KR20230112802A - Compositions for lowering blood sugar having the ability to inhibit alpha-glucoside activity using lactic acid bacteria, a health functional food, and a pharmaceutical composition - Google Patents

Abstract

The present invention provides a composition for lowering blood sugar comprising a fermented product obtained by fermenting safflower seeds with lactic acid bacteria as an active ingredient for inhibiting alpha-glucosidase activity.
In one aspect of the present invention, the lactic acid bacteria are Enterococcus faecium , Lactobacillus pentosus , Lactobacillus plantarum , Leuconostoc mesenteroides , lacto Bacillus fermentum ( Lactobacillus fermentum ), Lactobacillus brevis ( Lactobacillus brevis ), Lactobacillus sakei ( Lactoba cillus sakei ), and Leuconostoc holzapfelii strains .

Description

Compositions for lowering blood sugar having the ability to inhibit alpha-glucoside activity using lactic acid bacteria, a health functional food, and a pharmaceutical composition composition}

The present invention relates to a composition for lowering blood sugar derived from a natural product, and more particularly, to a composition comprising a fermented product obtained by fermenting a natural product using lactic acid bacteria as an active ingredient for lowering blood sugar and its use.

The fermented product of lactic acid bacteria according to the present invention can be used as a medicine or health functional food (health supplement food) for the prevention, improvement, or treatment of diabetes by reducing blood sugar by inhibiting α-glucosidase activity.

Diabetes mellitus is a metabolic disease caused by higher than normal levels of glucose (blood sugar) in the blood. Glucose, which is supposed to be used as energy in the body, is not secreted normally (type 1 diabetes), or even if secreted, it does not function properly. (Type 2 Diabetes) As the cells are unable to enter, glucose continues to build up in the blood, causing hyperglycemia. Diabetes mellitus and hyperglycemia cause damage to blood vessel walls rather than themselves, resulting in more serious diabetic complications such as myocardial infarction, stroke, retinopathy, and renal failure. In the prevention, delay, improvement, and treatment of these diabetic complications, the most important goal is to control blood glucose levels as close to normal as possible.

Methods for controlling blood sugar vary according to their mechanism of action, but above all, it is important to suppress or inhibit the activity of α-glucosidase in order to suppress a rapid increase in blood sugar after food intake. Since alpha-glucosidase is an enzyme that breaks down carbohydrates in the diet and converts them into glucose, alpha-glucosidase inhibitors (inhibitors) compete with alpha-glucosidase in the small intestine to delay the absorption of carbohydrates, thereby increasing postprandial blood sugar. acts to reduce.

Alpha-glucosidase inhibitors that are currently used clinically include acarbose, voglibose, and miglitol, but some have side effects such as abdominal distension, vomiting, and diarrhea due to long-term use. being reported

On the other hand, efforts to treat various diseases using natural products such as plants and herbal medicines have been introduced for a long time as herbal treatments or folk remedies, and various methods of hypoglycemic agents using natural products are also known.

As an active ingredient that inhibits or inhibits alpha-glucosidase activity in relation to blood sugar lowering, for example, Korean Patent Publication No. 10-2004-0052398 discloses grape seed extract, and Korean Patent Publication No. 10-2004-0077976 discloses pine needles. Or a hot water extract or alcohol extract of pine bark, Korean Patent Publication No. 10-2016-0126715 discloses a hot water extract or alcohol extract of cinnamon or loquat leaves.

On the other hand, various techniques are known for lowering blood sugar using beneficial bacteria such as lactic acid bacteria or cultures thereof. For example, Korean Patent Publication No. 10-2012-0119555 discloses a composition for inhibiting α-glucosidase activity using Pediococcus sp. JNU534 strain or its culture as an active ingredient, there is.

The inventors of the Republic of Korea Patent Publication No. 10-2021-0041681 (Patent Registration No. 10-2284112) use seomoktae as a natural raw material, such as Saccharomyces servazzii or Saccharomyces cerevisiae. A method of using a culture obtained by culturing a yeast strain as a composition for lowering blood sugar has been disclosed.

Republic of Korea Patent Publication No. 10-2004-0052398 Republic of Korea Patent Publication No. 10-2004-0077976 Republic of Korea Patent Publication No. 10-2012-0119555 Republic of Korea Patent Publication No. 10-2016-0126715 Republic of Korea Patent Publication No. 10-2021-0041681

An object of the present invention is to provide a composition for lowering blood sugar that can inhibit and inhibit the activity of alpha-glusidase using natural products. Such a composition for lowering blood sugar can be usefully used as a health functional food (or health supplement food) for preventing or improving diabetes or a pharmaceutical composition for preventing or treating diabetes.

The present invention provides a composition for lowering blood sugar comprising a fermented product obtained by fermenting safflower seeds with lactic acid bacteria as an active ingredient for inhibiting alpha-glucosidase activity.

In one aspect of the present invention, the lactic acid bacteria are Enterococcus faecium , Lactobacillus pentosus , Lactobacillus plantarum , Leuconostoc mesenteroides , lacto Bacillus fermentum ( Lactobacillus fermentum ), Lactobacillus brevis ( Lactobacillus brevis ), Lactobacillus sakei ( Lactoba cillus sakei ), and Leuconostoc holzapfelii strains .

In one aspect of the present invention, the lactic acid bacteria is Enterococcus faecium ( Enterocuccus faecium ) It is preferable that the strain, the lactic acid bacteria may be Enterococcus faecium Ceb-ch-001 (KCCM 11909P) strain.

In one aspect of the present invention, there is provided a health functional food or health supplement for preventing or improving diabetes comprising the composition for lowering blood sugar as an active ingredient.

In one aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating diabetes comprising the composition for lowering blood sugar as an active ingredient.

On the other hand, the present invention provides a method for preparing a composition for lowering blood sugar, comprising culturing pulverized safflower seeds mixed with a medium and lactic acid bacteria, and preparing a composition for lowering blood sugar from the fermented product obtained after the culture.

In one aspect of the present invention, the medium may be water substantially free of nutrients, preferably sterilized water, distilled water, or purified water.

In one aspect of the present invention, the nutrient source may be 0.5% by weight (w / v) or less based on the total medium volume.

In one aspect of the present invention, the safflower seed may be 1 to 4% by weight (w / v) based on the total medium volume.

According to the present invention, the fermented product obtained by fermenting safflower seeds with lactic acid bacteria had a significantly superior blood sugar lowering effect compared to safflower seeds not treated with lactic acid bacteria. It was found that the blood sugar lowering effect was higher than that of

Therefore, the fermented product of safflower seeds using lactic acid bacteria according to the present invention can be usefully used in natural foods such as health functional foods and health supplements for lowering blood sugar, and can also be usefully used as natural medicines.

1 is a graph showing the difference in α-glucosidase inhibitory ability (%) according to the type of lactic acid bacteria in Examples of the present invention.
Figure 2 is a graph showing the difference in α-glucosidase inhibitory ability (%) according to the type of medium and culture time in Examples of the present invention.
Figure 3 is a graph showing the difference in α-glucosidase inhibitory ability (%) according to the change in safflower seed content during fermentation in Examples of the present invention.
Figure 4 is a graph showing the difference in α-glucosidase inhibitory ability (%) according to the change in pH of the fermented safflower seed according to the present invention.
5 is a graph showing the difference in α-glucosidase inhibitory ability (IC ₅₀ , mg/mL) of fermented safflower seeds according to the present invention.

In order to develop a composition for lowering blood sugar derived from natural products, the present inventors conducted research for a long time and used seomoktae (black soybean) as Saccharomyces servazzii strain or Saccharomyces cerevisiae strain It was confirmed that the culture obtained by culturing with the same yeast had an excellent hypoglycemic effect, and was disclosed in Korean Patent Publication No. 10-2021-0041681 (Patent Registration No. 10-2284112).

As a result of continuous research on various natural product candidates that can be used as hypoglycemic agents, the present inventors have confirmed that the fermented product obtained by fermenting safflower seeds with lactic acid bacteria has a remarkably excellent hypoglycemic effect. In particular, Enterococcus faecium ( Enterocuccus faecium ) The present invention was completed by confirming that the fermented product of safflower seeds using the strain had a higher hypoglycemic effect compared to acarbose, which is known as an excellent alpha-glucosidase inhibitor in clinical practice.

Safflower (Carthamus tinctorious L.) is an annual plant belonging to the Asteraceae family. The medicinal parts of safflower include flowers and seeds. do. In particular, safflower oil obtained by milking roasted safflower seeds contains a large amount of unsaturated fatty acids and tocopherols, including linoleic acid, and is known to have antioxidant, antithrombotic and antihypertensive actions.

The general components of safflower seeds are shown in Table 1 below.

moisture crude fiber crude fat views crude protein N-free extract 7.7±0.2 41.2±0.3 17.3±0.1 3.0±0.1 17.5±0.5 13.3±0.4

In addition, safflower seeds contain phenolic compounds such as serotonin, lignans, and flavonoids, so they have strong antioxidant and antihyperlipidemic effects. In particular, as a phytoestrogen component with estrogen-like activity, bone formation similar to components known as existing phytoestrogen compounds can prevent osteoporosis in menopausal women and form new bone in case of bone defect. Toshiyuki Takahashi et al. reported that safflower seeds were solvent-extracted, and serotonin derivatives contained in safflower seeds had the effect of inhibiting the activity of α-glucosidase ( Phytother Research . 2012 May; 26(5): 722- 6). However, these phenolic compounds are water-soluble and physiologically active components, so when hot water extraction is performed using safflower seeds containing a large amount of oil and fat as it is, as well as when hot water extraction is performed using safflower seed powder obtained after degreasing through a conventional solvent extraction method The extraction yield is low, and a complicated purification process is required, resulting in problems such as residual organic solvent, solvent removal, and low separation effect of oil components.

The present inventors primarily select 8 species with excellent pharmacological and physiological activities for 50 or so lactic acid bacteria directly isolated and identified from kimchi and cheonggukjang, and each of these lactic acid bacteria has a fermented product obtained by fermenting safflower seeds using As a result of testing the ability to inhibit alpha-glucosidase activity, it was confirmed that the hypoglycemic effect was very good.

The present invention provides a composition for lowering blood sugar comprising a fermented product obtained by fermenting safflower seeds with lactic acid bacteria as an active ingredient for inhibiting alpha-glucosidase activity.

In one aspect of the present invention, the lactic acid bacteria for fermentation of safflower seeds are Enterococcus faecium , Lactobacillus pentosus , Lactobacillus plantarum , Leuconostoc mesenteroides ( Leuconostoc mesenteroides ), Lactobacillus fermentum ( Lactobacillus fermentum ), Lactobacillus brevis ( Lactobacillus brevis ), Lactobacillus sakei ( Lactobasa cillus sakei ), and Leuconostoc holzapfelii ). no. However, the lactic acid bacteria are more preferably Enterococcus Faecium .

All of the lactic acid bacteria used in the present invention were isolated and identified by the present inventors and are as follows.

Enterococcus faecium Ceb -ch-001 is a strain isolated from Cheonggukjang, and on October 07, 2016, the Korean Culture Center of Microorganisms (Hongje-2-ga, Seodaemun-gu, Seoul) -Gil 45) was deposited under the accession number KCCM 11909P. The Enterococcus Faecium Ceb-ch-001 strain is known to be effective in lowering cholesterol according to Korean Patent Registration No. 10-1790548.

Lactobacillus pentosus Ceb-kc-009 ( Lactobacillus pentosus Ceb-kc-009) strain was isolated from kimchi and was deposited with the Korea Microorganism Conservation Center on October 16, 2017 under the accession number KCCM 12129P.

Lactobacillus plantarum subspecies Plantarum Ceb-kc-007 ( Lactobacillus plantarum subsp. plantarum Ceb-kc-007) is a strain isolated from kimchi and was deposited with the Korea Microorganism Conservation Center on October 16, 2017 under the accession number KCCM 12127P. has been deposited The Lactobacillus plantarum Ceb-kc-007 strain is known to be effective in lowering cholesterol in Korean Patent Registration No. 10-1790548, and is known to be effective in anti-obesity in Patent Registration No. 10-2274678.

Leuconostoc mesenteroides Ceb-kc-001 ( Leuconostoc mesenteroides Ceb-kc-001) strain was isolated from kimchi and was deposited with the Korea Microorganism Conservation Center on April 6, 2016 under the accession number KCCM 11827P. The Leuconostoc mesenteroides Ceb-kc-001 is known to be effective in decomposing fat in Korean Patent Registration No. 10-1790548 to prevent hair loss or promote hair growth.

The Lactobacillus fermentum Ceb-kc-005 strain was isolated from kimchi and was deposited with the Korea Microorganism Conservation Center on March 30, 2017 under the accession number KCCM 12003P. The Lactobacillus fermentum Ceb-kc-005 strain or culture is known to be effective in preventing hair loss, promoting hair growth, or improving sexual function according to Korean Patent Registration No. 10-1791088.

Lactobacillus brevis Ceb-kc-006 ( Lactobacillus brevis Ceb-kc-006) strain is a strain isolated from kimchi by the present inventor and deposited with the Korea Microorganism Conservation Center on October 16, 2017 under accession number KCCM 12126P.

Lactobacillus sakei Ceb-kc-002 ( Lactobacillus sakei Ceb-kc-002) strain was isolated from kimchi and was deposited with the Korea Microorganism Conservation Center on June 1, 2016 under the accession number KCCM 11841P. The Lactobacillus Sakei Ceb-kc-002 strain is known to be effective in lowering cholesterol in Korean Patent Registration No. 10-1790548, thereby preventing hair loss or promoting hair growth.

Leuconostoc holzapfelii Ceb-kc-003 ( Leuconostoc holzapfelii Ceb-kc-003) strain was isolated from kimchi and was deposited with the Korea Microorganism Conservation Center on April 11, 2016 under the accession number KCCM 11830P. The Leuconostok Holjappeli Ceb-kc-003 strain or culture is disclosed as effective in preventing hair loss, promoting hair growth, or improving sexual function in Korean Patent Registration No. 10-1734960.

Safflower seeds may be used both dry and non-dried, and may be ground and used for contact area with lactic acid bacteria (safflower seed powder, etc.).

In one aspect of the present invention, fermentation of safflower seeds with similar bacteria may be performed in the presence of water (sterilized water, distilled water, purified water, etc.) that does not or does not contain additional nutrients other than safflower seeds. As a result of the actual experiment described later, when fermented in MRS medium, it showed relatively low alpha-glucosidase inhibitory ability, and dextrose medium (2% w/v) showed higher inhibitory ability than MRS medium, but additional The fermented product fermented in distilled water without nutrients showed the highest inhibitory activity. This is because in the analysis of the general components of dried safflower seeds, crude fiber belonging to carbohydrates as a carbon source is very high at 41.2%, and crude protein, a nitrogen source, is also high at 17%, so safflower seeds themselves act as a sufficient source of nutrients. It is judged to be

The fact that the nutrient source is not substantially included in the above means that the fermentation activity of lactic acid bacteria is not inhibited, and the nutrient source is preferably 0.5% by weight (w / v) or less based on the total medium volume.

In one aspect of the present invention, the amount of safflower seeds added to the medium is preferably 1 to 4% by weight (w/v) based on the total volume of the medium.

The lactic acid bacteria fermented product of safflower seeds may be in a liquid state immediately after completion of fermentation or in a purified liquid state, or the fermented product may be in a dry state, preferably in a dried or freeze-dried powder state.

The hypoglycemic composition according to the present invention may be in a liquid or dry state, or in a powder form.

The composition may be formulated into various oral or parenteral formulations, but may be preferably oral formulations.

Health functional food (health supplement) according to the present invention, the pharmaceutical composition may further include conventionally known additives that are pharmaceutically acceptable or food-related. For example, antioxidants, buffers, bacteriostatic agents, diluents, dispersants, surfactants, binders, and the like may be used as additives, if necessary. The health functional food (health supplement) and pharmaceutical composition according to the present invention may be formulated into liquid formulations such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets.

The daily dose of the composition or the fermented safflower seed lactic acid bacteria may vary depending on the weight, age, sex, health condition, treatment, prevention, or improvement of the main symptoms, administration time, administration method, and severity of the subject, but about 0.01 It may be mg/kg to about 10 g/kg, preferably about 0.05 mg/kg to about 500 mg/kg, and may be administered or applied 1 to 6 times a day, such as 1 to 4 times a day.

The present invention will be described in detail through the following examples. The following examples are only examples of the present invention, and the technical spirit of the present invention is not limited to the following examples.

Example 1: Preparation of lactic acid bacteria fermented product of safflower seeds ( Enterococcus Passium Ceb-ch-001 strain)

The dried safflower seeds were finely pulverized with a grinder to prepare a powder form.

에서 15분간 멸균하였다.30 g of safflower seed powder was added to 1000 mL of distilled water, 3% by weight (w / v) was added, and after mixing, about 121

sterilized for 15 minutes.

배양기에서 120rpm을 유지하면서 48시간 동안 배양하였다. 배양 종료 후 121

/15min 동안 멸균한 다음, 여과한 후 액상을 동결건조 후 -20

에 보관하였다.After adding and mixing 3% by weight (w / v) of the dextrose to the safflower seed solution, the liquid seed of the Enterococcus faecium Ceb-ch-001 strain was 5% (v / v) based on the safflower seed solution After inoculation (the viable cell count of the liquid seed of lactic acid bacteria is constantly adjusted to 1.0 × 10 ⁹ CFU/mL),

It was cultured for 48 hours while maintaining 120 rpm in an incubator. After incubation 121

/ After sterilizing for 15min, filtering and freeze-drying the liquid at -20

stored in.

Example 2: Preparation of lactic acid bacteria fermented product of safflower seeds ( Lactobacillus Pentosus Ceb-kc-009 strain)

In Example 1, Lactobacillus fermented product of safflower seeds was obtained in the same manner as in Example 1, except that Lactobacillus pentosus Ceb-kc-009 strain was used instead of Enterococcus faecium Ceb-ch-001 strain. .

Example 3: Preparation of lactic acid bacteria fermented product of safflower seeds ( Lactobacillus Plantarum Ceb-kc-007)

Lactobacillus fermented product of safflower seeds was obtained in the same manner as in Example 1, except that the Lactobacillus plantarum Ceb-kc-007 strain was used instead of the Enterococcus faecium Ceb-ch-001 strain in Example 1. did

Example 4: Preparation of lactic acid bacteria fermented product of safflower seeds ( Leuconostoc mesenteroides Ceb-kc-001)

Lactobacillus fermented product of safflower seeds was prepared in the same manner as in Example 1, except that the Leuconostoc mecenteroides Ceb-kc-001 strain was used instead of the Enterococcus faecium Ceb-ch-001 strain in Example 1. obtained.

Example 5: Preparation of lactic acid bacteria fermented product of safflower seeds ( Lactobacillus Fermentum Ceb-kc-005)

Lactobacillus fermented product of safflower seeds was obtained in the same manner as in Example 1, except that the Lactobacillus fermentum Ceb-kc-005 strain was used instead of the Enterococcus faecium Ceb-ch-001 strain in Example 1. .

Example 6: Preparation of lactic acid bacteria fermented product of safflower seeds ( Lactobacillus brevis Ceb-kc-006)

Lactobacillus fermented product of safflower seeds was obtained in the same manner as in Example 1, except that the Lactobacillus brevis Ceb-kc-006 strain was used instead of the Enterococcus faecium Ceb-ch-001 strain in Example 1.

Example 7: Preparation of lactic acid bacteria fermented product of safflower seeds ( Lactobacillus sake Ceb-kc-002)

Lactobacillus fermented product of safflower seeds was obtained in the same manner as in Example 1, except that the Lactobacillus Sakei Ceb-kc-002 strain was used instead of the Enterococcus faecium Ceb-ch-001 strain in Example 1. .

Example 8: Preparation of lactic acid bacteria fermented product of safflower seeds ( Leukonostok Holjappelli Ceb-kc-003)

Lactic acid bacteria fermented product of safflower seeds was obtained in the same manner as in Example 1, except that the Leuconostok Holjappeli Ceb-kc-003 strain was used instead of the Enterococcus Faecium Ceb-ch-001 strain in Example 1. did

Experimental Example 1: Evaluation of alpha-glucosidase inhibitory ability

Alpha-glucosidase inhibitory ability (activity inhibition) was evaluated for the fermented safflower seed lactic acid bacteria (freeze-dried) prepared in Examples 1 to 8. As a positive control for comparing the hypoglycemic effect of the composition according to the present invention, acarbose (Control 1) used as an alpha-glucosidase inhibitor and a safflower seed powder solution not treated with lactic acid bacteria (Control 2) α-glucosidase inhibitory ability was measured in the same manner.

α-glucosidase inhibition was measured by the following method:

Each sample was dissolved in 100mM sodium phosphate buffer (pH 6.8), centrifuged at 13,000rpm/2min, and the supernatant was used as a sample solution.

에서 10분간 가온 후 5mM 기질(P-NPG) 20μL를 넣고 37

에서 20분간 반응시켰다. 그리고 0.1M Na₂CO₃ 50μL를 넣어 효소 반응을 종료시키고 405nm에서 흡광도를 측정하여 그 결과를 하기 표 2 및 도 1에 나타내었다.20μL of sample solution of a certain concentration dissolved in 40μL of 100mM sodium phosphate buffer (pH 6.8) and 20μL of α-glucosidase (1U/mL) were added to a 96-well plate, respectively, and 37

After warming for 10 minutes, 20 μL of 5 mM substrate (P-NPG) was added and 37

reacted for 20 minutes. Then, 50 μL of 0.1M Na ₂ CO ₃ was added to terminate the enzyme reaction, and absorbance was measured at 405 nm. The results are shown in Table 2 and FIG. 1 below.

[Equation 1]

Inhibition (%) = [1-(A _sample /A _control )]×100

(In Equation 1 above,

A _control is the control absorbance, which is the absorbance in the absence of an inhibitor,

A _sample is the absorbance of the sample and is the absorbance in the presence of an inhibitor.)

Example sample name material name badge strain name α-glucosidase
Inhibition (%) control 1 A Acarbose - - 86.4±0.3 control 2 3 safflower seed
3.0%
(w/v) Distilled water - 67.9±0.2 Example 1 3EfD safflower seed
3.0%
(w/v) Dextrose
3.0%
(w/v) E. faecium
Ceb-ch-001 83.8±0.8 Example 2 3LpeD L. pentosus Ceb-kc-009 77.2±0.5 Example 3 3LplD L. plantarum
Ceb-kc-007 75.9±0.3 Example 4 3LmD L. mesenteroides
Ceb-kc-001 78.9±0.3 Example 5 3LfD L. fermentum
Ceb-kc-005 75.2±0.4 Example 6 3LbD L. brevis
Ceb-kc-006 78.0±0.3 Example 7 3LsD L. sakei
Ceb-kc-002 74.7±0.8 Example 8 3LhD L. holzapfelii
Ceb-kc-003 77.8±0.4 *Acarbose: 50 mg/mL
* Sample concentration: 20mg/mL
* Glucosidase: 1U/mL
* Incubation time: 35℃, 48h

As shown in Table 2 and FIG. 1, the unfermented safflower seeds (Control 2) showed 67.9% inhibition, whereas the fermented product fermented with the lactic acid bacteria of the present invention showed more than 75% inhibition, indicating that the fermentation of lactic acid bacteria inhibited α-glucosidase. It can be seen that there is a great effect on activity inhibition. Among the eight lactic acid bacteria , Enterococcus faecium Ceb-ch-001 showed the highest inhibition of 83.8%, which was not significantly different from that of acarbose (control 1).

Examples 9 to 13: Preparation of lactic acid bacteria fermented product of safflower seeds (type of medium and culture time)

The same method as in Example 1 was performed using the Enterococcus faecium Ceb-ch-001 strain showing the best alpha-glucosidase inhibitory activity in Examples 1 to 8, but an addition for optimization of safflower seed culture in Example 1 In order to confirm the nutrient source and culture time, dextrose as a carbon source, MRS as a medium optimized for lactic acid bacteria culture, and distilled water were fermented for 24 hours and 48 hours, respectively, to prepare fermented products of safflower seeds.

Then, α-glucosidase inhibitory ability was measured for the fermented safflower seeds prepared in the above examples in the same manner as in Experimental Example 1, and the results are shown in Table 3 and FIG. 2.

sample name Material name/
strain name badge incubation time
(35℃) After incubation
pH α-glucosidase
Inhibition (%) control 2 Con Safflower seeds 3% (w/v)/
E. faecium
Ceb-ch-001 Distilled water - 6.43 67.9±0.3 Example 1 ED48 Dextrose
2% (w/v) 48h 3.92 83.8±0.8 Example 9 ED24 Dextrose
2% (w/v) 24h 4.06 82.2±1.5 Example 10 E48 Distilled water 48h 4.97 87.1±0.8 Example 11 E24 Distilled water 24h 4.98 88.5±0.7 Example 12 EM48 MRS 48h 4.31 60.1±0.3 Example 13 EM24 MRS 24h 4.42 59.8±0.8 * Sample concentration: 20mg/mL
* Glucosidase: 1U/mL

As shown in Table 3 and FIG. 2, the sample fermented using MRS, which is an optimized medium for lactic acid bacteria culture, showed the lowest inhibition of all samples at 59.8 to 60.1%, followed by 2% dextrose as a carbon source The added sample showed 82.2~83.8% of inhibition. On the other hand, in the case of the sample fermented with lactic acid bacteria without any additional nutrient source, it was the highest at 87.1 ~ 88.5%, and the difference according to the fermentation time was not large.

This means that safflower seeds contain 41.2% of crude fiber and do not require a separate carbon source.

It contains 17.5% of crude protein, which means that it is supplied as a nitrogen source for lactic acid bacteria cultivation.

Examples 14 to 19: Preparation of lactic acid bacteria fermented product of safflower seeds (fermented safflower seeds content)

에서 24시간 동안 배양하였다.In order to optimize the α-glucosidase inhibitory ability according to the fermentation content of safflower seeds, the same method as in Example 1 was performed, but 1.0% by weight (w / v) of safflower seeds and 1.5% by weight (w / v) of safflower seeds were used instead of 3.0% by weight (w / v) of safflower seeds. / v), 2.0% by weight (w / v), 2.5% by weight (w / v), 3.5% by weight (w / v), 4.0% by weight (w / v) were changed to each, and the medium was added Only distilled water without nutrient source, incubation time was 35

incubated for 24 hours.

Thereafter, α-glucosidase inhibitory ability was measured for the fermented safflower seeds prepared in the above examples in the same manner as in Experimental Example 1, and the results are shown in Table 4 and FIG. 3.

Example sample name material name strain name After incubation
pH α-glucosidase
Inhibition (%) control 2 1.0 Safflower seeds 1.0% (w/v) - 6.23 68.6±0.3 Example 14 1.0E Safflower seeds 1.0% (w/v) E. faecium
Ceb-ch-001 4.49 69.8±1.1 Example 15 1.5E Safflower seeds 1.5% (w/v) 4.61 76.3±0.8 Example 16 2.0E Safflower seeds 2.0% (w/v) 4.75 78.7±0.2 Example 17 2.5E Safflower seeds 2.5% (w/v) 4.83 82.5±0.9 Example 11 3.0E Safflower seeds 3.0% (w/v) 4.98 88.5±0.7 Example 18 3.5E Safflower Seed 3.5% (w/v) 5.11 80.1±1.3 Example 19 4.0E Safflower seeds 4.0% (w/v) 5.15 78.5±0.5 * Sample concentration: 20mg/mL
* Glucosidase: 1U/mL
* Culture medium: distilled water
* Incubation time: 35℃, 24h

As shown in Table 4 and FIG. 3, the highest inhibition was shown when 3.0 wt% (w/v) of safflower seeds was used.

Experimental Example 2: Evaluation of α-glucosidase inhibitory ability before and after adjusting the pH of the sample

In order to compare the α-glucosidase inhibitory ability of fermented safflower seeds before and after pH adjustment, the pH of the fermented safflower seed-like bacteria prepared in Example 11 was changed from pH 4.98 to pH 6.80 to measure α-glucosidase inhibitory ability, as shown in Table 5 and Figures below. 4.

Example sample name material name strain name pH α-glucosidase
Inhibition (%) control 2 Con Safflower seeds 3% (w/v) - 6.43 67.9±0.3 Example 11 3E4.98 Safflower seeds 3% (w/v) E. faecium Ceb-ch-001 4.98 88.5±0.7 Example 20 3E6.80 Safflower seeds 3% (w/v) E. faecium
Ceb-ch-001 6.80 87.7±0.5 * Culture medium: distilled water
* Incubation time

As shown in Table 5 and FIG. 4, it was found that there was no significant difference in inhibition ability from the sample whose pH was adjusted to 6.8.

Experimental Example 3: α-glucosidase inhibitory ability of dried fermentation broth (IC ₅₀ , mg/mL) evaluation

As in Example 11, 3% by weight (w / v) of safflower seeds was fermented for 24 hours in distilled water without a separate nutrient source with Enterococcus faecium Ceb-ch-001 strain, and freeze-dried fermented product and acarbose (control 1), α-glucosidase inhibitory activity (IC ₅₀ , mg/mL) of fermented safflower seeds (Control 2) was analyzed. After setting the sample to various concentrations, the α-glucosidase inhibitory ability was analyzed, and the concentration at which 50% inhibition was confirmed using the SPSS statistical program was shown in Table 6 and FIG. 5 below.

Example sample name IC ₅₀ (mg/mL) control 1 Acarbose 3.3±0.2 control 2 Untreated Safflower Seed 25.5±0.5 Example 11 Safflower seeds 3% (w/v) 2.5±0.3 * Nutrient medium: distilled water
* Incubation time: 35℃, 24h

As shown in Table 6 and FIG. 5, the IC ₅₀ of fermented safflower seeds fermented with lactic acid bacteria was 2.5 mg/mL, which was lower than 3.3 mg/mL of acarbose as a positive control. Therefore, fermented safflower seeds using lactic acid bacteria can be usefully used as natural food (health functional food, health supplement food, etc.) know that it can.

Korea Microbial Conservation Center (Overseas) KCCM11909P 20161007 Korea Microbial Conservation Center (Overseas) KCCM12129P 20171016 Korea Microbial Conservation Center (Overseas) KCCM12127P 20171016 Korea Microbial Conservation Center (Overseas) KCCM11827P 20160406 Korea Microbial Conservation Center (Overseas) KCCM12003P 20170330 Korea Microbial Conservation Center (Overseas) KCCM12126P 20171016 Korea Microbial Conservation Center (Overseas) KCCM11841P 20160601 Korea Microbial Conservation Center (Overseas) KCCM11830P 20160411

Claims (10)

A composition for lowering blood sugar, comprising a fermented product obtained by fermenting safflower seeds with lactic acid bacteria as an active ingredient for inhibiting alpha-glucosidase activity.
According to claim 1,
The lactic acid bacteria are Enterococcus faecium , Lactobacillus pentosus , Lactobacillus plantarum , Leuconostoc mesenteroides , Lactobacillus fermentum , Lactobacillus brevis ( Lactobacillus brevis ), Lactobacillus sakei ( Lactoba cillus sakei ) and Leuconostoc Holzapfelii ( Leuconostoc holzapfelii ) At least one member selected from the group consisting of strains, a composition for lowering blood sugar.
According to claim 1,
The lactic acid bacteria Enterococcus faecium ( Enterocuccus faecium ) strain, blood sugar lowering composition.
According to claim 1,
The lactic acid bacteria is Enterococcus Faecium Ceb-ch-001 (KCCM 11909P) strain, blood sugar lowering composition.
A health functional food or health supplement food for preventing or improving diabetes, comprising the composition for lowering blood sugar according to any one of claims 1 to 4 as an active ingredient.
A pharmaceutical composition for preventing or treating diabetes, comprising the composition for lowering blood sugar according to any one of claims 1 to 4 as an active ingredient.
A method for preparing a composition for lowering blood sugar, comprising culturing pulverized safflower seeds in a mixture with a medium and lactic acid bacteria, and preparing a composition for lowering blood sugar from a fermented product obtained after the culture.
According to claim 7,
The method of claim 1 , wherein the medium is water containing substantially no nutrient source.
According to claim 8,
The nutrient source is 0.5% by weight (w / v) or less based on the total medium volume, method for producing a composition for lowering blood sugar.
According to claim 7,
The safflower seed is 1 to 4% by weight (w / v) based on the total medium volume, method for producing a composition for lowering blood sugar.