Monascus purpureus and application thereof

Abstract

The invention provides a monascus strain ZX-99 capable of expressing multiple beneficial active substances and application of the monascus strain ZX-99, and belongs to the technical field of microorganisms. The preservation number of the monascus strain ZX-99 provided by the invention is CGMCC No.40962, the strain does not produce citrinin (the detection limit is 25 mg / kg), and compared with other monascus cheese and commercially available mould cheese, the prepared cheese sample has the advantages that the content of citrinin in the cheese sample is greatly reduced, and the content of citrinin in the cheese sample is greatly reduced. Substances such as oxymatrine, bufalin, azadirachtin, gamma-aminobutyric acid, hydroxybenzylamine, maleic acid, maltotriose, maltotetraose and diacetyl are enriched in the monascus cheese ZX-99, and the monascus cheese ZX-99 is expected to be used for developing biological functional dairy products or compounding flavor strains.

Description

Technical Field

[0001] This application relates to the field of microbial technology, and in particular to a Monascus purpureus and its applications. Background Technology

[0002] Cheese is a delicious food that provides consumers with a variety of nutrients, including protein, vitamins, calcium, and other minerals, which have a positive impact on the immune system, bone health, and overall physiological functions. Besides being eaten directly, it can be added to various dishes, pizzas, sandwiches, etc., to enhance the texture and flavor of food. There are thousands of types of cheese, which can be classified according to the type of mold used to mature them: bacterial, mold, and yeast-matured. Currently, Penicillium and white mold are commonly used to make mold-matured cheeses, such as blue cheese, Brie, and Camembert. Cheese made from Penicillium often has a pungent odor and spicy taste due to the higher degree of protein hydrolysis and fat decomposition. White mold cheese is a soft cheese covered with white fungus; it has a rich milky flavor but also a certain musty or mushroom taste, which differs somewhat from the tastes of traditional Chinese cuisine.

[0003] Therefore, it is necessary to try using other types of molds in cheese making to develop mold-based cheeses that are more suitable for Chinese tastes. my country has many long-standing edible bacterial strains, among which Monascus purpureus is one. Monascus purpureus has wide applications in the food, health product, and pharmaceutical fields. Asian countries such as China, Japan, and South Korea use Monascus purpureus for food fermentation or as a food additive to enhance food color, improve texture, and impart unique flavors. Common Monascus purpureus fermented foods include red yeast rice, red yeast wine, and red yeast fermented bean curd. Furthermore, some metabolites of Monascus purpureus possess certain biological activities and can be used in the manufacture of health products and pharmaceuticals to alleviate problems such as hypertension, hyperlipidemia, and cardiovascular diseases. Therefore, Monascus purpureus has the potential to be used in cheese processing.

[0004] However, during the screening of Monascus purpureus strains, it was found that only a few strains do not produce citrinin, thus meeting the prerequisite for food safety. Furthermore, Monascus purpureus strains with cheese-maturation capabilities and the ability to produce excellent flavor are extremely rare. Therefore, the limited number of Monascus purpureus strains suitable for cheese maturation results in a very limited selection of flavor-blending strains for the industrial-scale production of Monascus purpureus cheese, making it difficult to solve the problem of a single flavor profile in the product.

[0005] To address the above issues, it is crucial to screen Monascus strains that do not produce citrinin, have good cheese-maturation capabilities, and produce excellent flavor, and to stockpile more Monascus strains suitable for industrial cheese production. Summary of the Invention

[0006] This application provides a Monascus purpureus (Monascus)Monascus sp (), its accession number is CGMCC No.40962.

[0007] This application also provides the use of the aforementioned Monascus purpureus in the preparation of products with probiotic functions and / or improved flavor.

[0008] This application also provides a method for preparing mature cheese, in which the above-mentioned Monascus purpureus liquid is applied to the surface of immature cheese, fermented, and matured.

[0009] This application also provides a mature cheese prepared by the above method.

[0010] The preservation information of the strain applied for is as follows: Strain name: Monascus purpureus Monascus sp The accession number is: CGMCC No. 40962; Date of deposit: October 26, 2023; Name of the depository: China General Microbiological Culture Collection Center, China Committee on the Preservation and Management of Microbial Cultures; Abbreviation of depositary institution: CGMCC; Address of the depository: No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0011] One object of the present invention is to provide a new strain of Monascus purpureus and its uses.

[0012] This invention provides a *Monascus fumeus* strain that possesses cheese-maturation capability and produces cheese products with superior flavor. This provides a wider selection of mature strains for *Monascus* cheese production and allows for the possibility of strain blending. The *Monascus fumeus* strain provided by this invention is named ZX-99.

[0013] Some Monascus purpureus strains exhibit citrinin-producing characteristics. Even some Monascus purpureus strains already used in traditional fermented foods can produce citrinin despite having excellent flavor. The smoky gray Monascus purpureus strain provided in this invention (…) Monascus fumeus The strain was selected from 25 food-derived Monascus strains. Among them, only 8% of the Monascus strains possessed the combined properties of not producing citrinin (detection limit of 25 mg / kg), cheese ripening ability, and producing excellent flavor. The remaining 16% of the Monascus strains, which possessed both cheese ripening ability and excellent flavor, produced citrinin at levels as high as 6702~65345 mg / kg, and were not suitable for cheese ripening.

[0014] Some Monascus purpureus fermentation products (red yeast rice) use high-temperature inactivation treatment to prevent the production of citrinin during fermentation, but this results in some loss of beneficial substances in the finished product, such as Monacolin K. The present invention provides a smoky gray Monascus purpureus strain (… Monascus fumeus After long-term fermentation (28 days) in various substrates, no citrinin was detected (detection limit was 25 mg / kg). High-temperature inactivation treatment was not required to prevent the production of citrinin during fermentation, thus avoiding the loss of beneficial substances.

[0015] The research of this invention has found that when the *Monascus purpureus* strain ZX-99 of this invention is used in mature cheese, some beneficial substances are enriched compared to mature cheese products made with other molds.

[0016] The *Monascus purpureus* strain ZX-99 of this invention can be cultured aerobically in commonly used *Monascus purpureus* culture media (such as rice culture medium, rice flour culture medium, YES medium, PDA medium, milk, etc.). The optimal fermentation temperature is 30-33℃, and the optimal fermentation time is 4-5 days.

[0017] In summary, this invention provides a smoky gray Monascus strain ZX-99 and its products. This Monascus strain has the ability to mature cheese, and the finished product has an excellent flavor. The strain does not produce citrinin during fermentation in various substrates. Compared with cheeses matured by other molds, the cheese matured by this strain has a variety of beneficial substances enriched. Attached Figure Description

[0018] This application will be further described by way of exemplary embodiments, which will be described in detail with reference to the accompanying drawings. These embodiments are not limiting, wherein: Figure 1 This is a colony morphology diagram of the *Monascus purpureus* strain ZX-99 provided by this invention.

[0019] Figure 2 Microscopic features of the *Monascus purpureus* strain ZX-99 provided by this invention.

[0020] Figure 3 The image shows a hierarchical clustering heatmap based on the significantly enriched metabolites found in Monascus purpureus cheese ZX-99 (compared to commercially available Penicillium and Erythromyces cheeses). In the image: A is Monascus purpureus cheese BC20, B is Monascus purpureus cheese ZX-99, C is Penicillium cheese (blue stripe), and D is Erythromyces cheese (Bri).

[0021] Figure 4The image shows a hierarchical clustering heatmap based on the significantly enriched metabolites found in Monascus purpureus cheese ZX-99 (compared to commercially available Penicillium and Erythromyces cheeses). In the image: A is Monascus purpureus cheese BC20, B is Monascus purpureus cheese ZX-99, C is Penicillium cheese (blue stripe), and D is Erythromyces cheese (Bri). Detailed Implementation

[0022] To more clearly illustrate the technical solutions of the embodiments in this specification, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are merely some examples or embodiments of this specification. For those skilled in the art, these drawings can be applied to other similar scenarios without creative effort. Unless obvious from the context or otherwise specified, the same reference numerals in the drawings represent the same structures or operations.

[0023] As indicated in this specification and claims, unless the context clearly indicates otherwise, the words "a," "an," "an," and / or "the" do not specifically refer to the singular and may also include the plural. Generally speaking, the terms "comprising" and "including" only indicate the inclusion of expressly identified steps and elements, which do not constitute an exclusive list, and the method or apparatus may also include other steps or elements.

[0024] Flowcharts are used in this specification to illustrate the operations performed by the system according to embodiments of this specification. It should be understood that the preceding or following operations are not necessarily performed in exact order. Instead, the steps can be processed in reverse order or simultaneously. Furthermore, other operations can be added to these processes, or one or more steps can be removed from them.

[0025] This application provides a Monascus purpureus (Monascus) Monascus sp (), its accession number is CGMCC No.40962.

[0026] In some embodiments, the rRNA coding sequence of Monascus purpureus may include a nucleotide sequence as shown in SEQ ID NO.1.

[0027] In some embodiments, the red mold may be *Monascus purpureus* (smoke gray red mold). Monascus fumeus ).

[0028] This application also provides the use of the aforementioned Monascus purpureus in the preparation of products with probiotic functions and / or improved flavor.

[0029] Probiotic function refers to the positive regulatory function of beneficial microorganisms (such as probiotics) on the host's health by colonizing the host's body, regulating the balance of intestinal flora, enhancing the function of the immune system, and assisting in nutrient metabolism. Its core mechanisms include inhibiting the proliferation of harmful bacteria, promoting digestion and absorption, and reducing inflammatory responses.

[0030] In some embodiments, the Monascus purpureus does not produce citrinin.

[0031] Citrulline, also known as citrulline penicillin, is an organic compound with the chemical formula C. 13 H 14 O5. Citrinin is a common foodborne fungal toxin and a secondary metabolite produced by fungi. It can contaminate long-stored foods and cause various toxic effects, such as nephrotoxicity, liver damage, and cell damage. Citrinin is mainly found in stored grains, but it is sometimes also found in fruits and other plant products.

[0032] In some embodiments, the product may include at least one of food, medicine, or health products.

[0033] In some embodiments, the product may contain Monascus fermentation products and / or culture products.

[0034] In some embodiments, the fermentation products may include beneficial active substances and flavor substances, which improve the flavor of fermented foods and endow them with probiotic functions.

[0035] In some embodiments, the fermented food may be a fermented dairy product. In some embodiments, preferably, the fermented dairy product may be cheese. In some embodiments, more preferably, the cheese may be matured cheese.

[0036] Fermented dairy products are dairy products made from animal milk such as cow's milk and sheep's milk through fermentation by microorganisms such as lactic acid bacteria. During fermentation, microorganisms convert lactose into metabolic products such as lactic acid, causing significant changes in the product's flavor, texture, nutritional value, and shelf life.

[0037] In some embodiments, fermented dairy products may include, but are not limited to, fermented milk, cheese, mare's milk wine, curd, and fermented buttermilk.

[0038] In some embodiments, the beneficial active substance may include any one or more of oxymatrine, bufotalin, neem salsaline, γ-aminobutyric acid, hydroxybenzylamine, or maleic acid.

[0039] In some embodiments, the flavor substance may include any one or more of maltotriose, maltotetraose, or diacetyl.

[0040] This application also provides a method for preparing mature cheese, in which the above-mentioned Monascus purpureus liquid is applied to the surface of immature cheese, fermented, and matured.

[0041] Unripened cheese, also known as fresh cheese, refers to cheese that has not undergone a ripening (maturation) process. It is made from raw milk (skim milk or cream), with the addition of starter cultures and coagulants to coagulate the proteins. After the whey is removed, the cheese is pressed. It has a high water content, a smooth and refreshing texture, and lower protein and calcium content than ripened cheese but higher than yogurt. It contains live probiotics and is easily digestible. Representative varieties include Fromage Frais (French white cheese), mozzarella (soft and juicy, often used in pizza), and mascarpone (the base for tiramisu).

[0042] Ripened cheese (also known as Aged cheese) refers to cheese made by aging fresh cheese in a specific temperature and humidity environment for several weeks, months, or even years.

[0043] Unlike fresh cheese, mature cheese is not "ready to eat after solidification," but rather requires a controlled "fermentation and aging" process during which the cheese's flavor, texture, aroma, and appearance undergo fundamental changes.

[0044] The maturation process is essentially a controlled decomposition process. Under the influence of enzymes, bacteria (lactic acid bacteria, propionic acid bacteria, etc.) or molds (monascus, penicillium, white mold, etc.) remaining inside and on the surface of the cheese, the following three major changes occur: Protein breakdown (protein hydrolysis): The proteins in cheese are broken down into polypeptides and amino acids. This is not only the source of umami (glutamate), but also makes the cheese soft or crumbly, and creates unique layers of flavor.

[0045] Lipolysis: Fat is broken down into free fatty acids. This directly contributes to the spiciness, fruitiness, or creamy flavor of cheese, and some short-chain fatty acids also produce the signature "cheesy" taste or even a slightly pungent odor.

[0046] Moisture evaporation and structural remodeling: Over time, moisture gradually evaporates, making the cheese texture denser; at the same time, gases such as carbon dioxide produced by microbial metabolism may form pores inside (such as the large pores in Emmental cheese).

[0047] Mature cheeses, due to their long fermentation process, possess complex, rich, and varied flavors (such as the spiciness of blue cheese and the creaminess of Brie). Unripened cheeses have a milder, more delicate flavor. Common types of matured cheeses include those matured by bacteria (such as washed-in cheeses) and those matured by mold (such as blue cheeses and white mold cheeses). Examples include cheddar, Emmental, Brie, and Roquefort. Common unripened cheeses include: Quark, Cream Cheese, Cottage, Ricotta, Feta, and some Mozzarella cheeses. In some embodiments, the fermentation temperature can be 26–30°C. In some embodiments, the fermentation temperature can be 27–29°C. In some embodiments, preferably, the fermentation temperature can be 28°C.

[0048] In some embodiments, the fermentation humidity can be 45-55%. For example, the fermentation humidity can be 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55%. Any range characterized by combinations of the above values ​​is also included, which will not be elaborated here. In some embodiments, preferably, the fermentation humidity can be 50%.

[0049] In some embodiments, the fermentation time can be 12 to 16 days. In some embodiments, the fermentation time can be 13 to 15 days. In some embodiments, preferably, the fermentation time can be 14 days.

[0050] This application also provides a mature cheese prepared by the above method.

[0051] Unless otherwise specified, the experimental methods used in the following examples are conventional methods. Unless otherwise specified, the experimental materials used in the following examples were all purchased from conventional biochemical reagent companies. All quantitative experiments in the following examples were performed in triplicate, and the results were averaged.

[0052] Unless otherwise specified, all raw materials used in the following embodiments were purchased commercially.

[0053] Cheese used for inoculation with Monascus purpureus ZX-99, Monascus purpureus cheese BC20 (different from Monascus purpureus cheese ZX-99 only in the inoculated strain): Bright Dairy Research Institute Kilesa Blue Cheese (Production Date: November 16, 2021): Alphard, Denmark Ile de France Brie cheese (production date: August 26, 2021): Sinodis Foods (Shanghai) Co., Ltd. Example 1: Ash Red Monascus strain ZX-99 The grayish-brown Monascus strain ZX-99 of this invention was obtained from the Dairy Research Institute of Bright Dairy Co., Ltd., and was isolated from red yeast rice wine lees.

[0054] Screening process: Multiple Monascus purpureus fermented foods from different regions were collected for Monascus purpureus strain isolation and purification, resulting in 25 food-derived Monascus purpureus strains. These strains were used to prepare inoculum for Monascus purpureus cheese production. Suitable Monascus purpureus strains for cheese maturation were screened, excluding strains susceptible to contamination by other microorganisms and those with low maturation efficiency. The flavor of the Monascus purpureus cheese produced from the initially selected strains was then evaluated to identify Monascus purpureus strains suitable for cheese production with superior flavor. Subsequently, the production of citrinin by the strains in different substrates was tested, and strains with food safety were selected, ultimately yielding Monascus purpureus strain ZX-99.

[0055] 1. *Monascus purpureus* strain ZX-99 (Fumigant Grey) The bacterial strain was cultured on PDA medium at 25°C in the dark for 7 days. The colonies were 50 mm in diameter and gray in color. Fluffy texture, darker color on the reverse side ( Figure 1 ).

[0056] The hyphae are septate, much-branched, and 3–5 μm in diameter. The cleistothecia are spherical, brown, and 30–50 μm in diameter. Figure 2 a. Ascospores are colorless and transparent, elliptical, 4.5~6.5 × 3~4 μm ( Figure 2 b. Small to medium-sized conidia). Conidia are nearly spherical or obpyriform, 10~12.5 × 7~10 μm ( Figure 2 b (the larger ones).

[0057] 2. rRNA gene sequence determination results (including ITS1-5.8S-ITS2 region sequence fragments) Example 2: Determination of citrinin production by Monascus purpureus strain ZX-99 Some Monascus purpureus strains possess the characteristic of producing citrinin. To ensure the food safety of Monascus purpureus fermented products, the citrinin content in the final product must be controlled within an appropriate range. In this embodiment, the citrinin content produced by the Monascus purpureus strain ZX-99 of the present invention was tested in eight different substrates.

[0058] Preparation of inoculum: A small amount of *Monascus purpureus* mycelium was inoculated from PDA medium into a 250 mL Erlenmeyer flask containing 120 mL of PDB medium (Qingdao Haibo Biotechnology Co., Ltd.) and cultured at 170 r / min and 32℃ for 7 days. Subsequently, the culture solution was filtered using a 5 mL syringe filled with approximately 1 cm of absorbent cotton. Plate counting showed that the viable cell count in the filtrate was 10-1. 6 CFU / L. Store the filtrate at room temperature and prepare fresh before use.

[0059] Method for preparing culture medium: (1) Rice flour culture medium (liquid): Weigh rice and water into a soymilk maker to make rice milk with a rice concentration of 5%. Take 200 ml of the rice milk and sterilize it in a 500 ml conical flask for later use.

[0060] (2) Rice flour inorganic salt culture medium (liquid): Based on the culture medium in (1) above, add 0.5% soybean peptone, 0.05% KH2PO4, 0.1% K2HPO4 and 0.05% MgSO4.

[0061] (3) Rice culture medium (solid): Weigh 50 g of rice and 100 ml of water into a 500 ml beaker, sterilize, and set aside.

[0062] (4) Yeast extract sucrose medium (liquid): Weigh 4% yeast extract and 16% sucrose to prepare a solution, take 200 ml and sterilize it in a 500 ml conical flask for later use.

[0063] (5) Potato glucose medium (liquid): Weigh 5.2 g potato glucose powder (6 g / L potato extract powder, 20 g / L glucose), prepare 200 ml culture medium, sterilize in a 500 ml conical flask and set aside.

[0064] (6) Whole milk (liquid): 200 ml is sterilized in a 500 ml conical flask and then used for later use.

[0065] (7) Skim milk (solid): Take an appropriate amount of skim milk powder and dry heat sterilize it at 160℃ for 2 hours. After sterilization, mix it with sterile water at a ratio of 1:1.5.

[0066] (1) to (7) Take 200 μl of inoculum and incubate at 170 r / min and 32℃ for 28 days.

[0067] (8) Unripe cheese (solid): Unripe cheese was provided by the Dairy Research Institute of Bright Dairy Co., Ltd. (For the preparation process of unripe cheese, please refer to patent ZL202211704622.3 "A kind of red yeast cheese and its preparation method"). The surface of the unripe cheese was evenly dipped in the inoculation liquid and ripened for 14 days at a temperature of 28℃ and a humidity of 50%.

[0068] Methods for detecting citrinin: Refer to GB 5009.222—2016 "Determination of penicillin in food by immunoaffinity column purification-high performance liquid chromatography".

[0069] The results of the detection of citrinin content of Monascus purpureus strain ZX-99 in eight different substrates are shown in Table 1. In the eight samples fermented by Monascus purpureus strain ZX-99 of the present invention, the quantitative detection results of citrinin were all less than the detection limit of 25 μg / kg in the above-mentioned national standard, which is far lower than the requirement of ≤50 μg / kg in the light industry standard (QB / T 2847-2007 "Functional Red Yeast Rice (Powder)"). It has high food safety and meets the needs of industrialization.

[0070] Table 1. Citrulline content in fermentation products Example 3: Monascus purpureus strain ZX-99 used for cheese ripening Red yeast cheese ZX-99 was prepared according to the method in Example 2 (8). After evaluation, it was found to have a delicious taste and a moderate saltiness.

[0071] Penicillium and white mold cheeses are the most representative mold-matured cheeses. Therefore, this example uses Monascus purpureus cheese ZX-99 as the experimental sample, and a commercially available Penicillium cheese (blue cheese) and a commercially available white mold cheese (Bri cheese) with high acceptance as control samples. Low-molecular-weight metabolites in the three cheeses were characterized using non-targeted metabolomics. Differential analysis was used to preliminarily infer the metabolite characteristics in Monascus purpureus cheese ZX-99. Simultaneously, the screened metabolites were compared between Monascus purpureus cheese ZX-99 and Monascus purpureus cheese BC20, which is also a Monascus purpureus cheese, to analyze the advantages of the Monascus purpureus strain ZX-99.

[0072] For the qualitative results provided by the testing company, multidimensional statistical analysis was performed using SIMCA 14.1 software (data preprocessed with Pareto-scaling), and t-tests were used for significance analysis in SPSS 26.0. Heatmaps were generated using the bioinformatics platform (https: / / bio-cloud.aptbiotech.com / ). Unidimensional statistical analysis and other data were processed using Excel. Based on the variable importance for the projection (VIP) values ​​obtained from the OPLS-DA model, metabolites with P < 0.05, VIP > 1, and fold change (FC) > 2 or < 0.5 were defined as significantly different metabolites. Each sample was tested six times.

[0073] The significantly enriched metabolites of Monascus purpureus cheese ZX-99 compared to those of Penicillium and Erythromyces cheese were screened (VIP). BC、BD >1, FC BC、BD >2, P BC、BD <0.05), 112 and 67 significantly enriched metabolites were qualitatively identified in positive ion mode (POS) and negative ion mode (NEG), respectively. Hierarchical clustering heatmaps were performed, and the results are shown below. Figure 3 As shown, the names of the compounds are labeled on the right side of the heatmap, and the different cheese samples are labeled at the bottom. The color blocks at different positions represent the relative expression levels of the corresponding metabolites, with red indicating relatively high expression levels and purple indicating relatively low expression levels. Metabolites with similar expression patterns are clustered together on the left side under the same cluster.

[0074] Depend on Figure 3 and Figure 4 ( Figure 3 , 4 As shown in both positive and negative ion modes, the two Monascus purpureus cheeses were clustered into the same group, exhibiting certain chemical similarities but also some differences. This indicates that Monascus purpureus cheeses share certain common characteristics. Furthermore, the Monascus purpureus strain ZX-99 provided by this invention has significant value for future blending of Monascus purpureus strains and is expected to alleviate the problem of monotonous flavor in products fermented with single strains.

[0075] Figure 3 and Figure 4 Among them, there are 26 POS and 4 NEG metabolites, and the concentration of Monascus purpureus cheese ZX-99 is higher than that of Monascus purpureus cheese BC20. These 30 enriched metabolites are the value that Monascus purpureus ZX-99 brings to cheese compared to Monascus purpureus BC20. Detailed material information is shown in Table 2.

[0076] Table 2 shows that Monascus purpureus cheese ZX-99 exhibits significantly enriched metabolites compared to Monascus purpureus cheese BC20, Penicillium cheese, and Evodia rutaecarpa cheese. Note: A is Monascus purpureus cheese BC20, B is Monascus purpureus cheese ZX-99, C is Penicillium cheese (blue stripe), and D is white mold cheese (Bri).

[0077] Table 2 shows that some substances have certain beneficial activities, such as oxymatrine, bufotalin, neem salsaline, γ-aminobutyric acid, hydroxybenzylamine, and maleic acid; some substances have a certain positive impact on the flavor of the product, such as maltotriose, maltotetraose, and diacetyl.

[0078] Oxymatrine has anti-inflammatory, anti-fibrotic, and anti-tumor effects, and can also treat diabetic cardiomyopathy. Bufotalin has good anti-tumor activity and a certain inhibitory effect on cancer cells. Neem alpinia is one of the main active ingredients of neem, and its limonoid alkaloids have inhibitory activity against human breast cancer cells. Gamma-aminobutyric acid (GABA) is a neurotransmitter, one of the most important inhibitory neurotransmitters in the central nervous system, with sedative, anti-anxiety, and sleep-promoting effects. Hydroxybenzylamine can relieve peptic ulcers and has antispasmodic and anti-secretive effects on the gastrointestinal tract. Maleic acid is a dicarboxylic acid with good preservative effects on oily substances. Maltotriose and maltotetraose are oligosaccharide organic compounds with low sweetness, good taste, and strong moisturizing properties. Diacetyl has a creamy flavor and is commonly found in fermented foods, dairy products, and beverages, such as yogurt and cheddar cheese.

[0079] In addition to the aforementioned beneficial active substances and flavor compounds, Monascus purpureus cheese ZX-99 is also rich in anthraquinones, organic acids, and heterocyclic compounds. Anthraquinones have a certain influence on the cheese's color and possess some anti-cancer activity. Organic acids are commonly found in fruits and vegetables, primarily providing fruity and sour flavors, and are also safe anti-thrombotic factors, playing a positive role in preventing cardiovascular and cerebrovascular diseases, enhancing the cheese's flavor and giving it probiotic properties. Heterocyclic compounds are highly recognized in the flavor industry; most possess characteristic aromas and have extremely low thresholds. Differences in these substances within cheese may be key to flavor variations. The enrichment of these organic acids and heterocyclic compounds gives Monascus purpureus strain ZX-99 a unique flavor advantage.

[0080] Therefore, the Monascus purpureus strain ZX-99 provided by this invention has flavor advantages and potential probiotic advantages. When used in cheese making, it can enrich beneficial substances such as oxymatrine, γ-aminobutyric acid, and diacetyl to a high degree.

[0081] The basic concepts have been described above. Obviously, for those skilled in the art, the detailed disclosure above is merely illustrative and does not constitute a limitation of this specification. Although not explicitly stated herein, those skilled in the art may make various modifications, improvements, and corrections to this specification. Such modifications, improvements, and corrections are suggested in this specification and therefore remain within the spirit and scope of the exemplary embodiments described herein.

[0082] Furthermore, this specification uses specific terms to describe embodiments thereof. For example, "an embodiment," "one embodiment," and / or "some embodiments" refer to a particular feature, structure, or characteristic associated with at least one embodiment of this specification. Therefore, it should be emphasized and noted that references to "an embodiment," "one embodiment," or "an alternative embodiment" in different locations throughout this specification do not necessarily refer to the same embodiment. Moreover, certain features, structures, or characteristics in one or more embodiments of this specification can be appropriately combined.

[0083] In some embodiments, numbers describing the quantity of components and attributes are used. It should be understood that such numbers used in the description of embodiments are modified in some examples with the terms "approximately," "approximately," or "generally." Unless otherwise stated, "approximately," "approximately," or "generally" indicates that the numbers are allowed to vary by ±20%. Accordingly, in some embodiments, the numerical parameters used in the specification and claims are approximate values, which may be changed depending on the characteristics required by individual embodiments. In some embodiments, numerical parameters should take into account specified significant digits and employ a general method of digit reservation. Although the numerical ranges and parameters used to confirm their breadth of range in some embodiments of this specification are approximate values, in specific embodiments, such values ​​are set as precisely as feasible.

[0084] Finally, it should be understood that the embodiments described in this specification are merely illustrative of the principles of the embodiments described herein. Other variations may also fall within the scope of this specification. Therefore, alternative configurations of the embodiments described herein are intended to be illustrative rather than limiting, and should be considered consistent with the teachings of this specification. Accordingly, the embodiments described herein are not limited to those explicitly introduced and described herein.

Claims

1. A type of Monascus purpureus ( Monascus sp (), its accession number is CGMCC No.40962.

2. The Monascus purpureus as described in claim 1, characterized in that, The rRNA coding sequence of the *Monascus purpureus* includes the nucleotide sequence shown in SEQ ID NO. 1; And / or, the red mold is *Monascus fumaroides*.

3. The use of Monascus purpureus as described in claim 1 or 2 in the preparation of products with probiotic functions and / or improved flavor.

4. The use as described in claim 3, characterized in that, The aforementioned Monascus purpureus does not produce citrinin; And / or, the product includes at least one of food, medicine or health products; And / or, the product contains Monascus ferment and / or culture products.

5. The use as described in claim 4, characterized in that, The fermentation products include beneficial active substances and flavor substances, which improve the flavor of fermented foods and endow them with probiotic functions.

6. The use as described in claim 5, characterized in that, The fermented food is a fermented dairy product, preferably a cheese, and more preferably a mature cheese.

7. The use as described in claim 5, characterized in that, The beneficial active substances include any one or more of the following: oxymatrine, bufotalin, neem salsaline, γ-aminobutyric acid, hydroxybenzylamine, or maleic acid. And / or, the flavor substance includes any one or more of maltotriose, maltotetraose, or diacetyl.

8. A method for preparing mature cheese, characterized in that, The Monascus purpureus liquid described in claim 1 or 2 is applied to the surface of immature cheese for fermentation and maturation.

9. The method as described in claim 8, characterized in that, The fermentation temperature is 26-30℃, preferably 28℃; And / or, the fermentation humidity is 45-55%, preferably 50%; And / or, the fermentation time is 12 to 16 days, preferably 14 days.

10. A mature cheese prepared by the method of claim 8 or 9.

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