KR102559546B1 - Method for enhancing the content of functional ingredients in seeds - Google Patents

Abstract

The present invention relates to a method of increasing the content of a desired functional component contained in a sprout crop through plasma treatment during seed germination, a method of manufacturing a sprout crop having an increased functional component content, and a sprout crop produced by the method. If this is used, it can be widely used in the development of functional health foods.

Description

Method for enhancing the content of functional ingredients in seeds}

The present invention relates to a method of increasing the content of a desired functional component contained in a sprout crop through plasma treatment during seed germination, a method of manufacturing a sprout crop having an increased functional component content, and a sprout crop produced by the method.

Plasma is referred to as the fourth state of matter after solid, liquid, and gas according to the size of the internal energy of the particles constituting the material. When electrical energy is applied to an atom or molecule in a gaseous state, the nucleus composed of electrically positive charge and the electrons distributed around it are separated, resulting in a gaseous state composed of ions and electrons. In the plasma created at an ultra-high temperature, molecules are completely dissociated and become atoms, and all electrons bound to the atoms are released from electron orbits as free electrons, resulting in a gaseous state composed of ions and electrons. However, when the plasma temperature is not high, highly reactive chemical species (radicals) are generated from incompletely dissociated molecules, incompletely ionized ion molecules, or ion atoms. That is, in the case of low-temperature plasma, since it is complexly composed of a wide variety of chemical species including ions and electrons, the applicable fields are very wide.

Plasma has been initially applied mainly to surface treatment or coating, and has been developed along with the growth of the semiconductor and display industries. More than 70% of ultra-high-density semiconductor manufacturing processes are performed by plasma processes. However, in the semiconductor and display fields, the current plasma technology has matured and saturated, and the plasma technology has moved in search of new applications. In other words, plasma application fields have been expanded to new energy production fields such as solar cells and low-grade coal gasification, environmental improvement fields such as air and water purification, and the like. Recently, interest in plasma application technology is increasing in the agricultural and food field.

The inventors of the present invention have introduced plasma technology to improve the functional component content and production efficiency of sprout crops, and as a result, the functional component content of sprout crops can be increased and the desired specific functional component content can be increased by adjusting plasma treatment conditions. The present invention was completed.

An object of the present invention is to provide a method for enhancing the functional component content of sprout crops through plasma treatment.

Another object of the present invention is to provide a method for producing sprout crops with enhanced content of functional components through plasma treatment.

Another object of the present invention is to provide sprout crops with enhanced functional component content prepared by the above production method.

A first aspect of the present invention is a first step of immersing the seeds in water to produce seeds at an early stage of germination; And a second step of cultivating sprouts by treating the seed in the early stage of germination with plasma, wherein the plasma has an output of 40 W to 60 W, and the atmospheric pressure plasma treatment is carried out in the range of 4 minutes to 8 minutes per time. It provides a method for enhancing the functional component content of sprout crops.

The present invention is characterized in that the content of the desired functional component among the functional components contained in sprout crops can be adjusted by controlling the number of times of plasma treatment.

The seed of the present invention may be any one selected from the group consisting of rice, barley, oats and wheat.

The term "sprout crop" of the present invention refers to a crop in which leaves and roots can be harvested after about 7 to 30 days after seed germination.

The term “functional component” of the present invention refers to a component containing “functionality” contained in the seed. The functional component of the present invention is specifically at least one selected from the group consisting of GABA, avenacoside a and avenacoside b.

The term of the present invention, "GABA (γ-aminobutyric acid)" is a kind of non-proteinaceous amino acid distributed in nature and is an essential component of the central nervous system such as the brain or spinal cord, and is known as a major inhibitory neurotransmitter. GABA is involved in many physiological regulation. In the case of animals, it activates blood flow to the brain, increases oxygen supply to the brain, promotes metabolic function of brain cells, reduces neutral fat in the blood and improves liver function, which is effective in preventing adult diseases such as hypertension, and has a nerve stabilizing function for pain relief, depression, and anxiety. In the case of modern people suffering from stress, especially students, excessive alcohol consumption, etc., it is reported that the concentration of GABA in the brain and blood is low, and the extreme lack of concentration in the body of GABA causes seizures, convulsions, and epilepsy. Due to this physiological action, GABA is used as an intravenous injection to improve cerebral blood flow and promote brain metabolism in stroke, head trauma, and cerebral artery sequelae, and is also sold as an oral prescription drug.

As used herein, "Avenacoside" is a steroidal saponin present in grains (particularly oats) or leaves. Abenacoside a is known to have a cholesterol-lowering effect, and avenacoside b is known to have detoxification and antifungal activity.

The first step of the present invention refers to the step of generating seeds in the early stages of germination.

Specifically, the seed may be immersed in tap water at a temperature of 20 ° C to 30 ° C in about 4 to 5 times the weight of the seed for about 16 to 24 hours. The initial germination rate of seeds may be the highest in the immersion conditions.

Next, the second step of the present invention means a step of increasing the functional component content of the seed through plasma treatment of the seed in the early stage of germination.

The term "plasma" is generated by applying energy to a gas, and the applied energy includes heat, current, and electromagnetic radiation. The present invention may use plasma generated by electrical energy.

The present invention is to treat the surface of the seed using the plasma, specifically atmospheric pressure plasma. One surface or several surfaces of the seed surface may be treated, and preferably, a better effect can be obtained than when two or more surfaces are treated at the same time. The atmospheric pressure plasma uses various methods of electric discharge under atmospheric pressure, but the intensity of electron energy is maintained higher than that of ions and neutral particles, that is, the plasma is in a non-equilibrium state. While the temperature of the entire plasma is room temperature to 1000K, the temperature of electrons is maintained at a temperature of 10,000K to 100,000K, enabling discharge suitable for various plasma chemical reactions and surface treatment applications.

As a method of implementing the plasma, there are technologies such as dielectric barrier discharge (DBD), corona discharge, microwave discharge, and arc discharge. Among them, dielectric barrier discharge is suitable for surface modification of low melting point materials as it can realize the concentration of reactive active species (radicals) 100 to 1000 times higher than conventional vacuum plasma, but the temperature is low to room temperature ~ 150 ℃. In addition, there is an advantage in that it is stable because there is no emission of harmful gases or substances in terms of environment.

In addition, the plasma treatment may mean direct plasma treatment of initially germinating seeds in a wet state after being immersed, or direct plasma treatment of initially germinating seeds in a dry state after immersion.

The plasma can be used as a source of stimulating the metabolism of seeds and can remove harmful substances on the surface. Therefore, it is possible to increase the content of the functional component, which is a metabolite of the seed.

The plasma may be generated by a power supply having a frequency of 10 KHz to 50 KHz.

The power supply output may be 40 W to 60 W. Plasma treatment under the power condition may be performed for 4 minutes to 8 minutes per time, specifically about 6 minutes. If the treatment time is less than 4 minutes, the functional component is not sufficiently increased, and if the treatment time is more than 10 minutes, there is a risk of damage to the seeds due to heat and gas.

As the plasma discharge gas, an inert gas such as argon or helium or nitrogen may be used, and nitrogen gas is preferably used. In the case of using the nitrogen gas, there is an advantage in that process costs are reduced at a low price.

The plasma treatment may be performed 1 to 5 times, specifically 1 to 3 times, and more specifically, 1 or 2 times. The content of the functional component may be different depending on the number of plasma treatments.

Specifically, when the plasma is treated twice or more, it may be treated once a day. This prevents the temperature of the germinated seeds from rising due to continuous plasma treatment and prevents the metabolism of the germinated seeds from being inhibited by setting a constant time interval of about 22 to 26 hours.

The present invention is characterized in that, when the plasma is treated once, the contents of γ-aminobutyric acid (GABA), avenacoside a, and avenacoside b extracted from sprout crop leaves are higher than when the plasma is treated twice or more times.

The increase in the GABA content is because when the content of glutamic acid and alanine is increased by treating the plasma, glutamic acid decarboxylase (GAD) is activated to increase GABA production. However, since the increase in the contents of glutamic acid and alanine is the highest when plasma treatment is performed once, the GABA content also shows the maximum value when plasma treatment is performed once.

It can be inferred that the increase in the contents of avenacosides a and b is because germinated seeds recognize plasma treatment as an external stress and produce avenacosides as metabolites to counteract it.

A second aspect of the present invention includes a first step of immersing seeds in water to produce seeds in an early stage of germination; And a second step of cultivating sprouts by treating the seed in the early stage of germination with plasma, wherein the power of the plasma is 40 W to 60 W, and the plasma treatment is carried out in the range of 4 minutes to 10 minutes per time. It provides a method for producing sprout crops with enhanced functional component content.

The present invention is characterized by providing a method for manufacturing sprout crops with increased functional component content while enhancing the functional component content of sprout crops.

The “seed”, “sprout crop”, “functional component” and “plasma” are as described above in the first aspect.

A third aspect of the present invention provides a sprout crop with increased functional component content prepared by the method of the second aspect.

1 to 3, the functional component is specifically at least one selected from the group consisting of GABA, avenacoside a, and avenacoside b, and the production method of the second aspect of the present invention is used. It is possible to provide sprout crops with increased content of the functional component.

The “seed”, “sprout crop” and “plasma” are as described above in the first aspect.

According to the plasma treatment method of the present invention, the GABA content of sprout crops can be increased.

According to the plasma treatment method of the present invention, the content of avenacoside in sprout crops can be increased.

According to the plasma treatment method of the present invention, it is possible to increase the policosanol content of sprout crops.

According to an embodiment of the present invention, it is possible to select a suitable plasma treatment method in order to increase the desired functional component content of sprout crops.

1 is a bar graph showing the GABA content of oat sprout leaves prepared in one embodiment of the present invention according to the presence or absence of plasma treatment and the number of treatments.
Figure 2 is a bar graph showing the avenacoside a content of oat sprout leaves prepared in one embodiment of the present invention according to the presence or absence of plasma treatment and the number of treatments.
Figure 3 is a bar graph showing the avenacoside b content of oat sprout leaves prepared in one embodiment of the present invention according to the presence or absence of plasma treatment and the number of treatments.

Hereinafter, the present invention will be described in more detail through examples of the present invention. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited to these examples.

comparative example. Cultivation of oat sprouts without plasma treatment

After immersing the sea oat seeds for about 20 hours, the water was drained off, 2 sheets of filter paper were laid in a petri dish (12 × 12 cm), 25 mL of distilled water was poured, and 17 g of the soaked sea oat seeds were evenly spread and grown in an incubator at 15 ° C for 9 days. On the 9th day, the growth was investigated, and the leaves of oat sprouts were separated, freeze-dried, and used for component analysis.

Example 1. Cultivation of oat sprouts through plasma treatment

Oat seeds were immersed for 20 hours and treated with plasma for 3 days at the beginning of germination. Plasma treatment used Surface Dielectric Barrier Discharge (SDBD), plasma output was 51.7 W, frequency was 14.4 KHz, and treatment was performed at normal pressure and room temperature. That is, after laying 2 sheets of filter paper on a petri dish and pouring 10 mL of distilled water, the sea oat seeds were evenly spread and treated for 6 minutes. Plasma treatment was performed for 6 minutes each on the 1st (T-1), 2nd (T-2), and 3rd (T-3) days while cultivating in a 15° C. incubator for 9 days. On the 9th day, the growth was investigated, and the leaves of oat sprouts were separated, freeze-dried, and used for component analysis.

Example 2. Analysis of amino acid and GABA content in oat sprouts

GABA and amino acid contents of the sprout powder prepared in Example 1 were measured using an AccQ·Tag UPLC (Waters, Milford, MA) analysis system based on the method of Tahashi et al. (13). Sprout powder was filtered using a membrane filter, and the filtered sample was derivatized using the AccQ-Tag method for UPLC analysis. The UPLC analysis system used waters Acquity, the column was AccQ Tag Ultra Amino Acid Analysis column (3.9 X 150 mm; waters), the column temperature was 49 ℃, the sample temperature was 20 ℃, the detector was FLR Detector (waters) and the wavelength was EX. 266 nm, EM. It was 473 nm. Mobile phase A was 100% AccQ Tag ultra UPLC amino acid analysis eluent A, B was 10% AccQ Tag ulta UPLC amino acid analysis eluent B, C was 100% distilled water, and D was 100% AccQ Tag UPLC amino acid analysis eluent B. . Chromatogram data was analyzed using Empower personal software (waters), and amino acid content analysis was analyzed using waters amino acids standards (waters).

amino acid
mg/100g LT-con LT-1 LT-2 LT-3 his 8.0±4.2 10.1±4.7 6.5±0.6 6.5±1.0 ser 26.6±9.2 39.0±14.5 27.0±1.5 26.9±2.7 arg 121.2±34.3 169.8±49.7 131.3±6.0 96.1±6.1 gly 10.5±3.8 10.9±3.7 7.9±0.4 10.5±1.0 asp 10.2±1.7 12.5±3.6 12.1±0.6 11.9±1.7 glu 37.1±20.6 69.0±32.5 40.4±3.4 30.2±4.8 thr 16.4±7.1 26.0±11.7 16.4±1.2 17.6±2.6 ala 39.9±12.1 59.4±21.9 41.0±2.3 42.8±4.1 pro 20.8±10.0 31.1±12.5 23.3±1.4 23.3±2.7 cys 8.3±2.6 13.5±4.8 9.7±0.7 7.3±1.3 lys 22.9±11.7 56.2±28.6 32.6±2.4 31.4±4.6 tyr 260.7±66.1 305.8±86.8 240.7±7.0 303.9±20.9 met 14.0±6.3 22.2±9.5 14.7±1.1 12.7±1.7 val 54.1±17.0 77.9±27.0 55.7±2.7 53.3±4.7 ile 15.7±8.2 27.4±14.3 15.1±1.2 16.4±2.3 leu 39.1±14.4 57.3±22.2 38.5±2.1 40.1±4.2 phe 19.6±6.7 22.5±7.7 16.6±0.9 18.8±1.9 Sum 724.9 1010.5 729.3 749.5

In Table 1, LT-con, LT-1, LT-2, and LT-3 refer to oat sprout leaves treated with no plasma treatment (LT-CON), one plasma treatment (LT-1), two plasma treatments (LT-2), and three plasma treatments (LT-3).

Table 1 summarizes the amino acid content of leaves per 100 g of dry weight of oat sprouts according to the number of plasma treatments. In particular, it was confirmed that the contents of glutamic acid (glu) and alanine (ala) increased by about 1.5 to 2 times with one plasma treatment. Elevated levels of glutamic acid and alanine can activate glutamic acid decarboxylase (GAD) and increase GABA production.

As shown in Figure 1, the GABA content of the oat sprouts grown in the manner of Example 1 was analyzed. As a result, it was confirmed that the GABA content of LT-1, LT-2, and LT-3 treated with plasma increased compared to the GABA content of oat sprout leaves without plasma treatment. In particular, it was confirmed that the GABA content was the highest when plasma was treated once, in the same manner as the increase in the content of glutamic acid and alanine.

Example 3. Analysis of avenacoside content in oat sprouts

The sprout powder prepared in Example 1 was extracted by stirring with 20 times methanol for 5 hours, filtered using a membrane filter, and then UPLC analysis was performed. UPLC analysis was performed using a Waters UPLC detector using an ELSD detector, and analysis conditions are summarized in Table 2 below.

HPLC Waters UPLC system column Halo C18 2.1 x 100 mm 2.7 μm column temperature 35℃ sample temperature 15℃ menstruum A: 0.1% formic acid dissolved in distilled water B: acetonitrile (ACN) Injection 2 μl (Thermo 1.3 μl) Gradient time (min) flow (ml) %A %B 0 0.5 97 3 3 0.5 97 3 10 0.5 85 15 15 0.5 70 30 20 0.5 50 50 30 0.5 10 90 37 0.5 10 90 39.9 0.5 97 3 40 Solvent flow stop (stop run) Detector * ELS detector (Gain 300, gas pressure 40.0 psi, drift tube 60 ± 5℃),
*CAD (Data collect 10, Filter 10.0, Temperature high)

As shown in Figure 2, the change in the content of avenacosides a and b in oat sprout leaves according to the number of plasma treatments was analyzed. Through this, it was confirmed that both avenacosides a and b had the highest rate of increase in the content of LT-1.

Claims (8)

A first step of immersing oat seeds in water to produce seeds at an early stage of germination; and
A second step of cultivating oat sprouts by treating the germinated seeds with plasma;
The power of the plasma is 40 W to 60 W,
The plasma treatment is a one-time treatment,
The plasma treatment is performed in the range of 4 to 10 minutes, a method for enhancing the content of functional components of oat sprouts. According to claim 1,
The functional component is at least one selected from the group consisting of GABA, avenacoside a and avenacoside b, a method for enhancing the functional component content of oat sprouts. delete delete According to claim 1,
The plasma treatment is a method for enhancing the functional component content of oat sprouts, which is treated once per day. According to claim 1,
When the plasma is treated once, the content of GABA (γ-aminobutyric acid), avenacoside a, and avenacoside b extracted from oat sprout leaves is higher than when treated twice or more. A first step of immersing oat seeds in water to produce seeds at an early stage of germination; and
A second step of cultivating oat sprouts by treating the germinated seeds with plasma;
The power of the plasma is 40 W to 60 W,
The plasma treatment is a one-time treatment,
The plasma treatment is performed in the range of 4 to 10 minutes, a method for producing oat sprouts with increased functional component content. Oat sprouts with increased functional ingredient content, prepared by the method of claim 7.