KR20110052185A - Process of fermentated tomato using lactic acid bacteria and its product - Google Patents

Abstract

PURPOSE: Fermented tomato using lactobacillus, and a producing method thereof are provided to offer massive amount of lycopene, and to improve the physiological function activity of an eater. CONSTITUTION: A producing method of fermented tomato comprises the following steps: storing fresh tomato for ripening; washing the tomato, and dehydrating; removing damages portions from the dehydrated tomato; peeling the skin from the tomato; crushing the peeled tomato for separating flesh from seeds and the skin; filtering the tomato flesh to obtain flesh pulp containing tomato juice and tomato flesh, and cake containing the skins and the seeds; condensing the tomato flesh pulp, and sterilizing; fermenting the flesh pulp with lactobacillus; collecting solids from the fermented pulp; and inserting a stabilizing auxiliary agent before freeze-drying.

Description

Fermented tomato production method using lactic acid bacteria and its products {PROCESS OF FERMENTATED TOMATO USING LACTIC ACID BACTERIA AND ITS PRODUCT}

The present invention relates to a method for processing a tomato pulp obtained by physical treatment, concentrated and heat treated, followed by fermentation using lactic acid bacteria, and freeze-powdered tomatoes with a stabilizing aid and processed fermented tomatoes thereof.

Lycopene, a pigment that is involved in making tomatoes and watermelon red, is known to act as a powerful antioxidant that prevents cardiovascular diseases and exerts anti-cancer effects along with vitamin C, vitamin E and carotene. Lycopene, especially in tomatoes, has twice as much antioxidant activity as beta-carotene in carrots. In addition, anti-aging, anti-cancer effect (prostate cancer), cardiovascular disease prevention and hypoglycemic effect. In particular, it strongly inhibits IGF-1 (insulin like growth factors) factor, which is a major regulator of cancer cell growth, and stimulates protein 43 enzyme to express gap junctions. Is deeply linked to cancer suppression. Some experiments using animal or cancer cells have shown effectiveness in lung cancer, liver cancer, stomach cancer, breast cancer, cervical cancer, colon cancer, bladder cancer, pancreatic cancer, and clinical trials in humans showed excellent protection against breast cancer and prostate cancer. Reported as having. It is known to be effective against sarcoma and prostate cancer.

Lycopene is better absorbed by the body when heated. Lycopene has two types of isomers, trans and cis, which change to cis isomers that are better absorbed by the human body when heated. In this regard, FIG. 1 (quotation: John Shi et al. (2002) Neutraceutical & Foods, 17: 179-183), below 100 ℃ transforms the trans form into a cis form, a significant change in the total lycopene content However, it can be seen that at 190 ° C., the total lycopene content is significantly reduced, and the trans and cis-type contents are greatly reduced.

In addition, the lycopene content of the trans form is rapidly reduced according to the light intensity, it can be seen that the material sensitive to light. Lycopene is also slightly soluble in ether, petroleum ether and hexane, soluble in chloroform and benzene, hardly soluble in methanol and ethanol, and absorbed better when cooked in oil because it is fat-soluble.

Looking at domestic and foreign patent applications regarding the extraction method of lycopene in tomato, Korean Patent Application Publication No. 2006-0070846 discloses a supercritical extraction method of lycopene showing an excellent antioxidant effect by inhibiting the generation of free radicals in the process of oxygen respiration metabolism In extracting from tomato using the present invention, it discloses a useful oil antioxidant component that gives oleophilicity to be applied to makeup cosmetics in which solvent such as water or ethanol cannot be mixed using squalene as a co-solvent. It was.

In addition, Japanese Patent Application Laid-Open No. 10-276707 discloses a concentrated flop obtained by filtering a fine flop obtained by pectinase treatment and solid-liquid separation with a filter (0.3-0.7 mm) using a separation membrane (10 μm or less). It describes a low-viscosity high content of lycopene for use as a foodstuff manufacturing method.

US Patent No. 2005/0153038 also relates to a method for preparing lycopene extract from tomatoes, which are concentrated by heating the extracted tomato and extracted with water-saturated ethyl acetate.

In addition, the domestic patent 1991-0006937 sterilizes the tomato processing material obtained by crushing and juice the tomato, sterilized and cooled, and then the lactic acid fermentation beverage manufacturing method characterized in that the lactic acid fermentation is performed by adding the pre-cultured Lactobacillus Bulgarius. It is about.

In addition, Korean Laid-open Patent No. 10-1998-0053132 describes a method for producing a purple sweet potato pigment powder, which is a process of adding malto textrin to a purple sweet potato pigment concentrate and spray drying.

As such, the existing literatures describe chemical treatment with organic solvents, physical treatment with water or oil, and microbiological methods with Lactobacillus vulgaris lactobacillus. It does not describe a method of containing lycopene, nor does it describe a method of stably maintaining the content of lycopene by adding a stabilizing aid that performs better than maltotextrin during freeze drying.

Therefore, an object of the present invention is to stably contain a large amount of lycopene, so as to improve the activity of the physiological function of the intake, concentrating the tomato pulp obtained by physical treatment, heat treatment and fermentation using lactic acid bacteria, and adding a stabilizing aid Together with a method of processing a freeze-powdered tomato and a product thereof.

In order to achieve the above object, the present invention is to purchase a ripe tomato or to store the immature tomato for a certain period of time to complete;

A second step of washing the tomatoes and dehydrating to remove water from the surface;

A third step of removing the wounds or dents on the surface of the tomato;

A fourth step of peeling off the skin;

A fifth step of grinding the pulp to separate from the rind or seed, and separating the pulp from the pulp;

A sixth step of separating the juice pulp formed only from the pulp and separating the rind or seeds into a cake;

A seventh step of concentrating and sterilizing the juice pulp for the subsequent process;

An eighth step of fermenting the juice pulp with lactic acid bacteria; And

A ninth step of recovering the solids thereafter; And

Adding a stabilizing aid to the fermented tomato juice pulp, characterized in that it comprises a tenth step of lyophilized powder.

This will be described in detail below.

The first step of the method for producing a fermented tomato using the lactic acid bacteria of the present invention is to purchase ripe tomatoes or to put the unripe tomatoes into an indoor storage container to ripen.

Tomatoes become more red as they ripen, and the deeper the red, the richer the lycopene and beta-caroteine. Lycopene is more abundant in harvests that are fully red mature and harvested rather than harvested and ripened blue tomatoes. You can buy green less ripe and store it in a cool place at 15-18 ℃ to cook slowly and use it in the present invention.

In addition, in the second step of the present invention is a dehydration process to remove the foreign matter by washing the surface of the tomato in running water, to remove the moisture on the surface.

In addition, in the third step of the present invention, the wound on the surface is cut out with a knife, and the receded part is cut out with a knife to process only the fresh tomato pulp.

In the fourth step of the present invention, as a process of removing the skin of the tomato, a method of inducing the peel to peel off physically from the flesh by treating for 3 to 4 minutes in hot steam for mass production.

In the fifth step of the present invention, the pulp is pulverized so that the pulp is separated from the rind, the seed and the hard part by a pulverized pulp mill, and then separated from the rind, the seed, and the hard part which do not pass the pulp passing through the filter net using the filter net. to be. The filter network is not limited as long as it can separate the pulp, but preferably about 0.8 to 1.1 mm in size.

In addition, the sixth step of the present invention is a process for separating the juice pulp formed only of the pulp and the cake consisting of the rind, the seed and the hard portion separated in the fifth step.

The seventh step of the present invention is a process of concentrating the sterilized juice pulp pulp and heat treatment sterilization for 15 to 45 minutes at about 100 ℃. Concentration can concentrate the juice by membrane separation, centrifugation, or vacuum concentration process depending on the purpose.

In general, lycopene content of processed (concentrated) tomatoes is richer than fresh tomatoes. As can be seen in Table 1 below, 100 g of fresh red tomato is 3.1-3.7 mg level, 7.83 mg when processed into juice, and 30.0 mg in concentrated juice paste.

Lycopene content in 100 g of tomatoes Tomato form Lycopene Content Fresh red tomatoes 3.1-7.7mg Tomato processed juice 7.83mg Tomato concentrate paste 30.3 mg Source: Joseph Levy and Yoav Sgaroni (2004), Joseph Levy and Yoav Sgaroni (2004), The J. of the American Botanical Council

Therefore, the concentration of the juice pulp pulp can be obtained by a high content of improved cis-type lycopene with good absorption in the process including heat treatment.

In addition, the eighth step of the present invention is a process of fermenting the tomato juice using lactic acid bacteria.

The lactobacillus used in the present invention is not particularly limited, but Lactobacillus. L. acidophillus, Lactobacillus .Frantanum (L. plantarum), Lactobacillus. L.bulgaris, Streptococcus.St.thermophillus, Bifiidobacterium breve, Bifiidobacterium longum, Bifiidobacterium infantis, And leuconostoc mesentroide.

In addition, the pure culture of lactic acid bacteria is activated by incubating overnight at about 35 to 37 ° C by inoculating 100 ml of the stored strain, inoculated into 100 ml cap tube containing MRS broth using MRS medium. Then, it can be used for the main culture by culturing as a seed in a sealed Erlenmeyer flask containing 50 ~ 100ml MRS culture.

In addition, when fermenting tomatoes in an Erlenmeyer flask, use an Erlenmeyer flask incubator from 100 ml to 500 ml of the tomato concentrate sample, and after inoculating the lactic acid bacteria, seal the inlet to prevent air from entering and prevent exposure to light during the incubation. And, using a shaking incubator (Shaking incubator) is incubated at 35-37 ℃, 120rpm.

In case of using 5L Jar fermentor, sterilize by adjusting total volume to 4L to ferment tomato concentrate to lactic acid bacteria, inoculate lactic acid bacteria and incubate at 200rpm to prevent air from entering do.

The ninth step of the present invention is a process of recovering the solids.

Finally, a method of freeze drying the lactic acid bacteria fermented tomato in the powdering of the tenth step may be used. Freeze-drying (FD) method for obtaining a dry sample may be useful for the survival of heat-sensitive samples or beneficial bacteria than the heat drying method.

In particular, in the freeze powder drying process of the tenth step, a stabilizing aid is added. In general, maltodextrin is used as an additive in the freezing process, but in order to improve the stabilization of the lycopene component of the fermented tomato, it is preferable to use a stabilizing composition composed of vitamin C, vitamin E, and cyclodextrin. The composition of the stabilizing composition may be appropriately adjusted by those skilled in the art, but about 0.05 to 0.2 weight of vitamin C, about 0.01 to 0.03 weight of vitamin E, and cyclodextrin, based on 100 weight of cake (solid content 10%) obtained after tomato fermentation About 2 to 4 weights are preferred, in particular, vitamin C is preferably 0.1 weight, vitamin E is 0.02 weight, and cyclodextrin is 3 weight.

The present invention concentrates the tomato pulp obtained by the compression physical treatment, heat treatment and fermentation using lactic acid bacteria, and freeze-powdered it with the addition of a stabilizing aid, so that the processed fermented tomato is a functional ingredient lycopene By stably contained in the content, it can improve the physiological activity function of the body ingested the processed fermented tomato. Although it is inappropriate to use tomatoes with reduced product value as foods, fermentation processing using lactic acid bacteria improves the intrinsic odor of tomatoes, increases the content of lycopene as a functional ingredient, and improves the utilization of functional products by improving stabilization technology. By linking primary producers and secondary processors with the health food industry, it is possible to produce useful food raw materials and functional raw materials.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. Configurations shown in the drawings and embodiments described herein are only one of the most preferred embodiments of the present invention and do not represent all of the technical idea of the present invention, various modifications that can be substituted for them at the time of the present application It should be understood that there may be

Example  One. Lycopene  Content and Stabilization Experiment

(1) Tomato fermentation and powdering process

Washing and dehydrating the tomato to remove the wound and the tap, and using a juice pulper pulverized the flesh to separate the cake (cake), to obtain a tomato juice pulp. The tomato juice pulp was concentrated by a centrifugal separator to obtain a tomato pulp concentrate. Sterilization of the tomato pulp concentrate was heat-treated at 100 ° C. for 15 to 30 minutes, and during fermentation, Bifidobacterium breve was inoculated with the seed and fermented at 35 ° C. in a 5 L Jar Fermentor. After that, the sample containing the fermented tomato pulp was lyophilized at -50 ° C. or below using a freeze dryer to be powdered.

(2) Lycopene content analysis method

Lycopene analysis was measured by spectrophotometer measurement. To 2 g of the homogenized sample, 6% pyrogallo was added to 10 mL of ethanol, and filled with nitrogen gas. Then, 8 mL of 60% KOH solution was added and refilled with nitrogen gas to connect the cooling tube. This was saponified for 50 minutes in a 70 ° C. water bath, cooled, and the ethanol concentration was adjusted to 30% using 2% NaCl solution. 15 mL of extraction solvent (hexane: acetone: methanol) containing BHT (0.01%) was added thereto, followed by extraction three times. The extract was removed with anhydrous MgSO ₄ and applied to the final 50 mL. The hexane layer was collected by filtration using a 0.22um membrane filter (Sartorius, Goettingen, Germany), centrifuged, and absorbance was measured using a spectrophotometer (Perkin Elmer UV / VIS Lambd35) at 470 nm and 502 nm. The standard reagent lycopene was calculated by the standard curve and the standard formula.

(3) Change of Lycopene Content Before and After Fermentation of Tomato

1 kg of fresh tomatoes were washed with water, steamed at 100 ° C. for 5 minutes, processed with a tomato juicer to remove cakes, and the solids content of the concentrate obtained by concentrating the obtained tomato juice pulp was adjusted to 15-20%. After incubation for 30 days and then inoculated with lactic acid bacteria Bifidobacterium brave heat-treated for 30 minutes at 100 ℃ after fermentation and recovery of the samples after fermentation and freeze-drying for 6 months, respectively, the content of lycopene is measured in Table 2 As 32.6 mg and 96.0 mg per 100 g dry weight, respectively.

Lycopene content (mg / 100g) before and after fermentation of tomatoes Before fermentation After fermentation 32.6mg / 100g 96.0mg / 100g

It is more effective to carry out lyophilization in order to increase the viability of the metabolite formed during fermentation and the lactic acid bacteria itself by the addition of lactic acid bacteria of fermented tomato. The fermented tomato juice concentrate (non-fermented tomato: heat treatment 100 ℃, 30 minutes) and the fermented tomato juice concentrate (fermented tomato: fermented by Bifidobacterium brave), respectively, into a container and put inert gas After four months of storage experiments after 6 months of storage in the refrigerated state, the relative contents of lycopene were observed to obtain the results shown in Table 3.

When the content of lycopene during storage for 4 months was measured, the lycopene content of the fermented tomato lyophilized powder was relatively stable, and the lycopene content was maintained relatively high in the fermented tomato freeze powder.

Changes of Lycopene Contents in Storage of Tomato Lyophilized Powders after Fermentation of Tomato Lyophilized Powders Before Fermentation Retention period 0 Month 1 month 4 months Average Lycopene Ratio 2.94 3.43 3.10 3.16 Storage condition: unfilled upper inert gas in the container, storage temperature: refrigeration temperature

(4) Stability Change of Lycopene by Freeze-Drying Stabilizing Composition

In order to measure the effect of the adjuvant composition added to improve the stability of lycopene, an antioxidant substance in fermented tomato during freeze drying, experiments were conducted under accelerated storage conditions at 40 ° C. and nitrogen gas. Table 4 and FIG.

process Composition composition type
Relative Content of Lycopene in Accelerated Tests Before the experiment February April Reduction ratio (%) Control group - One 0.84 0.47 53% Test I Maltotextrin One 0.89 0.49 51% Test II Vitamin c
Vitamin E
Cyclodextrin One 0.89 0.53 47%

It is shown that the composition of Test II is more effective in lycopene stabilization due to the relatively smaller decrease in lycopene content than when no composition is added (control) or when maltodextrin is commonly used (Test I). .

Example  2. Biological activity experiment

(1) experimental animals

After purchasing 5-week-old ICR male mice from Orient Bio Co., Ltd., the lighting time was adjusted to 12 hours from 7 am to 7 pm and the room temperature was adjusted to 22 ~ 24 ° C. It was used for the experiment. Feed provided AIN-76 feed and drinking water was used, but feed and water supply were not limited.

(2) Treatment of experimental animals

As shown in Table 5, 25 mice were fermented tomato lyophilized powder from NC (control), EC treated group (ethanol-only group), vitamin C treated group (ethanol and vitamin C-treated group), EC treated group. Divided into groups (FTE) ingested for 1 week, the experiment was carried out in a dark and dark condition repeated in temperature (22 ~ 24 ℃), humidity (50%), 12 hours cycle. Each ethanol, vitamin C, tomato lyophilized powder was orally administered for a week.

Treated group Treatment method NC Water dosing EC 5 g / kg (50:50 (v / v)) ethanol administration Vitamin c 5 g / kg (50:50 (v / v)) ethanol with vitamin C (0.1 g / kg body weight) FTE 5 g / kg (50:50 (v / v)) ethanol administration including fermented tomato freeze-dried powder (1 g / kg / weight)

 (3) Measurement of body weight and tissue weight of laboratory animals

The weight gain was calculated by measuring the weight before and after the experiment and at 12 hours after the final oral administration, the mice in each group were dissected to determine the amount of liver, kidney, spleen, testis, kidney fat and testis fat. Measured.

Experimental results are shown in Table 6 below, but did not show a significant difference between each other, in the case of liver FTE group was relatively good recovery, long-term ingestion seems to help normal recovery.

Treatment group
Weight
Tracheal weight increase (%; ㅎ /b.w.x100) liver kidney Visa Testicle Kidney fat Testicular NC 3.63 ± 1.05 4.41 ± 0.08 1.57 ± 0.11 0.25 ± 0.02 0.65 ± 0.03 0.50 ± 0.04 1.56 ± 0.10 EC 2.65 ± 0.62 5.30 ± 0.26 1.77 ± 0.05 0.23 ± 0.01 0.76 ± 0.04 0.50 ± 0.04 1.38 ± 0.08 Vit c 2.40 ± 0.32 4.95 ± 0.50 1.52 ± 0.04 0.23 ± 0.02 0.63 ± 0.07 0.57 ± 0.15 1.57 ± 0.35 FTE 2.25 ± 0.34 4.80 ± 0.19 1.57 ± 0.04 0.28 ± 0.01 0.73 ± 0.06 0.57 ± 0.04 1.59 ± 0.10

(4) In In vivo antioxidant activity assessment

Liver tissue was homogenized with PBS (pH 7.4) and used for the evaluation of antioxidant activity.

1) Catalase (CAT) Analysis

Catalase activity was analyzed by Spectrophotometric method of Aebi. A 96 well plate was prepared, and 290 μl of 20 mM H ₂ O ₂ solution was added to 10 μl of the tissue solution, and the change in absorbance was measured at 240 nm for 3 minutes using the kinetic method of the microplate reader. The activity of the catalase enzyme was converted to U / mg protein.

2) Glutathione-s-transferase (GST) analysis

Glutathione convertase activity was analyzed by spectrophotometric method of Habig. A 96-well plate was prepared, and a substrate solution containing 196 μl of Dulbecco's phosphate buffered saline, 2 μl of 200 mM glutathione, and 2 μl of 100 mM CDNB solution was prepared. 180 μl of substrate solution was added to 20 μl of the tissue solution, and the change in absorbance was measured at 340 nm for 5 minutes. Glutathione convertase enzyme activity was calculated by U / mg protein.

3) Glutathione peroxidase (GPx) assay

Glutathione peroxidase activity was analyzed by Thomson's spectrometric method. A 96 well plate was prepared, and 10 µl of the tissue solution was mixed with 120 µl of 3.5 mM glutathione, 20 µl of 2.5 mM NADPH and 20 µl of 0.5 U glutathione reductase, followed by 30 mM tert-butyl hydroperoxide 20 The reaction was induced by the addition of [mu] l and the change in absorbance was measured at 340 nm for 5 minutes. Glutathione peroxidase activity was calculated by U / mg protein.

4) Glutathione reductase (GR) analysis

Glutathione reductase activity was analyzed by spectrometric method. Prepare a 96 well plate, add 30 μL of 100 mM potassium phosphate buffer (pH 7.0), 100 μL of 2 mM oxidized glutathione, and 50 μL of 3 mM DTNB to 20 μl of the tissue solution, and mix well. Then, add 10 μL of 2 mM NADPH to react. Induction and GR enzyme activity was measured at 412 nm for 5 minutes. Glutathione reductase enzyme activity was calculated as U / mg protein.

5) Reduced glutathion (GSH) analysis

Glutathione amounts were analyzed by Griffith's spectrometric method. A 96 well plate was prepared, and a substrate solution was prepared by mixing 7.54 mL of 100 mM potassium phosphate buffer (pH 7.0), 228 μl of 6 unit glutathione reductase, and 228 μl of 3 mM DTNB. After mixing 150 μl of the prepared substrate solution to 10 μl of the tissue solution, 50 μl of 50 mM NADPH was added to induce the reaction, and the change in absorbance was measured at 412 nm for 5 minutes. The amount of GSH in tissues was calculated using the GSH standard curve and expressed as umoles / mg protein.

(5) experimental results

The mice were inoculated with a small phase with alcohol, the fermented tomato of the present invention was ingested into the mice, and tissues were extracted to measure changes in health indicators between the mice.

CAT (catalase) (FIG. 3), GST (glutathione-S-transferase) (FIG. 4), GPx (glutathione-peroxidase) (FIG. 5), GHS (glutathione) (FIG. 6), which are liver health indicators It was confirmed that the activity of GR (glutathione reductase) (FIG. 7) was enhanced.

Figure 1 is a graph showing the change in lycopene content under accelerated storage conditions by the addition of the adjuvant.

Figure 2 is a graph showing the change in lycopene content under accelerated storage conditions during nitrogen treatment of Test B.

Figure 3 is a graph showing the change in activity of CAT (catalase) in mouse liver experiments ingested processed tomatoes.

Figure 4 is a graph showing the change in activity of GST (glutathione-S-transferase) in mouse liver experiments ingested processed tomato.

5 is a graph showing the change in activity of GPx (glutathione peroxidase) in mouse liver experiments ingested processed tomatoes.

Figure 6 is a graph showing the change in activity of GSH (reduced glutathione) in mouse liver experiments ingested processed tomatoes.

Figure 7 is a graph showing the change in the activity of GR (glutathione reductase) in mouse liver experiments ingested processed tomato.

Claims (6)

Purchasing the ripe tomatoes or storing the immature tomatoes for a period of time to ripen; A second step of washing the tomatoes and dehydrating to remove water from the surface; A third step of removing the wounds or dents on the surface of the tomato; A fourth step of peeling off the skin; A fifth step of pulverizing the pulp to separate the pulp, the seed, or the hard portion, and separating the pulp from the pulp; A sixth step of separating the juice pulp formed only from the pulp using a filter and separating the rind or seeds into a cake; A seventh step of concentrating and sterilizing the juice pulp for the subsequent process; An eighth step of fermenting the juice pulp with lactic acid bacteria; And A ninth step of recovering the solids thereafter; And Adding a stabilizing aid to the fermented tomato pulp, and comprising a tenth step of freezing and powdering, stably contains a high content of lycopene, the method of processing fermented tomato using lactic acid bacteria having good physiological activity. The method according to claim 1, wherein in the fermentation of lactic acid bacteria in the eighth step, L. acidophillus, L. acidophillus, L. plantarum, L. plantarum, L. bulgaris , Streptococcus thermophillus, Bifiidobacterium breve, Bifiidobacterium longum, Bifiidobacterium infantis, and loconostock mecentroid (Leuconostoc mesentroide) A method for processing fermented tomatoes using lactic acid bacteria, characterized in that one or more selected from the group consisting of lactic acid bacteria. The method of processing fermented tomatoes using lactic acid bacteria according to claim 1, wherein the stabilizing aid added in the freeze-powdering of the tenth step comprises vitamin C, vitamin E and cyclodextrin. According to claim 3, Vitamin C is about 0.05 to 0.2 weight, Vitamin E is about 0.01 to 0.03 weight, Cyclodextrin is about 2 to 4 weight with respect to 100 weight of the cake obtained after the tomato fermentation is contained in the stabilizing aid Processing method of fermented tomato using lactic acid bacteria to make. 5. The fermented tomato of claim 4, wherein the vitamin C is 0.1 weight, vitamin E is 0.02 weight, and cyclodextrin is 3 weight based on 100 weight of the cake obtained after the tomato fermentation. Processing method. Fermented tomato processed according to claim 1.

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