KR102216618B1 - Fermentation method of lactic acid bacteria using artichoke and its use - Google Patents

Abstract

The present invention relates to a method of fermenting lactic acid bacteria using artichoke and a use thereof, and the fermented artichoke manufactured by using lactic acid bacteria of the present invention has improved cynarin content and antioxidant activity compared to an existing artichoke extract, thereby being able to be more usefully used in health functional foods for lowering cholesterol, improving liver/kidney/physiological metabolism functions, or relieving hangover.

Description

Fermentation method of lactic acid bacteria using artichoke and its use

The present invention relates to a method for fermenting lactic acid bacteria using artichoke and a use thereof.

Oxidative apoptosis is a phenomenon defined as an imbalance of antioxidant and oxidative systems in vivo, and occurs due to an increase in accumulation of reactive oxygen species (ROS) in cells. Free radicals are removed by antioxidants in the human body, and free radicals are removed to maintain the balance of oxidation-oxidation, thereby preventing DNA damage, protein modification and lipid peroxidation. However, the generation of excessive reactive oxygen species causes damage to normal cells, apoptosis, and cell necrosis, and is involved in various diseases such as cancer, arthritis, chronic inflammation, diabetes, lung disease, and stroke. In particular enemy cell damage oxidation is hydrogen peroxide _{(hydrogen peroxide, H 2 O 2} ) non-free radical, as well as excess negative ion oxide, including ^- there is mediated by reactive oxygen species including free radicals, such as, which (superoxide anion, O ₂₎ It attacks various biological polymers such as DNA, proteins, and lipids in the human body to destroy cell functions, resulting in cell death. Such oxidative cell death can be reduced by ingesting antioxidants, but synthetic antioxidants such as butylated hydroxy anisol (BHA) and butylated hydroxy toluene (BHT) have side effects such as cancer and lipid metabolism imbalance. Are doing. Therefore, there is a need to develop a safe natural product-derived antioxidant.

On the other hand, Artichoke is a thistle-like perennial flower belonging to the Asteraceae family, and it is a herb characterized by a large thistle-like flower that is called the giant thistle. It is said that it has been cultivated since ancient Greece and Roman times and used for food. Unlike other vegetables and spices, large buds are used for food. Artichoke contains sugars that act like insulin, so it is highly valued for medicinal purposes, and leaves and roots are known to have cholesterol-lowering effects. In addition, it has strong antioxidant activity, protective function for hepatocytes, blood sugar control, and other effects of preventing adult diseases and promoting digestion.

On the other hand, Korean Patent No. 1725638 discloses'Artichoke fermented tea using artichoke, fermented powder tea and fermented pill manufacturing method', and Korean Patent No. 1854444 discloses'artichoke leaf extract as an active ingredient. It discloses a cosmetic composition for preventing and improving stretch marks, including. However, the'lactic acid bacteria fermentation method using artichoke and its use' of the present invention has not yet been disclosed.

The present invention was derived from the above requirements, the inventors of the artichoke extract Lactobacillus plantarum CBG-C21 ( Lactobacillus plantarum CBG-C21, KACC92083P) strain, Lactobacillus rhamnosus CBG-C14 ( Lactobacillus rhamnosus CBG- C14, KACC91981P) strain or Bifidobacterium animalis CBG-C10 ( Bifidobacterium animalis CBG-C10, KCTC11978BP) strain was respectively inoculated to prepare an artichoke fermented product, or cellulase, pectinase, or inulinase in artichoke extract When the artichoke fermented product was prepared by inoculating each of the three strains and inoculating each of the three strains, it was confirmed that the content of sinarine and the antioxidant activity were significantly enhanced compared to the artichoke extract before fermentation, thereby completing the present invention.

In order to solve the above problems, the present invention provides a method for producing an artichoke fermented product with improved content of sinarine and antioxidant activity, including fermenting by inoculating an artichoke extract with lactic acid bacteria.

In addition, the present invention provides an artichoke fermented product with improved content of sinarine and antioxidant activity prepared by the above production method.

In addition, the present invention provides a health functional food comprising an artichoke fermented product with improved content of sinarine and antioxidant activity.

The artichoke fermented product prepared using the lactic acid bacteria of the present invention has increased content of sinarine and antioxidant activity compared to conventional artichoke extract, reducing cholesterol, improving liver, kidney, physiological metabolism, or relieving hangovers. It can be used more usefully in food.

1 is an artichoke extract Lactobacillus plantarum CBG-C21 ( Lactobacillus plantarum CBG-C21) strain, Lactobacillus rhamnosus CBG-C14 ( Lactobacillus rhamnosus CBG-C14) strain and Bifidobacterium animalis CBG-C10 ( Bifidobacterium animalis CBG-C10) strains were each inoculated to confirm the change in the content of sinarine in fermented artichokes. ** and * mean that the effect of increasing the content of sinarine is statistically significant compared to before fermenting the artichoke extract, and means that the p values are less than 0.01 and 0.05.
Figure 2 is a Lactobacillus plantarum CBG-C21 strain, Lactobacillus rhamnosus CBG-C14 strain and Bifidovac after each treatment of an artichoke extract with cellulase (A), pectinase (B) and inulinase (C) This is the result of confirming the change in the content of sinarine of fermented artichokes prepared by inoculating each of Therium animalis CBG-C10 strains. ** and * mean that the effect of increasing the content of sinarine is statistically significant compared to before fermenting the artichoke extract, and means that the p values are less than 0.01 and 0.05.

In order to achieve the object of the present invention, the present invention provides a method for producing an artichoke fermented product with improved content of sinarin and antioxidant activity, including fermenting by inoculating lactic acid bacteria in an artichoke extract.

In the manufacturing method of the artichoke fermented product of the present invention, the lactic acid bacteria are Lactobacillus plantarum CBG-C21 ( Lactobacillus plantarum CBG-C21) (KACC92083P) strain, Lactobacillus rhamnosus CBG-C14 ( Lactobacillus rhamnosus CBG-C14) (KACC91981P) The strain, Bifidobacterium animalis CBG-C10 (KCTC11978BP) strain, or a mixed strain thereof, but is not limited thereto. The Lactobacillus plantarum CBG-C21 (KACC92083P) strain is a strain deposited as of July 10, 2015 at the Agricultural Biotechnology Research Institute, and the Lactobacillus rhamnosus CBG-C14 ( Lactobacillus rhamnosus CBG- C14) (KACC91981P) strain is a strain deposited with the Institute of Agricultural Biotechnology on September 30, 2014, and the Bifidobacterium animalis CBG-C10 ( Bifidobacterium animalis CBG-C10) (KCTC11978BP) strain is the Korea Research Institute of Bioscience and Biotechnology. It is a strain deposited on July 6, 2011.

In addition, the fermentation may be performed at 32 to 42° C. for 20 to 28 hours, preferably at 37° C. for 24 hours, but is not limited thereto.

In addition, the artichoke extract may further include a step of pretreating a glycolytic enzyme, but is not limited thereto. The enzyme pretreatment may be cellulase, pectinase, inulinase, or a mixed treatment thereof, and preferably 0.005 to 0.03% of cellulase, pectinia or inulinase may be treated, and most preferably It may be treated with 0.02% of cellulase, 0.01% of pectinase, and 0.025% of inulinase, but is not limited thereto.

The manufacturing method according to the present invention is more specifically,

1) sterilizing the artichoke extract;

2) fermenting by inoculating lactic acid bacteria in the artichoke extract sterilized in step 1); And

3) lyophilizing the artichoke fermented product of step 2) and pulverizing it; may include, and preferably

1) sterilizing the artichoke extract at 100 to 120° C. for 20 to 40 minutes;

2) inoculating the lactic acid bacteria of 1.0×10 ⁷ ~1.0×10 ⁹ CFU/ml to the artichoke extract sterilized in step 1) and fermenting at 32-42°C for 20-28 hours; And

3) freeze-drying and powdering the artichoke fermented product of step 2); may include, and most preferably

1) sterilizing the artichoke extract at 105° C. for 30 minutes;

2) 1.0×10 ⁸ CFU/ml of Lactobacillus plantarum CBG-C21 (KACC92083P) strain, Lactobacillus rhamnosus CBG-C14 ( Lactobacillus rhamnosus ) in the artichoke extract sterilized in step 1) CBG-C14) (KACC91981P) strain, Bifidobacterium animalis CBG-C10 ( Bifidobacterium animalis CBG-C10) (KCTC11978BP) strain or a mixed strain thereof inoculated and fermented at 37° C. for 24 hours; And

3) lyophilizing the artichoke fermented product of step 2) to powder it, but is not limited thereto.

In addition, the artichoke extract used to prepare the artichoke fermentation product may be extracted using various extraction methods such as solvent extraction, physical extraction, and enzymatic extraction, but is not limited thereto.

In addition, the artichoke extract is intended to contain any one of an extract obtained by an extraction treatment, a dilute or concentrate of the extract, a dried product obtained by drying the extract, or a crude product or a purified product.

The present invention also provides an artichoke fermented product with improved content of sinarine and antioxidant activity prepared by the above production method.

The present invention also provides a health functional food comprising an artichoke fermented product with improved content of sinarine and antioxidant activity.

There is no particular limitation on the kind of the health functional food. Examples of health functional foods containing the artichoke fermentation products include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, rice cakes, dairy products including ice cream, various soups, and beverages. , Tea, drinks, alcoholic beverages and vitamin complexes, and includes all health functional foods in the usual sense.

The artichoke fermented product according to the present invention can be used as a health functional food because the content of sinarin is increased and DPPH and ABTS radical scavenging activity is excellent compared to the artichoke extract.

The health functional food may be prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, effervescent tablet, and beverage, but is not limited thereto.

When the fermented artichoke of the present invention is used as a food additive, the fermented artichoke may be added as it is, or may be used with other foods or food ingredients, and may be appropriately used according to a conventional method. The active ingredient can be appropriately used depending on the purpose of use. In general, in the manufacture of food or beverage, the fermented artichoke of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less based on the raw material. However, in the case of long-term intake for the purpose of health control, it can be used in an amount within a range without any problems in terms of safety.

The health functional food of the present invention includes ingredients that are commonly added during food production, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when prepared as a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates are monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrins, cyclodextrins, etc.) or sugar alcohols (eg , Xylitol, sorbitol, erythritol, etc.). The flavoring agent may be a natural flavoring agent (eg, taumatin, stevia extract, etc.) and a synthetic flavoring agent (eg, saccharin, aspartame, etc.). In addition to the health functional food composition, various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonic acid It may further contain a carbonation agent used in beverages.

Although the ratio of the added ingredients is not very important, it is generally selected from 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food of the present invention.

Hereinafter, it will be described in detail by examples of the present invention. However, the following examples are only illustrative of the present invention, and the contents of the present invention are not limited to the following examples.

Materials and methods

1. Artichoke fermentation production

The artichoke was washed with purified water, dried, and then steamed under pressure at 121°C for 30 minutes under a pressure of 1.5 atm. The artichoke steamed under pressure was dried with hot air, pulverized and pulverized using a high-speed grinder, and 20 times of purified water was added to the artichoke powder, followed by hot water extraction at 100°C for 3 hours. The prepared extract was filtered through a filter paper and then concentrated with a reduced pressure concentrator to finally obtain an artichoke extract. The obtained artichoke extract (85 brix) was diluted with 10 brix using purified water, sterilized at 105° C. for 30 minutes, and then Lactobacillus plantarum CBG-C21 ( Lactobacillus plantarum CBG-C21, KACC92083P) strain, lacto Bacillus rhamnosus CBG-C14 ( Lactobacillus rhamnosus CBG-C14, KACC91981P) strain and Bifidobacterium animalis CBG-C10 ( Bifidobacterium animalis CBG-C10, KCTC11978BP) strain were inoculated at a concentration of 1.0×10 ⁸ CFU/ml, respectively. Fermentation was performed at °C for 24 hours to obtain each fermented artichoke (hereinafter referred to as'sample'). The obtained fermentation product was powdered by adding dextrin and lyophilization.

2. Enzyme-treated artichoke fermentation production

After diluting the artichoke extract (85 brix) prepared as described above to 10 brix using purified water, 0.02% cellulase (Celluclast ^® 1.5L, Novozymes, Denmark), 0.01% pectinase (Pectinex ^® Ultra SP) -L, Novozymes, Denmark) and 0.025% inulinase (Nocozym ^® 960, Novozymes, Denmark) were reacted and sterilized at 105°C for 30 minutes, and then Lactobacillus plantarum CBG-C21 ( Lactobacillus plantarum CBG -C21, KACC92083P) strain, Lactobacillus rhamnosus CBG-C14 ( Lactobacillus rhamnosus CBG-C14, KACC91981P) strain and Bifidobacterium animalis CBG-C10 ( Bifidobacterium animalis CBG-C10, KCTC11978BP) strain of 1.0×10 ⁸ CFU Each inoculated at a concentration of /ml and fermented at 37°C for 24 hours to obtain each fermented artichoke (hereinafter referred to as'sample'). The obtained fermented product was powdered by adding dextrin and then lyophilized.

3. Measurement of cinnarine content

100 mg of the artichoke extract and the artichoke fermented product prepared above were each taken, 1 ml of 70% methanol was added to mix well, and then centrifuged at 12,000 rpm for 5 minutes to obtain a supernatant. The obtained supernatant was filtered through a filter and then analyzed by HPLC using 15 µl. HPLC analysis was performed at a flow rate of 0.5 ml/min at 320 nm using a Shiseido CapCellPAK MGII C18 column at 40° C. using a multiwavelength detector (MWD). At this time, a concentration gradient was applied using 0.1% aqueous formic acid and acetonitrile as analysis solvents.

4. DPPH radical scavenging activity analysis

20 µl of sample and 80 µl of DPPH (2,2-Diphenyl-1-picrylhydrazyl, Sigma-Aldrich, St Louis, MO, USA) solution were mixed in a 96-well plate and reacted in the dark for 30 minutes at room temperature, It was measured at 517 nm using a spectrophotometer Multiskan ELISA (Thermo scientific Co. Ltd). For the activity comparison, ascorbic acid (Sigma-Aldrich) was used as a positive control.

5. ABTS radical scavenging activity measurement

80mM ABTS[2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonatic acid)] solution and 30mM potassium persulfate (K ₂ S ₂ O ₈ ) solution are mixed and approximately 1 at 30℃ in the dark. After reacting for a period of time, an ABTS solution diluted so that the absorbance at 734 nm became 1±0.1 was used. 90 µl of diluted ABTS solution and 10 µl of sample were mixed and reacted in a dark place at 37°C for 30 minutes to measure absorbance at 734 nm. At this time, the positive control was Trolox ^® (Sigma-Aldirch), a water-soluble vitamin E derivative. I did.

Example 1. Changes in Sinarine Content Before and After Artichoke Fermentation

To confirm the change in the content of cinnarine before and after fermentation of the artichoke extract, Lactobacillus plantarum CBG-C21 ( Lactobacillus plantarum CBG-C21, KACC92083P) strain, Lactobacillus rhamnosus CBG-C14 ( Lactobacillus ) in the artichoke extract rhamnosus CBG-C14, KACC91981P) strain and Bifidobacterium animalis CBG-C10 ( Bifidobacterium animalis CBG-C10, KCTC11978BP) strains were respectively inoculated and fermented. As a result, as disclosed in Table 1 below, it was confirmed that the content of sinarine was increased by 20 to 25% in the artichoke fermentation product compared to the artichoke extract before fermentation (FIG. 1).

Changes in Sinarine Content of Artichoke Extract Before and After Fermentation by Strains division Before fermentation CBG-C10
After fermentation CBG-C14
After fermentation CBG-C21
After fermentation Sinarine content (mg/g) 392.32±30.12 468.82±12.24 484.52±5.49 490.99±22.10 Sinarine increase rate (%) - 19.5 23.5 25.1

Example 2. Changes in sinari content before and after fermentation of artichoke treated with glycolytic enzyme

In order to confirm the change in the content of sinarine due to the decomposition of the sugar component in the artichoke extract, the artichoke extract was treated with cellulase, pectinase, and inulinase, which are representative glycolytic enzymes, respectively, and then Lactobacillus plantarum CBG- C21 ( Lactobacillus plantarum CBG-C21, KACC92083P) strain, Lactobacillus rhamnosus CBG-C14 ( Lactobacillus rhamnosus CBG-C14, KACC91981P) strain and Bifidobacterium animalis CBG-C10 ( Bifidobacterium animalis CBG-C10, KC1978 strain). Each was inoculated and fermented. As a result, as disclosed in Table 2 below, it was confirmed that in the artichoke fermented product treated with enzymes and strains, the content of sinarine increased by 23-28% compared to the artichoke extract before treatment (FIG. 2).

Changes in Sinarine Content of Artichoke Extract Before and After Fermentation by Enzyme and Strains Processing enzyme division Sinarine content (mg/g) Sinarine increase rate (%) Cellulase Before fermentation 369.97±30.12 - After CBG-C10 fermentation 462.09±24.77 24.9 After CBG-C14 fermentation 470.23±19.25 27.1 After CBG-C21 fermentation 463.57±31.28 25.3 Pectinase Before fermentation 351.22±22.41 - After CBG-C10 fermentation 443.56±12.93 26.2 After CBG-C14 fermentation 450.26±39.17 28.2 After CBG-C21 fermentation 433.41±25.55 23.4 Inulinase Before fermentation 379.38±19.99 - After CBG-C10 fermentation 469.29±26.18 23.7 After CBG-C14 fermentation 466.26±32.86 22.9 After CBG-C21 fermentation 478.40±17.82 26.1

Example 3. Antioxidant Activity of Artichoke Fermentation According to Lactic Acid Bacteria Fermentation

In order to confirm the antioxidant efficacy of the artichoke fermentation product prepared by enzyme and strain of the present invention, 100 mg of the artichoke fermentation product under each condition was taken, and then extracted with 1.0 ml of ethanol to prepare a sample. Take 20 µl (DPPH analysis) and 10 µl (ABTS analysis) of samples, respectively, and DPPH (2,2-Diphenyl-1-picrylhydrazyl) and ABTS[2,2'-azino-bis(3) of the artichoke fermentation under each condition. -ethylbenzthiazoline-6-sulfonatic acid)] radical scavenging ability was measured. As a result, it was confirmed that, as disclosed in Table 3 below, each artichoke fermentation product significantly increased DPPH and ABTS radical scavenging activity compared to the artichoke extract before fermentation.

DPPH and ABTS radical scavenging activity by enzymes and strains of artichoke fermentation Processing enzyme division DPPH radical scavenging activity (%) ABTS radical scavenging activity (%) Ascorbic acid(0.1mg/ml) 91.84±0.19 - Trolox (0.1mg/ml) - 95.79±0.18 - Before fermentation 18..44±0.60 19.19±0.94 After CBG-C10 fermentation 35.53±2.50 28.86±0.19 After CBG-C14 fermentation 28.71±3.10 32.04±2.53 After CBG-C21 fermentation 42.19±0.18 16.55±1.82 Cellulase Before fermentation 17.96±2.53 24.72±0.09 After CBG-C10 fermentation 29.33±1.97 51.05±0.25 After CBG-C14 fermentation 15.42±0.25 24.09±1.04 After CBG-C21 fermentation 33.76±1.10 61.98±3.36 Pectinase Before fermentation 19.17±0.72 21.04±0.25 After CBG-C10 fermentation 37.52±1.74 51.05±2.55 After CBG-C14 fermentation 46.18±1.93 35.56±1.60 After CBG-C21 fermentation 64.29±2.62 38.21±3.36 Inulinase Before fermentation 18.97±0.40 18.19±0.47 After CBG-C10 fermentation 28.11±1.77 42.72±2.11 After CBG-C14 fermentation 39.58±0.19 28.86±0.25 After CBG-C21 fermentation 64.20±0.17 33.55±2.50

Institute of Agricultural Biotechnology KACC92083P 20150710 Institute of Agricultural Biotechnology KACC91981P 20140930 Korea Research Institute of Bioscience and Biotechnology KCTC11978BP 20110706

Claims (7)

Artichoke extract Lactobacillus plantarum CBG-C21 ( Lactobacillus plantarum CBG-C21) (KACC92083P) strain, Lactobacillus rhamnosus CBG-C14 ( Lactobacillus rhamnosus CBG-C14) (KACC91981P) strain, Bifidobacterium animalis CBG- C10 ( Bifidobacterium animalis CBG-C10 (KCTC11978BP) strain or a mixed strain thereof, comprising the step of inoculating and fermenting the artichoke fermented product with improved content of sinarine and antioxidant activity. delete The method of claim 1, wherein the fermentation is performed at 32 to 42°C for 20 to 28 hours. The method of claim 1, further comprising the step of pretreating a glycolytic enzyme to the artichoke extract. The method of claim 4, wherein the pretreatment of the glycolytic enzyme comprises cellulase, pectinase, inulinase, or a mixture of them. An artichoke fermented product with improved content of sinarine and antioxidant activity prepared by the method of any one of claims 1, 3 to 5. A health functional food containing artichoke fermented product with improved content of sinarine and antioxidant activity of claim 6.

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