KR20190040738A - Novel microorganism, starter composition including the same and fermented product by the same - Google Patents

Abstract

The present invention relates to a Tetragenococcus halophilus KUD23 KCCM12026P strain. The present invention, specifically, relates to a strain which is a dominant species in soybean paste, can produce acids under high salt, has excellent protease and lipase activities, can be used as a seed for high salt fermentation, and does not produce biogenic amines, thereby having no harmful effects on human body such as hypertension, headache, diarrhea and localized inflammation; a starter composition for fermentation comprising the same; and a product fermented thereby.

Description

TECHNICAL FIELD [0001] The present invention relates to a novel microorganism, a starter composition containing the microorganism, and a fermented fermented product thereof.

The present invention relates to a novel microorganism, a starter composition containing the same, and fermented fermented product.

The types of miso are conventional doenjang using meju, modified miso soup using starch raw materials, and companion doenjang which compromises of two methods.

Conventional doenjang is made by fermenting meju into soybean and brick or spherical shape, fermenting it naturally, immersing it in saline, aging for 6 to 12 months, separating the soy sauce and aging the solid.

Improved doenjang using starch raw materials increased starch raw materials such as wheat flour and Aspergillus oryzae ), followed by emulsification without mixing with salt, salt water, expanded starch raw materials, and soybean seeds.

It is designed to protect traditional flavor ingredients and nutrient spills containing soybean paste with soy sauce. Grind roughly ground meju to low salt and turn it into salt water. Then, add soy sauce and stir gently to mix with remaining meju.

There are various traditional fermented foods such as kimchi, doenjang, and salted fish in Korea, and they are made by characteristic production methods based on local fermentation and local families.

The traditional fermented foods made in the home began to be industrialized in the 1970s due to the increase of various industries and the increase in the number of meals. The changes in the housing environment due to economic growth, the development of the processed food industry, , And the increasing interest in health, the industry size is continuously growing.

Especially, the effects of anticancer and immunity enhancement of traditional doenjang have been reported and commercial production has been increasing due to difficulty in manufacturing at home unit. However, the necessity of securing the safety required for commercial distribution of conventional doenjang has been raised.

Biogenic amines, one of the metabolites produced by fermentation microorganisms during fermentation, are responsible for the physiological functions of growth regulation, inflammation regulation and neurotransmission in the human body. However, biogenic amines Is ingested through food, it is known that harmful symptoms occur. Detection of biogenic amines in commercially produced doenjang has been reported, and various studies for the reduction of biogenic amines are underway.

Although various types of biogenic amines have been detected in fermented foods, histamine has been reported to cause diarrhea, abdominal pain and headache, tyramine causes vasoconstriction and hypertension, and histamine is regulated internationally . Therefore, selection of a fermentation strain that does not produce a biogenic amine can greatly contribute to the reduction of the biogenic amine of the fermented food.

Korean Patent No. 0369841

It is an object of the present invention to provide a strain which is a dominant species of miso and can be used as a seed for high salt fermentation and does not produce a biogenic amine.

It is an object of the present invention to provide a starter composition comprising the strain.

It is an object of the present invention to provide a fermented product fermented with the above composition.

1. Tetragenococcus halofilus KUD23 ( Tetragenococcus halophilus KCC23) KCCM12026P strain.

2. The strain of 1 above, wherein the strain is isolated from miso.

3. The strain of 1 above, wherein the strain does not produce histamine.

4. The strain according to item 1 above, wherein the strain does not produce tyramine.

5. A fermentation starter composition comprising the strain of 1 above.

6. A fermented product fermented with the composition of the above 5.

7. The fermented product of claim 6, wherein the fermented product is a fermented product of legumes.

8. The fermented product according to 7 above, wherein the fermented soybean is at least one selected from the group consisting of meju, soy sauce, miso, kochujang, chunjang and chungkukjang.

The strain of the present invention is a dominant species in soybean paste and exhibits high salt resistance and can be used as a seed for high salt fermentation.

The strains of the present invention do not produce biogenic amines and thus are not harmful to humans such as hypertension, headache, diarrhea and localized inflammation.

The strain of the present invention is capable of acid production even with high salt, and is excellent in protease and lipase activity.

Brief Description of the Drawings Figure 1 shows the amount of biogenic amine produced by the test strain Tetragonococcus halofilus KCCM 40823 and Tetragenococcus halophilus KUD23 under laboratory conditions.

Hereinafter, the present invention will be described in detail.

The present invention relates to a novel microorganism Tetragenococcus halophilus KUD23 KUD23) KCCM12026P strain.

The strain of the present invention was isolated from doenjang prepared by fermenting soybean beans and deposited at the Korean Microorganism Conservation Center on May 19, 2017, and received the deposit number KCCM12026P.

Tetragenococcus halophilus belonging to the genus Tetragenococcus, made separately from the lactic acid bacterium Pediococcus , Gram-positive, tetrad-forming, nonmotile, catalase-negative, and horseradish lactic acid bacteria.

Tetragenococcus halophilus has been detected as a dominant in high salt foods, including salted and fermented fish products, miso, and soy, and can be selected as a potential starter candidate for their production.

The strain of the present invention is dominant in miso and exhibits appropriate physiological characteristics and is suitable for use as a seed bacterium.

The strain of the present invention exhibits high salt resistance and can be particularly preferably used for fermentation of a conventional fermented food fermented in high salt. It is possible to survive even at salt concentrations of, for example, 7% (w / v), 14% (w / v) and 21% (w / v).

In addition, acid production is possible even under high salt, and protease and lipase activity are excellent.

The strain of the present invention does not produce biogenic amines (biogenic amine non-producers) and has no human health hazards such as hypertension, headache, diarrhea and localized inflammation.

Generally, fermentation starter strains produce a large amount of biogenic amines such as histamine and tyramine, which can be used as a starter to cause the above problems in food production. However, the strain of the present invention does not produce histamine.

In addition, the strain of the present invention does not produce tyramine. In the present invention, production of biorenic amines does not mean only 0 mg / L of production, but also includes production of substantially 0 mg / L.

The present invention also provides a fermentation starter composition comprising the strain.

The fermentation product can be produced by fermenting with the starter composition of the present invention.

The starter composition of the present invention may further include, but is not limited to, Aspergillus oryzae, Bacillus subtilis and the like which are widely used in the art in addition to the above-mentioned strains.

The present invention also provides a fermented product fermented with the starter composition.

The fermented product of the present invention may be, for example, a fermented product of legumes. Specific examples thereof include, but are not limited to, meju, soy sauce, miso, kochujang, chunjang, and chungkukjang.

Soybeans to be fermented are, for example, soybean, soybean, kidney bean, pea, peanut, peanut, diagonal, and the like, and may be specifically soybeans.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to examples.

Example

Materials and methods

Strain and culture conditions

Tetragenococcus halofilars separated by the dominant species of miso KUD23, a test strain Tetragenococcus halophilus KCCM 11377, a published strain of tetrazenococarbinephylline KCCM 40823 was used. The strain was cultivated at 30 ° C for 48 hours in a MRS (Difco, USA) medium (NaCl) supplemented with 3% (w / v) of NaCl for maintenance and preservation of activity. The incubation used a sealable container to maintain the possible anaerobic conditions.

Biogenic Amine  Production measurement

Tetrazanococus Hollow Pillars for Biogenic Amine Production Evaluation After incubation, the absorbance was measured at 600 nm, and the concentration of the cells in the culture solution was adjusted to be the same. Then, in a MRS liquid medium supplemented with 3% of NaCl and 4 kinds of biogenic amine precursors, 1 strain of KCCM40823 % (v / v). L-tyrosine monohydrochloride monohydrate, L-lysine monohydrochloride and L-ornithine monohydrochloride (pH 5.8) were added to the final concentration of 0.25% (w / v) and pyridoxal- Was added at a final concentration of 0.0005% (w / v) and incubated for 3 days with possible anaerobic conditions at 30 ° C. (Hwang, DF, Chang, SH, Shiua, CY, Chai, T., 1997. High-performance liquid chromatographic determination of biogenic amines in fish poisoning in food poisoning. Chromatogr. B Biomed. Sci. Appl. 693, 23-29.). The dansyl derivatized material of the biogenic amines was dissolved in 5 mL of acetonitrile and 10 μL was taken and analyzed by HPLC (high performance liquid chromatography). The calibration curve for quantification was the standard of biogenic amines (cadaverine, histamine, putrescine, Materials were purchased and prepared.

Agilent Technologies HPLC 1200 series system (Palo Alto, CA, USA) was used for the analysis of the biogenic amines produced by the strains, and biogenic amines were identified by UV detector at 254 nm. Nova-Pak C18 column (4 μm, 150 × 4.6 mm, Waters, Milford, Mass., USA) was used as the stationary phase.

50: 50 (v / v) acetonitrile: 0.1 M ammonium acetate solution was used as a fluidized bed and elution started at a flow rate of 1 mL / min. And then changed to 50:50 for 2 minutes. The mean and standard deviation of the experimental results are shown as quantitative values.

Identification of salt tolerance, acid production, protease and lipase activity

Tetragenococcus halofilars KUD23, Tetragenococcus halophilus KCCM 11377, tetrazenococarb < / RTI > The salt tolerance of KCCM 40823 strain was confirmed by growth on MRS agar medium supplemented with NaCl concentration up to 21% (w / v).

Growth was confirmed by culturing for 4 days, 6 days, and 8 days in media containing NaCl concentrations of 7%, 14%, and 21%, respectively.

Acid production was confirmed on MRS agar medium supplemented with CaCO ₃ 0.7% (w / v).

Protease activity was confirmed on MRS agar medium supplemented with skim milk at 2% (w / v), and lipase activity was confirmed by tributyrin agar (Sigma) supplemented with 1% (v / v) tributyrin.

Tributyrin supplemented medium was emulsified by sonication and then autoclaved.

Colonies cultured on MRS agar medium supplemented with 3% NaCl were inoculated on a medium supplemented with substrate for measurement of each activity and cultured at 30 ° C for 48 hours. The enzyme activity was determined by the size of the transparent circle around the colonies appearing after the visible colonies appeared. The effect of NaCl on each activity was determined by adding up to 14% (w / v) NaCl to each medium.

result

Biogenic Amine  produce

Biogenic amines produced by the enzymatic decarboxylation of amino acids are known to stimulate the nervous system and vascular system to cause toxicity when overdosed. The production of four types of biogenic amines (cadaverine, histamine, putrescine, and tyramine) that are frequently detected in soybean fermented foods was examined.

Tetragenococcus halophilus KCCM 40823 did not produce histamine but produced tyramine, whereas tetrazenococarbinephylline It was confirmed that KUD23 did not produce histamine and almost no tyramine (Fig. 1).

The production of low concentrations of both catarrhine and putrescine was detected in both strains, but it was judged to be insignificant considering the amount of tyramine produced by Tetranyanococcus halophiles KCCM 40823. Considering that various types of biogenic amines have been detected from foods, and that they are globally regulating detection only for histamine, The production of cadaverine and putrescine by KUD23 is unlikely to cause problems in terms of food safety.

Also, given the amount of tyramine produced by Tetranyanococus halophilus KCCM 40823, Cadaverine, histamine, putrescine, and tyramine produced by KUD23 are not expected to be produced by decarboxylase, an enzyme involved in these production.

Salt resistance, acid production, protease and lipase activity

Functional characteristics were evaluated to assess their applicability as a starter for high-quality miso production (Table 1).

Since doenjang is aged at 12 ~ 15% NaCl concentration, salt tolerance is the most important functional requirement as miso starter. Tetragenococcus halophilus KCCM 11377, Tetranychococcus halophilus KUD23 strains grew on MRS agar medium containing 21% NaCl and produced acid at a concentration of 14% NaCl. These results suggest that fermentation by the strain may occur during fermentation of doenjang.

Considering the high protein and fat content of soybean beans, the protease and lipase activity of starter strains may contribute to changes in the flavor of meju and doenjang.

Protease and lipase activity assays showed that Tetranygenococus halophilus KUD23 was active at 3% and 4% NaCl concentrations, respectively, which would be involved in the fermentation of doenjang.

Effect of NaCl on functional properties and activity of starter candidate strains in MRS agar medium supplemented with substrates capable of measuring each activity

Strain Salt resistance Acid production Protease Lipase 7% 14% 21% 0% 7% 14% 0% 3% 0% 2% 4% T. halophilus KUD23 + + + + + + + + + + + T. halophilus KCCM 11377 + + + + + + - - - - - T. halophilus KCCM 40823 + + - + + - - - - - -

+: positive activity, -: negative activity.

Lipase activity was measured on tributyrin agar supplemented with 1% tributyrin (v / v), and the maximum concentration of NaCl that can be added to the medium is 4% (w / v).

Claims (8)

Tetragenococcus KUD23 ( Tetragenococcus halophilus KCC23) KCCM12026P strain. The strain according to claim 1, wherein the strain is isolated from miso. The strain according to claim 1, wherein the strain does not produce histamine. The strain according to claim 1, wherein the strain does not produce tyramine. A starter composition for fermentation comprising the strain of claim 1. A fermented product fermented with the composition of claim 5. The fermented product according to claim 6, wherein the fermented product is a fermented product of legumes. [Claim 7] The fermented product according to claim 7, wherein the fermented soybean is at least one selected from the group consisting of meju, soy sauce, soybean paste, kochujang, chunjang and chungkukjang.

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