 10g of red yeast rice raw material, soak it in 75% alcohol for sterilization, filter out the alcohol after sterilization, then wash it 5 times with sterile water and drain it.

[0018] Step 2: Activation of Monascus purpureus.

[0019] Preparation of seed activation solution: The seed activation solution is prepared from 75% alcohol and maltose aqueous solution, the amount of which is... Take 100ml of pure Add 3ml of 75% alcohol and 2g of maltose to purified water, mix well, and the activation solution is ready.Prepare;

[0020] Red yeast rice activation: Immerse the cleaned red yeast rice raw material in seed activation solution; then place it in an incubator, control the temperature at 35℃ and humidity at 70%, and incubate for 12 hours. A white, fluffy mycelial film will form on the surface of the red yeast rice raw material, thus completing the red yeast rice activation.

[0021] Step 3: Purification of Monascus purpureus.

[0022] Preparation of solid culture medium: Mix 70 g glucose, 15 g beef extract, 2 g NaNO3, 0.5 g MgSO4·7H2O, 1.5 g KH2PO4, and 2 g agar powder into 1000 ml sterile water and stir well. Sterilize the mixture at 0.1 MPa, 121 °C for 30 min. Dispense into Petri dishes and let cool and solidify. The preparation of solid culture medium is complete.

[0023] Purification of Monascus purpureus: Take one activated Monascus purpureus raw material, crush it in a 1.5ml sterile centrifuge tube, add 1ml sterile water to dilute it, and use a sterile disposable pipette to take a small amount and inoculate it onto a solid culture medium and incubate for 8 hours. After Monascus purpureus grows on the solid culture medium, pick out the part with pure color and transfer it to a new culture medium. Repeat this process several times until the colonies that grow are regular in shape and uniform in color, thus completing the purification of Monascus purpureus.

[0024] Step 4: Propagation and Culture of Monascus purpureus

[0025] Preparation of seed culture medium: 70 g glucose, 15 g beef extract, 2 g NaNO3, 0.5 g MgSO4·7H2O, and 1.5 g KH2PO4 were mixed with 1000 ml sterile water and stirred evenly. The mixture was then dispensed into 250 ml Erlenmeyer flasks, wrapped with 8 layers of clean gauze, and sterilized under high pressure at 0.1 MPa and 121℃ for 30 min to complete the preparation of seed culture medium.

[0026] Monascus purpureus propagation culture: Under sterile conditions, a small number of Monascus purpureus colonies purified in step 3 were picked up with sterilized tweezers and inoculated into the liquid seed culture medium in an Erlenmeyer flask. The temperature was controlled at 35℃ and the humidity at 70% for propagation culture. The culture was carried out for 72 hours until the color of the seed culture medium gradually changed from clear to orange-yellow and a large number of mycelial clusters grew, thus completing the propagation culture of Monascus purpureus.

[0027] Step 5 uses alkali-treated oats as raw material to produce oat red yeast rice.

[0028] Oat selection and washing: Select oats that are plump and free from pests and diseases, put them in a basin and submerge them in water to remove oats that float to the surface.

[0029] Alkali solution preparation: Add 2g NaOH to every 100ml of sterile water to complete the preparation of alkali solution;

[0030] Oat soaking: After rinsing the oat seeds, soak the oats in an alkaline solution for 2 hours;

[0031] Adjust pH: Soak alkaline oats in an acetic acid solution with a pH of 4 for 2.5 hours;

[0032] Preparation of oat solid culture medium: After adjusting the pH, filter out the oat with gauze and drain the water. Add oat solid culture medium to control the moisture content to within 70%. Weigh 50g and put it into an Erlenmeyer flask. Sterilize at 0.1MPa, 121℃ for 30min. Cool to room temperature to obtain oat solid culture medium.

[0033] Fermentation of oat red yeast rice: In a sterile environment, add 3 ml of 75% alcohol to the culture medium, measure 10 ml of the bacterial solution cultured in step 4 using a graduated cylinder, mix well, seal the bottle, and place it in an incubator for culture, controlling the temperature at 35℃ and the humidity at 70%; after 240 hours of culture, the fermentation of oat red yeast rice is complete.

[0034] Preparation of oat red yeast rice: Transfer the oat red yeast rice fermentation material to a drying oven for drying at a temperature of 50°C; dry until the humidity is below 25%, which is the finished oat red yeast rice product.

[0035] Beneficial effects

[0036] High safety: The Monascus purpureus strain purified and cultured by the method of this invention has no harmful components detected, which fundamentally solves the problem of harmful bacteria accompanying Monascus purpureus during production and storage, greatly improves the safety of Monascus purpureus products, and provides consumers with healthy and reliable products.

[0037] Significant advantages of raw materials: The cultivation material uses oats from Qinghai. The Qinghai Plateau has cool summers (annual average temperature 5.5℃ to -0.6℃), resulting in oat seedlings with strong cold resistance, tolerance to low temperatures during tillering, and avoidance of high-temperature sensitive periods during flowering and grain-filling, which is conducive to the growth and nutrient accumulation of oats. The total water requirement for oats throughout their entire growth cycle is approximately 29.1-33.3 m³. 3 / hm 2 Qinghai experiences simultaneous rainfall and heat (82%-96% of annual precipitation occurs during periods ≥5℃), meeting the peak water demand after the jointing stage. The arid climate of the eastern foothills of Qinghai, coupled with scarce rainfall during the oat harvest season, allows for rapid dehydration and drying of harvested oats. This also reduces the risk of contamination by fungi such as Aspergillus flavus, giving Qinghai a natural advantage as a high-quality raw material base. Using Qinghai oats as a culture medium not only ensures the quality and safety of the raw materials but also fully utilizes local resources, contributing to local economic development.

[0038] Reduced allergy risk: This invention uses oats as the sole raw material to produce oat red yeast rice, avoiding the use of other raw materials that may cause allergies, thus reducing the risk of allergies and expanding the product's target population.

[0039] Optimized production process: Precise control of oat processing conditions ensures moderate oat moisture content, eliminating the need for water replenishment during fermentation. This reduces external interference with the fermentation process, lowers the risk of contamination, simplifies production equipment and processes, reduces production costs, and improves production efficiency.

[0040] Thorough strain purification: Through a systematic purification process of Monascus purpureus strains, harmful substances produced by other fungi such as penicillin, penicillic acid, and aflatoxin are fundamentally prevented, ensuring the quality and stability of Monascus purpureus products and laying a solid foundation for the further development and application of Monascus purpureus products.

[0041] Microscopic images of oat-based red yeast rice cultured in alkali-treated oats. Oat-based red yeast rice was cultivated using a solid culture medium made from alkali-treated oats. Because the red yeast mold can more easily penetrate the oat epidermis and develops faster, alkali-treated oats grow faster than ordinary oats, shortening the fermentation time by 120 hours. It also reduces the risk of contamination by other microorganisms and ensures the surrounding environment is free of other microorganisms and impurities. Attached Figure Description

[0042] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below.

[0043] Figure 1 This is a comparison image of Example 1;

[0044] Figure 2 This is the effect diagram for Comparative Example 2;

[0045] Figure 3 This is the effect diagram for Comparative Example 3;

[0046] Figure 4 This is a microscope image of Comparative Example 3;

[0047] Figure 5 This is a microscope image of Example 1;

[0048] Figure 6 This is a diagram showing the purification effect of Monascus purpureus in step 3.

[0049] Figure 7 This is a diagram showing the results of step 4, Monascus purpureus propagation culture.

[0050] Figure 8 This is a process flow diagram of the present invention. Detailed Implementation

[0051] The steps in the technical solution will be described one by one below through four comparative examples and three embodiments, providing a clear and complete description of the technical solution in the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0052] Comparative Example 1

[0053] Using commercially available red yeast rice as the strain source, weigh out an appropriate amount of red yeast rice raw material, and in step 1, use oats as raw material to produce oat red yeast rice.

[0054] The specific method is as follows:

[0055] Oat selection and washing: Select oats that are plump and free from pests and diseases, put them in a basin and submerge them in water to remove oats that float to the surface.

[0056] After rinsing the oats, soak the seeds in purified water at 25°C for 48 hours.

[0057] The moisture content of oats was controlled at 70%. 50g of oats were weighed and placed into an Erlenmeyer flask. The flask was sterilized at 0.1MPa and 121℃ for 30min and then cooled to room temperature to obtain oat solid culture medium.

[0058] Production of oat red yeast rice: In a sterile environment, 10g of red yeast rice raw material is added to the culture medium, mixed evenly, the bottle is sealed, and placed in an incubator for cultivation, with the temperature controlled at 35℃ and the humidity at 70%; contamination occurs after 24 hours of cultivation.

[0059] Figure 1 A photograph of the appearance of the contaminated oat red yeast rice provided in Comparative Example 1 of this application, such as Figure 1 As shown, the contaminated culture medium in the conical flask contains yellow-green bacteria.

[0060] Comparative Example 2

[0061] According to Comparative Example 1, the red yeast rice raw material was disinfected by soaking in 75% alcohol before use.

[0062] Step 1: Cleaning the red yeast rice raw materials.

[0063] Specifically:

[0064] Using commercially available red yeast rice as the strain, weigh an appropriate amount of red yeast rice raw material, soak it in 75% alcohol for sterilization, filter out the alcohol after sterilization, then wash it 5 times with sterile water and drain it.

[0065] Step 2 involves producing oat red yeast rice using oats as the raw material.

[0066] Specifically:

[0067] Oat selection and washing: Select oats that are plump and free from pests and diseases, put them in a basin and submerge them in water to remove oats that float to the surface.

[0068] After rinsing the oats, soak the seeds in purified water at 25°C for 48 hours.

[0069] The moisture content of oats was controlled at 70%. 50g of oats were weighed and placed into an Erlenmeyer flask. The flask was sterilized at 0.1MPa and 121℃ for 30min and then cooled to room temperature to obtain oat solid culture medium.

[0070] Production of oat red yeast rice: In a sterile environment, 10g of washed red yeast rice raw material is added to the culture medium, mixed evenly, the bottle is sealed, and placed in an incubator for cultivation, with the temperature controlled at 35℃ and the humidity at 70%; contamination occurs after 72 hours of cultivation.

[0071] Figure 2 A photograph of the appearance of the contaminated oat red yeast rice provided in Comparative Example 2 of this application, such as Figure 2 As shown, the contaminated culture medium in the conical flask contains black and yellow-green portions indicating miscellaneous bacteria. Compared to Comparative Example 1, the red yeast rice raw material, after being disinfected by soaking in 75% alcohol, was contaminated two days later.

[0072] Comparative Example 3

[0073] According to Comparative Examples 1 and 2, the red yeast rice raw material was disinfected by soaking in 75% alcohol and then used after activation culture.

[0074] Step 1: Cleaning the red yeast rice raw materials.

[0075] Specifically:

[0076] Using commercially available red yeast rice raw materials as the source of microorganisms, weigh an appropriate amount of red yeast rice raw materials, soak them in 75% alcohol to sterilize them, filter out the alcohol after sterilization, and then wash them 5 times with sterile water and drain them.

[0077] Step 2: Activation of Monascus purpureus.

[0078] Specifically:

[0079] The cleaned red yeast rice raw materials were immersed in seed activation solution; then placed in an incubator and cultured for 12 hours. A white, fluffy mycelial film formed on the surface of the red yeast rice raw materials, completing the red yeast rice activation.

[0080] Step 3 involves producing oat red yeast rice using oats as the raw material.

[0081] Specifically:

[0082] Oat selection and washing: Select oats that are plump and free from pests and diseases, put them in a basin and submerge them in water to remove oats that float to the surface.

[0083] After rinsing the oats, soak the seeds in purified water at 25°C for 48 hours.

[0084] The moisture content of oats was controlled at 70%. 50g of oats were weighed and placed into an Erlenmeyer flask. The flask was sterilized at 0.1MPa, 121℃ for 30min and then cooled to room temperature to obtain oat solid culture medium.

[0085] Production of oat red yeast rice: In a sterile environment, 10g of cleaned and activated red yeast rice raw material is added to the culture medium, mixed evenly, the bottle is sealed, and placed in an incubator for cultivation, with the temperature controlled at 35℃ and the humidity at 70%; contamination occurs after 168h of cultivation.

[0086] Figure 3 A photograph of the appearance of the contaminated oat red yeast rice provided in Comparative Example 3 of this application, such as Figure 3 As shown, the contaminated culture medium in the conical flask contains miscellaneous bacteria, with the white part of the oat skin being contaminated.

[0087] Figure 4 A photomicrograph of the contaminated oat red yeast rice provided in Comparative Example 3 of this application, such as Figure 3 The image shows a microscopic photograph of the contaminated culture medium in the conical flask, where the black lollipop-shaped spores on the oat surface indicate contamination by bacteria. Compared to Comparative Example 2, this method added a Monascus purpureus activation step, resulting in a final contamination time 4 days later than the Comparative Example.

[0088] Comparative Example 4

[0089] According to Comparative Examples 1, 2, and 3, the red yeast rice raw material was disinfected by soaking in 75% alcohol, then activated and cultured, and finally purified and propagated before use.

[0090] Step 1: Cleaning the red yeast rice raw materials.

[0091] Specifically:

[0092] Using commercially available red yeast rice as the strain, weigh 10g of red yeast rice raw material, soak it in 75% alcohol for sterilization, filter out the alcohol after sterilization, then wash it 5 times with sterile water and drain it.

[0093] Step 2: Activation of Monascus purpureus.

[0094] Specifically:

[0095] Preparation of seed activation solution: The seed activation solution is prepared by 75% alcohol and maltose aqueous solution. Take 100ml of purified water, add 3ml of 75% alcohol and 2g of maltose, mix well, and the activation solution is ready.

[0096] Red yeast rice activation: Immerse the cleaned red yeast rice raw material in seed activation solution; then place it in an incubator, control the temperature at 35℃ and humidity at 70%, and incubate for 12 hours. A white, fluffy mycelial film will form on the surface of the red yeast rice raw material, thus completing the red yeast rice activation.

[0097] Step 3: Purification of Monascus purpureus.

[0098] Specifically:

[0099] Preparation of solid culture medium: Mix 70 g glucose, 15 g beef extract, 2 g NaNO3, 0.5 g MgSO4·7H2O, 1.5 g KH2PO4, and 2 g agar powder into 1000 ml sterile water and stir well. Sterilize the mixture at 0.1 MPa, 121 °C for 30 min. Dispense into Petri dishes and let cool and solidify. The preparation of solid culture medium is complete.

[0100] Purification of Monascus purpureus: Take one activated Monascus purpureus raw material, crush it in a 1.5ml sterile centrifuge tube, add 1ml sterile water to dilute it, and use a sterile disposable pipette to take a small amount and inoculate it onto a solid culture medium and incubate for 8 hours. After Monascus purpureus grows on the solid culture medium, pick out the part with pure color and transfer it to a new culture medium. Repeat this process several times until the colonies that grow are regular in shape and uniform in color, thus completing the purification of Monascus purpureus.

[0101] Step 4: Propagation and Culture of Monascus purpureus

[0102] Specifically:

[0103] Preparation of seed culture medium: 70 g glucose, 15 g beef extract, 2 g NaNO3, 0.5 g MgSO4·7H2O, and 1.5 g KH2PO4 were mixed with 1000 ml sterile water and stirred evenly. The mixture was then dispensed into 250 ml Erlenmeyer flasks, wrapped with 8 layers of clean gauze, and sterilized under high pressure at 0.1 MPa and 121℃ for 30 min to complete the preparation of seed culture medium.

[0104] Monascus purpureus propagation culture: Under sterile conditions, a small number of Monascus purpureus colonies purified in step 3 were picked up with sterilized tweezers and inoculated into the liquid seed culture medium in an Erlenmeyer flask. The temperature was controlled at 35℃ and the humidity at 70% for propagation culture. The culture was carried out for 72 hours until the color of the seed culture medium gradually changed from clear to orange-yellow and a large number of mycelial clusters grew, thus completing the propagation culture of Monascus purpureus.

[0105] Step 5 involves producing oat red yeast rice using ordinary oats as raw material.

[0106] Specifically:

[0107] Oat selection and washing: Select oats that are plump and free from pests and diseases, put them in a basin and submerge them in water to remove oats that float to the surface.

[0108] Oat soaking: After rinsing the oat seeds, soak the oats in water for 48 hours;

[0109] Preparation of oat solid culture medium: After adjusting the pH, filter out the oat with gauze and drain the water. Add oat solid culture medium to control the moisture content to within 70%. Weigh 50g and put it into an Erlenmeyer flask. Sterilize at 0.1MPa, 121℃ for 30min. Cool to room temperature to obtain oat solid culture medium.

[0110] Fermentation of oat red yeast rice: In a sterile environment, add 3 ml of 75% alcohol to the culture medium, measure 10 ml of the bacterial solution cultured in step 4 using a graduated cylinder, mix well, seal the bottle, and place it in an incubator for culture, controlling the temperature at 35℃ and the humidity at 70%; after 360 hours of culture, the fermentation of oat red yeast rice is complete.

[0111] Preparation of oat red yeast rice: Transfer the oat red yeast rice fermentation material to a drying oven for drying at a temperature of 50°C; dry until the humidity is below 25%, which is the finished oat red yeast rice product.

[0112] Since Comparative Example 4 successfully cultivated oat red yeast rice, but the process was lengthy, the following two examples are based on Comparative Example 4, adjusting different parameters in the cultivation process of Comparative Example 4 to obtain the optimal cultivation conditions for oat red yeast rice.

[0113] In Example 1, oats were treated with alkali; in Example 2, different cultivation temperatures were set; and in Example 3, different alcohol contents were set.

[0114] Example 1

[0115] Step 1: Cleaning the red yeast rice raw materials.

[0116] Specifically:

[0117] Using commercially available red yeast rice as the strain, weigh 10g of red yeast rice raw material, soak it in 75% alcohol for sterilization, filter out the alcohol after sterilization, then wash it 5 times with sterile water and drain it.

[0118] Step 2: Activation of Monascus purpureus.

[0119] Specifically:

[0120] Preparation of seed activation solution: The seed activation solution is prepared by 75% alcohol and maltose aqueous solution. Take 100ml of purified water, add 3ml of 75% alcohol and 2g of maltose, mix well, and the activation solution is ready.

[0121] Red yeast rice activation: Immerse the cleaned red yeast rice raw material in seed activation solution; then place it in an incubator, control the temperature at 35℃ and humidity at 70%, and incubate for 12 hours. A white, fluffy mycelial film will form on the surface of the red yeast rice raw material, thus completing the red yeast rice activation.

[0122] Step 3: Purification of Monascus purpureus.

[0123] Specifically:

[0124] Preparation of solid culture medium: Mix 70 g glucose, 15 g beef extract, 2 g NaNO3, 0.5 g MgSO4·7H2O, 1.5 g KH2PO4, and 2 g agar powder into 1000 ml sterile water and stir well. Sterilize the mixture at 0.1 MPa, 121 °C for 30 min. Dispense into Petri dishes and let cool and solidify. The preparation of solid culture medium is complete.

[0125] Purification of Monascus purpureus: Take one activated Monascus purpureus raw material, crush it in a 1.5ml sterile centrifuge tube, add 1ml sterile water to dilute it, and use a sterile disposable pipette to take a small amount and inoculate it onto a solid culture medium and incubate for 8 hours. After Monascus purpureus grows on the solid culture medium, pick out the part with pure color and transfer it to a new culture medium. Repeat this process several times until the colonies that grow are regular in shape and uniform in color, thus completing the purification of Monascus purpureus.

[0126] Step 4: Propagation and Culture of Monascus purpureus

[0127] Specifically:

[0128] Preparation of seed culture medium: 70 g glucose, 15 g beef extract, 2 g NaNO3, 0.5 g MgSO4·7H2O, and 1.5 g KH2PO4 were mixed with 1000 ml sterile water and stirred evenly. The mixture was then dispensed into 250 ml Erlenmeyer flasks, wrapped with 8 layers of clean gauze, and sterilized under high pressure at 0.1 MPa and 121℃ for 30 min to complete the preparation of seed culture medium.

[0129] Monascus purpureus propagation culture: Under sterile conditions, a small number of Monascus purpureus colonies purified in step 3 were picked up with sterilized tweezers and inoculated into the liquid seed culture medium in an Erlenmeyer flask. The temperature was controlled at 35℃ and the humidity at 70% for propagation culture. The culture was carried out for 72 hours until the color of the seed culture medium gradually changed from clear to orange-yellow and a large number of mycelial clusters grew, thus completing the propagation culture of Monascus purpureus.

[0130] Step 5 uses alkali-treated oats as raw material to produce oat red yeast rice.

[0131] Specifically:

[0132] Oat selection and washing: Select oats that are plump and free from pests and diseases, put them in a basin and submerge them in water to remove oats that float to the surface.

[0133] Alkali solution preparation: Add 2g NaOH to every 100ml of sterile water to complete the preparation of alkali solution;

[0134] Oat soaking: After rinsing the oat seeds, soak the oats in an alkaline solution for 2 hours;

[0135] Adjust pH: Soak alkaline oats in an acetic acid solution with a pH of 4 for 2.5 hours;

[0136] Preparation of oat solid culture medium: After adjusting the pH, filter out the oat with gauze and drain the water. Add oat solid culture medium to control the moisture content to within 70%. Weigh 50g and put it into an Erlenmeyer flask. Sterilize at 0.1MPa, 121℃ for 30min. Cool to room temperature to obtain oat solid culture medium.

[0137] Fermentation of oat red yeast rice: In a sterile environment, add 3 ml of 75% alcohol to the culture medium, measure 10 ml of the bacterial solution cultured in step 4 using a graduated cylinder, mix well, seal the bottle, and place it in an incubator for culture, controlling the temperature at 35℃ and the humidity at 70%; after 240 hours of culture, the fermentation of oat red yeast rice is complete.

[0138] Preparation of oat red yeast rice: Transfer the oat red yeast rice fermentation material to a drying oven for drying at a temperature of 50°C; dry until the humidity is below 25%, which is the finished oat red yeast rice product.

[0139] Figure 5 Microscopic images of oat red yeast rice cultured with alkali-treated oats provided in this application. Oat red yeast rice is cultivated using oat solid culture medium made from alkali-treated oats. Because red yeast rice can penetrate the oat epidermis more easily and develop faster, alkali-treated oats grow faster than ordinary oats, which can shorten the fermentation time by 120 hours, while reducing the risk of contamination by other microorganisms, and there are no other microorganisms or impurities in the surrounding environment.

[0140] Example 2

[0141] Step 1: Cleaning the red yeast rice raw materials.

[0142] Specifically:

[0143] Using commercially available red yeast rice as the strain, weigh 10g of red yeast rice raw material, soak it in 75% alcohol for sterilization, filter out the alcohol after sterilization, then wash it 5 times with sterile water and drain it.

[0144] Step 2: Activation of Monascus purpureus.

[0145] Specifically:

[0146] Preparation of seed activation solution: The seed activation solution is prepared by 75% alcohol and maltose aqueous solution. Take 100ml of purified water, add 3ml of 75% alcohol and 2g of maltose, mix well, and the activation solution is ready.

[0147] Red yeast rice activation: Immerse the cleaned red yeast rice raw material in seed activation solution; then place it in an incubator, control the temperature at 35℃ and humidity at 70%, and incubate for 12 hours. A white, fluffy mycelial film will form on the surface of the red yeast rice raw material, thus completing the red yeast rice activation.

[0148] Step 3: Purification of Monascus purpureus.

[0149] Specifically:

[0150] Preparation of solid culture medium: Mix 70 g glucose, 15 g beef extract, 2 g NaNO3, 0.5 g MgSO4·7H2O, 1.5 g KH2PO4, and 2 g agar powder into 1000 ml sterile water and stir well. Sterilize the mixture at 0.1 MPa, 121 °C for 30 min. Dispense into Petri dishes and let cool and solidify. The preparation of solid culture medium is complete.

[0151] Purification of Monascus purpureus: Take one activated Monascus purpureus raw material, crush it in a 1.5ml sterile centrifuge tube, add 1ml sterile water to dilute it, and use a sterile disposable pipette to inoculate a small amount onto a solid culture medium and incubate for 24h at 35℃ and 70% humidity for 8h. After Monascus purpureus grows on the solid culture medium, pick out the part with pure color and transfer it to a new culture medium. Repeat this process several times until the colonies grow out of a regular shape and have a uniform color, thus completing the purification of Monascus purpureus.

[0152] Step 4: Propagation and Culture of Monascus purpureus

[0153] Specifically:

[0154] Preparation of seed culture medium: 70 g glucose, 15 g beef extract, 2 g NaNO3, 0.5 g MgSO4·7H2O, and 1.5 g KH2PO4 were mixed with 1000 ml sterile water and stirred evenly. The mixture was then dispensed into 250 ml Erlenmeyer flasks, wrapped with 8 layers of clean gauze, and sterilized under high pressure at 0.1 MPa and 121℃ for 30 min to complete the preparation of seed culture medium.

[0155] Monascus purpureus propagation culture: Under sterile conditions, a small number of Monascus purpureus colonies purified in step 3 were picked up with sterilized tweezers and inoculated into the liquid seed culture medium in an Erlenmeyer flask. The temperature was controlled at 35℃ and the humidity at 70% for propagation culture. The culture was carried out for 72 hours until the color of the seed culture medium gradually changed from clear to orange-yellow and a large number of mycelial clusters grew, thus completing the propagation culture of Monascus purpureus.

[0156] Step 5 uses alkali-treated oats as raw material to produce oat red yeast rice.

[0157] Specifically:

[0158] Oat selection and washing: Select oats that are plump and free from pests and diseases, put them in a basin and submerge them in water to remove oats that float to the surface.

[0159] Alkali solution preparation: Add 2g NaOH to every 100ml of sterile water to complete the preparation of alkali solution;

[0160] Oat soaking: After rinsing the oat seeds, soak the oats in an alkaline solution for 2 hours;

[0161] Adjust pH: Soak alkaline oats in an acetic acid solution with a pH of 4 for 2.5 hours;

[0162] Preparation of oat solid culture medium: After adjusting the pH, filter out the oat with gauze and drain the water. Add oat solid culture medium to control the moisture content to within 70%. Weigh 50g and put it into an Erlenmeyer flask. Sterilize at 0.1MPa, 121℃ for 30min. Cool to room temperature to obtain oat solid culture medium.

[0163] Fermentation of oat red yeast rice: In a sterile environment, add 3 ml of 75% alcohol to the culture medium, measure 10 ml of the bacterial solution cultured in step 4 with a graduated cylinder, mix well, seal the bottle, and place it in an incubator for culture. Control the temperature at 25, 30, 35, and 40℃, the humidity at 70%, and culture for 240 hours. The fermentation of oat red yeast rice is then complete.

[0164] Preparation of oat red yeast rice: Transfer the oat red yeast rice fermentation material to a drying oven for drying at a temperature of 50°C; dry until the humidity is below 25%, which is the finished oat red yeast rice product.

[0165] Table 1 shows the effect of different culture temperatures on the fermentation time of oat red yeast rice.

[0166] Incubation temperature (°C) Step 5: Oat solid culture medium color change time (h) 25 268 30 252 35 240 40 265

[0167] Based on Table 1, under different cultivation temperatures, apart from the different cultivation times, there is no significant difference in the final results of each cultivation step. When the temperature is 35℃, the fermentation time of oat red yeast rice is the shortest. Therefore, the cultivation temperature in each step is set at 35℃.

[0168] Example 3

[0169] Step 1: Cleaning the red yeast rice raw materials.

[0170] Specifically:

[0171] Using commercially available red yeast rice as the strain, weigh 10g of red yeast rice raw material, soak it in 75% alcohol for sterilization, filter out the alcohol after sterilization, then wash it 5 times with sterile water and drain it.

[0172] Step 2: Activation of Monascus purpureus.

[0173] Specifically:

[0174] Preparation of seed activation solution: The seed activation solution is prepared by 75% alcohol and maltose aqueous solution. Take 100ml of purified water, add 3ml of 75% alcohol and 2g of maltose, mix well, and the activation solution is ready.

[0175] Red yeast rice activation: Immerse the cleaned red yeast rice raw material in seed activation solution; then place it in an incubator, control the temperature at 35℃ and humidity at 70%, and incubate for 12 hours. A white, fluffy mycelial film will form on the surface of the red yeast rice raw material, thus completing the red yeast rice activation.

[0176] Step 3: Purification of Monascus purpureus.

[0177] Specifically:

[0178] Preparation of solid culture medium: Mix 70 g glucose, 15 g beef extract, 2 g NaNO3, 0.5 g MgSO4·7H2O, 1.5 g KH2PO4, and 2 g agar powder into 1000 ml sterile water and stir well. Sterilize the mixture at 0.1 MPa, 121 °C for 30 min. Dispense into Petri dishes and let cool and solidify. The preparation of solid culture medium is complete.

[0179] Purification of Monascus purpureus: Take one activated Monascus purpureus raw material, crush it in a 1.5ml sterile centrifuge tube, add 1ml sterile water to dilute it, and use a sterile disposable pipette to inoculate a small amount onto a solid culture medium and incubate for 24h at 35℃ and 70% humidity for 8h. After Monascus purpureus grows on the solid culture medium, pick out the part with pure color and transfer it to a new culture medium. Repeat this process several times until the colonies grow out of a regular shape and have a uniform color, thus completing the purification of Monascus purpureus.

[0180] Step 4: Propagation and Culture of Monascus purpureus

[0181] Specifically:

[0182] Preparation of seed culture medium: 70 g glucose, 15 g beef extract, 2 g NaNO3, 0.5 g MgSO4·7H2O, and 1.5 g KH2PO4 were mixed with 1000 ml sterile water and stirred evenly. The mixture was then dispensed into 250 ml Erlenmeyer flasks, wrapped with 8 layers of clean gauze, and sterilized under high pressure at 0.1 MPa and 121℃ for 30 min to complete the preparation of seed culture medium.

[0183] Monascus purpureus propagation culture: Under sterile conditions, a small number of Monascus purpureus colonies purified in step 3 were picked up with sterilized tweezers and inoculated into the liquid seed culture medium in an Erlenmeyer flask. The temperature was controlled at 35℃ and the humidity at 70% for propagation culture. The culture was carried out for 72 hours until the color of the seed culture medium gradually changed from clear to orange-yellow and a large number of mycelial clusters grew, thus completing the propagation culture of Monascus purpureus.

[0184] Step 5 uses alkali-treated oats as raw material to produce oat red yeast rice.

[0185] Specifically:

[0186] Oat selection and washing: Select oats that are plump and free from pests and diseases, put them in a basin and submerge them in water to remove oats that float to the surface.

[0187] Alkali solution preparation: Add 2g NaOH to every 100ml of sterile water to complete the preparation of alkali solution;

[0188] Oat soaking: After rinsing the oat seeds, soak the oats in an alkaline solution for 2 hours;

[0189] Adjust pH: Soak alkaline oats in an acetic acid solution with a pH of 4 for 2.5 hours;

[0190] Preparation of oat solid culture medium: After adjusting the pH, filter out the oat with gauze and drain the water. Add oat solid culture medium to control the moisture content to within 70%. Weigh 50g and put it into an Erlenmeyer flask. Sterilize at 0.1MPa, 121℃ for 30min. Cool to room temperature to obtain oat solid culture medium.

[0191] Fermentation of oat red yeast rice: In a sterile environment, add 0, 1, 2, 3 and 4 ml of 75% alcohol to the culture medium respectively. Measure 10 ml of the bacterial solution cultured in step 4 with a graduated cylinder, mix well, seal the bottle, and place it in an incubator for culture. Control the temperature at 25, 30, 35 and 40℃ and the humidity at 70% respectively. Culture for 240 hours to complete the fermentation of oat red yeast rice.

[0192] Preparation of oat red yeast rice: Transfer the oat red yeast rice fermentation material to a drying oven for drying at a temperature of 50°C; dry until the humidity is below 25%, which is the finished oat red yeast rice product.

[0193] Table 2 shows the effect of different alcohol contents on the color change time of the oat solid culture medium in step 5.

[0194]

[0195] Monascus purpureus exhibits resistance to acetic acid and alcohol. As shown in Table 2, in step 5, when 0, 1, 2, and 3 ml / 50g of 75% alcohol were added to produce oat red yeast rice, the maximum difference in growth time was 4 hours. However, considering that 75% alcohol has the effect of inhibiting the growth of other microorganisms, it can reduce the aseptic requirements of the culture environment and simplify the production equipment. Therefore, without significantly affecting the growth of Monascus purpureus, the amount of 75% alcohol added in step 5 when using alkali-treated oats as raw material to produce oat red yeast rice is 3 ml.

[0196] Final conclusion: The optimal culture conditions were those in the culture process of Example 1, which yielded the best quality oat red yeast rice.

Claims

1. A method for cultivating oat red yeast rice, characterized in that, The steps include the following: Step 1: Cleaning the red yeast rice raw materials: Using commercially available red yeast rice raw materials as the source of bacteria, clean the red yeast rice raw materials several times with 75% alcohol and sterile water in sequence, and then drain the water. Step 2: Activation of Monascus purpureus: Add the cleaned Monascus purpureus raw material to the prepared seed activation solution for activation culture for 12 hours; Step 3: Purification of Monascus purpureus: The activated Monascus raw material is inoculated into a solid culture medium and cultured to obtain a single Monascus purpureus colony; Step 4: Propagation and culture of Monascus purpureus: Inoculate the single Monascus purpureus colony described in Step 3 into the seed culture medium for culture. When the color of the seed culture medium gradually changes from clear to orange-yellow and a large number of mycelial clumps grow, the propagation and culture of Monascus purpureus is completed. Step 5: Production of oat red yeast rice using alkali-treated oats as raw material: Seed culture solution containing red yeast mold clusters is inoculated into a pre-prepared oat solid culture medium and cultured for 240-360 hours, with the temperature controlled at 25-40℃ and humidity at 70%, to obtain oat red yeast rice fermentation product. The oat red yeast rice fermentation product is dried to a humidity of less than 25% to obtain oat red yeast rice; wherein, the oat solid culture medium is a culture medium prepared by alkalizing oats and adjusting the pH.

2. The method for cultivating oat red yeast rice according to claim 1, characterized in that, The specific process and method for cleaning the red yeast rice raw materials in step 1 are as follows: Using commercially available red yeast rice as the source of microbial culture, weigh 10g of red yeast rice raw material, soak it in 75% alcohol for sterilization, filter out the alcohol after sterilization, and then wash it 5 times with sterile water and drain it.

3. The method for cultivating oat red yeast rice according to claim 1, characterized in that, The specific process and method for activating Monascus purpureus in step 2 are as follows: Preparation of seed activation solution: The seed activation solution is prepared by 75% alcohol and maltose aqueous solution. Measure 100ml of purified water, add 3ml of 75% alcohol and 2g of maltose, mix well to complete the preparation of the activation solution. Red yeast rice activation: Immerse the cleaned red yeast rice raw material in seed activation solution; then place it in an incubator, control the temperature at 35℃ and humidity at 70%, and incubate for 12 hours. A white, fluffy mycelial film will form on the surface of the red yeast rice raw material, thus completing the red yeast rice activation.

4. The method for cultivating oat red yeast rice according to claim 1, characterized in that, The specific process and method for purifying Monascus purpureus in step 3 are as follows: Preparation of solid culture medium: Mix 70g glucose, 15g beef extract, 2g NaNO3, 0.5g MgSO4·7H2O, 1.5g KH2PO4, and 2g agar powder into 1000ml sterile water and stir well. Sterilize the mixture at 0.1MPa, 121℃ for 30min. Dispense into Petri dishes and allow to cool and solidify. The preparation of solid culture medium is complete. Purification of Monascus purpureus: Take one activated Monascus purpureus raw material, crush it in a 1.5ml sterile centrifuge tube, add 1ml sterile water to dilute it, and use a sterile disposable pipette to inoculate a small amount onto a solid culture medium and incubate for 8 hours. After Monascus purpureus grows on the solid culture medium, pick out the part with pure color and transfer it to a new culture medium. Repeat this process several times until the colonies that grow are regular in shape and uniform in color, thus completing the purification of Monascus purpureus.

5. The method for cultivating oat red yeast rice according to claim 1, characterized in that, The specific process and method for step 4, Monascus purpureus propagation and culture, are as follows: Preparation of seed culture medium: Mix 70g glucose, 15g beef extract, 2g NaNO3, 0.5g MgSO4·7H2O, and 1.5g KH2PO4 into 1000ml sterile water and stir well. Then, dispense the mixture into 250ml Erlenmeyer flasks, wrap each flask with 8 layers of clean gauze, and autoclave at 0.1MPa and 121℃ for 30 minutes to complete the preparation of seed culture medium. Monascus purpureus propagation culture: Under sterile conditions, a small number of Monascus purpureus colonies purified in step 3 are picked up with sterilized tweezers and inoculated into the liquid seed culture medium in an Erlenmeyer flask. The temperature is controlled at 35℃ and the humidity at 70% for propagation culture. The culture is carried out for 72 hours until the color of the seed culture medium gradually changes from clear to orange-yellow and a large number of mycelial clusters grow, thus completing the propagation culture of Monascus purpureus.

6. The method for cultivating oat red yeast rice according to claim 1, characterized in that, The specific process and method for producing oat red yeast rice using alkali-treated oats as raw material in step 5 are as follows: Oat selection and washing: Select oats that are plump and free from pests and diseases, put them in a basin and submerge them in water to remove oats that float to the surface. Alkali solution preparation: Add 2g NaOH to every 100ml of sterile water to complete the preparation of alkali solution; Oat soaking: After rinsing the oat seeds, soak the oats in an alkaline solution for 2 hours; Adjust pH: Soak alkaline oats in an acetic acid solution with a pH of 4 for 2.5 hours; Preparation of oat solid culture medium: After adjusting the pH, filter out the oat with gauze and drain the water. Add oat solid culture medium to control the moisture content to within 70%. Weigh 50g and put it into an Erlenmeyer flask. Sterilize at 0.1MPa, 121℃ for 30min. Cool to room temperature to obtain oat solid culture medium. Fermentation of oat red yeast rice: In a sterile environment, add 3 ml of 75% alcohol to the culture medium, measure 10 ml of the bacterial solution cultured in step 4 with a graduated cylinder, mix well, seal the bottle, and place it in an incubator for culture, controlling the temperature at 35℃ and the humidity at 70%; after 240 hours of culture, the fermentation of oat red yeast rice is complete. Preparation of oat red yeast rice: Transfer the oat red yeast rice fermentation material to a drying oven for drying at a temperature of 50°C; dry until the humidity is below 25%, which is the finished oat red yeast rice product.

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