KR102284251B1 - Method for producing red yeast rice with enhanced γ-oryzanol and Monacolin K content by controlling moisture content - Google Patents

Abstract

The present invention comprises the steps of (1) immersing brown rice of Hangaru variety in water and then taking it out and sterilizing it; (2) adding water to the sterilized brown rice of step (1); (3) inoculating the brown rice with water in step (2) and then culturing it; and (4) sterilizing and drying the cultured red yeast rice in step (3). It relates to a method of enhancing the γ-oryzanol and monacolin K content of the prepared γ-oryzanol and monacolin K content of enhanced red rice and hongguk rice.

Description

Method for producing red yeast rice with enhanced γ-oryzanol and monacolin K content by controlling moisture content {Method for producing red yeast rice with enhanced γ-oryzanol and Monacolin K content by controlling moisture content}

The present invention comprises the steps of (1) immersing brown rice of Hangaru variety in water and then taking it out and sterilizing it; (2) adding water to the sterilized brown rice of step (1); (3) inoculating the brown rice with water in step (2) and then culturing it; and (4) sterilizing and drying the cultured red yeast rice in step (3). It relates to a method of enhancing the γ-oryzanol and monacolin K content of the prepared γ-oryzanol and monacolin K content of enhanced red rice and hongguk rice.

Rice ( Oryzae sativa ) is one of the most important grains in the Korean diet, and is used as an important food resource in Southeast Asia and other countries. However, per capita rice consumption has decreased by more than half over the past 30 years, and edible consumption has decreased faster than rice production. In addition, about 90% of the rice consumed is used for cooking, and most of the rice used for processing is used for making rice cakes.

Among the recently developed cultivars for processing, Hangaru, a rice flour-only variety, has an opaque grain like glutinous rice, but contains 18% or more of amylose. It is also known to be suitable for brewing due to its low protein and fat content and high alcohol yield due to its high reducing sugar content. Samgwang rice is the highest quality rice variety cultivated in Korea and is used for cooking because it has a sweet taste and a soft texture. It is known that the amylose content is 18% or more, and it has polygonal starch particles and has a hardness of 5.85 kg, which is higher than the 3.62 kg of snow crab.

Red yeast rice (Monascus) is a fungus that produces polyketide-type secondary metabolites such as monacolin K or red yeast pigment, and has been mainly used in the form of inoculation and culturing in rice in China, Japan, and Korea. The secondary metabolites produced by red yeast rice have been reported to have various functions such as anti-inflammatory, neurotransmission, blood pressure lowering effect, and antioxidant action, and are used not only as general food but also as health functional food and medicine.

The traditional manufacturing method of red rice is mainly performed in the order of washing and immersing the rice, draining, steaming, inoculating and culturing bacteria, and the culture is fermented at 25-32°C for 2-4 weeks, so it is complicated and takes a long time. The role of water activity and water content of the substrate is important in culturing red yeast rice. The higher the moisture content, the smoother the culture of red yeast rice, but there are problems such as bacterial contamination, adhesion between rice grains, and non-uniform culture.

Korean Patent No. 1557028 discloses a method for producing red rice using glutinous rice, and Korean Patent Publication No. 2017-0139789 discloses a method for producing red rice using anchovy extract, but γ-oryzanol and It is different from the manufacturing method of hongguk rice with enhanced monacolin K content.

The present invention has been derived from the above needs, and an object of the present invention is to optimize the manufacturing conditions such as selection of rice varieties, milling, and addition of water to produce red yeast rice with enhanced functional ingredients and antioxidant activity to achieve quality and preference An object of the present invention is to provide a method for producing improved red yeast rice.

In order to solve the above problems, the present invention comprises the steps of (1) immersing the brown rice of the Hangaru variety in water and then taking it out and sterilizing it; (2) adding water to the sterilized brown rice of step (1); (3) inoculating the brown rice with water in step (2) and then culturing it; and (4) sterilizing and drying the cultured red yeast rice in step (3).

In addition, the present invention provides γ-oryzanol and monacolin K content of improved red yeast rice prepared by the above method.

In addition, the present invention comprises the steps of (1) immersing the brown rice of Hangaru variety in water, then taking it out and sterilizing it; (2) adding water to the sterilized brown rice of step (1); (3) inoculating the brown rice with water in step (2) and then culturing it; and (4) sterilizing and drying the cultured red rice in step (3).

Red rice rice using brown rice of the Hangaru variety of the present invention can provide high-quality red rice rice with excellent antioxidant activity while increasing gamma oryzanol and monacolin K contents.

1 is a microscopic observation of a cross section of raw rice and rice immersed in water.

In order to achieve the object of the present invention, the present invention

(1) sterilizing the brown rice of the Hangaru variety after immersion in water;

(2) adding water to the sterilized brown rice of step (1);

(3) inoculating the brown rice with water in step (2) and then culturing it; and

(4) It provides a method for producing hongguk rice with enhanced γ-oryzanol and monacolin K content, characterized in that it comprises the step of sterilizing and drying the cultured red yeast rice in step (3).

In the method for producing red yeast rice of the present invention, the step (1) is preferably after immersing the brown rice of the Hangaru variety in water at 2 to 6° C. for 20 to 28 hours, and then taking it out at 110 to 130° C. for 10 to 20 minutes. It can be sterilized during the period, and more preferably, brown rice of the Hangaru variety was immersed in water at 4° C. for 24 hours, and then taken out and sterilized at 121° C. for 15 minutes to prepare brown rice suitable for fermentation of red yeast rice.

In a preliminary experiment, the present inventors found that preparing hongguk rice using brown rice of the Hangaru variety can further enhance gamma oryzanol and monacolin K content compared to other varieties of rice (Sangju glutinous rice, Chilbo, Samgwang, etc.) could check

In addition, in the method for producing red yeast rice of the present invention, the addition of step (2) may preferably add 50 to 70% (v/w) of water to sterilized brown rice compared to brown rice, more preferably sterilized. 60% (v/w) of water can be added to one brown rice compared to brown rice. Adding water under the above conditions could further enhance the functional ingredient content and antioxidant activity compared to adding water under other conditions.

In addition, in the method for producing red yeast rice of the present invention, the step (3) is preferably inoculated with 8 to 12% (v/w) of hongguk bacteria compared to brown rice to brown rice to which water is added, and then 6 to at 28 to 32 ° C. It can be cultured for 8 days, and more preferably, 10% (v/w) of honggukgyun compared to brown rice is inoculated into brown rice to which water is added, and then cultured at 30° C. for 7 days. Culturing under the same conditions as described above was able to smoothly ferment red yeast rice.

In addition, in the method for producing red yeast rice of the present invention, the step (4) preferably sterilizes the cultured red yeast rice at 110 to 130 ° C. for 10 to 20 minutes, and then the water content is 8 to 12 at 55 to 65 ° C. % (v/w) may be dried, and more preferably, after sterilizing the cultured red yeast rice at 121°C for 15 minutes, it may be dried to a moisture content of 10% (v/w) at 60°C . Sterilization and drying under the same conditions as described above could improve the storability while maintaining the inherent characteristics of red rice.

The method of the present invention for producing red yeast rice with enhanced γ-oryzanol and monacolin K content, more specifically

(1) immersing the brown rice of the Hangaru variety in water at 2~6℃ for 20~28 hours, then taking it out and sterilizing it at 110~130℃ for 10~20 minutes;

(2) adding 50 to 70% (v/w) of water compared to brown rice to the sterilized brown rice of step (1);

(3) inoculating 8 to 12% (v/w) of honggukgyun compared to brown rice into brown rice to which water was added in step (2), followed by culturing at 28 to 32° C. for 6 to 8 days; and

(4) After sterilizing the cultured red yeast rice in step (3) at 110~130℃ for 10~20 minutes, drying it at 55~65℃ so that the moisture content becomes 8~12% (v/w) may include

more specifically

(1) immersing the brown rice of Hangaru variety in water at 4°C for 24 hours, then taking it out and sterilizing it at 121°C for 15 minutes;

(2) adding 60% (v/w) of water to the sterilized brown rice of step (1) compared to the brown rice;

(3) inoculating 10% (v/w) of honggukgyun compared to brown rice into brown rice to which water was added in step (2), followed by culturing at 30° C. for 7 days; and

(4) After sterilizing the cultured red yeast rice in step (3) at 121°C for 15 minutes, it may include drying at 60°C so that the water content becomes 10% (v/w).

The present invention also provides γ-oryzanol and monacolin K content of improved red yeast rice prepared by the above method.

The present invention also

(1) sterilizing the brown rice of the Hangaru variety after immersion in water;

(2) adding water to the sterilized brown rice of step (1);

(3) inoculating the brown rice with water in step (2) and then culturing it; and

(4) It provides a method for enhancing γ-oryzanol and monacolin K content of red rice, including the step of drying after sterilizing the cultured red rice in step (3).

The method of enhancing the γ-oryzanol and monacolin K contents of red yeast rice of the present invention, more specifically

(1) immersing the brown rice of the Hangaru variety in water at 2~6℃ for 20~28 hours, then taking it out and sterilizing it at 110~130℃ for 10~20 minutes;

(2) adding 50 to 70% (v/w) of water compared to brown rice to the sterilized brown rice of step (1);

(3) inoculating 8 to 12% (v/w) of honggukgyun compared to brown rice into brown rice to which water was added in step (2), followed by culturing at 28 to 32° C. for 6 to 8 days; and

(4) After sterilizing the cultured red yeast rice in step (3) at 110~130℃ for 10~20 minutes, drying it at 55~65℃ so that the moisture content becomes 8~12% (v/w) may include

more specifically

(1) immersing the brown rice of Hangaru variety in water at 4°C for 24 hours, then taking it out and sterilizing it at 121°C for 15 minutes;

(2) adding 60% (v/w) of water to the sterilized brown rice of step (1) compared to the brown rice;

(3) inoculating 10% (v/w) of honggukgyun compared to brown rice into brown rice to which water was added in step (2), followed by culturing at 30° C. for 7 days; and

(4) After sterilizing the cultured red yeast rice in step (3) at 121°C for 15 minutes, it may include drying at 60°C so that the water content becomes 10% (v/w).

Hereinafter, production examples and examples of the present invention will be described in detail. However, the following Preparation Examples and Examples only illustrate the present invention, the content of the present invention is not limited to the following Preparation Examples and Examples.

manufacturing example One. red rice manufacturing

(1) Brown rice of Hangaru variety was washed 5 times to remove foreign substances and dried at 60°C for one hour before use. The dried brown rice was immersed in water at 4°C for 24 hours, then taken out and sterilized at 121°C for 15 minutes.

(2) The sterilized brown rice of step (1) was cooled to 25° C., and water was added 60% (v/w) compared to brown rice.

(3) After inoculating 10% (v/w) of a culture solution of red yeast rice (dry cell weight about 3.6 mg/mL) compared to brown rice to brown rice to which water was added in step (2), it was cultured at 30° C. for 7 days.

(4) After sterilizing the cultured red yeast rice in step (3) at 121° C. for 15 minutes, it was dried and ground at 60° C. so that the water content was 10% (v/w).

Materials and Methods

1) Experimental materials

For the rice varieties used in this experiment, Samgwang and Hangaru grown in 2017 were used. Japan) to prepare white rice and use it in the experiment. In addition, Monascus pilosus 305-9 (KFCC 11410P) was provided by F&P Co., Ltd. and was used.

2) Cultivation of red yeast

The culture of red yeast rice has excellent production capacity of monacolin K and does not produce the neurotoxin citrinin ( Monascus). pilosus ) 305-9 (KFCC 11410P) was purchased from F&P Co., Ltd. and used. Hong Guk-gyun cut the seeds cultured on potato agar medium (potato dextrose agar, PDA, Difco laboratories, Sparks, MD, USA) with a cork borer with a diameter of 0.5 cm and inoculated it again on the PDA, which was inoculated at 30 ℃, humidity A seed plate was prepared by culturing at 70% for 7 days. Liquid culture was performed to inoculate rice with Hong Guk bacteria, and the above PDA medium was cut into a 0.5 cm diameter cork borer and 1 piece per 50 mL of potato extract medium (potato dextrose broth, PDB, Difco laboratories, Sparks, MD, USA) was added and cultured for 7 days in a shaking incubator at 150 rpm and 30°C.

3) Rice preparation and inoculation

Raw rice was washed with water 5 times to remove the husk and foreign substances, and dried at 60° C. for one hour before use. Double distilled water was added to the washed rice, immersed at 4°C for 24 hours, the water was discarded, the external moisture was removed, and then sterilized at 121°C for 15 minutes. At the same time, distilled water was sterilized and prepared, and after sterilization, it was cooled to 25°C. Water was added to the sterilized rice in 20% increments from 0 to 100% of the weight of the raw rice. Red yeast rice cultured in liquid for 7 days on sterilized rice was homogenized with a homogenizer (Ultra-Turrax, model T25, Janke & Kunkel, Staufen, Germany). cultured. After completion of the incubation, sterilization was carried out at 121°C for 15 minutes, dried at 60°C, and the dried product was crushed with a grinder (SMFP-30000, Hanil electric Co., Seoul, Korea) and used in the experiment.

4) Cross-section observation of raw rice and rice after pretreatment

After washing the raw rice 5 times, the cross section was cut and the shape of the rice starch was observed by magnifying 5,000 times with a scanning electron microscope (Scanning Electron Microscopy, ULTRA PLUS, Carl Zeiss AG, Oberkochen, Germany). also. After immersion and dehydration for 24 hours, the sterilized rice was freeze-dried, and the lyophilized rice was cut and the cross section was observed at 10,000 times magnification using a scanning electron microscope.

5) Measurement of Red Guk Rice Ingredients

The components of red rice rice were measured for protein (Micro Kjeldahl method), crude fat (soxhlet extraction method), ash (550℃ ashing method), and reducing sugar (DNS method). The maltose content decomposed by amylase treatment was expressed as the enzymatic hydrolysis rate.

6) Preparation of extract and measurement of chromaticity

To prepare the extract for measuring the active ingredient content, 40 mL of 75% ethanol was added to 4 g of red yeast rice powder, followed by ultrasonic extraction for 1 hour, centrifugation at 3,000 × g for 20 minutes, and 40 mL of the supernatant was used. (Uvikon 933, Kontron Instruments, Neufahrn, Germany) was used to measure at 400, 470, and 500 nm.

7) Measurement of antioxidant activity (DPPH radical scavenging ability)

DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging ability was measured by adding 0.8 mL of 0.2 mM DPPH (Sigma-Aldrich Co.) solution to 0.2 mL of the extract, leaving it at room temperature for 30 minutes, and then measuring the absorbance at 520 nm, It was expressed as AEAC (ascorbic acid equivalent antioxidant capacity, mg AA eq/g).

8) γ-oryzanol content

Extract preparation for gamma oryzanol content analysis was performed by adding 40 mL of an extraction solvent in which dichloromethane and methanol were mixed in a ratio of 2:1 per 2 g of sample, followed by ultrasonic extraction for 30 minutes and then 3,000 rpm for 10 minutes. After centrifugation, the supernatant was filtered with a filter paper, and the precipitate was sequentially treated with 30 mL of an extraction solvent and 20 mL, and the above method was repeated twice, and the final filtrate was concentrated with a nitrogen concentrator. The concentrated precipitate was dissolved in 1 mL of isopropyl alcohol and filtered with a syringe filter (PVDF, 0.45 μm) to use as a sample. The γ-oryzanol content was analyzed using HPLC (ACME9000 system, Younglin, Anyang, Korea), and a C-18 column (Mightysil RP-18 GP column, 4.6×250 mm, Kanto Chemical, Tokyo, Japan) was used. The mobile phase is methanol: acetonitrile: dichloromethane: acetic acid = 50:44:3:3 (v/v/v/v) by injecting 20 μl of the sample with isocratic elution, eluting at a flow rate of 1.4 mL/min, and UV Measurements were made at 325 nm using a detector.

9) Analysis of active and inactive monacolin K content

The standard material of monacolin K was prepared by accurately weighing 10 mg of inactive monacolin K (Mevinolin, Sigma-Aldrich, St. Louis, MO, USA) and dissolving it in 50 mL of 75% ethanol. Of this, 2 mL was taken, 0.5 mL of 0.05N sodium hydroxide solution was added, and left at room temperature for 30 minutes to prepare. The extract was filtered through a 0.45 μm membrane filter and then analyzed using HPLC.

The monacolin K content was analyzed using HPLC (ACME9000 system, Younglin, Anyang, Korea), and a C-18 column (Mightysil RP-18 GP column, 4.6×250 mm, Kanto Chemical, Tokyo, Japan) was used. The mobile phase is acetonitrile as A and 0.2% phosphoric acid as B, from 35:65 to 75:25 at 20 minutes, 100:0 at 21 minutes, 100:0 at 35 minutes, 35 at 36 minutes. :65, 35:65 gradient elution was performed at 45 minutes, and 20 μl of the sample was injected, eluted at a flow rate of 1 mL/min, and measured at 237 nm using a UV detector.

Example One. raw rice and cross-sectional observation of rice after pretreatment

The results of observation at 5,000 times magnification with a scanning electron microscope for cross-sectional observation of raw rice are as shown in FIG. As a result of observing a sample of freeze-dried rice sterilized by water immersion and dehydration for 24 hours with a scanning electron microscope at 10,000 times magnification, the pore size of Hangaru was small and the number of pores was very large. has large and few cross-sections. When culturing honggukgyun, it is thought that hangaru has an advantage in culturing honggukgyun compared to Samkwang because the surface area where nutrients of raw rice can be used increases and oxygen supply is easy.

Example 2. red rice moisture after production, crude protein , george content

Table 1 shows the results of measuring the moisture, crude protein, and crude fat content of Hongkuk rice prepared by controlling the water addition amount in two varieties (Hangaru and Samgwang) polished with brown rice and white rice. After soaking for 24 hours, the moisture content of Hangaru and Samgwang brown rice was 36.83% and 32.56%, respectively, but in polished rice, 40.85% and 34.73% were measured, respectively, indicating that Hangul had a higher water content than Samkwang. The moisture content was higher than that of brown rice. It is thought that soft rice, Hangaru, has a difference in moisture absorption due to loose space between starches, and hard rice, Samgwang, has endosperm cells in which protein bodies are tightly bound between starch particles, making it difficult to absorb moisture. As the amount of water added increased, the crude protein and crude fat content increased. This is probably because the growth of Monascus mycelium increases and as the culture progresses, fat-soluble secondary metabolites such as lactone-type monacolin or red yeast pigment are produced. However, when the amount of water added was 100% of the weight of the rice, the crude protein and crude fat content decreased. I think.

Example 3. of red rice amylose, reducing sugar content and enzymatic hydrolysis rate

Table 2 shows the results of measuring the amylose, reducing sugar content, and enzymatic hydrolysis rate of red yeast rice prepared by controlling the amount of moisture added to two varieties (Hangaru and Samgwang) polished with brown rice and white rice. The amylose content of the existing raw rice was 18.62% and 15.29% in brown rice, respectively, in Hangaru and Samkwang, and 18.92% and 16.93% in white rice, respectively. The reducing sugar content of raw rice was 7.66 and 7.52 mg/g in Hangaru and Samgwang, respectively, in brown rice, and 3.91 and 3.20 mg/g in white rice, which was higher in brown rice than in white rice, and there was no significant difference between varieties. The enzymatic hydrolysis rates of brown rice were 6.07 and 4.07% in Hangaru and Samgwang, respectively, and 5.81 and 5.33% in white rice. When culturing red rice, the content of amylose and reducing sugar increased according to the amount of water added, and when 80% of water was added to white rice, the highest values were 25.84% and 30.13 mg/g. However, the enzymatic hydrolysis rate was measured to be more than 20% for Hangaru compared to raw rice, but 9~15% for Samgwang, which showed a low enzymatic hydrolysis rate.

Example 4. red rice chromaticity of the extract

Table 3 shows the chromaticity of the 75% ethanol extract of red yeast rice prepared by controlling the amount of water added to two varieties (Hangaru and Samgwang) polished with brown rice and white rice. The overall chromaticity of red rice rice was measured to be high in Hangaru, and it was found that the amount of pigment produced increased as the amount of water added increased.

Example 5. of red rice gamma oryzanol content

Tables 4 and 5 show the gamma oryzanol content of red-guk rice prepared by controlling the amount of moisture added to two varieties (Hangaru and Samgwang) polished with brown rice and white rice. Before culturing red yeast, the gamma-oryzanol content was measured to be 491.73 μg/g and 435.95 μg/g, respectively, of Samgwang and Hangaru in brown rice. It was. After culturing, the total gamma oryzanol content increased compared to before culturing, and as the amount of water added increased, the gamma oryzanol content increased as the culture of red yeast became more active. was shown.

Example 6. of red rice monacolin K content

Table 6 shows the monacolin K content of hongguk rice prepared by controlling the amount of water added to two varieties (Hangaru and Samgwang) polished with brown rice and white rice. The total monacolin K content was measured to be the highest at 749.34 mg/kg when 60% moisture was added to powdered brown rice, and at this time, inactive monacolin K 405.54 mg/kg and activated monacolin K 343.80 mg/kg were measured. . Similarly, the total monacolin K content was somewhat low at 705.99 mg/kg, but the active type monacolin K content was measured as high as 362.96 mg/kg at 80% of powdered white rice-water added. The monacolin K content of red rice according to the amount of water added was higher in the Hangaru variety than in the Samkwang, regardless of the degree of milling, so it is considered that the growth of Monascus is active.

Example 7. of red rice antioxidant capacity

Table 7 shows the results of measuring the antioxidant activity of hongguk rice prepared by controlling the amount of water added to the powdered brown rice. DPPH radical scavenging ability showed a tendency to increase as the amount of water added to rice increased.

DPPH radical scavenging activity of red yeast rice (mg AA eq/g) raw rice Water addition rate (%) 0 20 40 60 80 100 0.19±0.00 ^e 0.24±0.01 ^d 0.11±0.00 ^e 0.28±0.00 ^c 0.76±0.00 ^a 0.57±0.01 ^b 0.57±0.00 ^b

Different superscripts in the same row mean significant difference ( p <0.05)

Claims (5)

(1) immersing brown rice of Hangaru variety dried at 60°C for 1 hour in water at 4°C for 24 hours, then taking it out and sterilizing it at 121°C for 15 minutes;
(2) cooling the sterilized brown rice of step (1) to 25° C., and adding 60% (v/w) water compared to the brown rice;
(3) 10% (v/w) inoculation of a red yeast culture solution having a dry cell weight of 3.6 mg/mL into the brown rice to which the water of step (2) has been added, followed by culturing at 30° C. for 7 days; and
(4) sterilizing the cultured red yeast rice in step (3) at 121°C for 15 minutes, followed by drying at 60°C so that the water content becomes 10% (v/w) , cycloartenyl ferulate, 2,4-methylenecycloartanyl ferulate, campesteryl ferulate, sitosteryl ferulate and A method for producing red yeast rice with enhanced γ-oryzanol content, including campestanyl ferulate. delete delete delete (1) immersing brown rice of Hangaru variety dried at 60°C for 1 hour in water at 4°C for 24 hours, then taking it out and sterilizing it at 121°C for 15 minutes;
(2) cooling the sterilized brown rice of step (1) to 25° C., and adding 60% (v/w) water compared to the brown rice;
(3) 10% (v/w) inoculation of a red yeast culture solution having a dry cell weight of 3.6 mg/mL into the brown rice to which the water of step (2) has been added, followed by culturing at 30° C. for 7 days; and
(4) Cycloartenyl of Red Guk Rice, including the step of sterilizing the cultured red yeast rice in step (3) at 121°C for 15 minutes and then drying it at 60°C so that the moisture content becomes 10% (v/w) cycloartenyl ferulate, 2,4-methylenecycloartanyl ferulate, campesteryl ferulate, sitosteryl ferulate and campestenyl ferulate A method for enhancing γ-oryzanol content comprising campestanyl ferulate.

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