阅读说明：本技术 一种紫色红曲菌my-21菌株及其应用 (Monascus purpureus MY-21 strain and application thereof ) 是由 洪厚胜 于卓然 陈程鹏 于 2019-08-14 设计创作，主要内容包括：本发明公开了一种紫色红曲菌MY-21菌株及其应用,利用该红曲菌通过发酵,能够获得Monacolin K含量较高的功能性红曲产品,从而能够应用于生产降脂药物如血脂康和保健功能性产品。本发明的紫色红曲菌MY-21菌株,该菌种已在中国微生物菌种保藏管理委员会普通微生物中心保藏,保藏编号为：CGMCC No.18110,保藏日期为：2019年7月19日。(The invention discloses a purple monascus MY-21 strain and application thereof, and a functional red yeast product with higher Monacolin K content can be obtained by fermenting the monascus MY-21 strain, so that the monascus MY-21 strain can be applied to production of lipid-lowering medicines such as blood lipid-lowering medicines and health-care functional products. The monascus purpureus MY-21 strain of the invention is preserved in the common microorganism center of China Committee for culture Collection of microorganisms with the preservation number as follows: CGMCC No.18110, the preservation date is: 7/19/2019.)

1. A monascus purpureus MY-21 strain is preserved in the China general microbiological culture Collection center of the Committee for culture Collection of microorganisms with the preservation number as follows: CGMCC No. 18110.

2. an application of the purple red rice MY-21 strain of claim 1 in preparing lipid-lowering drugs and health functional foods.

3. The use according to claim 2, characterized by being in red yeast rice fermentation.

4. The use according to claim 3, characterized by being in increasing the Monacolin K content in the solid state fermentation of functional red yeast rice.

5. The use according to claim 4, characterized in that the method for preparing the red yeast product by fermentation comprises the following steps:

A. Preparing seed solution, namely inoculating the monascus purpureus MY-21 strain on a PDA culture medium, and culturing at 30-32 ℃ for 7-8 days for activation; eluting the spores on the culture medium with sterile water, and shaking up to break up spore aggregates to obtain spore suspension; inoculating the spore suspension into a liquid seed culture medium, and culturing to obtain a purple red yeast strain seed liquid;

B. Solid state fermentation: inoculating purple red yeast strain seed liquid into the culture medium under aseptic condition, sealing, and transferring into a culture chamber for fermentation culture to obtain a red yeast product with high Monacolin K content.

6. The use of claim 5, wherein the spore suspension of step A is inoculated into a liquid seed medium at an inoculum size of 5% by volume of the medium, and the liquid seed medium is subjected to shaking culture at 30-32 ℃ and 180r/min for 48H, wherein the liquid seed medium comprises glucose 60g/L, peptone 25g/L, corn flour 10g/L, NaNO ₃ 2g/L, K ₂ HPO ₄ 1g/L, MgSO ₄.7H ₂ O1g/L, and the pH is adjusted to 5.0-6.0.

7. The use according to claim 6, wherein the spore suspension is at a concentration of 10 ⁶ cfu/mL.

8. The use of claim 5, wherein the solid fermentation in step B comprises two stages of normal temperature fermentation and low temperature fermentation, wherein the normal temperature fermentation is performed first, and then the low temperature fermentation is performed, and the culture conditions of the normal temperature fermentation stage are as follows: culturing for 2-3 days at 30-32 ℃; the culture conditions in the low-temperature fermentation stage are as follows: culturing for 18-20 days at 18-22 ℃ and the humidity is 60-75%.

9. The application of claim 8, wherein after the solid state fermentation is finished, the culture medium is dried at 45-55 ℃, and then crushed and sieved by a 40-mesh sieve to obtain the final functional red yeast rice finished product.

10. The use according to claim 5, wherein the fermentation substrate of step B is prepared by a process comprising: soaking rice for 1-2 h, draining, steaming, adding 2.5-7.5% of glycerol and 1-5% of soybean meal, and sterilizing at 121 ℃ for 20min, wherein the mass percentages are the total mass percentages.

Technical Field

The invention relates to a microorganism and application thereof, in particular to a monascus purpureus MY-21 strain and application thereof.

Background

The red yeast rice is prepared by taking non-sticky rice as a raw material and performing solid state fermentation on monascus. It is mainly produced in Fujian, so it is also called Fuqu and Danqu. Red yeast rice has at least two thousand years of history of dual purposes of medicine and food in China. Red yeast is mainly used for brewing wine, brewing vinegar, fermented bean curd and food pigment at home and used as a traditional Chinese medicine. In 1979, the Japanese scholars Yuancheng stamp found and extracted Monacolin K (Monacolin K) which is an active substance with the cholesterol synthesis inhibition effect from the culture solution of the red yeast rice, and the lipid lowering function of the red yeast rice becomes the main direction of modern red yeast rice research. Besides, the fermented red yeast rice also contains secondary metabolites such as pigment, choline, flavone, DHA and the like, and has the effects of reducing blood pressure, reducing blood sugar, resisting oxidation naturally and the like.

In recent years, with the improvement of living standard, diseases such as coronary heart disease, hyperlipidemia, cerebral thrombosis, prostate cancer and the like caused by high cholesterol threaten the health of human beings, the demand of lipid-lowering products is increasing, and functional red yeast rice which is naturally fermented by monascus and takes cholesterol-lowering substance Monacolin K as a main component gradually gets into the visual field of people. Research shows that compared with Monacolin K extracted from other fungi or artificially synthesized, Monacolin K in the red yeast rice product has less crystallization and high solubility, so the red yeast rice product has more obvious curative effect and less toxic and side effect. With the increasing demand of the domestic red yeast rice market and the European and American Japan on the red yeast rice products in China, how to effectively improve the yield of functional red yeast rice solid state fermentation Monacolin K becomes an important problem which must be solved in the development of the red yeast rice fermentation industry. The breeding of the monascus strains with high yield of lovastatin is the most direct method for improving the yield of MonacolinK.

Disclosure of Invention

The invention solves the defects in the prior art and provides a purple monascus MY-21 strain and application thereof, and a functional red yeast rice product with high Monacolin K content can be obtained by fermenting the monascus so as to be applied to producing lipid-lowering medicines such as Xuezhikang and health-care functional products.

Meanwhile, the invention also provides a method for preparing the red yeast product by fermenting the purple monascus MY-21 strain, which is simple in process.

The invention is realized by the following technical scheme:

The monascus purpureus MY-21 strain of the invention is preserved in the common microorganism center of China Committee for culture Collection of microorganisms at the preservation address: xilu No.1 Hospital No. 3, Beijing, Chaoyang, with the deposit number: cgmccno.18110, taxonomic nomenclature: monascus purpureus (Latin name: Monascus purpureus) has a preservation date of: 7/19/2019.

The morphological characteristics of the monascus purpureus MY-21 strain are as follows: the colony on the PDA culture medium grows in a film shape, the surface of the colony is provided with folds and aerial hyphae, the early-stage hyphae are white and then become red, the old colony is claret-colored, and the back surface of the colony is claret-colored; hyphae have transverse septa, multinucleate branches, and irregular branches, and granules are in cells; conidiophore sphere or pear shape, which grows on the top of hypha and its branch; spherical cyst shell, orange red; the ascosphere is spherical and disappears after being matured; the ascospores are oval. The ITS sequence analysis result shows that the similarity with the monascus purpureus is as high as 99%.

The purple red rice MY-21 strain is applied to the preparation of lipid-lowering medicines and health-care functional foods; the further technical proposal is the application in the fermentation of red yeast rice; the further technical proposal is the application in improving the Monacolin K content during the solid state fermentation of the functional red yeast rice.

The application of the purple red yeast MY-21 strain in improving the Monacolin K content in the solid state fermentation of functional red yeast rice has the further technical scheme that the method for preparing the red yeast rice product by fermentation comprises the following steps:

A. Preparing seed solution, namely inoculating the monascus purpureus MY-21 strain on a PDA culture medium, and culturing at 30-32 ℃ for 7 days for activation; eluting the spores on the culture medium with sterile water, and shaking up to break up spore aggregates to obtain spore suspension; inoculating the spore suspension into a liquid seed culture medium, and culturing to obtain a purple red yeast strain seed liquid;

B. Solid state fermentation: inoculating purple red yeast strain seed liquid into the culture medium under aseptic condition, sealing, and transferring into a culture chamber for fermentation culture to obtain a red yeast product with high Monacolin K content.

The method for preparing the red yeast rice product by fermenting the purple red yeast rice MY-21 strain has the further technical scheme that the spore suspension in the step A is inoculated into a liquid seed culture medium according to the inoculation amount of 5% of the volume percentage of the culture medium, and is subjected to shaking culture for 48 hours at the temperature of 30-32 ℃ and under the condition of 180r/min, wherein the liquid seed culture medium comprises 60g/L of glucose, 25g/L of peptone, 10g/L of corn flour, ₃ 2g/L of NaNO, 2g/L of K ₂ HPO ₄ 1g/L of MgSO ₄, 7H ₂ O1g/L of MgSO 5.0-6.0, and the concentration of the spore suspension is 10 ⁶ cfu/mL.

The further technical scheme of the method for preparing the red yeast rice product by fermenting the purple red yeast rice MY-21 strain can be that the solid state fermentation in the step B comprises two stages of normal temperature fermentation and low temperature fermentation, wherein the normal temperature fermentation is carried out firstly, and then the low temperature fermentation is carried out, and the culture conditions in the normal temperature fermentation stage are as follows: culturing for 2-3 days at 30-32 ℃; the culture conditions in the low-temperature fermentation stage are as follows: culturing for 18-20 days at 18-22 ℃ and the humidity is 60-75%. The further technical scheme is that after the solid state fermentation is finished, the culture medium is dried at the temperature of 45-55 ℃, and then is smashed and sieved by a 40-mesh sieve, so that the final functional red yeast finished product is obtained.

The further technical scheme of the method for preparing the red yeast rice product by fermenting the purple red yeast rice MY-21 strain can also be that the preparation method of the fermentation culture substrate in the step B comprises the following steps: soaking rice for 1-2 h, draining, steaming, adding 2.5-7.5% of glycerol and 1-5% of soybean meal, and sterilizing at 121 ℃ for 20min, wherein the mass percentages are the total mass percentages.

The invention has the beneficial effects that:

the invention adopts the special excellent purple monascus which is separated and purified from various monascus products and screened out by low-temperature culture as a production strain, and the functional monascus with higher Monacolin K content is fermented by a scientific and reasonable fermentation process. The Monacolin K content of the functional red yeast rice obtained by the invention is far higher than the quality standards of red yeast rice in the national light industry standards of the people's republic of China, the technical standards for health food inspection and evaluation and the Chinese pharmacopoeia, so the functional red yeast rice can be used as raw material medicine of Chinese patent medicine Xuezhikang and health functional food, the process is scientific and simple, the Monacolin K yield is high, and the Monacolin K has obvious economic benefit and academic research value. Meanwhile, the purple red yeast MY-21 strain is simple in fermentation process and low in production cost when used for preparing a red yeast product through fermentation.

drawings

FIG. 1 is a schematic diagram of a monascus phylogenetic tree established based on ITS sequences

FIG. 2 is a standard curve of acid form Monacolin K detection by HPLC

FIG. 3 is a standard curve of HPLC detection lactone type Monacolin K

FIG. 4 is a Monacolin K HPLC chromatogram of example 2, note: the retention time is 13.285min, acid form Monacolin K is obtained, and the peak area is 2919.18mAU s; the retention time is 24.365min, the lactone Monacolin K is shown, and the peak area is 2092.48mAU s

FIG. 5 is a Monacolin K HPLC chromatogram of example 3, note: the retention time is 13.294min, acid form Monacolin K is obtained, and the peak area is 310.78mAU s; the retention time is 24.372min, the lactone Monacolin K is shown, and the peak area is 4593.61mAU s

FIG. 6 is a Monacolin K HPLC chromatogram of example 4, noting: the retention time is 13.291min, acid form Monacolin K is obtained, and the peak area is 3381.76mAU s; the retention time is 24.361min, the lactone type Monacolin K is shown, and the peak area is 3592.17mAU s

Detailed Description

The monascus purpureus MY-21 strain of the invention is preserved in the common microorganism center of China Committee for culture Collection of microorganisms with the preservation number as follows: CGMCC No. 18110.

In the following examples, unless otherwise specified, all methods are conventional; the reagents and raw materials used in the following examples were all purchased from conventional sources unless otherwise specified; in the following examples,% is by mass unless otherwise specified.