KR101732729B1 - Process for preparing fermented white ginseng extrudate containing Compound-K - Google Patents
Process for preparing fermented white ginseng extrudate containing Compound-K Download PDFInfo
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- KR101732729B1 KR101732729B1 KR1020150137026A KR20150137026A KR101732729B1 KR 101732729 B1 KR101732729 B1 KR 101732729B1 KR 1020150137026 A KR1020150137026 A KR 1020150137026A KR 20150137026 A KR20150137026 A KR 20150137026A KR 101732729 B1 KR101732729 B1 KR 101732729B1
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- white ginseng
- ginseng
- extrudate
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/06—Enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P30/00—Shaping or working of foodstuffs characterised by the process or apparatus
- A23P30/20—Extruding
-
- A23Y2220/37—
-
- A23Y2260/35—
Abstract
(A) obtaining white ginseng extrudate by extrusion molding white ginseng; (b) reacting the white ginseng extrudate obtained in step (a) with a carbohydrase to obtain an enzyme-treated product; And (c) inoculating the enzyme-treated product obtained in step (b) with a fermentation strain to obtain a fermented product, and to provide a fermented product of white ginseng extruded fermented product produced thereby .
The method of the present invention for producing a fermented product of white ginseng extract can be effectively used as a processing method of ginseng by achieving the effect of converting ginsenoside into ginsenoside, particularly, compound K, which is excellent in efficacy in ginseng such as white ginseng.
Description
The present invention relates to a method for producing fermented white ginseng extrudate, and more particularly, to a fermented fermented product obtained by extrusion molding, carbohydrase treatment and fermentation.
Ginseng (Panax ginseng C. A. Meyer) is a perennial semi-synthetic osteopteran belonging to the plant of Panax ginseng (Araliaceae). It has been used for thousands of years as a traditional medicinal herb with health promotion function in Korea and other Asian countries. The ginseng root is composed of white sap, red ginseng, and tae guk gins (Kim & Ryu, 2005. J Korean Soc Food Sci Nutr 34: 544-548) The main components of the ginsenosides are the polysaccharides, the nitrogenous compounds, the free sugars, the free acids, the vitamins and the inorganic components, among which the ginsenosides have important pharmacological effects (Lee et al. 2004. Korean J Med Crop Sci 12: 237-242, Jeon et al. 2005. J Ginseng Res 29: 138-144).
Saponin is one of the main substances showing the pharmacological effects of ginseng. Up to now, about 30 kinds of saponin have been isolated from ginseng and its chemical structure has been identified and named ginsenoside. Among the saponins of ginseng, it has been found that saponin produced by sugar hydrolysis of a major component saponin exhibits an excellent effect on absorption and pharmacological effects as compared with saponin as a main component (Jae-Gap Han, The Korean Society for Microbiology and Biotechnology).
Rg2, Rg3, Rh2, and Compound K, which are present in trace amounts in ginseng, are known to inhibit cancer cell metastasis and inhibit platelet aggregation. Ginseng is known as a substance having physiological activity such as action, and ginseng is used as food or medicine. As various effects of ginsenoside have been revealed, researches for converting ginsenoside contained in ginseng into ginsenosides having excellent efficacy have been actively conducted through various treatments (acid, heat, microorganism) . In particular, the fermentation of ginseng using lactic acid bacteria can supply a useful living microorganism as a probiotic, and by decomposing the sugar of ginsenoside, which is a glycoside structure, to change the structure of ginsenoside, (Kim et al. 2007. Korean J Microbiol, 43, 142-146), which has higher absorption efficiency in vivo than saponin.
On the other hand, the extrusion molding process controls pressure, viscosity, non-mechanical energy, etc., which are dependent variables, by changing independent variables such as moisture content, screw rotation speed, barrel temperature, sample input amount, (Gu & Ryu, 2011. J Food Eng Prog 15: 148-154). In addition, the extrusion molding process is an efficient process that can perform each unit operation such as mixing, grinding, heating, molding, etc. in a relatively short time. Extrusion molding has been used in the production of pasta and the like since it has been used in foods since the mid 1930s and has recently been applied in a variety of fields such as polymer plastics, food, feed, bio-industry, and pharmaceutical industry (Anderson RA, Conway HF, Pfeifer VF, Griffin EL, 1969. Cereal Sci Today 14: 4-7, 11-12). In the food, the above-mentioned independent variables (moisture content, screw rotation speed, structure of the discharge port, screw arrangement, etc.) can be adjusted according to the characteristics of the product, so that products having various desired properties can be produced (Govindasamy S, Campanella OH, Oates CG. 1997. J. Food Eng. 32: 403-426). It has been reported that the high temperature, high pressure and high shear force generated during the extrusion process efficiently change the molecular structure characteristics of the raw material. In the extrusion molding process, the composition of the raw materials and the process parameters are controlled, (Happer JM, 1989. AACC, St. Paul, MN, pp. 91-155).
In the study of extrusion molding of ginseng, the chemical composition and antioxidant activity of white ginseng extract by extrusion molding (Son & Ryu, 2009. J Korean Soc Food Sci Nutr 38: 946-950) (2, 2-diphenyl-1-picrylhydrazyl) measurement and saponin composition (measured by HPLC) were confirmed through the analysis of total phenol content, DPPH (DPPH) Especially, it was confirmed that Rg3s and Rg3r, which are peculiar components of red ginseng, are increased, so that the extrusion molding process can be used as a red ginseng change process of white ginseng.
(Han et al. 2007. J Food Eng Prog 11: 119-126), the paste viscosity, the water solubility index, the water absorption index and the alcohol fermentation efficiency of the fermentation broth of white ginseng, red ginseng and extruded white ginseng The white ginseng powder and extruded white ginseng powder showed the highest viscosity and the lowest viscosity was the lowest in white ginseng powder. When the ginseng was fermented by alcohol for 14 days, the pH was measured in the order of red ginseng fermentation liquid 4.03, extrusion fermentation liquid 3.24 and white ginseng fermentation liquid 3.12. The possibility of the development of ginseng fermentation products and materials was confirmed by extrusion molding. In addition, extrusion molding of white ginseng has been reported to have higher effects on temperature physicochemical properties and glycation characteristics than screw rotation speed, sample input amount and moisture content (Han et al. 2008. J Food Eng Prog 12: 36-43 ).
However, there is no report that the fermentation suitability and fermentation characteristics of white ginseng are influenced by the exit temperature of the extrusion process. Therefore, it is necessary to study the composition of white ginseng related to the extrusion process and fermentation.
The inventors of the present invention have found that a method of converting ginsenoside contained in ginseng such as white ginseng into ginsenoside having excellent efficacy such as Compound K which is known to have a physiological activity such as a cancer cell metastasis inhibiting action and a platelet aggregation inhibiting action , We found that ginsenosides such as compound K were detected in white ginseng by extrusion molding and carbohydrate decomposition and fermentation process.
Accordingly, it is an object of the present invention to provide a fermented product of extruded white ginseng, which comprises extrusion molding, carbohydrase degrading enzyme treatment and fermentation process, and a fermented product of white ginseng extruded by the fermentation.
According to one aspect of the present invention, there is provided a method for producing white ginseng, comprising: (a) extruding white ginseng at a discharge port temperature of 150 ° C to obtain a white ginseng extrudate; (b) reacting the white ginseng extrudate obtained in step (a) with a complex carbohydrate degrading enzyme comprising arabin hydrolytic enzyme, cellulase, beta-glucanase, hemicellulase and xylanase to obtain an enzyme-treated product; And (c) inoculating the enzyme-treated product obtained in step (b) with lactobacillus taxa to obtain a fermented processed product.
In one embodiment, the extrusion of step (a) may be performed at a discharge outlet temperature of 150 to 170 ° C, and the carbohydrase of step (b) may be an arabanase, a cellulase, , At least one enzyme selected from the group consisting of beta-glucanase, hemicellulase, xylanase, pectin lyase, and alpha-amylase. , And the fermentation strain of step (c) may be at least one strain selected from the group consisting of Lactobacillus gasseri and Leuconostoc mesenteroides subsp . Cremoris .
According to an aspect of the present invention, there is provided a method for producing white ginseng, comprising: (a) extruding white ginseng at a discharge port temperature of 150 ° C to obtain a white ginseng extrudate; (b) reacting the white ginseng extrudate obtained in step (a) with a complex carbohydrate degrading enzyme comprising arabin hydrolytic enzyme, cellulase, beta-glucanase, hemicellulase and xylanase to obtain an enzyme-treated product; And (c) inoculating the enzyme-treated product obtained in step (b) with lactobacillus taxa to obtain a fermented processed product.
Further, according to one aspect of the present invention, white germ extract extruded fermentations produced by the above production method are provided.
The ginsenosides Rg3s and Rg3r which were not detected in the white ginseng state were detected in the white ginseng extruded fermented product manufactured by the production method of the present invention. In the fermented product of white ginseng extruded through the carbohydrate decomposition process and the fermentation process, A compound K known as a substance having a physiological activity such as an inhibitory action and a platelet aggregation inhibitory action is detected and the ginsenoside contained in white ginseng according to the production method of the present invention is effectively used as a ginsenoside Respectively. Compound K was also found in extrudates at all temperatures of 150-170 ° C, which was extrusion-molded, but the most severe TLC results were obtained at 150 ° C. Furthermore, it was confirmed that the conversion of the compound K was most efficiently performed in the group treated with the fermentation strain, in particular, the lactobacillus taxa after treatment with the carbohydrase.
Therefore, the method for producing fermented product of white ginseng extrusion comprising extrusion molding, carbohydrase degrading enzyme treatment and fermentation process of the present invention has the effect of converting ginsenoside into ginsenoside having excellent efficacy in ginseng such as white ginseng, It can be used effectively as a processing method.
Figure 1 shows the screw arrangement used in the extrusion molding process of white ginseng.
FIG. 2 is a SEM photograph of white ginseng extruded at 150 ° C. (a), 160 ° C. (b), and 170 ° C. (c) by 150 × magnification of extruded white ginseng.
FIG. 3 is a graph showing the results of measurement of reducing sugar according to enzyme treatment time of white ginseng extrudate (A: 160 ° C EWG-V, B: 150 ° C EWG-V, C: 170 ° C EWG- W: WG-N, H: WG-N, I: 160 DEG C EWG-L, J: 170 DEG C EWG-N, E: 160 DEG C EWG- K: 150 占 폚 EWG-L, L: WG-L [WG: white ginseng, EWG: white ginseng extrudate, V: viscozyme L, N: Novozyme 33095, L:
4 is a graph showing the results of measurement of reducing sugar according to fermentation time using Lactobacillus gasseri (KCTC 3172) of white ginseng extrudate (A: 160 ° C EWG-V, B: 150 ° C EWG-V, C E: 170 DEG C EWG-V, D: WG-V, E: 150 DEG C EWG-N, F: 160 DEG C EWG-N, G: 170 DEG C EWG-N, H: WG- LW: WG, N: 150 DEG C EWG, O: 170 DEG C EWG, P: 160 DEG C EWG [WG: white ginseng, EWG: White ginseng extrudate, V: viscose L, N: Novozyme 33095, L: Liqui-Zyme supra)].
5 is a graph showing the results of measurement of reducing sugar according to fermentation time using Leuconostoc mesenteroides subsp. Cremoris (KCTC 3529) of white ginseng extrudate (A: 150 ° C EWG-V, B E: 160 DEG C EWG-N, G: WG-N, H: 170 DEG C EWG-N, F: 160 DEG C EWG-N, E: L: 150 DEG C EWG-L M: WG, N: 160 DEG C EWG, O: 150 DEG C EWG, L: 150 DEG C EWG-L, I: WG- EWG [WG: white ginseng, EWG: white ginseng extrudate, V: viscose L, N: Novozyme 33095, L: Liqui-Zyme supra]).
Figure 6 shows the results of TLC expansion of the butanol extraction supernatant by enzymatic hydrolysis and fermentation of extrudates at 150 ° C (a), 160 ° C (b), and 170 ° C (c) 2. White ginseng extruded product, 3. Viscose L-treated white ginseng extrudate, 4. Novozyme 39095 treated white ginseng extrudate, 5. Liqui-Zyme supra-treated white ginseng extrudate, 6. Viscozyme L-KCTC 3172 treated white ginseng extrudate, 7. Novozyme 39095-KCTC 3172 treated white ginseng extrudate, 8. Liquorice supra-KCTC 3172 treated white ginseng extrudate, 9. Viscozyme L-KCTC 3529 treated white ginseng extrudate, 10. Novozymes 39095-KCTC 3529 treated white ginseng extrusion Molded products, 11. Liquorice Supra-KCTC 3529 treated white ginseng extrudate, 12. F2, 13. Compound K).
(A) obtaining a white ginseng extrudate by extrusion molding white ginseng at a discharge port temperature of 150 ° C; (b) reacting the white ginseng extrudate obtained in step (a) with a complex carbohydrate degrading enzyme comprising arabin hydrolytic enzyme, cellulase, beta-glucanase, hemicellulase and xylanase to obtain an enzyme-treated product; And (c) inoculating the enzyme-treated product obtained in step (b) with lactobacillus taxa to obtain a fermented processed product.
In step (a), energy is applied to white ginseng at high temperature and high pressure to generate shearing force, thereby destroying the denseness of white ginseng tissue and treating the white ginseng so that elution of the components contained in white ginseng is facilitated. In step (a), the extrusion molding machine commonly used in the art can be used to carry out the process. For example, the screw diameter is 30.0 mm, the ratio of length to diameter (L / D ratio) is 23: 1, and the discharge port is And a diameter of 3 mm in a circular shape.
In the method for producing fermented white ginseng extrudate of the present invention, the extrusion molding of step (a) may be carried out at a temperature of 150 to 170 ° C, preferably 150 ° C. The extruded white ginseng obtained by extrusion molding can be subjected to post-treatment such as drying, crushing, and passing through a standard body to be used in the subsequent step.
Step (b) is a step of decomposing carbohydrates contained in white ginseng tissue by adding carbohydrase to the extrudate of white ginseng obtained by extrusion molding. In this step, the carbohydrate of the polymer contained in white ginseng is changed to a low molecular state by approaching carbohydrase to the dense white ginseng tissue obtained in the extrusion molding process.
The carbohydrase used in step (b) may be selected from the group consisting of arabanase, cellulase, beta-glucanase, hemicellulase, xylanase, pectin The enzyme may be at least one enzyme selected from the group consisting of pectin lyase and alpha-amylase, and preferably at least one enzyme selected from the group consisting of arabinhydase, cellulase, beta-glucanase, hemicellulase, It is a complex carbohydrate degrading enzyme comprising the kinase, and can be used commercially Visco atom TM L (Viscozyme TM L) sold by, Novozymes TM 33095 (Novozym TM 33095) or liquor atom TM Supra (Liquozyme TM Supra).
The carbohydrate decomposition process of step (b) can be suitably controlled by a person skilled in the art according to the state of the white ginseng extrudate to be degraded, and can be carried out, for example, by enzymatic reaction at 50 ° C for 1 hour or more, preferably 6 hours or more .
Step (c) is a step of inoculating and fermenting a fermentation strain into a carbohydrase-treated white ginseng extrudate.
The fermentation strains used in step (c) are selected from the group consisting of Lactobacillus gasseri and Leuconostoc mesenteroides species such as Leuconostoc mesenteroides subsp . cremoris ), and more preferably at least one strain selected from the group consisting of Lactobacillus cassia.
The fermentation process of step (c) can be suitably controlled by a person skilled in the art depending on the state of the fermented white ginseng extrudate. For example, the fermentation process is carried out at 27 to 37 ° C under anaerobic conditions for 1 hour or more, preferably 6 hours or more can do.
(A) obtaining white ginseng extrudate by extrusion molding white ginseng at a discharge port temperature of 150 ° C; (b) reacting the white ginseng extrudate obtained in step (a) with a complex carbohydrate degrading enzyme comprising arabin hydrolytic enzyme, cellulase, beta-glucanase, hemicellulase and xylanase to obtain an enzyme-treated product; And (c) inoculating the enzyme-treated product obtained in step (b) with lactobacillus taxa to obtain a fermented processed product.
The present invention also provides fermented white ginseng extrudates produced by the above method.
The ginsenosides Rg3s and Rg3r which were not detected in the white ginseng state were detected in the white ginseng fermented product manufactured by the above method. In the fermented product of white ginseng extruded through the carbohydrate decomposition process and the fermentation process, inhibition of cancer cell metastasis , Platelet aggregation inhibitory action, and the like. The compound K, which is known as a substance having a physiological activity, is detected. According to the production method of the present invention, the ginsenoside contained in white ginseng is converted into ginsenoside Respectively. The compound K was confirmed in the extrudates at all temperatures of 150 to 170 ° C. in which the extrusion molding was carried out, but the TLC was the most severe at 150 ° C. In addition, after confirming that the compound K was most effectively converted in the group treated with the fermentation strain, particularly the lactobacillus casei after the carbohydrase was treated, the fermented product of white ginseng extruded fermented product according to the present invention contained white ginseng Can be useful in converting ginsenosides into ginsenosides with excellent efficacy in ginseng.
Hereinafter, the present invention will be described in more detail by way of examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.
< Example >
1. Materials and reagents
The white ginseng powder was purchased from Dongmyung Ginseng Co., Seoul, Korea for 4 years and the enzymes were Viscozyme L, Novozym 33095 and Liquozyme Supra. The strains were selected from the group consisting of Lactobacillus gasseri [KCTC 3172], Leuconostoccepsin, Leuconostoc, mesenteroides subsp. cremoris [KCTC 3529]) were purchased from the Korean Center for Microbial Resources.
2. Extrusion molding
The screw diameter of the extruder was 30.0 mm and the ratio of length to diameter (L / D ratio) was 23: 1. The extruder was extruded using a biaxial extruder (THK 31T, Incheon Machinery Co., Incheon, Korea) , And a discharge port having a circular shape with a diameter of 3 mm was used, and the screw arrangement was as shown in Fig. A water content of 20%, a screw rotating speed of 200 rpm, a feed rate of 100 g / min, and extrusion temperature was 150 캜, 160 캜 and 170 캜. The extruded white ginseng samples were dried in a hot air dryer (DS-FCPO250, DongSeo Sci. Co., Seoul, Korea) at 50 ℃ for 6 hours. The obtained white ginseng extrudate was pulverized with a domestic pulverizer (FM-681, Hanil, Haman, Korea), and the powder passed through a 50 mesh standard (Chung-gye Sang-gong Co., Seoul, Korea) Water Soluble Index (WSI), Water Absorption Index (WAI), antioxidant activity by enzyme reaction or fermentation, reducing sugar level, and ginsenoside pattern including compound K were measured.
3. Enzyme reaction
The enzyme reaction was carried out as follows. After adding 30 mL of distilled water to 0.5 g of the sample, each enzyme (Viscozyme L, Novozymes 33095, Liquuzzyme Supra) was added at a ratio of 0.8%, and the reaction was allowed to proceed at 50 ° C at 150 rpm. The reaction was stopped in an autoclave (121 ° C, 15 minutes) after collecting a sample for 72 hours in an hour at the beginning of the reaction and at 6 hours after 6 hours. The antioxidative activity and reducing sugar were measured by TLC Ginsenoside pattern observation, and ginsenoside were measured.
4. Fermentation Process
The fermentation process was carried out as follows. After adding 30 mL of distilled water to 0.5 g of the sample, 1 mL of the strain (KCTC 3172 or KCTC 3529) was added to the enzyme-treated sample, and the sample was incubated at 27 to 37 ° C in anaerobic condition for 6 hours in total for 24 hours Were collected. The obtained samples were sterilized in an autoclave (121 ° C, 15 minutes), and then the antioxidative activity and reducing sugar were measured for the centrifuged samples of the fermentation filtrate, ginsenoside pattern observation by TLC, and ginsenoside were measured .
< Test Example >
One. Of non-mechanical energy Measure
The specific mechanical energy input (SME input) was performed according to the method described in the prior document (Patent Application No. 10-2015-0080656), and was expressed by the electric energy of the extruder consumed per unit mass of raw material (See equation 1). That is, the electric power applied to the actual raw material was obtained from the ratio between the electric power at the time of input of the raw material and the electric power at the idling of the motor and the production amount.
[Equation 1]
(However, SME input: the electrical energy input (kJ / kg), E: electric power (J / s), E 0 at the time of raw material input: Energy (J / s), P R at the time of motor idling: throughput rate (kg / s ).
2. Measurement of expansion rate
The diameter expansion ratio was determined by measuring the diameter of the
3. Measurement of water solubility index and water absorption index
In order to analyze the water-solubility of the sample, a water soluble index (WSI) and a water absorption index (WAI) were measured according to the method described in the prior art (Patent Application No. 10-2015-0080656) Respectively. 30 g of distilled water was added to 1.5 g of a dry weight standard sample, and the mixture was stirred in a constant temperature water bath (BF-45SB, Biofree Co., Seoul, Korea) for 30 minutes and centrifuged (H-1000-3, Hanil Science Industrial Co.). , Gangneung, Korea) at 3000 rpm for 20 minutes. The supernatant was poured into a weighing dish, and the weight of the precipitate was measured. The weighing dish was dried in a hot air drier (HB-502MP, Han Beak Co., Bucheon, Korea) at 105 ° C for 2 hours to measure the solid content of the supernatant, The water soluble index (WSI) and the water absorption index (WAI) were calculated by the following equations, respectively.
[Equation 2]
[Equation 3]
4. Chromaticity measurement of extrudate
The lightness, L, redness, a, and yellowness, b values were measured using a chromatic meter (Chroma Meter CR-300, Minolta Co., The maximum value and the minimum value were excluded one by one and the average value was shown. The values of the standard color plate were L = 98.07, a = 0.04, and b = 1.01.
5. Observation of microstructure of extrudate
The cross section of the extruded product was coated with platinum and the microstructure was observed with a high resolution scanning electron microscope (MIRA LMH, Tescan, Brno, Czech) at an accelerating power of 10 kV.
6. General composition measurement
The general components of the extruded samples were determined by the Association of Official Chemist (AOAC) method. The moisture was determined by the atmospheric pressure drying method at 105 ℃, the Soxhlet extraction method using crude fat, and the direct method using crude ash. Protein quantification was performed by the ninhydrin method as follows. After adding 3.5 mL of 6 N HCl to 0.5 g of sample, heat at 100 ° C for 24 hours and add 20 mL of distilled water. After centrifugation at 3000 rpm for 30 minutes, 5 mL of ninhydrin reagent was added to 0.15 mL of extract, reacted for 10 minutes at 100 ° C, and measured at 575 nm using a spectrophotometer. Standard curves were measured using gamma-globulin.
7. Reducing sugar measurement
The reducing sugar content was measured by the DNS method (3,5-dinitrosalicylic acid method) as follows. 3 mL of DNS (3,5-dinitrosalicylic acid) reagent was added to 1 mL of the fermentation broth and reacted at 100 ° C for 5 minutes, followed by cooling in ice water for 15 minutes. After 25 mL of the reaction solution was diluted with distilled water, absorbance was measured at 550 nm using a spectrophotometer (Libra S35, Biochrom Co., Cambridge, England). The standard curve of reducing sugar content was prepared using glucose.
8. Acid polysaccharide Measure
The acid polysaccharide was measured by 3-phenylphenol method. 5 mL of 0.0125 M sodium tetraborate / H 2 SO 4 was added to 1 mL of the fermentation broth and then heated in a constant temperature water bath (100 ° C.) for 5 minutes and cooled for 15 minutes. 1 mL of 0.15% 3-phenylphenol / 0.5% NaOH was added and measured at 520 nm using a spectrophotometer. The standard curve was measured at 0-100 ug / g using glacturonic acid.
9. Antioxidant activity measurement
To extract white ginseng and white ginseng extrudate, 6 mL of distilled water was added to 0.5 g of the sample, and 0.8% of each enzyme solution was added and reacted for 72 hours. 1 mL of lactic acid bacteria was added, fermented for 24 hours, added with 24 mL of ethanol, stirred at 100 rpm for 12 hours in a constant temperature water bath (30 ° C), and filtered to obtain total phenolic compound and DPPH radical scavenging ability .
The total phenolic content of the extract was determined by the Folin-Ciocalteu colorimetric method as follows. (1.5 mL, 2N [10-fold dilution]) and 10% Na 2 CO 3 (1.5 mL) were added to the extract (0.1 mL) and reacted in a dark room at room temperature for 60 minutes. Absorbance values were measured at 765 nm using a spectrophotometer. The standard curve used to calculate the total phenolic compound content was measured at 0-0.5 mg / g using gallic acid.
The radical scavenging ability of the extract was determined by the method of Brand-Williams using DPPH (2,2-diphenyl-1-picryhydrazyl radical). To the 0.5 mL extract, 3 mL of 0.1 mM DPPH solution was added, and the reaction was carried out in a dark room for 40 minutes. Then, the absorbance was measured at 517 nm using a spectrophotometer. The radical scavenging ability by DPPH was calculated using the following equation.
[Equation 4]
10. TLC observation
White ginseng and extrudates were enzymatically hydrolyzed and fermented. 0.5 ml of BuOH was added to 1 ml of the solution, and the mixture was shaken for 50 times. After fixing for 24 hours, the supernatant was extracted. This procedure was repeated three times, and then 40 μL (1 compound K, 2. white ginseng extrudate, 3. viscose L, 4.
11. Measurement of ginsenosides
UPLC (Acquity UPLC System; Water, Milford, Mass., USA) was used for ginsenoside measurement. The UPLC is equipped with two solvent delivery systems, an autosampler, an adjustable UV detector and a column (Acquity UPLC BEH C18 column, 1.7 uM, 2.1 x 100 mm). A: distilled water and B: acetonitrile were used as solvents. UPLC elution conditions were 0-0.5 min, A-B (85:15 v / v); 0.5-14.5 min, A-B (70:30 v / v); 14.5-15.5 min, A-B (68:32 v / v); 15.5-16.5 min, A-B (60:40 v / v); 16.5-20 minutes, A-B (45:55 v / v); 20-22 minutes, A-B (10:90); 22-27 minutes A-B (85:15 v / v) with a flow rate of 0.6 mL / min and a column temperature of 40 ± 2 ° C.
12. Statistical processing
Data were statistically analyzed using Duncan's multiple range test using SPSS (Ver. 12.0K, SPSS Inc., Chicago, IL, USA).
13. Results and discussion
(One) Non-mechanical energy input
The non-mechanical energy input is shown in Table 1 below. At the outlet temperature of 150 ℃, it was the highest at 463.75 ± 4.514 kJ / kg and the lowest at 350.98 ± 3.282 kJ / kg at the outlet temperature of 170 ℃. As the temperature increased, the amount of non-mechanical energy input of the extrusion molding decreased. This was due to the fact that the viscosity of the dough decreased as the extrusion temperature increased during extrusion molding at a temperature higher than 100 캜, Mechanical energy input decreases as the internal pressure and shear force increase). The inventors of the present invention analyzed the microstructure, viscosity and intrinsic viscosity of the suspension in the previous study (the effect of extrusion temperature on expansion of white ginseng and red ginseng), and found that the extrusion temperature was 115 ° C, It was reported that the molecular weight of the white ginseng decreased and the average melt velocity and viscosity decreased. The amount of non-mechanical energy input is an important dependent variable of the extrusion process and affects starch luxury, starch chain breaking, residence time, and pressure change with residence time.
(2) Expansion characteristics
The diameter expansion ratio, which is the expansion property of the extrudate, is an important factor affecting the texture properties of the extrudate. The expansion ratio and extinction ratio of the white ginseng extrudate are shown in Table 1 above.
The diameter expansion ratio was the highest at 0.943 ± 0.010 at the outlet temperature of 170 ℃ and the lowest at 0.812 ± 0.014 at the outlet temperature of 150 ℃. The expansion phenomenon refers to a phenomenon in which water evaporates due to a difference in pressure when raw materials pass through a discharge port at high temperature and high pressure in an extrusion molding machine, and rapidly evaporated water is generated as bubbles of the melt. The diameter expansion rate tended to increase with increasing temperature. The highest value was obtained at 262.40 ± 2.069 m / kg at the outlet temperature of 150 ℃, and 216.80 ± 3.244 m / kg at the outlet temperature of 170 ℃ And the results were inversely proportional to the expansion ratio and the resultant ratio. This was in accordance with the results of previous studies of the inventors of the present invention regarding the expansion ratio of brown rice and vegetable extrudate.
(3) Water absorption index and water dissolution index
The water absorption index (WAI) and water solubility index (WSI) of the white ginseng extrudate are shown in Table 2 below.
(° C)
As shown in Table 2, the WAI also increased as the outlet temperature increased. The highest value at 3.70 ± 0.064 g / g at 170 ° C. and the lowest at 3.84 ± 0.024 g / g at 150 ° C.
In contrast to WAI, WSI showed a tendency to decrease as the outlet temperature increased. It was the highest at 47.97 ± 0.057% at 150 ℃ and the lowest at 44.76 ± 0.479% at 170 ℃.
(4) Chromaticity
The chromaticity values of the white ginseng extrudate are shown in Table 2 above. The value of lightness (L) was highest at 85.38 ± 0.048 in white ginseng and lowest at 62.45 ± 0.157 at 170 ℃. L value decreased with increasing temperature. The redness and yellowness were the highest at 170 ℃, 8.40 ± 0.092 and 31.23 ± 0.115, respectively, and the lowest value was found at -0.10 ± 0.023 and 20.02 ± 0.111, respectively, for white ginseng. The redness (a) and yellowness (b) were increased by increasing the outlet temperature. The a and b values were estimated to be due to the change of white ginseng due to high temperature and high pressure during the extrusion process.
(5) Microstructure
The pore structure of the white ginseng extrudate by the extrusion temperature and the repeated extrusion molding was enlarged to 150 times as shown in FIG. 2, and the pore size of the pores formed at 150, 160, and 170 ° C. And the formation of porosity was found to be many. The pore was the largest at the outlet temperature of 170 ℃ and the smallest pore at the outlet temperature of 160 ℃. The pore size was large at each temperature, and especially at 170 ℃, the pore size and overall cross section were also increased.
(6) general components
The moisture content, crude ash, protein content and crude fat content of white ginseng, red ginseng and white ginseng extrudate are shown in Table 3. The content of crude fat was 1.70 ± 0.102% for white ginseng, but the crude fat content was about 0.3% for white ginseng extrudate regardless of the number of times. The protein and crude ash contents were 12.35 ± 0.037 ~ 13.37 ± 0.015% and 4.19 ± 0.080 ~ 4.38 ± 0.100%, respectively.
(° C)
(%)
(%)
(%)
(%)
(mg / mL)
(7) Evaluation of change of chemical properties according to extrusion molding of white ginseng
(7-1) Reducing sugar
Table 3 shows the results of measuring the reducing sugar of white ginseng and white ginseng extrudate. Reduced sugar content of white ginseng was 43.60 mg / mL. The highest reducing sugar content of extrudates was 68.03 mg / mL at the outlet temperature of 160 ℃ and the least amount of reducing sugar was 59.72 mg / mL at the outlet temperature of 150 ℃. Reducing sugar content of white ginseng extrudate was higher than that of white ginseng. It was presumed that pressure and shear force increased with increasing temperature during extrusion process, which affected the increase of reducing sugar.
(7-2) Acid polysaccharide
The measurement results of the acidic polysaccharide are shown in Table 4 below.
As shown in Table 4, the acid polysaccharide of white ginseng was measured to be 10.86 ± 0.160 mg / g, and in the case of white ginseng extrudate, the highest value was 31.78 ± 0.320 mg / g at the outlet temperature of 160 ° C., The formed white ginseng was found to have higher acid polysaccharide content than the untreated white ginseng. It is presumed that the extraction of acid polysaccharide is increased by the influence of pressure and shear force due to the introduction of mechanical energy during the extrusion process.
(7-3) Antioxidant activity
The antioxidants are shown in Table 4 as total phenolic compounds (total phenol) and DPPH radical scavenging activity (DPPH). It has been reported that cinnamic acid, a phenolic substance of white ginseng, is a major antioxidative substance in white ginseng and red ginseng. The total phenolic compound content of white ginseng was 3.03 ± 0.064 mg / g, and the content of total phenolic compounds in white ginseng extrudate was 3.86 ± 0.027 mg / g, which is the highest at the outlet temperature of 170 ℃. Total extruded white ginseng was found to have higher total phenolic compounds than white ginseng not extruded. The reason for the increase of the total phenolic compounds is presumed to be the increase of the measured content due to the easy dissolution due to the breaking of white ginseng tissue due to the shear force with high temperature during extrusion molding.
DPPH radical scavenging activity is used as an antioxidant index, and it is judged that the reducing power increases with an increase in DPPH radical scavenging ability. The DPPH radical scavenging ability was the highest at 44.18 ± 0.345% at the outlet temperature of 170 ℃.
(8) Evaluation of change of chemical properties according to enzymatic decomposition treatment of white ginseng extrudate
(8-1) Reducing sugar
Fig. 3 shows the results of reducing sugar when the white ginseng extrudate was subjected to enzymatic treatment. As shown in Fig. 3, in the enzyme treated product, the reducing sugar was measured to be higher than that of the untreated white ginseng. It was considered that the enzyme decomposed the sugar from white starch and increased the reducing sugar. When biscottin L was treated at 160 ℃, it showed 751.36 mg / g. On the other hand, the lowest concentration of 292.23 mg / g was obtained when treated with liquatime supra at 170 ° C.
Enzyme activity was higher in the order of 160, 150, and 170 ℃ in all enzymatic treatments. In the same extrusion conditions, enzyme activity was higher in the order of Viscozyme L, Novozyme 33095, and LiquiZime Supra. In the case of white ginseng which was not extruded, the reducing sugar content was increased during the enzyme treatment, and the highest reducing sugar content was found at 36 hours. Afterwards, the reducing sugar content was found to be constant, while in the case of white ginseng extrudate, , And thereafter showed a certain reducing sugar content. That is, when the enzyme reaction was carried out by extrusion molding of white ginseng, the time required for the reducing sugar content to reach the maximum point was 30 hours or more. This is because the high temperature, high pressure and shearing force of the extrusion molding process facilitates the accessibility of the enzyme to white ginseng It is presumed that it is because it is finished.
(8-2) Acid polysaccharide
The acid polysaccharide content of the white ginseng extrudate according to the enzyme treatment is shown in Table 5 below.
As shown in Table 5, when the enzyme was treated with biscottin L at 160 ° C, it was 42.59 ± 0.277 mg / g, which was the highest, and the lowest value was 26.50 ± 0.127 mg / g at 150 ° C . When the enzyme was added, the acid polysaccharide content increased. The acid polysaccharide also showed the same pattern as the reducing sugar. When the enzyme was treated regardless of the enzyme type, the white ginseng extrudate had a composition of 160, 150, And the enzyme activity was high. In addition, the activity of the enzyme was higher in the order of Biscozyme L, Novozyme 33095, and Liquozime Supra under the same extrusion conditions.
(8-3) Antioxidant activity
The antioxidant activity of the white ginseng extrudate according to the enzyme treatment is shown in Table 5 above. When the enzyme treatment was carried out, the highest value was 5.07 ± 0.032 mg / g when biscozyme L was added at 170 ℃, and the total phenolic content was increased when the enzyme was added as a whole.
When enzyme treatment was carried out, the highest value was 69.25 ± 0.172% when biscozyme L was added at 170 ℃. As a result, DPPH radical scavenging activity increased when the enzyme was added as a whole. In addition, the enzyme activity was higher in the order of 170, 160 and 150 ℃ for white ginseng extrudate when enzymes were treated regardless of enzyme type. In the same extrusion conditions, viscozyme L, Novozyme 33095, The activity of the enzyme was found to be higher in the order of supra.
(9) Evaluation of change of chemical properties according to fermentation treatment of white ginseng extrudate
(9-1) Reducing sugar
Changes in the reducing sugar content according to the fermentation treatment are shown in Figs. 4 to 5. In the case of some fermented products, the reducing sugar decreased until 0 to 6 hours, after which the reducing sugar increased. When fermented with Lactobacillus casei (KCTC 3172), the reducing sugar was the highest at 763.62 mg / g when treated with biscozyme L at 160 ℃ in white ginseng extrudate. In the 170 ℃ white ginseng extrudate, When the supra was treated, the reducing sugar content was the lowest at 335.15 mg / g. On the other hand, when fermented with Leuconostomycetoroidis subsp. Crmoris (KCTC 3529), the highest value was 747.62 mg / g when biscozyme L was treated in 150 ℃ extruded white ginseng, When the fermented product was extruded into Liquatzer supra, the lowest value was found as 322.88 mg / g of reducing sugar. The activity of the strain was highest at 160 ℃ and 150 ℃. The activity of the strain was higher than that of Lactobacillus casei (KCTC 3172) .
(9-2) Acid polysaccharide
The acidic polysaccharide content according to fermentation is shown in Table 6 below.
(° C)
(mg / g)
(%)
(mg / g)
As shown in Table 6, when fermented with Lactobacillus casei (KCTC 3172), the acidic polysaccharide content was highest at 46.04 ± 0.223 mg / g when treated with viscogen L in 160 ° C. white extrudate , And the lowest acidic polysaccharide was measured at 28.35 ± 0.187 mg / g when treated with 150 mg of white ginseng extrudate. When fermented with Ryukono Stock Mech. Toroidase subspecies crmoris (KCTC 3529), the highest value was 44.18 ± 0.159 mg / g when biscozyme L was treated in 160 ℃ extruded white ginseng, and 150 ℃ The acid polysaccharide was measured as 26.64 ± 0.353 mg / g in the case of treatment with Liquorice Supra in extruded white ginseng. Acid polysaccharides were also found to be higher in Lactobacillus casei (KCTC 3172) than in Ryukono Stokes mechantoroidase subsp. Crmoris (KCTC 3529).
(9-3) Antioxidant activity
The antioxidative activity following fermentation is shown in Table 6 above. When fermented with Lactobacillus casei (KCTC 3172), the highest value was 6.43 ± 0.043 mg / g when treated with viscogen L in the 170 ℃ white ginseng extrudate. The lowest total phenol content was measured as 4.91 ± 0.034 mg / g when the supra was treated. When fermented with Ryukono Stock Mech. Toroidase subspecies crmorris (KCTC 3529), the highest value was obtained when the biscottin L was treated in 170 ℃ white ginseng extrudate with 5.79 ± 0.054 mg / g, and 150 The lowest total phenol content was measured at 4.26 ± 0.012 mg / g when treated with. Lactobacillus gasseri (KCTC 3172) was found to have a higher total phenolic content than luukonosostomene toroidase subsp. Crmoris (KCTC 3529).
In the case of DPPH, when fermented with Lactobacillus casei (KCTC 3172), the highest value was 71.74 ± 0.359% when treated with viscogen L in 170 ℃ white ginseng extrudate, , The lowest DPPH radical scavenging activity was measured as 40.05 ± 0.359% when treated with LiquiZyme supra. In addition, when fermented with Ryukono Stock Mech. Toroidase subspecies crmorris (KCTC 3529), the highest value was 69.85 ± 0.456% when treated with viscogen L in the 170 ℃ white ginseng extrudate, The lowest DPPH radical scavenging activity was measured as 38.51 ± 0.603% when treated with Liquatizer Supra in white ginseng extrudate. In the case of DPPH, the lactobacillus gasseri (KCTC 3172) fermentation product showed higher DPPH radical scavenging activity than the fermented product of Lukonovostomene toroidase subsp. Crmoris (KCTC 3529) fermentation product, as did the reducing sugar and acid polysaccharide.
(10) Investigation of pattern of ginsenosides by TLC
The ginsenoside patterns of the enzyme hydrolyzate and fermentation products of white ginseng and 150, 160, and 170 ° C extrudates are shown in FIG. In the ginsenoside pattern at 150 占 폚, the compound K in the samples treated with viscogen L and Lactobacillus gasseri (KCTC 3172) (No. 6), novozyme 39095 and lactobacillus casei (KCTC 3172) treated group (No. 7) . At TLC of 160 and 170 ° C, compound K was observed in
(11) Measurement of ginsenoside
Table 7 shows changes in content of ginsenosides by extrusion of white ginseng and enzyme and fermentation.
As shown in Table 7, Rg3s and Rg3r, representative components of red ginseng, were not detected in white ginseng (WG) before extrusion molding, but Rg3s and Rg3r were found in white ginseng extrudate. Also, when the extrusion temperature was increased from 150 ° C to 170 ° C, Rg3s and Rg3r increased from 0.057 and 0.019 to 0.119 and 0.034 μg / ml, respectively. Detection of Rg3s and Rg3r of the extrudate can be said to mean red ginseng of white ginseng.
Rg3s and Rg3r in the hydrolysis of the extrudate with viscogen L increased to 1.793 and 0.559 μg / mL after the treatment with viscose L at 0.05 ° C and 0.019, respectively, at 150 ° C. extrudate (before enzyme treatment) And in the case of 170 캜 extrudate, the values of Rg3s and Rg3r decreased by viscose L treatment.
The content of ginsenosides Rb1, Rb2, Rc, and Rd was not significantly changed in the white and white ginseng extrudates, but decreased by hydrolysis and fermentation by enzyme. Compound K was not detected in white ginseng and white ginseng extrudates, but was detected in the fermented group. Compound K is generally produced by conversion of ginsenosides Rb1, Rb2, Rc and Rd in both fermentation groups of Lactobacillus gasseri (KCTC 3172) and Leuconostomycetoroids subsp. Crmoris (KCTC 3529) It was confirmed that compound K was generated. Particularly, it was confirmed that the compound K production was high in the Lactobacillus casei (KCTC 3172) treated group, and it was confirmed that Lactobacillus gasseri (KCTC 3172) was a more suitable strain. As in the pattern by TLC, Was found to be the best condition for the formation of this compound K.
Claims (6)
(b) reacting the white ginseng extrudate obtained in step (a) with a complex carbohydrate degrading enzyme comprising arabin hydrolytic enzyme, cellulase, beta-glucanase, hemicellulase and xylanase to obtain an enzyme-treated product; And
(c) obtaining a fermentation product by inoculating lactobacillus taxa into the enzyme-treated product obtained in step (b)
Of the fermented product.
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