KR101982680B1

KR101982680B1 - Black ginseng enriched ginsenoside Rd

Info

Publication number: KR101982680B1
Application number: KR1020170069562A
Authority: KR
Inventors: 고태훈; 윤일노; 임정수; 김종민; 최경임
Original assignee: 금산덕원인삼약초영농조합법인; 농업회사법인 금산흑삼주식회사
Priority date: 2017-06-05
Filing date: 2017-06-05
Publication date: 2019-05-27
Also published as: KR20170071457A; WO2018225972A1

Abstract

The present invention relates to black gins whose ginsenoside Rd content is increased and the degree of shortening is increased, and a process for producing the same. In the process of preparing black ginseng according to the present invention, first, hydrolyzing starch in ginseng and then hydrolyzing protopanaxadiol ginsenoside in ginseng to ginsenoside Rd. The present invention can increase the ginsenoside content of black ginseng from 1.00 to 7.00 mg / g and increase the sugar content from 5.00 to 7.00 Brix.

Description

Ginsenosides Rd enhanced black ginseng and methods for producing the same -

The present invention relates to a process for producing black ginseng having a high sugar content and a high ginsenoside Rd content, and can increase sugar content and fortify ginsenoside Rd in a steaming process, and thus is more efficient and economical than a conventional method using a concentrated liquid.

Ginsenoside, an indicator component of ginseng, is classified into dammarane type and oleanane type depending on the chemical structure of aglycone. Dammarane type is classified as protopanaxadiol and protopanaxatriol according to the hydroxy group attached to 3, 6, 12 carbon of aglycon. Protopanaxadiol ginsenoside is known as ginsenoside Rb1, Rd, Rc, Rg3 and protopanaxatriol ginsenoside is ginsenoside Rg1, Re, F1. The human metabolism of protopanaxadiol ginsenosides is as follows: Ginsenoside Rb1 is hydrolyzed to ginsenoside Rg3 by an enzyme produced by gastric or intestinal microorganisms in the stomach and intestines, or by Compound K through Ginsenoside Rd, Ginsenoside F2 And hydrolyzed. In this process, various ginsenoside metabolites are produced, but the main metabolic pathway is the pathway from ginsenoside Rb1 to ginsenoside Rg3, or from ginsenoside Rb1 to ginsenoside Rd, ginsenoside F2 to compound K. The metabolic pathways of the various ginsenosides are shown in Fig.

Korean Patent Laid-Open No. 10-2015-0088844 relates to a cosmetic composition or antioxidative health functional food composition containing ginsenosides Rg2, Rg3, Rh1, Rh2 and Rf, wherein the black ginseng extract, which is obtained by extracting black ginseng with water, Korean Patent Laid-Open No. 10-2015-0160977 discloses a process for processing ginseng fruit for increasing the ginsenoside content, wherein a saccharification fermentation process using malt was performed, but an increase in ginsenoside Rd was performed in the saccharification fermentation process Korean Patent No. 10-1120996 discloses a composition for promoting exercise capacity and fatigue recovery, which comprises ginsenoside Rg3 and Rg2. The present invention relates to a composition for promoting exercise capacity and fatigue recovery, which comprises pectinase, hemicellulase, A cellulase, and a polysaccharide degrading enzyme such as alpha-amylase. Korean Patent No. 10-1423100 relates to a process for producing fermented red ginseng which amplifies the content of ginsenosides Rg3 and Rb1, and Korean Patent No. 10-1308880 discloses a process for producing fermented red ginseng from a complex microorganism that the relates to a process for the preparation of fermented ginseng concentrate according to efficiently fermented to raw Red ginseng extract Lactis strains, L. Acidophilus strains and B. Animalis spp . In order to selectively increase the content of ginsenosides Rg3, Rh2 and Compound K by using the bacterium mixed with Lactis bacteria and the alpha amylase enzyme, Korean Patent No. 10-1018989 discloses a method for selectively increasing the content of ginsenosides Rg3, Rh2 and Compound K by using a ginseng polysaccharide In Korean Patent No. 10-1181515, an enzyme treatment step was carried out through alpha-amylase and cellulase to liberate the saponin bound to starch. However, the enzyme treatment with Ginsenoside Rd After the black ginseng extract was prepared, the enzyme treatment step was carried out.

The present invention relates to a process for the production of black gins which hydrolyze starch in ginseng to disaccharides and monosaccharides and hydrolyze protopanaxadiol ginsenosides to ginsenosides Rd in the course of producing black ginseng, Rd < / RTI >

Peeling the washed ginseng to prepare a peanut ginseng in which the wax layer of the ginseng is removed; Drying the peeled ginseng to a moisture content of 55 to 65% at 60 to 70 캜 to prepare dried ginseng; Immersing the dried ginseng in an enzyme diluting solution A; A first mashing step of hydrolyzing starch in dried ginseng to a disaccharide or a monosaccharide by mixing the enzyme diluted solution A with the dried ginseng immersed in the enzyme diluent A at 60 to 80 ° C for 5 to 8 hours; A second drying step of drying the ginseng having undergone the first steaming step to a moisture content of 55 to 65% to prepare a saccharified ginseng; A second boiling step of converting the protopanaxadiol-based ginsenoside into ginsenoside Rd by agitating the ginseng subjected to the second-stage drying step at 70 to 80 ° C for 6 to 8 hours while spraying the enzyme diluting solution B; A third drying step of drying the ginseng having undergone the second steaming step to a moisture content of 55 to 65% and a third steaming step of mixing the third dried ginseng at 60 to 70 캜 for 4 to 6 hours; A hot air drying step of drying the ginseng to a moisture content of 20%; Drying the ginseng with hot air drying to a moisture content of 15%, and dehumidifying the ginsenoside Rd, and a process for producing the same.

Preferably, the enzyme diluent A may be selected from alpha-amylase obtained from Bacillus licheniformis , alpha-amylase obtained from Bacillus amyloliquefaciens , or a mixture thereof.

Preferably, said enzyme diluent B is selected from the group consisting of glucoamylase obtained from Aspergillus niger , Trichoderma a combination enzyme of cellulase, beta-glucanase, hemisolurase , or a mixture thereof obtained from reesei can be selectively used.

More preferably, the enzyme diluent A may be alpha-amylase obtained from Bacillus licheniformis .

More preferably, the enzyme diluent B may be a glucoamylase obtained from Aspergillus niger .

In the present invention, the starch in ginseng is firstly hydrolyzed with disaccharides and monosaccharides to convert the protopanaxadiol ginsenosides in ginseng to ginsenosides Rd in a state where sugar content is high and hydrolysis interference of starch is minimized .

Fig. 1 shows the results of the dilution of the ginsenoside standard product treated with the enzyme FRED.
Figure 2 shows the results of the dilution of the Ginsenoside standard product treated with the enzyme LT300.
Fig. 3 shows the results of analysis of the diluted solution of the ginsenoside standard product treated with the enzyme BAG.
Fig. 4 shows the results of analysis of the diluted solution of the ginsenoside standard product treated with the enzyme RCL.
Fig. 5 shows the results of analysis of ginsenoside in the saccharified ginseng.
6 shows the result of measuring the sugar content of the ginseng extract.
FIG. 7 shows the results of analysis of ginsenosides in black ginseng.
8 shows the result of measuring the sugar content of the black ginseng extract.
9 shows the metabolic pathway of ginsenoside.

The method for preparing ginsenoside Rd enhanced ginsenoside Rd according to the present invention comprises the steps of: peeling ginseng ginseng to prepare peeled ginseng in which the wax layer of ginseng is removed; Drying the peeled ginseng to a moisture content of 55 to 65% at 60 to 70 캜 to prepare dried ginseng; Immersing the dried ginseng in an enzyme diluting solution A; A first mashing step of hydrolyzing starch in dried ginseng to a disaccharide or a monosaccharide by mixing the enzyme diluted solution A with the dried ginseng immersed in the enzyme diluent A at 60 to 80 ° C for 5 to 8 hours; A second drying step of drying the ginseng having undergone the first steaming step to a moisture content of 55 to 65% to prepare a saccharified ginseng; A second boiling step of converting the protopanaxadiol-based ginsenoside into ginsenoside Rd by agitating the ginseng subjected to the second-stage drying step at 70 to 80 ° C for 6 to 8 hours while spraying the enzyme diluting solution B; A third drying step of drying the ginseng having undergone the second steaming step to a moisture content of 55 to 65% and a third steaming step of mixing the third dried ginseng at 60 to 70 캜 for 4 to 6 hours; A hot air drying step of drying the ginseng to a moisture content of 20%; The ginseng having been subjected to the hot air drying step is dehumidified while being cooled and dried to a moisture content of 15% to prepare ginsenoside Rd enhanced black ginseng.

Each step will be described in detail below.

First, the washed ginseng is peeled and the waxy layer of the ginseng is removed to prepare the peeled ginseng. Generally, the process of mixing and drying ginseng without peeling is repeated three times to prepare black ginseng. In the present invention, the removal of the wax layer is carried out by mixing enzyme diluting liquid A which hydrolyzes starch in ginseng, protopanaxadiol ginsenoside So that the enzyme diluting solution B that hydrolyzes the enzyme can easily penetrate into the ginseng tissue.

The peeled ginseng is first dried at a temperature of 60 to 70 DEG C to a moisture content of 55 to 65% to prepare dried ginseng.

The dried ginseng is immersed in the enzyme diluting solution A. The preferred enzyme diluent A may be selected from alpha-amylase obtained from Bacillus licheniformis , alpha-amylase obtained from Bacillus amyloliquefaciens , or a mixture thereof. A more desirable enzyme diluent A is Bacillus alpha-amylase obtained from licheniformis can be used. Dipping of dried ginseng in enzyme diluent A enables hydrolysis of starch mainly distributed in the central part of ginseng tissue.

The enzyme dilution liquid A is sprayed on the dry ginseng soaked in the enzyme dilution liquid A and firstly mixed at 60 to 80 ° C for 5 to 8 hours. The starch in ginseng can be converted into a disaccharide or a monosaccharide in the first steaming stage.

The ginseng having been subjected to the first steaming step is secondarily dried to a moisture content of 55 to 65% to prepare a saccharified ginseng. The sugar content of the saccharified ginseng extract prepared through the secondary drying step may be increased to 5 to 7 Brix.

The enzyme diluted solution B is sprayed to the ginseng subjected to the second drying step and is secondly mixed at 70 to 80 캜 for 6 to 8 hours. A preferred enzyme diluent B is glucoamylase obtained from Aspergillus niger , Trichoderma a combination enzyme of cellulase, beta-glucanase, hemisolurase , or a mixture thereof obtained from reesei can be selectively used. A more preferred enzyme diluent B may be a glucoamylase obtained from Aspergillus niger . In this secondary steaming step, protopanaxadiol ginsenoside in ginseng is converted to ginsenoside Rd.

The ginseng having been subjected to the second steaming step is thirdly dried to a moisture content of 55 to 65%, and the third dried ginseng is thirdly boiled at 60 to 70 ° C for 4 to 6 hours.

A hot air drying step of drying the ginseng to a moisture content of 20%; The ginseng having been subjected to the hot air drying step is dehumidified while being cooled and dried to a moisture content of 15% to prepare ginsenoside Rd enhanced black ginseng. The content of ginsenoside in the black ginseng prepared by the above-described production method can be increased to 1.00 to 7.00 mg / g.

Hereinafter, the present invention will be described in detail by way of examples, but the scope of the present invention is not limited by these examples.

Preparation of ginseng starch diluted solution containing ginsenoside Rb1

The starch extracted from ginseng was diluted in distilled water to prepare a starch dilution, and the standard was diluted so that the concentration of ginsenoside Rb1 was 2.00 mg / mL.

Identification of the hydrolytic properties of ginsenoside Rb1 of spezyme FRED (Spezyme FRED)

The results of analyzing ginsenosides Rb1, Rd, F2 and Compound K while adding 2% (v / v) of speyizer FRED (hereinafter referred to as FRED) to the diluted solution of Example 1 and stirring the mixture at 70 DEG C for 24 hours are shown in Fig. Respectively. After 24 h bathing, the ginsenoside Rb1 content is 1.95 mg / mL, demonstrating that the enzyme FRED can not hydrolyze ginsenoside Rb1 to ginsenoside Rd, F2 or Compound K. It was confirmed that the sugar content of the ginseng starch increased from 0.01 Brix to 0.5 Brix, which proved that the ginseng starch hydrolyzed into disaccharides and monosaccharides.

Characterization of the hydrolysis of Ginsenoside Rb1 by Spzyme LT300 (Spezyme LT300)

The results are shown in FIG. 2, which is performed as in Example 2, but using LT300 instead of FRED. After 24 hours of bathing, the content of ginsenoside Rb1 was 1.37 mg / mL, and the ginsenosides Rd, F2 or Compound K were almost not detected and the sugar content was increased to 0.4 Brix. This demonstrates that LT300 can hydrolyze ginsenoside Rb1 but can not hydrolyze with ginsenosides Rd, F2 or Compound K and hydrolyzes ginseng starch.

Hydrolysis characterization of ginsenoside Rb1 of BioWin AG (BioWin AG)

The results are shown in FIG. 3, in which BAG is used instead of FRED. After 24 hours of bathing, the ginsenosides Rb1, F2 and Compound K were not detected. The ginsenoside Rd increased to 1.66 mg / mL and the sugar content did not increase to 0.01 Brix. This proves that BAG hydrolyzes ginsenoside Rb1 to ginsenoside Rd and can not hydrolyze it again with ginsenoside F2, Compound K, and proves that the starch can not hydrolyze.

Characterization of hydrolysis of ginsenoside Rb1 by Rohament CL

The results are shown in FIG. 4 using RCL instead of FRED. After 24 hours of bathing, the ginsenosides Rb1, F2 and Compound K were almost undetectable. The ginsenoside Rd increased to 1.17 mg / mL and the sugar content did not increase to 0.01 Brix. This proves that RCL hydrolyzes ginsenoside Rb1 to ginsenoside Rd and can not hydrolyze it again with ginsenoside F2, Compound K, and proves that the starch can not be hydrolyzed.

Preparation of glycosylated ginseng hydrolyzed with FRED enzyme

The peeled ginseng was prepared by removing the wax layer of the washed ginseng (10 kg) so that the enzyme solution could penetrate well, and the ginseng was dried at 65 ± 5 ° C to a moisture content of 60 ± 5% to prepare dry ginseng. The dried ginseng was immersed in a 30% (v / v) FRED enzyme dilution solution (enzyme dilution solution A) for 1 hour, and the enzyme dilution solution A was sprayed to the dried ginseng immersed in the enzyme dilution solution A at 70 ± 10 ° C The ginseng was boiled for 8 hours, and the steamed ginseng was dried to a moisture content of 60 ± 5%. The dried ginseng was hot-air dried to a moisture content of 20%, dehumidified by hot air drying and dried to a moisture content of 15% Respectively.

Production of Saccharified Ginseng Hydrolyzed with LT300 Enzyme

The same procedure as in Example 6 was carried out except that instead of the FRED enzyme, saccharose ginseng was prepared using the enzyme diluent A prepared from the LT300 enzyme.

Preparation of Saccharified Ginseng without Enzyme

The procedure of Example 6 was repeated except that the saccharified ginseng was prepared by using distilled water instead of the enzyme diluting solution A.

Analysis of Ginsenosides of Saccharogenic Ginseng

FIG. 5 shows the results of analysis of ginsenosides in the ginseng ginseng prepared in Example 6, Example 7 and Example 8 according to the ginsenoside analysis of the Health Functional Food Code. FRED and LT300 enzymes were found to be unable to hydrolyze ginsenoside Rb1.

Determination of sugar content of ginseng extract

The saccharified ginseng prepared in Example 6, Example 7 and Example 8 was extracted with purified water of 10 times, and the sugar content of the extract was measured with a sugar meter. The result is shown in FIG. The sugar content of Example 8, which did not use enzyme, was 2.45 Brix, while the sugar content of Example 6 and Example 7 were 6,56 Brix and 5.38 Brix, respectively. This proves that starch in ginseng is hydrolyzed to disaccharides and monosaccharides.

Production of black ginseng reinforced with ginsenoside Rd using BAG

The peeled ginseng in which the wax layer of the washed ginseng (10 kg) was removed so that the enzyme solution could penetrate well was prepared, and the ginseng was first dried at 65 ± 5 ° C to a moisture content of 60 ± 5% to prepare dry ginseng. The dried ginseng was immersed in a 30% (v / v) FRED enzyme dilution solution (enzyme dilution solution A) for 1 hour, and the enzyme dilution solution A was sprayed to the dried ginseng immersed in the enzyme dilution solution A at 70 ± 10 ° C And the first-boiled ginseng was secondary-dried to a moisture content of 60 ± 5%. The second dried ginseng was subjected to a second boiling for 8 hours at 70 ± 10 ° C while spraying a 30% (v / v) BAG enzyme diluent (enzyme diluent B) every 2 hours, and the second boiled ginseng was heated to 60 Followed by tertiary drying up to ± 5%. The third dried ginseng was tilted at 65 ± 5 ° C. for 6 hours for 3 hours. The third ginseng was heat-dried to a moisture content of 20%, and the ginseng was dehumidified by cold air drying to obtain a moisture content of 15% And dried.

Manufacture of black ginseng reinforced with ginsenoside Rd using RCL

The procedure of Example 11 was repeated except that BAG enzyme was replaced with Enzyme Diluent B prepared with RCL enzyme to prepare black ginseng.

Manufacture of black ginseng without enzymes

The procedure of Example 11 was repeated except that the enzyme dilution solution B was replaced with distilled water to prepare black ginseng.

Analysis of ginsenosides in black ginseng

The ginsenoside content analyzed using the ginsenoside method used in Example 9 is shown in Fig. The ginsenoside content of Example 13 without enzyme was not increased and only the ginsenoside Rg3 was increased to 1.37 mg / g. This is because a portion of ginsenoside Rb1 is converted to ginsenoside Rg3 through thermodynamic hydrolysis. In Example 11 using BAG enzyme, ginsenoside Rb1 was converted to ginsenoside Rd and increased to 6.52 mg / g. In Example 12 using RCL, ginsenoside Rb1 was hydrolyzed to ginsenoside Rd to yield 5.89 mg / g.

Determination of sugar content of black ginseng extract

The sugar content of the black ginseng extract prepared in the same manner as in Example 11 was measured and shown in Fig.

The enzyme used in the present invention is as follows.
BioWin AG is an enzyme containing glucoamylase obtained from Aspergillus niger and used in the present invention as "BAG". Rohament CL is an enzyme containing cellulase, beta-glucanase and hemicellulase obtained from Trichoderma reesei. "Spezyme FRED is an enzyme containing alpha-amylase obtained from Bacillus licheniformis and labeled with" FRED ". Spezyme LT300 is an enzyme containing alpha-amylase obtained from Bacillus amyloliquefaciens and labeled with" LT300 " Respectively.

Claims

Peeling the washed ginseng to prepare a peanut ginseng in which the wax layer of the ginseng is removed;
Drying the peeled ginseng to a moisture content of 55 to 65% at 60 to 70 캜 to prepare dried ginseng;
Immersing the dried ginseng in an enzyme diluting solution A;
A first mashing step of hydrolyzing starch in dried ginseng to a disaccharide or a monosaccharide by mixing the enzyme diluted solution A with the dried ginseng immersed in the enzyme diluent A at 60 to 80 ° C for 5 to 8 hours;
A second drying step of drying the ginseng having undergone the first steaming step to a moisture content of 55 to 65% to prepare a saccharified ginseng;
A second boiling step of converting the protopanaxadiol-based ginsenoside into ginsenoside Rd by agitating the ginseng subjected to the second-stage drying step at 70 to 80 ° C for 6 to 8 hours while spraying the enzyme diluting solution B;
A third drying step of drying the ginseng having undergone the second steaming step to a moisture content of 55 to 65% and a third steaming step of mixing the third dried ginseng at 60 to 70 캜 for 4 to 6 hours;
A hot air drying step of drying the ginseng to a moisture content of 20%;
Drying the ginseng to a moisture content of 15% while cooling and drying it,
Amylase obtained from Bacillus licheniformis, α-amylase obtained from Bacillus amyloliquefaciens, or a mixture thereof, is used as the enzyme diluting solution A in the primary steaming step to hydrolyze the glucoside bond of the starch,
In the second steaming step, using either glucoamylase obtained from Aspergillus niger as an enzyme diluent B, cellulase obtained from Trichoderma reesei, beta-glucanase, complex enzyme of hemicellulase, or a mixture thereof, The side Rb1 is hydrolyzed to ginsenoside Rd,
&Lt; / RTI > wherein the ginsenoside Rd is enhanced.

delete

The method according to claim 1,
Wherein the protopanaxadiol-based ginsenosides in the second steaming stage are ginsenosides Rb1, Rb2, Rb3 and Rc.

The method according to claim 1,
And the sugar content of black ginseng is 5.00 to 7.00 Brix.

The method according to claim 1,
Wherein the content of ginsenoside Rd in black ginseng is 1.00 to 7.00 mg / g.

A black ginseng prepared according to the production method of any one of claims 1 and 4 to 6.