Method for preparing lovastatin-containing functional food by fermenting glutenin and coix seeds through monascus

Abstract

The invention discloses a method for preparing lovastatin-containing functional food by fermenting glutenin and coix seeds through monascus. Mixing the vital wheat gluten with a 65% ethanol solution, stirring and extracting for 2 hours at 35 DEG C, separating supernate from the precipitate, and continuously and repeatedly extracting the precipitate twice by using 65% ethanol; and centrifugally precipitating the extracted solution to obtain glutenin. The method comprises the following steps: soaking coix seeds in tap water at 30-40 DEG C for 2-4 hours, draining, mixing with glutenin according to a mass ratio of 7: 3, and sterilizing at 110 DEG C for 40 minutes to obtain a solid culture medium to be fermented; the method comprises the following steps: dissolving spores in a purified Monascus purpureus slant culture medium by using sterile water, inoculating the spores to a solid culture medium, uniformly mixing, and carrying out constant-temperature culture in a constant-temperature incubator at 32 DEG C; wherein the inoculation amount of the spore liquid is 0.5-2.0% of the mass of the culture medium, and the fermentation time is 20-30 days.

Description

[0001] The invention relates to a method for preparing functional food containing lovastatin by fermenting glutenin and coix seed with Monascus. Background Art

[0002] Monascus is a filamentous fungus that can be used for both medicine and food. It has been used for medicine and food fermentation in China, Japan and other Southeast Asian countries for more than 2,000 years. The poet Wang Can described red yeast rice with the poem "Traveling to Liang in the west, staying at the imperial residence for vegetarian meals, Guazhou red yeast, mixed with ginseng, soft and smooth, flowing into the mouth". The production process and medical uses of red yeast rice are recorded in detail in the Compendium of Materia Medica. Red yeast rice is widely used as a raw material for the production of Chinese red wine and fermented tofu, a food colorant and a meat preservative. Lovastatin is one of the secondary metabolites of Monascus. There are two main structural forms, namely lactone type and open-ring acid type. Lactone type lovastatin can be converted to open-ring acid type in the body, and it is the open-ring acid type that really exerts its drug effect in the body. Since the open-ring acid form of statins is similar to the structure of hydroxymethylglutaryl coenzyme A, it can competitively inhibit the synthesis of cholesterol in the body, block the intracellular mevalonate metabolic pathway, reduce intracellular free cholesterol, and play a role in reducing the body's blood lipid content.

[0003] High blood lipids can cause hyperlipidemia, hypertension, fatty liver, coronary atherosclerotic heart disease, etc., which has become one of the main factors endangering human health. In the treatment of such diseases, patients mostly control the disease by taking lovastatin drugs, which are expensive and have limited efficacy. At present, lovastatin products on the market mainly exist in the form of high-purity pharmaceutical preparations, and functional foods with high lovastatin content are rare. According to literature reports, the lipid-lowering function of Monascus fermented products containing lovastatin is significantly stronger than that of pure lovastatin at the same concentration, indicating that there are other substances in Monascus fermented products that synergize with lovastatin to lower blood lipids. The development of Monascus fermented foods with high lovastatin content can effectively control the occurrence and development of human high blood lipid-related diseases and play a role in protecting the health of the Chinese people. Summary of the invention

[0004] The present invention aims to overcome the deficiencies in the prior art and provides a method for preparing functional food containing lovastatin by fermenting glutenin and coix seed with Monascus.

[0005] The present invention is achieved through the following technical scheme: a method for preparing a functional food containing lovastatin by fermenting glutenin and coix seed with Monascus, comprising the following steps:

[0006] The gluten powder and 65% ethanol solution were mixed at a mass ratio of 1:30, stirred and extracted at 35°C for 2 hours, the supernatant was separated from the precipitate, and the precipitate was repeatedly extracted with 65% ethanol twice. The extracted solution was centrifuged at 3500r / min for 5 minutes to obtain the precipitate, which was glutenin.

[0007] Soak coix seeds in 30-40℃ tap water for 2-4h, drain the coix seeds, mix with glutenin in a mass ratio of 7:3 and place in a high pressure steam sterilizer, the sterilization temperature is 110℃, the sterilization time is 40min, and the solid culture medium to be fermented is obtained after sterilization. The spores in the purified Monascus purpureus slant culture medium are dissolved with sterile water and poured into a spray pot after high pressure sterilization, sprayed into the culture medium, mixed evenly and placed in a 32℃ constant temperature incubator for constant temperature culture. The spore liquid inoculation amount is 0.5%-2.0% of the culture medium mass, and the fermentation time is 20-30d.

[0008] The standard curve of lovastatin was determined using a mixed solvent of ethanol and sodium bisulfite. A 2 mg volumetric flask of standard lovastatin sample was taken and the volume was adjusted to 100 ml. Lovastatin solutions with concentrations of 16 ug / ml, 12 ug / ml, 8 ug / ml, 4 ug / ml, and 2 ug / ml were prepared. The wavelength was set at 237 nm. The absorbance (OD) was measured at a wavelength of 237 nm using the mixed solvent as the standard sample. 237 ).

[0009] After the culture medium is fully grown, it is placed in a 65°C oven for drying, 1 g is weighed, crushed and placed in a test tube, 1 ml of deionized water and 9 ml of 70% ethanol are added and shaken, and then extracted at 55°C, 130 r / min for 1 h; 4200 r / min, after appropriate dilution, the OD is measured. 237 .

[0010] Compared with the prior art, the present invention has the following beneficial effects:

[0011] The method of the present invention does not use the traditional culture medium with maltose as the carbon source and peptone as the nitrogen source, but only uses a culture medium in which glutenin and coix seeds are mixed to provide the carbon source and nitrogen source. Monascus directly uses the nutrients in the protein coix seeds to grow and secretes beneficial secondary metabolites such as lovastatin, monascus pigment, and gamma-aminobutyric acid. Therefore, this Monascus fermented food with a high content of lovastatin has a stronger lipid-lowering effect.

[0012] Instruction Manual

[0013] Figure 1 This is a picture of Monascus purpureus fermented in glutenin and coix seed medium for 20 days;

[0014] Figure 2 This is a picture of Monascus purpureus fermented in glutenin and coix seed medium for 30 days;. DETAILED DESCRIPTION

[0015] The present invention is further described in detail below in conjunction with specific embodiments.

[0016] Example 1

[0017] 100g of gluten powder and 3L of 65% ethanol solution were mixed by weight, stirred and extracted at 35℃ for 2h, the supernatant and the precipitate were separated, and the precipitate was extracted twice with 65% ethanol. After extraction, the solution was centrifuged at 3500r / min for 5min to obtain 43g of precipitate, which was glutenin.

[0018] Weigh 70g of coix seed and 30g of glutenin, add 35ml of deionized water to the coix seed, soak it in 30℃ tap water for 2h, stir it with glutenin to make it fully mixed and transfer it to a conical flask and seal it with cotton plugs and newspaper; measure 40ml of deionized water in a beaker and seal it with newspaper; at the same time, wrap the medicine spoon, glass rod, culture dish, spray bottle, etc. with newspaper; put the above items in a high-pressure steam sterilizer and sterilize them at 110℃ for 40min. The clean bench was opened for ultraviolet sterilization for 30 minutes, the cooled sterile water and the conical flask were placed in the clean bench after being disinfected with alcohol, the alcohol lamp was turned on, sterile water was added to the purified Monascus purpureus slant, the spores were evenly distributed by stirring with a glass rod, the supernatant was poured into a spray bottle after high-pressure sterilization, the spray bottle was adjusted to spray mist, 0.5% of the mass of the culture medium was inoculated with spore liquid, and the mixture was evenly mixed by stirring, and the mixture was placed in a 32°C constant temperature incubator for constant temperature culture for 20 days, and then dried at 60°C to constant weight, crushed through a 100-mesh sieve, and the lovastatin content was determined.

[0019] Weigh 0.1g, crush it and place it in a test tube, add 1ml of deionized water and 9ml of 70% ethanol, shake well, extract at 55℃, 130r / min for 1h; 4200r / min, dilute it 20 times and measure its absorbance at a wavelength of 237nm. According to the standard curve, the lovastatin content is 32.64mg / g.

[0020] Example 2

[0021] 1000g of gluten powder and 30L of 65% ethanol solution were mixed by mass, stirred and extracted at 35℃ for 2h, the supernatant and the precipitate were separated, and the precipitate was extracted twice with 65% ethanol. The extracted solution was centrifuged at 3500r / min for 5min to obtain 446g of precipitate, which was glutenin.

[0022] Weigh 700g of coix seeds and 300g of glutenin, add 350ml of deionized water to the coix seeds, soak them in 40℃ tap water for 4h, stir with glutenin to mix them thoroughly and transfer them to a conical flask and seal it with cotton plugs and newspaper; measure 40ml of deionized water in a beaker and seal it with newspaper; at the same time, wrap the medicine spoon, glass rod, culture dish, spray bottle, etc. with newspaper; put the above items in a high-pressure steam sterilizer and sterilize them at 110℃ for 40min. The clean bench was opened for ultraviolet sterilization for 30 minutes, the cooled sterile water and the conical flask were placed in the clean bench after being disinfected with alcohol, the alcohol lamp was turned on, sterile water was added to the purified Monascus purpureus slant, the spores were evenly distributed by stirring with a glass rod, the supernatant was poured into a spray bottle after high-pressure sterilization, the spray bottle was adjusted to spray mist, spore liquid was inoculated according to 2.0% of the mass of the culture medium, and the mixture was evenly mixed by stirring, and the mixture was placed in a 32°C constant temperature incubator for constant temperature culture for 30 days, and then dried at 60°C to constant weight, crushed through a 100-mesh sieve, and the lovastatin content was determined. Weigh 0.1g, crush it and place it in a test tube, add 1ml of deionized water and 9ml of 70% ethanol, shake well, extract at 55℃, 130r / min for 1h; 4200r / min, dilute it 20 times and measure its absorbance at a wavelength of 237nm. According to the standard curve, the lovastatin content is 40.22mg / g.

Claims

1. A method for preparing functional food containing lovastatin by fermenting glutenin and coix seed with Monascus, characterized in that: The steps include: (1) Wheat gluten powder and 65% ethanol solution were mixed at a mass ratio of 1:30, stirred and extracted at 35°C for 2 hours, the supernatant and the precipitate were separated, and the precipitate was repeatedly extracted with 65% ethanol twice. The extracted solution was centrifuged at 3500 r / min for 5 minutes to obtain the precipitate, which was glutenin; (2) Using glutenin and coix seed as raw materials, high-pressure sterilization was used to prepare a solid fermentation medium. The spores in the purified Monascus slant medium were dissolved with sterile water and poured into a high-pressure sterilized spray bottle, which was sprayed into the medium, mixed evenly, and placed in a 32°C constant temperature incubator for constant temperature culture; (3) After the fermentation, the solid culture medium is dried and crushed, and then the lovastatin content is determined by spectrophotometry.

2. The method for preparing functional food containing lovastatin by fermenting glutenin and coix seed with Monascus according to claim 1, characterized in that: The Monascus is classified and named as Monascus purpureus, with a preservation number of 2023J8250, and is deposited in the Institute of Microbiology, Chinese Academy of Sciences.

3. The method for preparing functional food containing lovastatin by fermenting glutenin and coix seed with Monascus according to claim 1, characterized in that: The coix seeds are soaked in 30-40°C tap water for 2-4 hours, and after being drained, the coix seeds are mixed with glutenin in a mass ratio of 7:3 and placed in a high-pressure steam sterilizer. The sterilization temperature is 110°C, the sterilization time is 40 minutes, and a solid culture medium to be fermented is obtained after sterilization.

4. The method for preparing functional food containing lovastatin by fermenting glutenin and coix seed with Monascus according to claim 1, characterized in that: The spore liquid inoculation amount is 0.5% to 2.0% of the culture medium mass, and the fermentation time is 20 to 30 days.

5. The method for preparing functional food containing lovastatin by fermenting glutenin with Monascus according to claim 1, characterized in that: The specific method for determining the lovastatin content in step (3) is to take 0.1 g of dried glutenin culture medium, add 1 ml of deionized water, and shake well in 9 ml of ethanol (70%), extract at 55° C., 130 r / min for 1 hour; centrifuge at 4200 r / min for 5 minutes, and after appropriate dilution, determine the absorbance value (OD237) at 237 nm, and determine the lovastatin content according to the lovastatin standard curve.

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