KR20030061127A - The production method of chitosan oligo saccharide which is reinforcing inhibitory hypertension and thereof food containing above chitosan oligo saccharide - Google Patents

Abstract

PURPOSE: Provided are a preparation method of chito oligosaccharides having reinforced hypertension inhibition effect and foods, such as kimchi and turnips pickled in salt water, containing the same for the treatment of hypertension by inhibiting angiotensin converting enzyme(ACE). CONSTITUTION: A preparation method of chito oligosaccharides comprises the steps of: dissolving chitosan which is deacetylated more than 90% into acetic acid solution to make 2.0%-6.0% solution; adding 1.5-3.5unit/g(chitosan) of chitosanase and reacting at 30-40deg.C for 15-30 hours; and obtaining chito oligosaccharide with more than 20% of dimeric and trimeric oligosaccharide.

Description

TECHNOLOGY METHOD OF CHITOSAN OLIGO SACCHARIDE WHICH IS REINFORCING INHIBITORY HYPERTENSION AND THEREOF FOOD CONTAINING ABOVE CHITOSAN OLIGO SACCHARIDE}

The present invention relates to a method for producing chitooligosaccharide enhanced hypertension and food containing the chitooligosaccharide, more specifically kimchi, Dongchimi.

In general, chitosan oligo sugar (Chitosan Oligo Saccharide) is not easily dissolved in an aqueous solution, but the high absorption of the body in the chitosan, which is a high molecular material with low absorption rate in the body is made to absorb the human body.

In addition, chitooligosaccharide refers to a sugar having a structure of chitosan and glucosamine hexavalent sugar (monosaccharide) is bound to two to twelve.

In addition, chitooligosaccharide has a high physiological activity such as cholesterol lowering effect, tumor suppression effect, immune regeneration effect, antibacterial effect, and intestinal action.

Such chitooligosaccharides are being developed in the form of capsules and concentrates, and in addition to chitooligosaccharides, tocopherols, vitamins, nutritional yeasts, etc., have been developed in the form of processed foods.

The chitooligosaccharide processed food is the main food group of health supplements because of the various effects such as the removal of heavy metals in the body, anti-cancer, antibacterial action and aging of chitooligosaccharides.

However, such studies on chitooligosaccharides correspond to higher oligosaccharides and studies on chitooligosaccharides containing dimers or trimer oligosaccharides are poor.

In addition, hypertension is a chronic degenerative disease that causes all adult diseases, especially the circulatory system, and is the most common and poorly managed adult disease. As a result, urbanization, industrialization, complicated life expectancy, and extended life expectancy lead to aging population. Increasingly, the frequency and distribution of hypertension, or prevalence, is increasing.

Accordingly, the present inventors have confirmed that chitooligosaccharides having increased oligosaccharides of dimers and trimers inhibit angiotensin converting enzymes, which are the cause of hypertension, and provide a method for producing chitooligosaccharides having enhanced hypertension inhibitory ability.

In addition, the present inventors add the produced chitooligosaccharide to kimchi or Dongchimi to provide kimchi or Dongchimi with enhanced hypertension inhibitory ability.

1 is a manufacturing process diagram of the chitooligosaccharides with increased oligosaccharide content of dimers and trimers.

Figure 2 is a manufacturing process of kimchi containing chitooligosaccharides with increased oligosaccharide content of dimers and trimers.

Figure 3 is a manufacturing process of the Dongchimi containing chitooligosaccharides with increased oligosaccharide content of dimers and trimers.

The present invention relates to a method for producing chitooligosaccharide enhanced hypertension and food containing the chitooligosaccharide, more specifically kimchi, Dongchimi.

In the present invention, chitosan having a deacetylation degree of 90% or more and a viscosity in the range of 10 cps to 40 cps is dissolved in a 1.0% to 4.0% solution of acetic acid so as to be 2.0% to 6.0%. A method of producing dimers and trimers of increased chitooligosaccharides by reaction at 40 ° C. and chitosanase at 1.5 to 3.5 units / g chitosan for at least 15 hours.

In another aspect, the present invention is characterized by kimchi or Dongchimi containing the chitooligosaccharide.

Hereinafter, the configuration of the present invention will be described in more detail based on Examples, Experimental Examples or Comparative Examples and drawings or diagrams. However, the scope of the present invention is not limited to the claims of the present invention by the embodiments or the drawings.

Example 1

Preparation of Chitooligosaccharides with Increased Dimers and Trimers of Oligosaccharides

Chitosan having a deacetylation degree of 90% or more and a viscosity ranging from 10 cps to 40 cps was added to a 1% to 4% solution of acetic acid and allowed to be used at 30 ° C to 40 ° C for 3 hours. Chitosanase was added to the solubilized solution at 1.5 to 3.5 units per g of chitosan and reacted for 15 to 30 hours. Chitosan to which chitosanase was added was heated at 70 ° C. for 30 minutes to inactivate chitosanase, thereby producing chitooligosaccharides with increased oligosaccharide content of dimers and trimers. In addition, the manufacturing process of the chitooligosaccharide with the oligosaccharide of the dimer and trimer is shown in FIG.

Experimental Example 1

The amount of dimer and trimer oligosaccharides in chitooligosaccharides over time

TSK produced by the chitosan having a viscosity of 10 cps, 17 cps, 20 cps, and 40 cps produced by Example 1 with the dimers and trimers of the oligosaccharides of four oligosaccharides increased by 15 hours, every 30 hours. The oligosaccharide content of dimer and trimer was measured and analyzed by HPLC system using gel amide-80 column.

Degradation Effect of Chitosanase on Chitosan by Concentration Chitosan Response time (hours) Oligosaccharide (% equivalent) Dimer Trimer Tetramer Pentamer Hexamer Hexamer 8-mer or more system(%) 10cps 15 9.1 11.8 22.6 18.3 8.1 3.2 5.0 78.1 30 10.5 23.0 24.8 17.1 5.4 2.4 3.3 86.5 17cps 15 9.8 15.7 20.6 17.5 9.0 5.3 5.8 83.7 30 10.4 22.6 24.7 17.0 5.6 2.6 2.0 84.9 20 cps 15 9.5 16.7 21.3 17.6 8.7 4.7 4.8 83.3 30 11.0 23.2 24.3 15.5 4.5 2.2 2.9 83.6 40cps 15 10.3 18.3 22.1 16.7 6.2 2.7 1.9 78.2 30 11.3 22.9 23.5 14.3 3.5 2.1 0.1 77.7

As a result, the oligosaccharide content of dimers and trimers is 9.1% to 10.3% and 11.8% to 18.3%, respectively, when chitosan is decomposed for 15 hours, whereas 10.4% to 11.3% and 22.6 are decomposed for 30 hours to decompose chitosan, respectively. As a result, it was confirmed that the oligosaccharide content of the dimers and trimers and the viscosity of chitosan were not significantly related and were proportional to time.

In addition, the use of chitosan having a deacetylation degree of 90% or more as a material material indicates that chitosan has an amino group in the deacetylation process, so that it is easily soluble in acid, and thus chitosan has an effect of chitosan according to the degree of deacetylation. Because it is determined.

Experimental Example 2

Production of chitooligosaccharides with increased oligosaccharides of dimers and trimers according to chitosanase reaction temperature

HPLC apparatus using a TSK gel amide-80 column was inactivated chitosanase of dimers and trimers of chitooligosaccharides prepared from chitosan having a 20 cps viscosity produced by Example 1 as a raw material system).

Chitooligosaccharide Formation of Dimers and Trimers by Chitosanase Reaction Amount of chitosan Reaction temperature (℃) Response time (hours) 15 20 25 30 3unit / chitosan 30 30.11 35.25 32.73 30.14 35 35.32 40.00 36.22 32.85 40 37.35 42.75 37.34 37.74

As a result, the dimer and trimer content showed the maximum yield when reacted for 20 hours, resulting in 35.25% at 30 ° C, 40% at 35 ° C, and 42.75% at 40 ° C. It was confirmed that the production amount increased.

Experimental Example 3

ACE Inhibitory Effect and Dimer and Trimer Contents of Chitooligosaccharides

The dimer and trimer content of dimers and trimers of chitooligosaccharides prepared as raw materials of chitosan having a 20 cps viscosity produced by Example 1 was measured and analyzed by HPLC system using TSK gel amide-80 column. It was. In addition, the manufacturing process of the chitooligosaccharide containing the oligomer of the dimer and trimer is shown in FIG. In addition, the effects of ACE inhibition were measured using the method of Cushman and Cheung. In other words, ACE 0.004 unit and chitooligosaccharide were added to 0.1M sodium borate buffer, and 50 μl of Hypuryl Histidine-Leucine solution as a substrate was added thereto, followed by reaction at 37 ° C. for 10 minutes. The reaction was stopped by adding 0.2N hydrochloric acid (HCl). 3 ml of ethyl acetate was added thereto, 2.5 ml of the supernatant was taken, dried, dissolved in 3 ml of distilled water, and absorbance was measured at 228 nm. ACE inhibition activity was calculated according to the following formula.

ACE inhibitory activity (%) = (1-A / B) × 100

A: Absorbance of Sample Addition

B: Absorbance of Sample-Free Furniture

ACE Inhibitory Effect and Dimer and Trimer Contents of Chitooligosaccharides division Response time (hours) 15 20 25 30 ACE inhibitory effect ^a) 17.18 45.35 33.18 22.83 Dimer and trimer content ^b) 37.35 42.75 37.34 37.74 ^a) ACE inhibitory effect was measured by diluting the liquid sample with distilled water 40 times. ^b) HPLC analysis on TSK gel amide-80 column (relative to standard, see Table 2)

As a result, when the reaction was 20 hours, the dimer and trimer content was 42.75%, the maximum, and the ACE inhibitory effect was 45.35%, the highest among the treatments. In the above results, the content of dimer and trimer was the maximum content of dimer and trimer when the reaction time was 20 hours, resulting in the maximum ACE inhibitory effect, and the content of dimer and trimer Increasing ACE inhibitory effect was found to increase the oligosaccharides of dimers and trimers of the oligosaccharides were confirmed to inhibit the ACE. The red light (inhibition of angiotensin converting enzyme activity of chitosan oligosaccharide and hypertension inhibition in SHR, Sang-pil Hong et al., Korean Journal of Food Science and Technology (30) 6, 1998) showed that the amino group of chitooligosaccharide (C ₂ -NH ₂ group) was tested. Because it shows nucleophile at pH8.3, it can bind with zinc ions, ionic bonds with active sites containing c-terminal cations, and hydroxyl groups (-OH group) or amino groups of chitooligosaccharides. The possibility that the (C ₂ -NH ₂ group) could form hydrogen bonds with the central H ⁺ active site and the conformation of three-dimensional enzyme-substrate reactions could possibly result in the highest ACE inhibitory activity of the chitooligosaccharides of the chitooligosaccharides. Considering this high report, chitooligosaccharide is considered to be a very effective substance for inhibiting ACE activity of Kimchi and Dongchimi.

Example 2

Kimchi production containing chitooligosaccharide

Chitosan having a viscosity of 20 cps produced by Example 1 was prepared using various dimers and trimers of chitooligosaccharides as raw materials, and various kimchis were prepared at the compounding ratios shown in Table 4 below. The main material (cabbage) was selected and cut as shown in FIG. After the dyeing, washing and dehydration to mix kimchi oligosaccharides with seasoning material to mix to prepare kimchi. The kimchi was placed in a container and fermented at 10 ° C. until the acidity reached 0.6%.

Mixing Ratio of Kimchi Ingredients Containing Chitooligosaccharides material weight(%) A B C D E F cabbage 100 100 100 100 100 100 radish 10 10 12 11 12 13 chili powder - 1.2 3.5 1.1 2.5 1.5 garlic 3.4 3.5 3.6 1.5 2.5 3.5 ginger 0.2 0.1 0.2 0.2 0.1 0.1 Green onion 3.3 - 1.5 2.3 2.6 2.8 Sect - 1.5 - - - 1.2 Shrimp - 3.5 - 3.6 1.3 3.5 Anchovy Sauce 3.2 - 3.1 1.3 - 1.1 ear - - - - 0.2 1.2 Sugar 1.0 - - 1.5 - - Buttercup - - - - 2.6 2.4 oyster - - - - - 3.1 chives - - - 1.5 - - Quitooligosaccharides 0.5 0.5 0.5 0.5 0.5 0.5

Comparative Example 1

Sensory Evaluation of Kimchi Prepared by Mixing Ratio

Sensory tests on the appearance, flavor, texture (crunching), color, overall symbolism, etc. of 30 aged kimchi prepared by blending ratio according to Example 2 were carried out by a five-point evaluation method. Indicated.

Sensory test of kimchi according to compounding ratio (5-point standard method) Item Kimchi Test Group A B C D E F Exterior 4.2 4.2 4.3 4.5 4.2 4.5 Flavor 4.2 4.3 4.2 4.4 4.3 4.5 Texture (Crunch) 4.2 4.1 4.4 4.5 4.3 4.4 color 4.1 4.5 4.7 4.2 4.0 4.5 Overall preference 4.2 4.3 4.6 4.3 4.2 4.4

The kimchi produced by the blending ratio according to Table 4 showed relatively high evaluation scores, from 4.2 to 4.5, in terms of appearance and flavor. In the kimchi texture (crunching) category, the panels showed a difference, and in the color category, the C treatment group that added the most red pepper powder was the highest with 4.7 points and the C treatment group with the highest degree of overall preference at 4.6 points.

Comparative Example 2

Comparison of ACE Inhibitory Effect, pH, and Acidity in the Ripening Period of Kimchi with Chitooligosaccharide and Kimchi without Chitooligosaccharide

Kimchi prepared using Treatment C in the blending ratio of Example 2 was used as an experimental group, and kimchi prepared by excluding chitooligosaccharide in the blending ratio of Treatment C was used as a control. In addition, in order to measure the ACE, pH and acidity of other kimchi in addition to the control group and the experimental group used in this comparative example, four commercially available kimchi was selected and measured by the same method. The control, experimental group, commodity kimchi 1, 2, 3, 4 were sampled in the ripening period and the ACE, pH and acidity of kimchi by treatment groups were measured and shown in Table 6.

ACE Inhibitory Effect, pH, and Acidity in the Ripening Season of Kimchi Added with Chitooligosaccharides Item ACE (%) pH Acidity (%) Control 45.49 4.07 0.61 Kimchi with Chitooligosaccharides (Experimental Group) 71.75 4.11 0.66 Commodity Kimchi 1 49.34 4.42 0.64 Commodity Kimchi 2 19.51 3.88 1.30 Commodity Kimchi 3 50.07 4.07 0.89 Commodity Kimchi 4 53.56 3.93 0.95

As a result, the ACE inhibitory effect of the control group was 45.49%, whereas the chitooligosaccharide-added experimental group was 71.75%.

In addition, the acidity was decreased in the experimental group compared to the control group, which was considered to be the preparation of chitooligosaccharide by dissolving chitosan in 1% ~ 4% acetic acid solution.

Comparative Example 3

Antihypertensive Effects of Kimchi Containing Chitooligosaccharides

The mice used in the experiment were supplied with male SHR / NCrj (Charles River Japan Inc.), 8 weeks old, and subjected to a week of adaptive training on the blood pressure measuring holder prior to the test. Measured. The SHR used for the experiment was about 10 weeks old, and 6 animals were selected as systolic blood pressure showing 180 mmHg to 200 mmHg.

The kimchi broth prepared by fermentation ripening at the maturity level was prepared using the blending weight treatment C of Example 2, and the systolic blood pressure was fed to the selected six SHRs as drinking water three times a day for two weeks. The change is measured and shown in Table 7.

Antihypertensive Effects of Kimchi Containing Chitooligosaccharides Treatment group (n = 6) 3 days 7 days 10 days 14 days Control 202.3 ± 11.4 205.0 ± 14.6 200.0 ± 10.5 195.0 ± 19.2 Experimental group (kimchi containing chitooligosaccharide) 195.0 ± 13.4 197.0 ± 14.2 190.4 ± 13.4 184.6 ± 17.0 * Average blood pressure before test (n = 12): 192.8 ± 15.3mmHg

As a result, the experimental group as a whole showed a tendency to decrease compared to the control group. The reference blood pressure before the test was 192.8 ± 15.3mmHg. At 14 days after drinking water, the control group had 195.0mmHg, which was not significantly changed compared to the initial reference blood pressure. It showed a tendency to decrease significantly in the initial blood pressure, and it was confirmed that chitooligosaccharides containing dimers and trimer oligosaccharides lowered blood pressure.

Example 3

Manufacture of Dongchimi Containing Chitooligosaccharide

Chitosan having a viscosity of 20 cps produced by Example 1 was prepared using various dimers and trimers of chitooligosaccharides prepared as raw materials at various mixing ratios as shown in Table 8 below. The main material (beet) was selected and washed according to Figure 3, and the salt was placed on the bottom of the jar, and the whole radish was put and soaked for 2 days. The dried radish was pressed with stone, and a bag made by selecting and cutting seasoning materials was placed in the center of the radish. The ratio of salt and water in the radish and seasoning material is 1:18, and brine containing chitooligosaccharide was poured and fermented and aged until the acidity was 0.6% at 10 ° C.

Mixing ratio of Dongchimi containing chitooligosaccharide material weight(%) A B C D E F radish 100 100 100 100 100 100 Coarse salt 10 10 12 11 12 1.3 garlic - 1.2 3.5 1.1 2.5 1.5 ginger 3.4 3.5 3.6 1.5 2.5 3.5 Green onion 0.2 0.1 0.2 0.2 0.1 0.1 Sect 3.3 - 1.5 2.3 2.6 2.8 Silpa - 1.5 - - - 1.2 Green pepper - 3.5 - 3.6 1.3 3.5 Red pepper 3.2 - 3.1 1.3 - 1.1 ear - - - - 0.2 1.2 lampshade 1.0 - - 1.5 - - ship - - - - 2.6 2.4 Quitooligosaccharides 0.5 0.5 0.5 0.5 0.5 0.5

Comparative Example 1

Sensory Evaluation of Dongchimi Prepared by Mixing Ratio

Sensory evaluation was carried out by the five-point evaluation method for the appearance, flavor, texture (crunching), sour taste, general taste, etc. of 30 aged panels of aged Dongchimi prepared by blending ratio according to Example 3, and the result In average, it is shown in Table 9.

Sensory Evaluation of Dongchimi Containing Chitooligosaccharides According to the Mixing Ratio (5-point Evaluation Method) division Dongchimi Test Group A B C D E F Exterior 4.7 4.2 4.3 4.5 4.4 4.5 Flavor 4.5 4.2 4.1 4.4 4.5 4.3 Texture (Crunch) 4.7 4.3 4.5 4.1 4.4 4.7 Sour taste 4.6 4.5 4.1 4.6 4.0 4.5 Overall preference 4.7 4.4 4.2 4.4 4.5 4.5

The appearance and flavor of Dongchimi prepared by the above method were relatively high, with a score of 4.1 to 4.7, and the ratios of green pepper, red pepper and pear had a great influence on the appearance and flavor. It is produced by immersing in salt water of high salinity for a long time, and the crunchy items and palatability items are lowered because the degree of crunchyness of radishes is decreased. Overall, Dongchimi A had the highest overall acceptability.

Comparative Example 2

ACE inhibitory effect, pH, and acidity in the maturation period of Dongchimi containing chitooligosaccharide and Dongchimi not containing chitooligosaccharide

In the mixing ratio of Example 3, the treatment group A was used to prepare a control group, which was the same as Chichiol, except for the chitooligosaccharides. pH and acidity were measured and shown in Table 10.

ACE Inhibitory Effect, pH and Acidity in the Ripening Season of Dongchimi Containing Chitooligosaccharide Treatment group ACE (%) pH Acidity Control 39.88 4.88 0.30 Experimental group (dongchimi with chitooligosaccharides) 70.64 6.39 0.20

As a result, the activity of the control group was 39.98%, while the chitooligosaccharide-treated group was 70.64%.

In addition, the acidity was decreased in the experimental group compared to the control group, which was considered to be the preparation of chitooligosaccharide by dissolving chitosan in 1% ~ 4% acetic acid solution.

Comparative Example 3

Antihypertensive Effects of Dongchimi Containing Chitooligosaccharides

The mice used in the experiment were supplied with male SHR / NCrj (Charles River Japan Inc.), 8 weeks old, and subjected to a week of adaptive training on the blood pressure measuring holder prior to the test. Measured. The SHR used for the experiment was about 10 weeks old, and 6 animals were selected as systolic blood pressure showing 180 mmHg to 200 mmHg.

Dongchimi without chitooligosaccharide was prepared using Process A of the compounding ratio of Example 3, and Dongchimi with chitooligosaccharide was added, fermented and matured to a maturity level, and the equivalent Americans were selected from six SHRs. Table 11 shows the changes in systolic blood pressure while drinking three times a day for two weeks as drinking water.

Antihypertensive Effects of Dongchimi Containing Chitooligosaccharides Treatment group (n = 6) 3 days 7 days 10 days 14 days Control 189.4 ± 18.7 192.3 ± 15.9 196.5 ± 19.8 190.2 ± 15.8 Experimental group (dongchimi with chitooligosaccharides) 186.7 ± 12.7 186.7 ± 18.9 188.0 ± 10.5 180.0 ± 17.8 * Average blood pressure before test (n = 12): 192.8 ± 15.3mmHg

As a result, the experimental group showed a blood pressure reducing effect compared to the control group. The reference BP before the test was 192.8 ± 15.3mmHg, and the blood pressure of the control group was 190.2mmHg at 14 days after drinking water, but there was no significant change compared to the initial reference blood pressure. It was shown that chitooligosaccharides with dimer and trimer content increased by 12mmHg, which is effective for blood pressure control.

The present invention is not limited to the technical spirit described in the specific embodiments, diagrams or drawings described above, and those skilled in the art without departing from the gist of the invention claimed in the claims. As long as anyone can carry out various modifications, such changes are within the scope of the claims.

As described in detail above, the present invention relates to a method for preparing chitooligosaccharides to increase dimers and trimer oligosaccharides, and to kimchi and Dongchimi containing the chitooligosaccharides.

The present invention has the effect of inhibiting hypertension by inhibiting angiotensin converting enzymes that cause hypertension by preparing chitooligosaccharides with increased dimer and trimer oligosaccharide contents.

In addition, the present invention is a kimchi or Dongchimi containing the chitooligosaccharide produced by the addition of the chitooligosaccharide can lower blood pressure to prevent and treat hypertension, can develop kimchi and Dongchimi stayed in traditional foods as a functional food have.

Claims (7)

In the method for producing chitooligosaccharide, A method for producing high oligosaccharides with enhanced hypertension, characterized by dissolving chitosan in acetic acid solution and adding chitosanase to increase the dimer and trimer oligosaccharide contents. The method of claim 1, The chitosan was dissolved in acetic acid solution so that the content of chitosan was 2.0% to 6.0%, and chitosanase 1.5-3.5 unit / g (chitosan) was added at a temperature of 30 ° C to 40 ° C, followed by reaction for 15 to 30 hours. A method for producing hyperoligosaccharides with enhanced hypertension, characterized by increasing the content of dimers and trimer oligosaccharides. The method according to claim 1 or 2, The chitosan has a deacetylation degree of more than 90% high blood pressure inhibiting ability enhanced chito oligosaccharide production method. A chitooligosaccharide with enhanced hypertension-suppressing ability, comprising 20% or more of dimers and trimer oligosaccharides prepared by the method of any one of claims 1 to 3. Food containing the chitooligosaccharide of Claim 4. Kimchi containing the chitooligosaccharide of claim 4. Dongchimi containing the chitooligosaccharide of Claim 4.