Monascus strain capable of producing monascin and / or ankaflavin with high yield and use thereof

Abstract

Provided are a monascus strain capable of producing monascin and / or ankaflavin and the use thereof. Particularly, provided are a new monascus strain assigned with the accession number CGMCC NO. 41335 and the use thereof in the production of high-content monascin and / or ankaflavin and related products thereof. The strain is Monascus purpureus ZYXQ-2206, can produce potent and non-toxic lipid-lowering ingredients monascin and / or ankaflavin with a high yield, and does not produce harmful and toxic ingredients lovastatin (LOV) and citrinin (CIT). Since excessive intake of LOV can cause serious side effects such as liver and kidney injury and rhabdomyolysis, and excessive intake of CIT can cause toxic responses such as renal tubular dilation and degeneration and necrosis of epithelial cells, the monascus products produced by using the strain can effectively avoid these safety risks, and have extremely high application prospects and market potentials. By screening and optimizing a fermentation process, the monascus strain having high productivity, no toxicity and high functionality is provided, which has significant application value in the fields of development of lipid-lowering functional foods and dietary supplements.

Description

Monascus sp. producing monacolins and / or monascins and application thereof TECHNICAL FIELD

[0001] The present application relates to the field of microbial technology, in particular to a Monascus sp. producing monacolins and / or monascins and application thereof. BACKGROUND

[0002] Red koji is the mycelium of Monascus spp. parasitizing on grains such as rice and fermented, since the French scholar established the classification in 1884, 36 species have been identified, among which M. purpureus, M. anka, M. pilosus and M. ruber are more commonly used in industrial fermentation and food fermentation. In China, M. purpureus and M. anka are legally used in drugs, health foods and foods. Red koji is traditionally recorded as sweet, slightly warm, and belonging to the spleen, large intestine and liver meridians; it has the functions of promoting digestion and blood circulation, strengthening the spleen and nourishing the stomach, and is used for blood stasis abdominal pain, diarrhea, sprains and injuries, and postpartum lochia. The Book of Drinking and Eating records: "Red koji is sweet, flat, and non-toxic. It can strengthen the spleen, benefit qi and warm the middle". The Compendium of Materia Medica records: "Red koji is sweet and warm, non-toxic, and is good for women with blood and qi pain and postpartum lochia". Red koji is also often used in combination with other traditional Chinese medicines to treat digestive system diseases, metabolic diseases and cardiovascular diseases, etc. Modern research shows that red koji has multiple activities such as antioxidant, antibacterial, anti-atherosclerosis, anti-obesity, hypolipidemic, hypoglycemic, anti-tumor activity, etc.

[0003] In 1979, Japanese person Fumio Endo first extracted a functional active substance from the culture solution of some strains of M. pilosus, M. ruber and M. purpureus, which could significantly inhibit the synthesis of cholesterol, and named it Monacolin K. Subsequent studies have confirmed that Monacolin K is Lovastatin (LOV). Although LOV has good lipid-lowering activity by strongly inhibiting cholesterol synthesis, and was approved by the US FDA as a lipid-lowering drug in 1987, with the popularization of its application, its side effects such as liver and kidney damage and rhabdomyolysis have attracted more and more attention, and many countries in the European Union and the United States have issued restrictions on LOV in related products containing red koji.

[0004] Monascin and Ankaflavin both significantly reduced cholesterol levels by 16% (P<0.05) and 21.9% (P<0.05), respectively, by inhibiting the expression of acetyl-CoA transferase, microsomal triglyceride transfer protein, and apolipoprotein apoB-100. They also exhibited significant lipid-lowering activities, including significantly reducing low-density lipoprotein cholesterol and increasing high-density lipoprotein cholesterol (HDL-C) by 11% and 22%, respectively (J Food Drug). (Anal, 2018, 26(1):393.) and has excellent safety. Therefore, Monascin and Ankaflavin are used by the United States Pharmacopeia (USP) as quality control indicators for red yeast rice and red yeast rice extract, requiring that "Mon not less than 7.0 mg / g and Ankaflavin not less than 2.3 mg / g on a dry basis". LOV is also subject to a limit control as a toxic component, "not exceeding 0.2 μg / g". At the same time, citrinin (CIT), which can cause renal tubular dilation and epithelial cell degeneration and necrosis when exceeded, is also subject to a limit control, stipulating that "not exceeding 0.1 μg / g".

[0005] [Amended according to Rule 26, 23.01.2026] In recent years, the M. purpureus NTU568 strain screened by scholars in Taiwan, China, has produced red yeast rice containing lipid-lowering active ingredients Mon and Ank, and has been shown to inhibit cholesterol absorption and promote cholesterol decomposition. Its lipid-lowering, blood pressure-lowering and blood sugar-lowering pharmacological activities have been confirmed by animal and clinical studies, and its safety has also been confirmed and published (Toxicol Rep. 2022 Feb 22; 9:356-365. Food Funct. 2017 Jun 1; 8(6):2102-2109). The fermented product red yeast rice has been approved by the US FDA as a new dietary ingredient additive. However, the Mon and Ank content of its red yeast rice product is low (Mon 0.70% and Ank 0.23%).

[0006] [Revised from Detailed Rule 26 to 23.01.2026] Currently, most red yeast rice products in China contain high levels of LOV (Lower Oxygen Value). For example, the functional red yeast rice (powder) standard QB / T 2847-2007 stipulates LOV (acidic + lactone) ≥ 0.4%, and T_ZZB 1452-2019 stipulates LOV (acidic + lactone) ≥ 1.0%. The Taiwan standard for red yeast rice is 4.8–15.0 mg daily. The standard for Xuezhikang (a traditional Chinese medicine) specifies that red yeast rice raw materials contain LOV ≥ 0.22%. Thus, the LOV in domestic red yeast rice products is controlled at "no less than" for "effective / functional ingredients," which is completely contrary to the international standard of "no more than" (NMT) for "toxic ingredients."

[0007] [Revised from Detailed Rule 26 to 23.01.2026] Recently, a serious incident involving multiple deaths caused by Kobayashi red yeast rice health products was reported in Japan. Although the causes of death are varied, CIT (Cryogenic Tolerance) is almost universally acknowledged. This is confirmed by the fact that UPS and some Chinese red yeast rice standards have limits on CNT, such as: NMT 40μg / kg (GB 1886.181-2016), 50μg / kg (QB / T 2847-2007), 100μg / kg (USP), 1000μg / kg (GB 1886.66-2015), and 2000μg / kg. According to literature reports and our actual test data, the CIT of red yeast rice produced by many domestic manufacturers seriously exceeds the standard. In addition to the speculated origin of mold in some rice, the biosynthetic pathway of CIT is similar to that of LOV (Local Oxygen Demand), which is speculated to be the underlying cause.

[0008] [Revised from Detail 26 to 23.01.2026] Perhaps because many people "believe" in LOV, and the Chinese Pharmacopoeia also uses this indicator for quality control of red yeast rice medicines, red yeast rice lipid-lowering health foods, etc., before the "Kobayashi Red Yeast Rice Incident" in Japan, few people thought about what the effective / functional components of red yeast rice are. The M. purpureus NTU568 strain discovered by scholars in Taiwan, China, has only been recognized by the USP and included in the USP standard, but its content is low. "On a dry basis, Mon is not less than 7.0 mg / g, and Ank is not less than 2.3 mg / g."

[0009] Current literature has confirmed that Mon and Ank are effective / functional components of red yeast rice in lowering lipids, blood sugar, and blood pressure. LOV and CIT are internationally recognized limited toxic components. Therefore, it is urgent to find a new red yeast rice strain that produces high levels of Mon and Ank and does not contain LOV and CIT. Summary of the Invention

[0010] To address the shortcomings of existing technologies, this application screens stable strains of *M. purpureus* and provides a *Monascus* strain capable of high production of Mon and / or Ank. The *Monascus* strain provided in this application is named *Monascus purpureus* ZYXQ-2206, and was deposited on June 14, 2024, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, China, 100101, China, with accession number CGMCC NO.41335.

[0011] Specifically, this application adopts the following technical solution:

[0012] 1. A red mold, characterized in that the red mold (Monascus purpureus) has the preservation number: CGMCC NO.41335.

[0013] 2. The Monascus purpureus according to claim 1, characterized in that the DNA sequence of the Monascus purpureus is shown in SEQ ID NO:1.

[0014] 3. A culture containing the Monascus purpureus described in item 1.

[0015] 4. The use of the Monascus purpureus described in item 1 or the culture of item 3 in the culture of Monascin and / or Ankaflavin.

[0016] 5. A method for culturing Monascus rubra and / or Monascus xanthophyll, characterized in that it includes culturing Monascus rubra and / or Monascus xanthophyll using the Monascus mold described in item 1.

[0017] 6. The method according to item 5, characterized in that it further includes the step of:

[0018] The Monascus purpureus strain was activated and then inoculated into a seed culture medium to obtain a strain seed solution.

[0019] The seed culture of the strain was inoculated into a fermentation medium for fermentation culture;

[0020] The Monascus strain was activated before being inoculated into the seed culture medium.

[0021] 7. The method according to item 6, characterized in that the fermentation culture conditions are aerobic culture, the fermentation temperature is 25-50℃, the fermentation culture time is 2-20 days, and the fermentation culture method is solid-state fermentation or liquid-state fermentation.

[0022] 8. The method according to item 6, characterized in that the content of each component of the seed culture medium is: glucose 10-50 g / L, peptone 5-30 g / L, yeast powder 5-30 g / L, soybean powder 2-20 g / L, glycerol 20-80 mL / L, magnesium sulfate 0.2-2 g / L, sodium nitrate 0.5-5 g / L, zinc sulfate 0.5-5 g / L, potassium dihydrogen phosphate 0.2-2 g / L, maltodextrin 10-50 g / L; and / or

[0023] The raw materials for the fermentation medium are selected from one or more of rice, soybeans, corn, or yam.

[0024] 9. The method according to item 6, characterized in that the inoculum amount of the strain seed inoculated into the fermentation medium is 5-15% (V / V).

[0025] 10. The method according to item 6, characterized in that the plate culture conditions are cultured at 25-50°C for 2-20 days.

[0026] Beneficial effects: Compared with the prior art, this application has the following advantages:

[0027] This application provides a novel Monascus purpureus strain, namely Monascus purpureus ZYXQ-2206, which has significant advantages. During fermentation, it efficiently produces Mon and Ank without producing LOV and CIT. This breakthrough strain overcomes the problem of traditional Monascus purpureus strains producing high levels of toxic components such as LOV and CIT.

[0028] [Amended according to Rule 26, 23.01.2026] Through solid-state or liquid-state fermentation processes, the ZYXQ-2206 strain in this application can significantly increase the yield of Mon and Ank. Compared with strains in the prior art, such as the NTU568 strain from Taiwan, the Mon content obtained by the ZYXQ-2206 strain in this application is as high as 5.86%, and the Ank content is as high as 1.87%, which are 8.4 times and 8.1 times that of the NTU568 strain (Mon 0.70%, Ank 0.23%), respectively. This significantly increases the content of functional components of red yeast rice and solves the problem of insufficient Mon and Ank yield in traditional red yeast rice fermentation.

[0029] Furthermore, since LOV and CIT are recognized as toxic components, they are subject to strict limits internationally. The strain described in this application can avoid the formation of LOV and CIT, giving the fermentation product a significant advantage in safety and meeting stringent international and domestic quality standards for functional and toxic components. This characteristic makes the red yeast rice product provided in this application a safer and more effective natural dietary additive, reducing the risk of liver and kidney damage, rhabdomyolysis, and other complications caused by LOV and CIT exposure in consumers, and showing broad application prospects in pharmaceutical and functional food development.

[0030] Compared with traditional technologies, the red yeast rice products provided in this application are not only more functional but also free of toxic components, greatly improving product safety and market competitiveness, representing a major breakthrough in red yeast rice fermentation technology and product quality control. Attached Figure Description

[0031] The accompanying drawings are provided to better understand the content of this application, but do not constitute an undue limitation on this application. The specific content is as follows:

[0032] Figure 1 shows the colony morphology of Monascus purpureus 7-3 and ZYXQ-2206;

[0033] Figure 2 shows the microscopic morphological features of Monascus purpureus ZYXQ-2206;

[0034] Figure 3A shows the chromatogram of the reference standard determined by HPLC, where 1: citrinin (CIT); 2: lovastatin (LOV); 3: Monascin; 4: Ankaflavin;

[0035] Figure 3B shows the chromatogram of ZYXQ-2206 determined by HPLC, where 3: Monascin; 4: Ankaflavin.

[0036] Figure 3C shows the chromatogram of sample 7 determined by HPLC, where 2: lovastatin (LOV), 3: Monascin, and 4: Ankaflavin. Detailed Implementation

[0037] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; unless otherwise specified, the reagents and materials are commercially available.

[0038] It should be noted that certain terms are used in the specification and claims to refer to specific components. Those skilled in the art will understand that different terms may be used to refer to the same component. This specification and claims do not distinguish components based on differences in terminology, but rather on differences in function. The terms "comprising" or "including" used throughout the specification and claims are open-ended and should be interpreted as "comprising but not limited to." The following descriptions in the specification are preferred embodiments for carrying out this application; however, these descriptions are for the purpose of understanding the general principles of the specification and are not intended to limit the scope of this application. The scope of protection of this application shall be determined by the appended claims.

[0039] The technical terms mentioned in this specification have the same meanings as those commonly understood by those skilled in the art, and in case of any conflict, the definitions in this specification shall prevail.

[0040] In this application specification, "Monascin" refers to red yeast rice pigment, also known as red yeast rice coloring, CAS: 21516-68-7, which can be extracted from red yeast rice.

[0041] In this application specification, "Ankaflavin" refers to red yeast rice flavin, also known as Ankaflavin, CAS: 50980-32-0, which can be extracted from red yeast rice.

[0042] The red mold provided in this application is named Monascus purpureus ZYXQ-2206, and was deposited on June 14, 2024, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, China, 100101, China, with accession number CGMCC NO.41335.

[0043] The DNA sequence of Monascus purpureus provided in this application is shown in SEQ ID NO:1:

[0044] In one specific embodiment of this application, the screening method for Monascus purpureus includes the following steps:

[0045] Dissolve one portion of red yeast rice sample prepared according to traditional methods in 5-20 times its volume of sterilized purified water, and incubate at a constant temperature of 25-50℃ for 0.5-2 hours at 120-150 rpm / min. Dilute the supernatant after incubation with distilled water, and take appropriate amounts of the original solution... -1 10 -2 10 -3 The diluted solution was spread onto PDA plates and incubated in a constant temperature incubator at 25–50°C. The uniformity of the colony morphology was observed. Larger, redder colonies were selected, streaked, and purified to obtain the *Aspergillus purpureus* strain.

[0046] Inoculate *Aspergillus purpureus* onto a slant PDA medium. Scrape a small amount of mycelium and add it to an appropriate amount of sterilized distilled water, gently mixing by pipetting. Take an appropriate amount of bacterial solution and add 0.5–2 times the amount of distilled water. Irradiate the solution at a depth of 40 cm below a UV lamp for 1–5 minutes. After UV irradiation, place the solution in a 4°C refrigerator for 1–3 hours to stabilize. Then, spread an appropriate amount of the solution onto a PDA plate, making three replicates. Wrap the plates in aluminum foil and incubate at a constant temperature of 25–50°C for 5–7 days. Observe the colony growth. Select strains with larger colonies or significant morphological differences onto fresh PDA plates to obtain *Aspergillus purpureus* ZYXQ-2206.

[0047] This application also provides a culture containing the aforementioned *Monascus purpureus* ZYXQ-2206. In this application, the *Monascus purpureus* ZYXQ-2206 culture refers to the substance obtained by culturing the *Monascus purpureus* ZYXQ-2206 of this application; more specifically, it refers to a mixture of *Monascus purpureus* ZYXQ-2206 cells, the culture medium used for culturing *Monascus purpureus* ZYXQ-2206, and substances produced by the cultured *Monascus purpureus* ZYXQ-2206.

[0048] This application also provides the application of the above-mentioned Monascus purpureus ZYXQ-2206 or its culture in the cultivation of Monascus purpureus and Monascus xanthophyll.

[0049] This application also provides a method for producing monaxanthin and monaxanthin through solid-state or liquid-state fermentation, which uses the aforementioned Monascus purpureus ZYXQ-2206 or a culture containing the strain to cultivate monaxanthin and monaxanthin, and does not produce lovastatin or citrinin during the process.

[0050] In one specific embodiment of this application, the purple-red Aspergillus ZYXQ-2206 is placed on a plate culture medium, preferably potato glucose medium, with the carbon source being 6-18 g / L potato extract powder, 10-30 g / L glucose, 6-25 g / L agar powder, pH value 5.6±0.2, and the remainder being water.

[0051] In one specific embodiment of this application, the plate culture medium inoculated with Monascus purpureus has a Monascus purpureus culture temperature of 25-50°C, preferably 25-30°C, for example, 25°C, 26°C, 27°C, 28°C, 29°C, or 30°C.

[0052] In one specific embodiment of this application, the plate culture medium inoculated with Monascus purpureus has a Monascus purpureus culture time of 2 to 20 days, preferably 5 to 15 days.

[0053] In one specific embodiment of this application, the plate culture medium is selected from one or more of potato dextrose agar (PDA), potato dextrose water agar (PD), yeast extract sucrose agar (CYA), or malt extract agar (MEA).

[0054] In one specific embodiment of this application, the activated Monascus purpureus is inoculated into a seed culture medium containing: glucose 10-50 g / L, peptone 5-30 g / L, yeast powder 5-30 g / L, soybean powder 2-20 g / L, glycerol 20-80 mL / L, magnesium sulfate 0.2-2 g / L, sodium nitrate 0.5-5 g / L, zinc sulfate 0.5-5 g / L, potassium dihydrogen phosphate 0.2-2 g / L, and maltodextrin 10-50 g / L. The medium is then autoclaved at 121°C for later use.

[0055] In one specific embodiment of this application, the plate washing solution or the bacterial seed liquid is inoculated into the fermentation medium for solid-state fermentation or liquid-state fermentation.

[0056] In one specific embodiment of this application, the inoculum amount of seed culture medium inoculated into the fermentation culture medium is 5% to 15%, for example, it can be 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, or 15%.

[0057] In one specific embodiment of this application, the raw material in the fermentation culture medium is commercially available rice that has been sterilized at 121°C for 30 minutes.

[0058] In one specific embodiment of this application, the raw materials in the fermentation culture medium are selected from one or more of rice, soybeans, corn or yam, and further, the rice, soybeans, corn or yam are ground into powder.

[0059] Example

[0060] In the following examples, the seed culture medium used contained 10-50 g / L glucose, 5-30 g / L peptone, 5-30 g / L yeast powder, 2-20 g / L soybean powder, 20-80 mL / L glycerol, 0.2-2 g / L magnesium sulfate, 0.5-5 g / L sodium nitrate, 0.5-5 g / L zinc sulfate, 0.2-2 g / L potassium dihydrogen phosphate, and 10-50 g / L maltodextrin, and was autoclaved at 121°C before use.

[0061] Example 1: Isolation, screening and identification of purple monaxillary enzyme ZYXQ-2206

[0062] Dissolve 10-20g of red yeast rice prepared according to traditional methods in 100-150mL of sterilized purified water, and incubate at 25-50℃ in a constant temperature shaker at 120-150rpm / min for 0.5-2h. Collect the supernatant after incubation and serially dilute it with distilled water. Take 200μL of the original solution and dilute it with 10... -1 10 -2 10 -3The diluted solution was spread onto PDA plates and incubated at 20–50°C for 2–20 days. The uniformity of colony morphology was observed to obtain *Aspergillus purpureus* strain 7-3. *Aspergillus purpureus* strain 7-3 was inoculated onto PDA slant agar. A small amount of mycelium was scraped and added to 0.5–2 times the volume of sterilized distilled water, and gently mixed. 500 μL of the bacterial solution was added to 500 μL of distilled water, for a total of 5 tubes. The tubes were irradiated with a UV lamp at a depth of 40 cm for 1–5 minutes. After UV irradiation, the bacterial solution was stabilized at 4°C for 1–3 hours. 200 μL of the bacterial solution was then spread onto PDA plates (3 replicates), wrapped in aluminum foil, and incubated at 25–50°C for 2–20 days. Colony growth was observed. Strains with larger colonies or significant morphological differences were selected and placed on fresh PDA plates. One of the obtained strains was named Monascus purpureus ZYXQ-2206. The colony morphology of Monascus purpureus strain 7-3 and ZYXQ-2206 is shown in Figure 1.

[0063] Scrape 0.2–0.5 mg of ZYXQ-2206 mycelia and place them in a 2.0 mL centrifuge tube; add 2 glass beads (3–4 mm in diameter) and 100 μL of CTAB lysis buffer, and homogenize twice on a Retsch M400 homogenizer for 2 min each time; add 400 μL of CTAB lysis buffer preheated in a 65°C water bath, invert and mix well, then incubate at 65°C for 0.5–1 h; add an equal volume of chloroform:isoamyl alcohol (24:1), and gently invert and mix 50 times; centrifuge at 12000 rpm at room temperature for 5 min, and transfer the supernatant to a new centrifuge tube; add 1.5 times the volume of binding solution to the supernatant, mix well, transfer to an adsorption column, centrifuge at 12000 rpm at room temperature for 30 sec, and discard the waste liquid; add 500 μL of inhibitor removal solution, and centrifuge at 12000 rpm at room temperature for 30 s. ec, discard the waste liquid; add 600 μL of rinsing buffer, centrifuge at 12000 rpm at room temperature for 30 sec, discard the waste liquid; put the adsorption column back into the empty collection tube, centrifuge at 12000 rpm at room temperature for 2 min; transfer the adsorption column to a sterile 1.5 mL centrifuge tube, leave it open to dry at room temperature for 5 min; add 50–100 μL of elution buffer, place at room temperature for 3–5 min, centrifuge at 12000 rpm for 30 sec to collect the genomic DNA solution, use immediately or store at 20 °C for later use.

[0064] After template extraction, PCR amplification was performed using ITS primers. The PCR reaction system (50 μL) consisted of: 2 μL fungal genomic DNA, 25 μL 2×Easy Taq PCR Mix, 2 μL each of forward and reverse primers, and 19 μL sterile water. PCR conditions were: 95℃ pre-denaturation for 3 min, 94℃ denaturation for 1 min, 54℃ annealing for 40 s, 72℃ extension for 1 min, for 35 cycles; 72℃ repair extension for 10 min, and termination at 4℃. The product was then stored at 4℃. The PCR product was directly sent to a sequencing company for sequencing. The sequencing instructions required routine purification of the product. The sequencing results were compared with the NCBI database, and at the molecular level, strain ZYXQ-2206 was identified as *Monasus purpureus*. The DNA sequence of ZYXQ-2206 is shown in SEQ ID NO:1.

[0065] The microscopic morphological characteristics of the *Aspergillus violaceus* strain ZYXQ-2206 are shown in Figure 2. Colony characteristics are as follows: It grows rapidly on malt extract agar (MEA) medium; after 5 days in the dark at 25°C, the colony diameter is 20–25 mm. Aerial hyphae are abundant, initially white, later light orange; the reverse side of the colony is dark orange with a reddish-brown center, and contains no soluble pigment. The mycelial microscopic characteristics are: vegetative hyphae are septate, highly branched, and 2.0–4.0 μm wide. Conidia are produced in large quantities; conidiophores are not clearly specialized, straight or slightly curved; conidia are spherical, colorless, with slightly rough walls, truncate at the base, and arranged in chains, 4.5–12.8 μm in diameter. The pericarp primordia are spherical, up to 25 μm in diameter, light to brown, and immature; ascospores were not observed. On CYA, ZYXQ-2206 colonies are 15–18 mm in diameter, flat, sparse, and usually have a curly texture. Occasionally, a small number of aerial hyphae are present. The mycelium is white or pale red, with small and clear exudates, and secretes orange or pink pigment. The back is pale to pink or orange-red.

[0066] Example 2 Fermentation process and color scale determination

[0067] Rice was soaked in an acetic acid solution with pH 5.5 for 20–24 hours, drained, and sterilized at 121°C for 30 minutes to obtain a solid rice fermentation medium. *Aspergillus violaceus* strain ZYXQ-2206 was inoculated onto PDA slant agar and cultured for 4–5 days. Once the mycelium had completely covered the slant and the back of the tube was bright red, the spores on the slant were washed off with sterile water at an inoculation rate of 10-1. 5 ~10 6 One spore per g (dry weight of rice) was inoculated into the cooled rice fermentation medium.

[0068] Solid-state fermentation was used for cultivation. Seed culture medium was added at a ratio of 5%–15% and then poured into the rice fermentation medium and stirred thoroughly. The inoculated medium was placed in an artificial climate incubator (constant temperature and humidity) at 25–50℃ and 60%–80% relative humidity for 3 days, turning it over and adding 5%–8% sterile water every 24 hours. On the 4th day, the temperature was maintained at 25–35℃ and the relative humidity at 50%–60%, turning it over every 24 hours and continuing to add water (at a ratio of 5%–8%) until the 10th day. The fermentation time was 15–20 days, until the rice core turned red. The obtained red yeast rice was dried and stored at 55–60℃. Using the above solid-state fermentation method, two more batches were cultivated, dried, and stored for later use.

[0069] Red yeast rice powder was prepared using a liquid fermentation method. Raw materials (fermentation medium, such as rice powder) were dissolved in water and pumped into a fermenter, filling it to 45%–55% of the fermenter's volume. High-temperature sterilization was then performed using steam. Sterilization conditions: 120–130℃, 30–60 min; cooling to 40–45℃ for later use. The amplified seed culture medium was transferred into the fermenter at a volume fraction of 5%–30%. Initial fermentation conditions were set as follows: stirring speed 100–450 r / min, aeration rate 1.5–5 L / min, controlled temperature 25–35℃, tank pressure 0.04–0.08 MPa, initial pH 4.5–5.5, fermentation for 10–15 days. After fermentation, vacuum concentration was performed at 35–55℃ and a vacuum level of -0.08–-0.095 MPa. The concentrate is then spray-dried. The inlet temperature of the drying tower is 165-200℃ and the outlet temperature is 60-90℃. The liquid material is spray-dried to obtain red yeast rice powder.

[0070] The method for determining the color value of red yeast rice is as follows: The method for determining the color value content of red yeast rice refers to "GB 1886.19-2015 National Food Safety Standard for Food Additives: Red Yeast Rice". The red yeast rice sample is pulverized using a pulverizer and passed through a No. 7 mesh sieve. 0.2 g of the pulverized and uniformly mixed sample is accurately weighed, dissolved in 70% ethanol solution, and transferred to a 100 mL volumetric flask. The solution is then diluted to the mark and placed in a 60℃ water bath for 1 hour. After immersion, the sample is removed and cooled to room temperature. 70% ethanol solution is added to the mark, mixed well, filtered through filter paper, and the filtrate is collected in a stoppered cuvette for later use. Accurately pipette 2.0 mL to 5.0 mL of the above solution into a 50 mL volumetric flask (ensuring the final dilution absorbance falls within the range of 0.2 to 0.8), dilute to 50 mL with 70% ethanol solution, shake well, and use a 10 mm cuvette, with 70% ethanol solution as a reference, to determine the absorbance A of the sample soaking dilution solution at wavelengths of 420 nm, 470 nm, and 505 nm, representing the content of the three major pigments: yellow, orange, and red, respectively.

[0071] Color value X1 (U / g) is calculated using the following formula: X1 = A × 100 / m × 50 / V

[0072] Where A = absorbance of the sample soaking in the diluent;

[0073] m = the mass of the sample taken, in grams;

[0074] 100, 50 = conversion factor;

[0075] V = Volume of ethanol soaking solution taken, in mL;

[0076] The experimental results are based on the arithmetic mean of parallel measurements, and the absolute difference between two independent measurements obtained under repeatability conditions is no greater than 2%.

[0077] This *Aspergillus purpureus* strain ZYXQ-2206 exhibits strong pigment production capacity. Its solid-state fermentation yields a yellow pigment value of 3070.15±1.85 U / g, an orange pigment value of 2221.23±1.41 U / g, and a red pigment value of 3115.08±2.66 U / g. Its liquid-state fermentation yields a yellow pigment value of 2807.16±1.76 U / g, an orange pigment value of 2943.82±1.45 U / g, and a red pigment value of 2019.61±2.02 U / g. Therefore, the color value produced by *Aspergillus purpureus* strain ZYXQ-2206 fermenting red yeast rice is higher than that of commercially available red yeast rice (see Table 1).

[0078] Table 1. Determination of the color value of red yeast rice (U / g)

[0079] Example 3: Determination of the content of four components in fermentation products

[0080] Using the ZYXQ-2206 strain obtained from three batches of solid-state fermentation in Example 2, the ZYXQ-2206 strain obtained from liquid fermentation in Example 2, and other samples, with rice as the substrate, the contents of Monascin, Ankaflavin, Lovastatin (LOV), and Citrulline (CIT) in the fermentation substrate were measured.

[0081] The methods for determining Monascin, Ankaflavin, LOV, and CIT in the above-mentioned fermented red yeast rice are as follows:

[0082] HPLC detection conditions were: column: C 18(4.6mm×250mm), injection volume: 20μL, wavelength: 390nm, flow rate: 1mL / min, column temperature: 35℃; mobile phase: acetonitrile (A) and 0.1% phosphoric acid aqueous solution (B), gradient elution conditions: 0–10min 50% A, 10–25min 50–63% A, 25–35min 63% A, 35–40min 63%–72% A, 40–50min 72–75% A.

[0083] Preparation of test solution: After grinding the red yeast rice fermentation product into powder, pass it through a No. 7 sieve. Weigh 0.5g of the sieved sample, accurate to 0.001g, and place it in a 50mL Erlenmeyer flask. Add 10mL of 95% methanol, sonicate at 50Hz for 30min, filter it through a 0.22μm membrane, centrifuge at 12000rpm / min for 5min, and take 0.2mL of the supernatant for HPLC detection.

[0084] Preparation of standard solutions: Accurately weigh appropriate amounts of Monascin, Ankaflavin, LOV, and CIT standards, dissolve them in chromatographic grade methanol, and prepare a stock solution with a concentration of 1.000 mg / mL for later use. Using a serial dilution method, prepare standard concentrations of 20 μg / mL, 40 μg / mL, 80 μg / mL, 120 μg / mL, 160 μg / mL, and 200 μg / mL, respectively. Analyze the prepared series of standard solutions under HPLC conditions, and plot a standard curve with the standard concentration as the abscissa (X) and the peak area as the ordinate (Y).

[0085] The regression equation for Monascin is: Y = 0.8132X + 2.4375(R²). 2 =0.9992);

[0086] Ankaflavin's regression equation: Y = 0.428X + 4.6344(R) 2 =0.9992);

[0087] The regression equation for LOV is: Y = 0.0999X + 0.3861(R²). 2 =0.9993);

[0088] The regression equation for CIT is: Y = 0.2096X - 0.692(R²). 2 =0.9991).

[0089] The concentrations of Monascin and Ankaflavin in the sample were obtained by regression equation based on the standard curve. The amounts of LOV and CIT were below the limit of quantitation, as shown in Table 2.

[0090] The Monascus purpureus strain ZYXQ-2206 produced 58.643 mg / g of Monascin and 18.695 mg / g of Ankaflavin during solid-state fermentation, and 52.438 mg / g of Monascin and 17.776 mg / g of Ankaflavin during liquid-state fermentation. These levels are higher than those of the original Monascus purpureus strain 7-3 produced using traditional methods, and also higher than those found in commercially available red yeast rice (see Table 2). Furthermore, unlike existing technologies, no citrinin or lovastatin was detected in the fermentation products of strains 7-3 and ZYXQ-2206. The HPLC results for the reference standard, ZYXQ-2206, and red yeast rice sample 7 are shown in Figure 3.

[0091] Table 2. Chemical content of red yeast rice (mg / g) nd: Not detected

Claims

1. A type of Monascus purpureus, wherein, The preservation number of the red Monascus is: CGMCCNO.41335.

2. The Monascus purpureus according to claim 1, wherein, The DNA sequence of the red Monascus is shown in SEQ ID NO:

1.

3. A culture containing the Monascus purpureus as described in claim 1.

4. The use of the Monascus purpureus according to claim 1 or the culture according to claim 3 in the cultivation of Monascus purpureus and / or Monascus xanthophyll.

5. A method for culturing monaxioxin and / or monaxioxanthin, wherein, This includes using the Monascus purpureus according to claim 1 to cultivate Monascus purpureus and / or Monascus xanthophyll.

6. The method according to claim 5, wherein, It also includes the following steps: The Monascus purpureus strain was activated and then inoculated into a seed culture medium to obtain a strain seed solution. The seed culture of the strain was inoculated into a fermentation medium for fermentation culture; The Monascus strain was activated before being inoculated into the seed culture medium.

7. The method according to claim 6, wherein, The fermentation conditions are aerobic culture, fermentation temperature is 25-50℃, fermentation time is 2-20 days, and fermentation method is solid-state fermentation or liquid-state fermentation.

8. The method according to claim 6, wherein, The seed culture medium comprises the following components: glucose 10–50 g / L, peptone 5–30 g / L, yeast extract 5–30 g / L, soybean flour 2–20 g / L, glycerol 20–80 mL / L, magnesium sulfate 0.2–2 g / L, sodium nitrate 0.5–5 g / L, zinc sulfate 0.5–5 g / L, potassium dihydrogen phosphate 0.2–2 g / L, and maltodextrin 10–50 g / L; and / or The raw materials for the fermentation medium are selected from one or more of rice, soybeans, corn, or yam.

9. The method according to claim 6, wherein, The inoculation amount of the strain seed into the fermentation medium is 5-15% (V / V).

10. The method according to claim 6, wherein, The plate culture conditions are as follows: culture at 25–50°C for 2–20 days.

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