KR20220072474A - Medium Composition for Producing Monacolin K, Method for Preparing Fermented Substance Using Same and Fermented Substance Prepared Thereby - Google Patents

Abstract

The present invention relates to a medium composition for monacolin K production comprising brown rice, rice bran powder and sodium glutamate (MSG), wherein the medium composition according to the present invention is used for fermentation using a strain producing monacolin K. May increase the ability to produce monacolin K.
In addition, the present invention relates to a method for preparing a fermented product by inoculating and fermenting a strain producing monacolin K in the medium composition, a fermented product prepared by the above manufacturing method, and a health functional food comprising the same, the present invention According to the manufacturing method, it is possible to prepare a fermented product with an increased content of monacolin K, and since the fermented product has a high content of monacolin K, a substance that inhibits cholesterol synthesis, it can be effectively used as a health functional food for improving cholesterol can

Description

A medium composition for producing monacolin K, a method for preparing a fermented product using the same, and a fermented product prepared by the method {Medium Composition for Producing Monacolin K, Method for Preparing Fermented Substance Using Same and Fermented Substance Prepared Thereby}

The present invention relates to a medium composition for production of monacolin K, a method for producing a fermented product using the same, a fermented product prepared by the production method, and a health functional food including the same, and more particularly, to produce monacolin K A medium composition capable of increasing the monacolin K production ability of the strain by inoculating and fermenting the It relates to a fermented product having an increased content and to a health functional food for improving cholesterol comprising the fermented product.

Monacolin K (monacolin K) is a substance that exhibits an effect of inhibiting the biosynthesis of cholesterol, also known as lovastatin (lovastatin). HMG-CoA reductase (3-hydroxy-3-methylglutaryl coenzyme A reductase) is involved in the biosynthesis of cholesterol as a regulatory enzyme. Monacolin K has a structure very similar to HMG-CoA, a substrate of HMG-CoA reductase, and , it has a very high affinity for HMG-CoA reductase. Therefore, instead of HMG-CoA, monacolin K acts as an inhibitor of HMG-CoA reductase by pre-binding to HMG-CoA reductase. In particular, it has been reported that monacolin K preferentially lowers low-density lipoprotein (LDL) cholesterol, a major cause of cardiovascular disease, in the process of lowering blood cholesterol. Therefore, monacolin K can be used as a health functional food for lowering blood cholesterol levels, especially harmful LDL cholesterol levels.

An example of a food containing such a monacolin K may be red yeast rice. Honggukmi is red rice produced by fermenting Honggukgyun, a fungus of the genus Monascus sp., on white rice, and monacolin K is secreted during the fermentation of Honggukgyun. In order to increase the amount of monacolin K production, research on the types of strains and media used for fermentation of Hong Guk-gyun, research on optimization of fermentation conditions, etc. are in progress. For example, Korean Patent Laid-Open Publication No. 10-2005-0082694 describes that the production of monacolin K can be increased by inoculating and culturing honggukgyun in a solid medium obtained by mixing sodium chloride with rice. However, in the fermentation process of Hong Guk-gyun, not only monacolin K but also citrinin are produced, but citrinin has a problem in causing toxicity to the kidneys, so there is a limit in terms of safety.

As an alternative technique to this, Korean Patent No. 10-1547642 prevents the production of citrinin in the fermentation process by using a strain of the genus Aspergillus as a strain producing monacolin K, It describes a technique for increasing the production capacity of monacolin K through strain mutation. The strain of the Aspergillus genus is a yellowish-brown mycelium-forming fungus, and when it is used, citrinin is not produced in the fermentation process to produce monacolin K, so it can be used in food without fear of toxicity to the kidneys. However, the development of fermentation technology for increasing the production ability of monacolin K in the fermentation process using a strain of the genus Aspergillus is incomplete compared to the conventional fermentation technology of Hong Guk-gyun.

Therefore, there is a need for the development of a fermentation technology capable of increasing the production capacity of monacolin K without producing citrinin during the production of monacolin K.

An object of the present invention is to provide a medium composition for monacolin K production for increasing the monacolin K production ability of the strain.

Another object of the present invention is to provide a method for preparing a fermented product having an increased content of monacolin K using the medium composition.

Another object of the present invention is to provide a fermented product with an increased content of monacolin K prepared by the above production method.

Another object of the present invention is to provide a health functional food for improving cholesterol comprising the fermented product.

In order to achieve the above object, the present invention provides a medium composition for monacolin K production, comprising brown rice, rice bran powder, and sodium glutamate (MSG).

The medium composition of the present invention may include 88 to 94% by weight of brown rice, 4 to 10% by weight of rice bran powder, and 1 to 4% by weight of MSG based on the weight of the total medium composition.

In the present invention, the moisture content of the brown rice may be 30 to 40% by weight.

In the present invention, the rice bran powder may be roasted rice bran powder.

The medium composition of the present invention may be used for fermentation using a strain of the genus Aspergillus or a mutant thereof.

In the present invention, the Aspergillus sp. strain may be Aspergillus terreus .

The present invention also provides an inoculation step of (i) inoculating a strain producing monacolin K in a medium composition comprising brown rice, rice bran powder and sodium glutamate (MSG); And (ii) provides a method for producing a fermented product having an increased content of monacolin K, comprising a fermentation step of fermenting the culture medium composition inoculated with the strain to obtain a fermented product.

In the present invention, the fermentation step is preferably carried out at a humidity of 90%RH or more.

In the present invention, the fermentation step is preferably performed for 7 to 15 days at a temperature of 25 to 35 ℃.

In the present invention, the hydrotreating process may be performed 2-3 times in the fermentation step.

In the present invention, in the hydrotreating step, the amount of hydrolysis is preferably 1 to 2% by weight based on the total weight of the medium composition.

In the present invention, it is preferable that the entry moisture content at the start of fermentation of the strain-inoculated medium composition is 35 to 40% by weight.

In the present invention, the departure moisture content at the end of fermentation of the strain-inoculated medium composition is preferably 45 to 50% by weight.

The present invention also provides a fermented product with an increased content of monacolin K, prepared by the above manufacturing method.

The present invention also provides a health functional food for improving cholesterol comprising the fermented product having an increased content of monacolin K.

The medium composition for producing monacolin K according to the present invention is used for fermentation using a strain producing monacolin K, thereby increasing the monacolin K producing ability of the strain. In addition, according to the manufacturing method of the present invention, it is possible to prepare a fermented product having an increased content of monacolin K by using the medium composition and controlling fermentation conditions. Therefore, since the fermented product contains a large amount of monacolin K, the cholesterol synthesis inhibitory effect is excellent, and it can be effectively applied to health functional foods for improving cholesterol.

Hereinafter, specific implementation forms of the present invention will be described in more detail. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is those well known and commonly used in the art.

The present invention relates to a medium composition for monacolin K production, a method for producing a fermented product having an increased monacolin K content using the same, a fermented product prepared by the production method, and a health function for improving cholesterol comprising the fermented product provide food.

Monacolin K (IUPAC Name: [8-[2-(4-hydroxy-6-oxooxan-2-yl)ethyl]-3,7-dimethyl-1,2,3,7,8,8 a -hexahydronaphthalen- 1-yl] 2-methylbutanoate) is It is a material having a structure of the following formula (1). Monacolin K has the function of reducing blood cholesterol by inhibiting the biosynthesis of cholesterol by inhibiting the activity of HMG-CoA reductase in the cholesterol biosynthesis pathway.

[Formula 1]

According to the present invention, a fermented product having a very high content of monacolin K can be prepared by adjusting the composition, fermentation conditions, fermentation method, etc. of the medium for fermenting the strain producing monacolin K. Therefore, the fermented product of the present invention can be effectively applied to health functional foods for lowering blood cholesterol levels.

The present invention relates to a medium composition for monacolin K production, and the medium composition of the present invention includes brown rice, rice bran powder and sodium glutamate (monosodium glutamate, MSG). Preferably, the medium composition may include 82 to 98% by weight of brown rice, 1 to 15% by weight of rice bran powder, and 0.5 to 5% by weight of MSG based on the weight of the total composition.

In the present invention, the brown rice is used as a main raw material of the medium composition for monacolin K production. Thereby, the fermentation efficiency of the strain producing monacolin K is improved compared to the case of using white rice, and a fermented product having a high monacolin K content can be prepared.

The moisture content of the brown rice may be 20 to 50% by weight, preferably 30 to 40% by weight in terms of increasing the production ability of monacolin K, more preferably 33 to 35% by weight.

The brown rice may be included in 82 to 98% by weight based on 100% by weight of the total medium composition, preferably 88 to 94% by weight, more preferably 89 to 91% by weight is advantageous in terms of fermentation efficiency of the strain.

In the present invention, the rice bran powder is used as an auxiliary material for replenishing nutrients necessary for the production of monacolin K. Rice bran is a by-product separated in the process of milling brown rice into white rice, and has abundant nutrients such as vitamins, minerals, dietary fiber, protein, and lipids. In the present invention, by adding rice bran powder in the form of pulverized rice bran to the medium, the production rate of monacolin K by fermentation of strains that produce monacolin K, particularly strains of the genus Aspergillus, compared to the case where other grains are used as auxiliary materials. can be effectively increased.

In the present invention, the rice bran powder may be obtained by grinding rice bran. The grinding method is not particularly limited, and for example, a grain grinding method generally known in the art, such as ball milling, colloid milling, etc. may be used.

The rice bran powder may have a particle size of 10 to 50 mesh, preferably 20 to 40 mesh. When the average particle diameter of the rice bran powder is within the above range, nutrients contained in the rice bran may be smoothly mixed with the main ingredient of the medium. The mesh is a unit for grading particle size by the size of a sieve when sieving particles, and means the number of apertures or wires existing within 1 inch.

In the present invention, the rice bran powder may be roasted rice bran powder. When roasted rice bran powder is used as rice bran powder, fatty acids in the rice bran are rancid and nutrients are prevented from being lost, and microorganisms and foreign substances that may exist in the rice bran are inactivated to help improve the fermentation quality of the desired strain. can In addition, it is possible to improve the utilization of microorganisms by thermally decomposing nutrients in rice bran.

The roasted rice bran powder may be prepared by roasting rice bran at a temperature of 80 to 120° C., preferably 95 to 100° C., for 5 minutes to 30 minutes, preferably 10 to 20 minutes, and then pulverizing the rice bran.

The rice bran powder may be included in an amount of 1 to 15% by weight, preferably 4 to 10% by weight, more preferably 6 to 9% by weight based on 100% by weight of the total medium composition. If the content of the roasted rice bran powder is too low, the effect of improving the production ability of monacolin K may be insignificant. can be

In the present invention, the sodium glutamate (MSG) is a sodium salt of glutamic acid, an essential amino acid, and is used as an additive for replenishing a nitrogen source in the medium. The MSG contributes to an increase in the production of monacolin K by helping the fermentation of the strain effectively proceed.

The MSG may be included in an amount of 0.5 to 5% by weight, preferably 1 to 4% by weight, more preferably 2 to 3% by weight based on 100% by weight of the total medium composition. When the content of sodium glutamate is within the above range, the effect of increasing the amount of monacolin K production can be optimized.

In the present invention, the medium composition for monacolin K production may be used for fermentation using a strain producing monacolin K.

The strain producing the monacolin K may be a strain of the genus Aspergillus or a strain of the genus Monascus , and the scope of the strain is a concept including mutants thereof. The strain of the genus Aspergillus may be Aspergillus terreus , etc., and the strain of the genus Monascus is Monascus purpureus , Monascus pilosus , Monascus Anca. anka ), Monascus ruber ( Monascus ruber ), and the like.

In the present invention, the strain producing the monacolin K is preferably a strain of the genus Aspergillus or a mutant thereof, particularly Aspergillus terreus or a mutant thereof. In this case, there is an advantage in that the expression of genes involved in the biosynthesis pathway of citrinin, one of mycotoxins, is suppressed during the fermentation process.

Preferably, the strain producing the monacolin K may be Aspergillus teraeus DSMK01. The Aspergillus teraeus DSMK01 is a mutant strain of Aspergillus teraeus KCCM12225, does not produce citrinin during the fermentation process, and has superior ability to produce monacolin K compared to other Aspergillus teraeus strains. For the method of generating the Aspergillus teraeus DSMK01 mutant from Aspergillus teraeus KCCM12225, reference may be made to the contents described in other Patent No. 10-1547642 of the present applicant.

In addition, the present invention relates to a method for producing a fermented product having an increased content of monacolin K.

The method for producing a fermented product according to the present invention includes (i) an inoculation step of inoculating a strain producing monacolin K into a medium composition comprising brown rice, rice bran powder and sodium glutamate (MSG); and (ii) fermenting the culture medium composition inoculated with the strain to obtain a fermented product.

In the inoculation step, the medium composition may be prepared by adding rice bran powder and MSG to brown rice. Based on 100% by weight of the total weight of the medium composition, the content of brown rice may be 82 to 98% by weight, preferably 88 to 94% by weight, more preferably 89 to 91% by weight, and the content of the rice bran powder is 1 to 15% by weight, preferably 4 to 10% by weight, more preferably 6 to 9% by weight, and the content of MSG is 0.5 to 5% by weight, preferably 1 to 4% by weight, more preferably may be 2 to 3% by weight.

In the present invention, the brown rice of the medium composition may have a moisture content controlled through a moisture control process. The moisture control process may be performed before the rice bran powder and MSG are added to brown rice, thereby providing sufficient moisture necessary for the subsequent fermentation process to effectively ferment the strain. The moisture content of the brown rice is 20 to 50% by weight, preferably 30 to 40% by weight, more preferably 33 to 35% by weight is advantageous in terms of increasing the amount of monacolin K production.

In the present invention, the rice bran powder may be the stir-fried rice bran powder as described above.

Detailed descriptions of the technical significance and effects of each component of the medium composition are the same as those described above in the description of the medium composition, and thus, detailed descriptions are omitted to avoid duplication.

In the inoculation step, as the strain producing monacolin K, a strain of the genus Aspergillus, a strain of the genus Monascus, or a mutant thereof may be used, and their types are the same as those described above in the description of the medium composition, so Specific descriptions are omitted to avoid duplication.

In the present invention, the strain producing the monacolin K is preferably a strain of the genus Aspergillus or a mutant thereof, particularly Aspergillus terreus or a mutant thereof. In this case, there is an advantage in that the expression of genes involved in the biosynthesis pathway of citrinin, one of mycotoxins, is suppressed during the fermentation process. When a mutant strain is used as a strain, the mutant strain itself can be obtained and used, or a mutant strain can be prepared and used by mutating the parent bacteria.

Preferably, Aspergillus teraeus DSMK01 may be used as the strain producing the monacolin K. The Aspergillus teraeus DSMK01 is a mutant strain of Aspergillus teraeus KCCM 12225, and NTG (N-methyl-N'-nitro-N-nitrosoguanidine) was added to Aspergillus teraeus KCCM 12225 or irradiated with ultraviolet rays and then cultured. and can be obtained by selection. The Aspergillus teraeus DSMK01 does not produce citrinin during the fermentation process, and has a very excellent ability to produce monacolin K compared to other Aspergillus teraeus strains.

The inoculation amount of the strain is preferably 0.2 to 2.0% by weight, preferably 0.5 to 1.0% by weight based on the total weight of the medium in terms of fermentation efficiency.

The strain may be prepared in the form of an inoculum, that is, a seed, necessary for initiating fermentation. Specifically, the inoculation may be performed after inoculating the strain in a seed medium for culturing. The culture method is not particularly limited, and may be performed using methods known in the art.

In the fermentation step, a fermented product having an increased content of monacolin K is prepared by fermenting the culture medium composition inoculated with the strain.

In the present invention, the fermentation may be carried out at a temperature of 20 to 50 ℃, preferably 25 to 35 ℃. If the fermentation temperature is too low, the fermentation time may be prolonged, resulting in lower efficiency and a higher probability of contamination by bacteria. If the fermentation temperature is too high, the production of monacolin K by fermentation may decrease. In addition, the fermentation period may vary depending on the change in the content of monacolin K, but may generally be carried out for 5 to 20 days, and preferably may be carried out for 7 to 15 days.

In the present invention, the fermentation may be carried out in a humidity condition of 80%RH or more, preferably 90%RH or more, more preferably 95%RH or more. By maintaining the humidity during fermentation in the above range, it is possible to prevent the substrate from drying out in the fermentation process, and the fermentation of the strain can proceed effectively to significantly increase the production of monacolin K.

In a preferred embodiment of the present invention, a hydrotreating process may be performed in the fermentation step. The hydrotreating may be performed by mixing after adding sterilized process water to the upper part of the substrate during fermentation, so that the water content of the fermentation substrate reaches a level capable of increasing the amount of monacolin K production through hydrotreatment. can be adjusted

In the hydrotreatment process, the amount of hydrotreating may be 0.5 to 3% by weight based on the total weight of the medium, and the number of times of hydrotreatment may be 1 to 5 times. In particular, in order to control the moisture content of the fermentation substrate to a level capable of increasing the amount of monacolin K production, it is preferable to perform the hydrotreatment 2 to 3 times in an amount of 1 to 2% by weight based on the total weight of the medium. . If water evaporation in the fermentation substrate occurs during the fermentation process, the number of hydrotreatments can be increased.

In a preferred embodiment of the present invention, the production amount of monacolin K can be maximized by adjusting the entry and exit moisture content of the fermentation medium inoculated with the strain. In the present invention, the "entrance" refers to a state immediately after inoculation of the strain into the medium, that is, at the time when fermentation starts, and the "departure" refers to a state at the time when fermentation is terminated. The entry and exit moisture content can be adjusted by controlling fermentation humidity and hydrotreating process. In addition, if the entry moisture content is insufficient, the entry moisture content can be adjusted by adding sterilization process water at the time of seed inoculation, and the exit moisture content may vary depending on the steam generated during the fermentation process.

In the present invention, the entry moisture content may be 30 to 50% by weight, and the departure moisture content may be adjusted to 40 to 60% by weight. Preferably, when the entry moisture content is 35 to 40% by weight and the exit moisture content is 45 to 50% by weight, it is possible to prepare a fermented product having a very high content of monacolin K. If the entry moisture content is too high, the growth of other microorganisms (bacteria) present in the air as well as the strains inoculated into the medium may become active, and the amount of monacolin K production may decrease.

In one embodiment of the present invention, after the fermentation step, the method may further include drying the fermented product to secure preservation, stability, processability, and the like of the fermented product. At this time, it may be dried so that the moisture content of the fermented product is 10% by weight or less, preferably 8% by weight or less. In addition, a process of pulverizing the dried fermented product to an appropriate size according to the intended use may be additionally performed.

In addition, the present invention relates to a fermented product having an increased content of monacolin K produced by the above production method. The fermented product with an increased content of monacolin K of the present invention can be effectively used as a raw material for cholesterol-improving foods, health functional foods, feeds, or pharmaceuticals.

The fermented product prepared according to the production method of the present invention has a high content of monacolin K of 7.0 mg/g or more, and the monacolin K content of the fermented product is 8.0 mg/g or more, preferably through optimization of fermentation conditions and methods. It can be maximized with more than 10.0mg/g. As such, since the fermented product of the present invention has a very high content of monacolin K, it can be effectively used for cholesterol-lowering foods. In addition, when a strain of Aspergillus terreus genus or a mutant thereof is used as a strain for producing monacolin K during the production of a fermented product, there is an additional effect of inhibiting the production of citrine, so there is a risk of toxicity to the kidney The fermented product can be applied to food without

Since the fermented product may be added to food or feed to achieve a special purpose, in this aspect, it may be used in a food composition, a composition for food additives, a feed composition or a composition for feed additives. When the fermented product is used in feed or food, monacolin K inhibits the expression of the cholesterol synthetase HMG-CoA reductase (3-hydroxy-3-methylglutaryl-coenzyme A reductase) gene, thereby inhibiting the synthesis of cholesterol. may have an inhibitory effect.

When the fermented product is used for food, it may be used in the form of the fermented product itself prepared according to the manufacturing method of the present invention, or may be used in a processed form. For example, the fermented product may be used as a substitute for rice or mixed with rice to cook rice, or by drying, pulverizing, and concentrating the fermented product in various forms, such as bread, sikhye, makgeolli, vinegar, etc. It can be used as a raw material for food. The food may be a health functional food, and more specifically, may be a health functional food for improving cholesterol for lowering blood cholesterol concentration.

In the present invention, "for additives" is included as long as it is a component added to food or feed other than the main raw material, and specifically, the Ministry of Food and Drug Safety added for coloring, preservation, etc. It may be a food additive as defined in

In the present invention, the composition for feed may be prepared in the form of fermented feed, compound feed, pellet form, silage, and the like.

The composition is mixed with a carrier and fragrance commonly used in the food or pharmaceutical field, and is mixed with a tablet, a troche, a capsule, an elixir, a syrup, and a powder. , may be prepared and administered in the form of suspensions or granules. As the carrier, binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, and the like can be used. The method of administration may be oral, parenteral or application method, but preferably oral administration. In addition, the administration dose can be appropriately selected according to the absorption of the active ingredient in the body, the inactivation rate and excretion rate, the age, sex, condition of the subject, and the like. The pH of the composition can be easily changed according to the manufacturing conditions of the drug, food, etc. in which the composition is used.

Example

The present invention will be described in more detail with reference to the following examples. However, these Examples show some experimental methods and compositions for illustrative purposes of the present invention, and the scope of the present invention is not limited to these Examples.

Preparation Example: Preparation of Fermented Products Containing Monacolin K

After immersing brown rice (provided by Daesang Sunchang Factory), the main raw material of the medium, in a heating tank at 50°C for 3 hours, add process water at 20°C to the surface of the immersed brown rice to remove the residue, and under natural pressure The water content was adjusted to a level of about 35% by weight by dehydration treatment. Stir-fried rice bran powder is added to dehydrated brown rice, and MSG (provided by Daesang Co., Ltd. Gunsan Plant) is added so as to be 3% by weight based on the total weight of the medium composition, then uniformly mixed to prepare a medium, and a certain amount in three autoclaving chambers Each was subdivided.

After sterilization for about 15 minutes by inputting steam into the facility at a temperature of 121°C and a pressure of 1.5kgf/cm ³ in a state of being contained in a chamber, cold water is put into the jacket in the sterilizer and the temperature of the medium is increased to 30 through internal ventilation. It was cooled to below °C. For each of the three prepared fermentation media, the Aspergillus terreus ( Aspergillus terreus ) DSMK01 (provided by Daesang Co., Ltd.) strain was pre-cultured into the chamber and the prepared seed is evenly distributed in an amount of 0.6% by weight based on the total weight of the medium. inoculated. At this time, the moisture content of the substrate was measured, and it was confirmed that it was about 35% by weight.

After inoculation of the seed, the initial fermentation product temperature was maintained at 30° C. and the internal humidity was maintained at 100% RH during the fermentation period to ferment, and oxygen (air) was continuously supplied during the fermentation process. After checking the condition of the fermented product in the chamber on the 7th day of fermentation, sterilized distilled water of about 1% by weight relative to the weight of the fermentation substrate was evenly sprayed on the fermentation substrate, mixed uniformly, and hydrolyzed. After confirming that the moisture content of the fermentation substrate was 37% by weight or more, the fermentation process was continued, and hydrolysis was performed one more time in the same manner. Fermentation was terminated when the fermentation substrate was decomposed and transformed into a sponge-like tissue form. At the end of the fermentation, the moisture content of the substrate was measured, and it was confirmed that it was about 50% by weight.

After the fermentation was completed, the fermented product was placed in a drying room at 50° C., and then drying was performed so that the moisture content was about 8% by weight while minimizing contamination of the surrounding air. The dried fermented product was ground to a level of 30 mesh and stored refrigerated.

Experimental Example: Measurement of the content of monacolin K by HPLC analysis

High Performance Liquid Chromatography (HPLC) analysis was performed on the fermented product prepared according to the method of Preparation Example to be described later to measure the content of monacolin K.

First, to prepare a sample for HPLC analysis, 5 g of the fermented product was taken, 25 mL of methanol was added to the fermented product, and then extracted with shaking at a temperature of 27° C. and a speed of 180 rpm using a shaker for 3 hours. When the extraction was completed, it was filtered using filter paper, and centrifuged for 10 minutes in a centrifuge at a temperature of 4° C. and a speed of 3,000 rpm. After centrifugation was completed, the supernatant was taken and filtered with a syringe filter having a pore size of 0.45 μm, and HPLC analysis was performed using the filtrate as a sample.

On the other hand, in order to quantify the monacolin K content of the sample, HPLC analysis was performed on the monacolin K standard solution having a concentration in the range of 5 to 300 mg/L to prepare a calibration curve. By comparing the detection curve of the fermented sample with the calibration curve of the standard solution, it was confirmed that monacolin K was generated, and the peak of monacolin K from the sample detection curve was substituted into the calibration curve, and the monacolin K content for each sample (mg /g) was calculated.

The HPLC analysis was performed under the following conditions.

- Analytical instrument: Class-LC 10 (Shimadzu Corporation)

- UV detector: SPD-10A (Shimadzu Corporation)

- UV wavelength: 238nm

- Temperature: 40℃

- Column: Mightysil C18 reverse-phase column (4.6Х250mm) (Kanto Corporation)

- Mobile phase: methanol: 18 mM o -phosphoric acid = 77.5: 22.5 (weight ratio)

- Flow rate: 1.2mL/min

- Sample injection volume: 20 μl

Experimental Example 1: Experiment for comparing the content of monacolin K according to the main material of the medium

HPLC analysis was performed on each of the three fermented products prepared by adding 4% by weight of roasted rice bran powder in Preparation Example to measure the monacolin K content, and the results are shown in Table 1 below. In addition, it was prepared in the same manner as in Preparation Example, but the content of monacolin K was measured by performing HPLC analysis on a fermented product using white rice or wheat gluten instead of brown rice as the main raw material of the medium, and the results are shown in Table 1 below. indicated.

main raw material polished rice Brown rice wheat gluten Monacolin K content (mg/g) 5.2 7.2 4.8 5.1 8.1 4.3 4.9 7.6 4.9

From the results in Table 1, it was confirmed that by using brown rice as the main raw material of the fermentation medium, the ability to produce monacolin K could be improved compared to other cereal mediums. In particular, when brown rice was used as the main raw material of the fermentation medium, it was confirmed that the production ability of monacolin K was significantly superior compared to the case of using white rice, which is generally used as a substrate for the fermentation medium of Hong Guk-gyun.

Experimental Example 2: Comparative experiment of monacolin K content according to the auxiliary material of the medium

HPLC analysis was performed on each of the three fermented products prepared by adding 4.7 wt% of roasted rice bran powder in the preparation example to measure the monacolin K content, and the results are shown in Table 2 below. In addition, HPLC analysis was also performed on fermented products prepared in the same manner as in Preparation Example, but using stir-fried wheat (provided by Sunchang Plant of Daesang Co., Ltd.) or dandelion (provided by Osan Plant of Daesang Co., Ltd.) instead of stir-fried rice bran powder as an auxiliary material of the medium. to measure the content of monacolin K, and the results are shown together in Table 2 below.

auxiliary material stir-fried wheat fried rice bran powder Watermelon (starch syrup) Monacolin K content (mg/g) 6.3 8.3 7.9 6.6 7.9 7.5 6.1 8.1 7.6

From the results of Table 2, it was confirmed that when the roasted rice bran powder was added as an auxiliary material of the fermentation medium, the production ability of monacolin K was superior compared to other grain-based auxiliary materials.

Experimental Example 3: Comparative experiment of monacolin K content according to the amount of rice bran powder added

HPLC analysis was performed on each of the fermented products prepared by adding the roasted rice bran powder in an amount of 1, 2, 3, 5, 7, 9, 10, and 12% by weight based on the total weight of the medium composition in the above Preparation Example to obtain monacolin K The content was measured, and the results are shown in Table 3 below.

Addition amount (wt%) One% 2% 3% 5% 7% 9% 10% 12% Monacolin K content (mg/g) 6.8 7.0 7.1 8.3 10.5 8.5 8.1 7.5 6.3 6.9 7.0 8.1 10.6 8.3 8.0 7.3 6.2 6.9 7.2 8.4 10.6 8.3 8.0 7.6

From the results of Table 3, it was confirmed that the production amount of monacolin K varies according to the amount of added rice bran powder, which is an auxiliary raw material, and when the amount of rice bran powder added is 5 to 10% by weight based on the weight of the total medium composition, the production amount of monacolin K It was confirmed that it was excellent at 8.0 mg/g or more, and especially when the content of monacolin K was 7% by weight or more, the content of monacolin K was 10.5 mg/g or more.

Experimental Example 4: Comparison experiment of the content of monacolin K according to the amount of hydrolysis

HPLC analysis was performed on each of the three fermented products prepared by adding the roasted rice bran powder at 7.9% by weight in Preparation Example to measure the monacolin K content, and the results are shown in Table 4 below. In addition, but prepared in the same manner as in Preparation Example, HPLC analysis was performed on the fermented product prepared by controlling the amount of hydrolysis in the fermentation process as shown in Table 4 to measure the content of monacolin K, and the result is shown below. It is shown together in Table 4.

Hydrogen Throughput (%) 0.1% 0.5% One% 2% Monacolin K content (mg/g) 4.1 6.2 10.2 8.3 4.0 6.7 10.4 8.1 4,3 6.8 10.1 8.2

From the results of Table 4, it was confirmed that the production of monacolin K varies according to the amount of hydrolysis in the hydrotreatment process, and when the amount of hydrolysis is 1 to 2%, the production of monacolin K is excellent at 8.1 mg/g or more, In particular, when it was 1%, it was confirmed that the content of monacolin K was 10.1 mg/g or more, which was the most excellent.

Experimental Example 5: Comparison experiment of monacolin K content according to the number of times of hydrolysis

HPLC analysis was performed on each of the three fermented products prepared by adding roasted rice bran powder at 7.9% by weight in the above Preparation Example to measure the monacolin K content, and the results are shown in Table 5 below. In addition, but prepared in the same manner as in Preparation Example, as shown in Table 5 below, HPLC analysis was performed on the fermented product prepared by controlling the number of hydrotreating in the fermentation process to measure the content of monacolin K, and the result was It is shown together in Table 5 below.

Number of water treatments
(during 14-day incubation period) 1 time Episode 2 3rd time Monacolin K content (mg/g) 6.2 10.3 8.3 6.3 10.2 8.0 6.2 10.2 8.2

From the results in Table 5, it was confirmed that the production of monacolin K varies depending on the number of hydrotreating in the hydrotreatment process, and when hydrotreating 2 to 3 times, the production of monacolin K is excellent at 8.0 mg/g or more, In particular, it was confirmed that the content of monacolin K was the most excellent at 10.2 mg/g or more when the hydrotreatment was performed twice.

Experimental Example 6: Comparative experiment of monacolin K content according to fermentation humidity

HPLC analysis was performed on each of the three fermented products prepared by adding the roasted rice bran powder at 7.9% by weight in the Preparation Example to measure the monacolin K content, and the results are shown in Table 6 below. In addition, it was prepared in the same manner as in Preparation Example, but as shown in Table 6 below, the content of monacolin K was measured by performing HPLC analysis on the fermented product prepared by controlling the fermentation humidity, and the results are shown in Table 6 below. indicated.

Humidity (%RH) 30% 50% 70% 80% 90% 100% Monacolin K content (mg/g) 4.2 5.1 5.7 7.3 10.2 10.7 4.0 5.2 5.8 7.4 10.1 10.6 4.3 5.1 5.7 7.3 10.2 10.5

From the results of Table 6, it was confirmed that the production of monacolin K varies depending on the humidity conditions during the fermentation process of the spawn, and when the humidity is 90% or more, the production of monacolin K is excellent at 10.1 mg/g or more, especially 100 %, it was confirmed that the content of monacolin K was the most excellent as 10.5 mg/g or more.

Experimental Example 7: Comparison experiment of monacolin K content according to entry moisture content

HPLC analysis was performed on each of the three fermented products prepared by adding roasted rice bran powder at 7.9% by weight in the Preparation Example to measure the monacolin K content, and the results are shown in Table 7 below. Also, HPLC analysis was performed on fermented products prepared in the same manner as in Preparation Example, but prepared by adjusting the entry moisture content (moisture content at the time of fermentation start) in the fermentation process as shown in Table 7 below to determine the level of monacolin K. The content was measured, and the results are shown together in Table 7 below.

Entering moisture content (% by weight) 28% 30% 35% 40% Monacolin K content (mg/g) 5.7 6.4 10.6 9.1 6.0 6.8 10.5 9.3 5.9 6.7 10.3 9.3

From the results of Table 7, it was confirmed that the production of monacolin K varies according to the entry moisture content during the fermentation process of the spawn, and when the entry moisture content is 35 to 40%, the production of monacolin K is 9.1 mg/g or more It was confirmed that the content of monacolin K was excellent, especially when it was 35%, at 10.3 mg/g or more.

Experimental Example 8: Experiment for comparing the content of monacolin K according to the departure moisture content

HPLC analysis was performed on each of the three fermented products prepared by adding the roasted rice bran powder at 7.9% by weight in the Preparation Example to measure the monacolin K content, and the results are shown in Table 8 below. In addition, in each of the fermented products prepared in the same manner as in Preparation Example, but by adjusting the outbound moisture content (moisture content at the end of fermentation) as shown in Table 8 below by varying the amount or number of hydrotreatments in the fermentation process. For measuring the content of monacolin K by performing HPLC analysis, the results are shown together in Table 8 below. The content of monacolin K in the fermented product was calculated as (monacolin analysis value × 92)/(100 - departure moisture content) in terms of 8% moisture content after drying.

Departure moisture content (% by weight) 30% 40% 45% 50% Monacolin K content (mg/g) 7.3 7.8 10.6 10.0 7.1 7.9 10.4 10.2 7.1 7.8 10.4 10.3

From the results of Table 8, it was confirmed that the production of monacolin K varies depending on the departure moisture content in the fermentation process of the spawn, and when the departure moisture content is 45 to 50%, the production of monacolin K is 10.0 mg/g or more It was confirmed that the content of monacolin K was the most excellent, especially when it was 45% or more at 10.4 mg/g or more.

Experimental Example 9: Monacolin K content comparison experiment according to the moisture content change pattern

HPLC analysis was performed on each of the three fermented products prepared by adding the roasted rice bran powder at 7.9% by weight in the Preparation Example to measure the monacolin K content, and the results are shown in Table 9 below. In addition, it was prepared in the same manner as in Preparation Example, but by varying the number or number of sterilization steps in the seed inoculation step and the number or number of hydrotreating in the fermentation step, as shown in Table 9 below, the moisture content from the start of fermentation to 14 days later HPLC analysis was also performed on the fermented product prepared by controlling the change pattern to measure the monacolin K content, and the results are shown in Table 9 below.

Moisture content change pattern
(weight%) 28% → 40% 35% → 50% 45% → 50% Monacolin K content (mg/g) 8.2 10.3 6.3 8.3 10.5 6.6 8.2 10.5 6.4

From the results of Table 9, it was confirmed that the production of monacolin K was changed according to the change pattern of the moisture content, and when the moisture content was adjusted from 35% upon entry to 50% upon departure, the content of monacolin K was 10.3mg/ g or higher showed a high result.

On the other hand, if the entry moisture content is too low as 28% by weight, the production of monacolin K is at a maximum level of 8.3 mg/g. The results with limitations were confirmed. In addition, it was confirmed that when the entry moisture content is too high as 45% by weight, the production of monacolin K can be reduced to a maximum of 6.6mg/g even when the exit moisture content is adjusted to the level of 50% by weight, which is at the initial stage of fermentation. When the moisture content is high, the growth of not only the target spawn but also other microorganisms in the air becomes active, which is confirmed to be due to the cause of hindering the fermentation of the seed.

From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the claims to be described later and their equivalents.

Claims (15)

A medium composition for producing monacolin K, comprising brown rice, rice bran powder and sodium glutamate (MSG). The method of claim 1,
Based on the weight of the total medium composition, 88 to 94% by weight of brown rice, 4 to 10% by weight of rice bran powder, and 1 to 4% by weight of MSG, the medium composition for monacolin K production. The method of claim 1,
The medium composition for producing monacolin K, characterized in that the water content of the brown rice is 30 to 40% by weight. The method of claim 1,
The medium composition for producing monacolin K, characterized in that the rice bran powder is roasted rice bran powder. The method of claim 1,
The medium composition is Aspergillus ( Aspergillus ), characterized in that used for fermentation using a strain or a mutant thereof, Monacolin K production medium composition. 6. The method of claim 5,
The Aspergillus sp. strain is Aspergillus terreus ( Aspergillus terreus ), characterized in that the monacolin K production medium composition. (i) an inoculation step of inoculating a strain producing monacolin K in a medium composition comprising brown rice, rice bran powder and sodium glutamate (MSG); and
(ii) a fermentation step of fermenting the culture medium composition inoculated with the strain to obtain a fermented product
A method for producing a fermented product containing an increased content of monacolin K. 8. The method of claim 7,
A method for producing a fermented product having an increased content of monacolin K, characterized in that the fermentation step is performed at a humidity of 90%RH or higher. 8. The method of claim 7,
The method for producing a fermented product having an increased content of monacolin K, characterized in that the fermentation step is performed for 7 to 15 days at a temperature of 25 to 35 °C. 8. The method of claim 7,
A method for producing a fermented product with an increased content of monacolin K, characterized in that the hydrotreating process is performed 2-3 times in the fermentation step. 11. The method of claim 10,
In the hydrotreatment process, the method for producing a fermented product having an increased content of monacolin K, characterized in that the amount of hydrolysis is 1 to 2% by weight based on the total weight of the medium composition. 8. The method of claim 7,
In the fermentation step, the method for producing a fermented product with an increased content of monacolin K, characterized in that the entry moisture content at the start of fermentation of the strain-inoculated medium composition is 35 to 40% by weight. 8. The method of claim 7,
In the fermentation step, the method for producing a fermented product having an increased content of monacolin K, characterized in that the departure moisture content at the end of fermentation of the strain-inoculated medium composition is 45 to 50% by weight. The fermented product with an increased content of monacolin K, prepared by the method of any one of claims 7 to 13. A health functional food for improving cholesterol, comprising a fermented product having an increased content of monacolin K according to claim 14.