KR20190009879A - Producing method of rice bran extract with increased arabinoxylan content - Google Patents

Abstract

The present invention relates to a method of manufacturing a rice bran extract with increased arabinozylan content by using a strain of Penicillium roqueforti. The rice bran extract of the present invention has increased arabinoxylan content compared to a rice bran extract obtained by a traditional extraction method, and thus can be more advantageously used for a health functional food.

Description

Technical Field [0001] The present invention relates to a method for producing rice bran extracts with increased arabinoxylan content,

The present invention relates to a method for producing an extract of rice bran with increased arabinozylan content by using a strain of Penicillium roqueforti.

Rice bran is a by-product of rice bran and rice husk as a by-product of the rice milling process. Most of the nutrients in rice are known to be contained in rice bran. More specifically, the rice bran contains an abundant amount of lipid, vitamin B group and high-quality protein, and it is known that it contains abundant minerals such as fiber and phosphorus. However, despite the excellent nutritional value as described above, rice bran is used only as rice bran oil, feed and fertilizer, or is treated as agricultural waste, and therefore research on a wider industrial use method of rice bran is required.

Recently, it has been reported that a physiologically active substance having physiological activity such as immunostimulating activity, antidiabetic activity or anticancer activity is present in the rice germ. One of the physiologically active substances contained in the rice bran, arabinoxylan is a mixture of arabinose and xylose produced by decomposing hemicellulose.

The hemicellulose material may be selected from the group consisting of xylose, xylose, rhamnose, galactose, mannose, mannose, mannose, ) Or glucose (glucose).

The arabinoxylan is a component of hemicellulose, which can be obtained from the hemicellulose and has been reported to have various physiological activities as described above.

As described above, the rice bran contains a large amount of substances having excellent physiological activity, but the physiologically active substances contained in the rice bran are present in an insoluble form strongly bound to plant cell walls, . Therefore, in order to use the physiologically active substance in the rice gut, it is necessary to increase the bioavailability so that the physiologically active substance can be used in the living body.

In this connection, Korean Patent No. 0379587 discloses a method for producing a physiologically active substance by extrusion molding and complex enzyme treatment. Korean Patent Laid-Open Publication No. 2004-0026917 discloses a method for producing arabinosilane by decomposing hemicellulose contained in barley bran with hemicellulase. Korean Patent No. 10-1345729 discloses a process for increasing the content of arabinoxylan by enzyme treatment. Korean Patent No. 10-1677022 discloses a process in which the purity and content of arabinoxylan are increased by treating alkali during the rice bran extraction process.

However, the prior art methods have been pointed out to have poor purity and yield of extracting arabinosyllan from rice gangue, no economical efficiency due to cost problems of enzymes, and problems with bioavailability of alkali. Therefore, there is a growing need for studies on the extraction process of rice bran extracted from Arabidopsis thaliana.

As a result of intensive efforts to solve the above-mentioned problems, the inventors of the present invention found that, when a biological conversion process is carried out using a strain of Penicillium roqueforti, arabinose And the bioavailability was high through animal experiments. Thus, the present invention was completed.

One object of the present invention is to provide a method for producing arabinofuranosyl-enhanced bovine corn extract, which comprises culturing a medium containing rice bran by inoculating a strain of Penicillium roqueforti will be.

Yet another object of the present invention is to provide a method for producing an extract of rice bran with an increased arabinoxylan content by further comprising a step of adding an acid during the culture to hydrolysis.

In one aspect of the present invention, the present invention provides a method for producing an Arabidopsis thaliana extract, comprising the step of inoculating a culture medium containing rice bran with a strain of Penicillium roqueforti, Of the present invention.

Hereinafter, a method for producing the rice bran extract with increased arabinoxylan content in the present invention will be described in detail.

The term " rice bran " used in the present invention refers to the husked portion of brown rice produced by removing rice bran in the course of rice cultivation. Grain clusters are generally based on the bran layer and sometimes contain germs during the production process. An object of the present invention is to increase the content of arabinoxylan in the preparation of rice bran extract from rice bran.

In the present invention, rice bran can be obtained through all the methods known in the art, and it is preferable to prepare a sample by sterilizing and heat-treating the raw material of rice bran. Specifically, a part of the rice bran samples can be separated and used. In an embodiment of the present invention, after three layers of mash NO. 7, 10 and 20 are separated, the husks isolated in mash NO. 10 are sampled Respectively.

The term " arabinozaylan " used in the present invention is one of the physiologically active substances and is composed of arabinose and xylose prepared by decomposing hemicellulose. Arabinosaylan having arabinose in its side chain with xylose as a main chain is a structure in which arabinose, a pentabody, is partially bound to a polymer composed of xylose with an alpha - (1-4) bond.

The arabinoxylan of the present invention can be used in various forms such as arabinozylan in its own form, an extract in which the arabinozoyllan is dissolved in the liquid, or a dried product in which the arabinoxylan is dissolved.

In the present invention, Penicillium roqueforti strain was used to increase the content of arabinoxylan in the preparation of rice bran extract from rice bran. Specifically, arabinozylan can be contained in a high content by conducting bioconversion by inoculating a culture containing a sample of rice bran with a strain of Penicillium roqueforti. have.

The rice bran extract with increased arabinoxylan production according to the present invention can be mixed with the term 'arabinoxylan high content rice bran extract' or 'arabinozylan high rice bran extract.'

The extract of rice bran produced by the production method of the present invention may have an increase in the content of arabinoxylan compared to the rice bran extract prepared without inoculating the strain. In addition, the content of arabinoxylan may be increased compared with the case of inoculation with the comparative strain Pleurotus ostreatus or Aspergillus oryzae strain.

In one embodiment of the present invention, the extract of rice bran was prepared by inoculating various strains and the content of each arabinoxylan was determined. As a result, it was confirmed that the content of arabinoxylan was the highest when treated with penicillium roqueforti (Table 4).

At this time, the content of arabinoxylan may be 2% or more, specifically 2.5% or more, more specifically 2.9% or more. At this time, the content of arabinose may be 1% or more, specifically 1.3% or more, more specifically 1.5% or more. The content of xylose may be 1% or more, specifically 1.5% or more, more specifically 1.8% or more.

In another embodiment of the present invention, the content of arabinose-ylan was compared with that of the rice bran extract prepared without inoculation of the strain. As a result, the content of arabinose-ylan in the rice bran extract prepared by the method of the present invention (Fig. 2).

In the present invention, it is possible to further include a step of adding an acid during the culturing to hydrolysis. Further, it may further include a step of heating for hydrolysis after addition of the acid. The step of heating may further include cooling the reaction product after heating.

The acid may be a strong acid, specifically hydrochloric acid. The concentration of the hydrochloric acid is not particularly limited, but is preferably 5% to 10%, more specifically 5% hydrochloric acid.

In one embodiment of the present invention, only the acid treatment was carried out through a preliminary experiment before the inoculation of the strain to prepare the rice bran extract. As a result, it was found that the treatment with hydrochloric acid showed significantly increased arabinoxylan content, compared to the treatment with acetic acid. In particular, the treatment with 5% to 10% hydrochloric acid showed the highest level of arabinose (Table 2).

In the present invention, the medium for inoculating the strain of Penicillium roqueforti in the process of producing the rice bran extract is as long as it can achieve the object of the present invention for producing the rice bran extract with increased content of arabinoxylan from rice bran Any culture medium known in the art may be used without limitation. Specifically, in the step of biotransformation of rice bran of the present invention, the medium may be a potato extract medium (PDB), and the culture may be carried out at a temperature of 20 ° C to 30 ° C, specifically 22 ° C to 26 ° C, For 3 days to 5 days, more specifically for 4 days to 5 days, more specifically for 4 days in an agitation incubator.

On the other hand, as a final step for preparing the rice bran extract, it may further include adding an alkali solution for neutralization of the acidified medium by adding the acid. The alkali solution may be sodium hydroxide, potassium hydroxide, sodium carbonate or potassium carbonate, and may be sodium hydroxide, but is not limited thereto.

By the neutralization step, the pH can be adjusted to 6 to 7, specifically 6.5 to 6.8.

Further, it may further include a step of post-neutralization drying.

The drying step may be performed using conventional techniques known to those skilled in the art. For example, although not limited thereto, the solid content of the resultant hydrolyzate is dried through a drier, the liquid portion is sprayed onto a solid medium, After that, the dried extract can be finely ground using a mortar.

When the extract of rice bran with increased arabinoxylan content is prepared by the production method of the present invention, the bioavailability can be increased due to the availability of arabinoxylan, and it can be suitably applied to various foods and medicines.

According to the production method according to the present invention, arabinofuranoside can be provided with significantly increased rice bran extract. In addition, since the rice bran extract of the present invention has increased arabinoxylan content as compared with the rice bran extract obtained by the conventional extraction method, it can be more advantageously used for health functional foods.

Fig. 1 shows the process development process of the rice bran extract with increased arabinoxylan content.
Fig. 2 shows the contents and chromatograms of the rice bran extracts and arabinose-ylan increased rice bran extract.
FIG. 3 shows the results of forced swimming load test of rice bran extract with increased arabinofuranosid and arabinofoam.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrating the present invention, and the scope of the present invention is not limited to these examples.

Example  One. Arabi Noziran Increased Rice bran  For the preparation of the extract Rice bran Of the raw sample  Acid hydrolysis process

Rice bran was separated into three layers of mash NO.7, 10 and 20, and then rice bran extracted from mash NO.10 was used as a sample. 50 ml of acetic acid solution (25, 50, 75% acetic acid, 5, 10, 20% hydrochloric acid) having the same composition as that shown in Table 1 was added to each conical tube (Water bath) for 3 hours at 98 占 폚 for acetic acid and 90 占 폚 for hydrochloric acid. After heating, it was cooled at room temperature and the supernatant was separated. The supernatant was filtered, adjusted to pH 6.5-6.8 with 1.4 N sodium hydroxide, and concentrated at 50 < 0 > C.

NO. Experimental conditions Experiment 1 25% acetic acid treatment Experiment 2 50% acetic acid treatment Experiment group 3 75% acetic acid treatment Experiment group 4 5% hydrochloric acid treatment Experiment group 5 10% hydrochloric acid treatment Experiment group 6 20% hydrochloric acid treatment

Subsequently, the content of arabinoselanamine was analyzed by high performance liquid chromatography (HPLC, Shimadzu LC-20A HPLC instrument, Shimadzu, Japan) for the six concentrated extracts. The column used was a reversed phase column (Shimadzu VP? ODS, 250 L x 4.6, 5 μm) and a UV detector (245 nm) as the detector. The mobile phase used was 79% sodium phosphate buffer (40 mM) and 21% acetonitrile and the flow rate was 0.7 ml / min. The arabinose and xylose standards used in the experiments were used for the analysis using the derivative 1-phenyl-3-methyl-5-pyrazolone (PMP). 1 mg of each standard product was dissolved in water, and 100 μl of the solution was added. Then, 100 μl each of 0.3 M NaOH and 0.5 M PMP was added. The mixture was reacted at 70 ° C for 30 minutes, cooled, and then 100 μl of 0.3 M HCl was added to stop the reaction. 200 μl of chloroform was added, and the mixture was centrifuged. Then, 100 μl of the supernatant was collected, and this was repeated two more times. After filtration with 0.22 μm membrane, standard products and 10 μl of each sample were injected. The contents of arabinose and yilos were analyzed based on the contents of arabinose and xylose (Zheng, X., Li, L., & Wang, X. (2011) Molecules, 16 (4), 2743-2753.).

As a result, as shown in Table 2, it was confirmed that the content of arabinose and xylose was high in hydrochloric acid having a concentration of 5% and 10%, and 5% hydrochloric acid showed the highest content, In the subsequent experiments, 5% hydrochloric acid was used as the optimum acidic solution.

NO. Arabinose content (%) Xylose content (%) Arabian Nozawa Ilan ^a content (%) Experiment 1 0.035 0.004 0.034 Experiment 2 0.014 0.002 0.014 Experiment group 3 0.015 0.003 0.016 Experiment group 4 0.753 0.942 1.492 Experiment group 5 0.708 0.903 1.418 Experiment group 6 0.271 0.260 0.467

^a total arabinoxylan = 0.88 * (% arabinose +% xylose)

Example  2. Arabi Noziran Increased Rice bran  For the preparation of the extract Rice bran  Bioconversion process

Rice bran was separated into three layers of mash NO.7, 10 and 20, and then rice bran extracted from mash NO.10 was used as a sample. 10 g of rice bran was placed in a 250 ml Erlenmeyer flask, and 20 ml of potato extract medium (PDB) was added thereto and sterilized by autoclave using a high pressure steam sterilizer. The finished rice bran culture broth was sufficiently cooled at room temperature and then cultured. The culture of 100 占 페 Petri dish, Pleurotus ostreatus (16855), Penicillium roqueforti (11269) and Aspergillus oryzae, 11896) were dispersed (Table 3). When the surface is completely strained, add 20 ml of hydrochloric acid (5%), heat it in a water bath at 100 ° C, and heat it for 6 hours. After sufficiently cooling at room temperature, 20 ml of sodium hydroxide (1.4 N) was added to neutralize the mixture, the pH was adjusted to 6.5 to 6.8, and the mixture was filtered. The solid was transferred to a Petri dish and dried in a dryer (55 ° C). The liquid portion was appropriately concentrated and then sprayed onto a solid medium of a Petri dish and dried. The dried extract was finely ground with a mortar and then weighed and the yield was determined and stored at -20 ° C. The dried extract was analyzed for the content of arabinosyllan using the HPLC assay described in Example 1. [

NO. Experimental conditions Experiment group 7 The strain Flurotus ostreatus ( Pleurotus ostreatus treatment + 5% hydrochloric acid treatment Experiment group 8 The strain Penicillium roqueforti ( Penicillium roqueforti ) + 5% hydrochloric acid treatment Experiment group 9 Strain Aspergillus oryzae treatment + 5% hydrochloric acid treatment

As a result, as shown in Table 4, the content of arabinose and xylose was the highest in the experimental group 8 using Penicillium roqueforti (11269) strain. From these results, it was confirmed that the use of the bioconversion process by the strain of Penicillium roqueforti is most suitable for increasing the arabinoselan content.

NO. Arabinose content (%) Xylose content (%) Arabian Nozawa Ilan ^a content (%) Experiment group 7 0.980 1.030 1.769 Experiment group 8 1.510 1.810 2.922 Experiment group 9 1.130 1.050 1.918

^a total arabinoxylan = 0.88 * (% arabinose +% xylose)

Example  3. Arabi Noziran Increased Rice bran  Extract and existing Rice bran  Comparison of the content of arabinozylan in extracts

In order to confirm that the content of arabinoxylan of the present invention was increased, the content of arabinose-ylan was compared with that of the conventional rice bran extract and the rice bran extract prepared by the production method of the present invention.

As a comparative group, rice bran was separated into three layers of mash NO.7, 10 and 20, and then rice bran extracted from mash NO.10 was used as a sample. One gram of rice bran was put into a conical tube, and 50 ml of distilled water was added thereto. The mixture was heated to 90 ° C and then concentrated at 50 ° C to prepare a rice bran extract.

The above extract of rice bran was analyzed for the content of arabinose and arabinose and xylose by the HPLC method described in Example 1.

As a result, it was confirmed that the content of arabinose and xylose in the rice bran extract, which had increased arabinoxylan compared to rice bran extract, was remarkably high, and it was confirmed that the production method of the present invention significantly increased the content of arabinoselan (Fig. 2).

Example  4. Arabi Noziran Increased Rice bran  Forced swimming load test of extract

The forced swimming load test is an experiment in which when the animal is forced to swim, it measures the time that the animal remains immobile without movement. This experiment is an in vivo experiment using an experimental animal. It can be used to verify the effect of immunity enhancement by improving physical strength or tolerance, and it is a good model to grasp the effect of actual immune enhancement by observing changes in various body fluid components such as fatigue related substances . The decrease in immobility time is interpreted as a result of the immunity enhancement effect.

Seven-week-old rats (ICR mice) were placed in a circular cylinder at a temperature of 23-25 ° C and a height of 16 cm and observed for 6 minutes. The pre-swimming test was conducted for 6 minutes before oral administration. Afterwards, 100 mg / kg of arabinozoyllane was orally administered to the mice, and then the mice were placed in a circular cylinder for 24 hours. After 2 minutes of observation 6 minutes and then 4 minutes of immobility time, analysis was performed.

As a result, as shown in FIG. 3, the floating time of the rice bran extract-treated group produced by the production method of the present invention was significantly decreased as compared with the rice bran extract-treated rice group. Therefore, it was confirmed that the rice bran extract prepared by the production method of the present invention is superior to the rice bran extract (FIG. 3).

Formulation example  1. Preparation of tablets

Arabidopsis thaliana Extract 200 mg

Lactose 100 mg

Starch 100 mg

Magnesium stearate Suitable amount

The above components were mixed and tableted according to a conventional tablet preparation method to prepare tablets.

Formulation example  2. Liquid  Produce

Arabidopsis thaliana Extract 1000 mg

CMC-Na 20 g

Isomer 20 g

Lemon incense Suitable amount

Purified water was added to adjust the total volume to 1000 ml. The above components were mixed according to a conventional method for producing a liquid agent, and then filled in a brown bottle and sterilized to prepare a liquid agent.

Formulation example  3. Preparation of capsules

Arabidopsis thaliana Extract 300 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components were mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

Formulation example  4. Preparation of injections

Arabidopsis thaliana Extract 300 mg

Mannitol 180 mg

Sterile sterilized water for injection 2974 mg

Na2HPO412H2O 26 mg

(2 ml) per ampoule according to the usual injection preparation method.

Claims (8)

In a medium containing rice bran, Penicillium roqueforti ) inoculated with a strain of Arabidopsis thaliana.
The method according to claim 1, further comprising the step of adding an acid during the culture to hydrolyze.
3. The method of claim 2, further comprising the step of heating for hydrolysis after addition of said acid.
4. The method of claim 3, further comprising neutralizing and drying.
The process according to claim 2, wherein the acid is hydrochloric acid.
The process according to claim 5, wherein the hydrochloric acid has a concentration of 5% to 10%.
2. The method according to claim 1, wherein the arabinoxylan is composed of arabinose and xylose.
The method according to claim 1, wherein the rice bran extract has an increased content of arabinose triamine compared to rice bran extract prepared without inoculating the strain.

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