KR101531349B1 - Leuconostoc mesenteroides SK-1 isolated from octopus Baechu kimchi and method for preparing seafood kimchi using the same - Google Patents

Abstract

The present invention relates to an acid resistant Leuconostoc mesenteroides for actively generating dextran and uses thereof where the acid resistant Leuconostoc mesenteroides can be used as a food additive composition and a starter for kimchi. In the present invention, the acid resistant Leuconostoc mesenteroides is used for manufacturing seafood kimchi, which has less acidity and lactic acid compared with traditional kimchi, thereby long maintaining proper aging of kimchi and storing the same as well as distributing the same for a long time.

Description

Description: TECHNICAL FIELD The present invention relates to a leuconostoc mesenteroide SK-1 isolated from octopus Chinese cabbage kimchi, and a method for producing the leucomostoc mesenteroides SK-1 isolated from octopus kimchi using the same.

The present invention relates to a novel Ryukono Stokeshensteeroide SK-1 strain isolated from Chinese cabbage kimchi to which aquatic products such as octopus are added, and a method for producing the aquatic product kimchi using the same. More particularly, The present invention relates to a novel microorganism, Konostok mesenteroid SK-1 strain, and to a method for producing a kimchi having a lower sour taste and an appropriate maturity than a conventional kimchi using a starch.

The present invention can be used as a useful technique in the field of kimchi industry and fermented food.

Kimchi is a typical traditional fermented food in Korea that fermentes vegetables at low temperature by putting various ingredients such as seasoning and salted fish into the salt, and it can be varied according to the food production and geographical characteristics of each region. Depending on the immersion method, And has a taste. A record of Kimchi dipped with marine products from ancient times remained, and when we look at the "Collection of Rituals" written by Lee Hee-gang in 1809 in the Joseon Dynasty, Is a type of water kimchi that is immersed in soy sauce, cod, crayfish, early head and shellfish, using deep-seated soup, and 'stir-fried' is a kimchi dipped in seaweed and octopus. Is introduced. Based on these records, in modern times, mainly on the coastal area, besides seaweed, kimchi which has added a variety of marine products such as oysters, early, pollack, squid, shrimp and abalone is being eaten. Thus, the aquatic products added to kimchi are known to play a role not only in enhancing the taste of kimchi but also in supplementing nutritional value such as amino acid. Therefore, interest in aquatic products as a kimchi supplement has increased, and kimchi Kimchi, which is added with octopus and abalone, has started to be commercialized.

However, in the case of commercially available commodity kimchi, the kinds and number of lactic acid bacteria that lead to the fermentation of kimchi are different even when they are manufactured using the same raw materials and methods, and the difference becomes larger during aging, It is difficult to solve the problem. Furthermore, it is quite difficult to reproduce the taste and quality of aquatic products kimchi added with an animal protein source such as aquatic products as a raw material in a desired state in a desired state.

In order to solve these problems, there have been many attempts to control the taste and quality of kimchi by adding lactic acid bacteria derived from kimchi and adding it as a starter in the production of kimchi.

Korean Patent Registration No. 0181009 (Dec. 04, 1998) discloses that Kimchi is not only good in taste but also retarded rancidity and softness due to the addition of Ryukono Stockparamensenteroids and Ryukono Stokes Steroidis strain, And the microorganism added with Lulkonostomyces tenerais K8P4 or Leukonostomyces teneroides K2M5 and Lactobacillus casei K5M3 was added to the Korean patent registration 0519083 (2005.09.27) Korean Patent Registration No. 0536108 (Dec. 14, 2005) discloses a method for controlling the taste of kimchi by regulating the microbial composition of kimchi. A method of manufacturing a kimchi having improved functionality is disclosed. As such, it is important to reproduce the cool and clean taste of kimchi, which is the key to producing good taste and quality kimchi. Therefore, the use of leuconostomercenteroid, a heterofermentative lactic acid bacteria, as a starter of kimchi Research has been going on. In addition, Ryukonostok, a lactic acid bacterium that grows as a dominant species at the early stage of fermentation of kimchi, is a major microorganism involved in the fermentation and taste of food. It produces a wide variety of components such as lactic acid, acetic acid, carbon dioxide and mannitol during the fermentation process. In particular, the genus Leuconoosta secretes dextran sucrase in the presence of sucrose to produce fructose, a highly dense polysaccharide called dextran, and when dextran is ingested as a food, intestinal microorganisms It is effective to proliferate the growth of the plant. Using these characteristics of Ryukono-stock strain, Ryukono Stokes me- thelteroid (Korean Patent Publication No. 2008-0077390) and Ryukonosost Citrius (Korea patent registered 1302465), which are excellent in mannitol-producing ability, are added to Kimchi as a starter (Korean Patent Registration No. 0718344 (2007. 05. 08)) discloses a method for producing kimchi having excellent sensory characteristics, and Korean Patent Registration No. 0718344 (2007. 05. 08) relates to a method for producing kimchi, which is involved in the fermentation of kimchi and uses dextrane A method of producing a viscous functional fermented food product by inoculating the strain with a starter is disclosed.

[5] Meanwhile, in the conventional techniques described above, studies have been made on starches of the genus Lukonovost, which is a lactic acid bacterium derived from kimchi prepared without adding aquatic products, as a starter. However, even if the strain is added as a starter to kimchi prepared by adding aquatic products, There is a problem that the growth rate of the microorganism is changed due to the difference in the nutritional composition and the aging condition of the compounded raw material and the cool and fresh taste of the kimchi of the seafood can not be maintained or reproduced continuously. Accordingly, development of a lactic acid bacteria starter for aquatic products such as kimchi, which can reproduce and maintain a cool and clean taste of a seafood aquatic product for a long time, is desperately required. However, there is no known technology for producing lactic acid bacteria isolated from kimchi added with aquatic products and using the lactic acid bacteria as a starter for aquatic products.

1. Korean Patent Registration No. 0181009 (Dec. 04, 1998) 2. Korean Patent Registration No. 0519083 (September 27, 2005) 3. Korean Patent Registration No. 0536108 (Dec. 14, 2005) 4. Korean Patent Publication No. 2008-0077390 (Aug. 22, 2008) 5. Korean Patent Registration No. 1302465 (Feb. 6. Korean Patent Registration 0718344 (2007. 05. 08)

1. The Ritual Series (閨 閤 本 书): Written by Lee Hee-bang in the 1809 Joseon Dynasty

DISCLOSURE OF THE INVENTION The present invention has been made in view of the above needs, and an object of the present invention is to provide a novel lactic acid bacterium having an acid resistance and excellent dextran production ability from a cabbage kimchi prepared by adding aquatic products such as octopus, The present invention provides a method for producing aquatic product kimchi which can maintain a cool taste and a unique flavor for a long time by utilizing it as a starter of the added kimchi. It is expected that it will contribute greatly to activation of kimchi industry in Korea and diversification of quality of kimchi by utilizing this technology development, and it is expected to help increase consumption of kimchi.

In order to achieve the above-mentioned object, 5 to 10% (w / w) aquatic products such as octopus are added to the total weight of pickled Chinese cabbage (hereinafter, Chinese cabbage kimchi using octopus is specifically described as an example) Octopus, etc. Leuconostoc mesenteroides ( Leuconostoc), which has acid resistance at pH 3.2 to 5.0 and is superior in dextran production, mesenteroides ) subsp. suyonum SK-1 (Accession No .: KCCM11519P).

Further, the present invention provides a method for producing aquatic product kimchi characterized by adding the strain or a culture solution thereof.

In the present invention, a new lactic acid bacterium having acid resistance and excellent dextran production ability is provided in Chinese cabbage kimchi prepared by adding aquatic products such as octopus, and the result is added to aquatic product kimchi to maintain the titratable acidity of aquatic product kimchi for a long time, It is effective to reproduce a cool taste unique to a weak and seafood Kimchi. In addition, since the lactic acid bacteria have excellent dextran production ability, it is possible to produce aquatic product kimchi having a functionality.

Therefore, the lactic acid bacteria of the present invention can be usefully used as a starter of kimchi prepared by adding aquatic products.

Fig. 1 shows the results of comparing the amount of dextran produced by Ryukono Stokeshensteiron SK-1 of the present invention with that of a conventional lactic acid bacterium.
SK-1: Leukonostomycesenteroid SK-1 of the present invention
KCTC3505: Leuconostomesenteroid KCTC3505
KCTC13302: Leuconostomercenteroid KCTC13302
Fig. 2 is a graph showing the results of a comparison between the aquatic product kimchi prepared by adding the yeast strain SK-1 of the present invention and the conventionally known yeast KCTC3505 and KCTC13302 strains, And the pH change of the aquatic kimchi was shown.
?: Aquatic products prepared by adding a strain of Ryukono stokescensentero SK-1
≪ / RTI >< RTI ID = 0.0 >#:< / RTI > aquatic product kimchi prepared by adding strain KCTC3505
▲: aquatic product kimchi prepared by adding strain KCTC 13302
≪ RTI ID = 0.0 > x: < / RTI >
Fig. 3 is a graph showing the results of a comparison between the aquatic product kimchi prepared by adding the yeast strain SK-1 of the present invention and the conventionally known yeast KCTC3505 and KCTC13302 strains, The results are summarized as follows.
?: Aquatic products prepared by adding a strain of Ryukono stokescensentero SK-1
≪ / RTI >< RTI ID = 0.0 >#:< / RTI > aquatic product kimchi prepared by adding strain KCTC3505
▲: aquatic product kimchi prepared by adding strain KCTC 13302
≪ RTI ID = 0.0 > x: < / RTI >
FIG. 4 is a graph showing the results obtained by adding a kimchi of aquatic products prepared by adding the yeast strain SK-1 of the present invention and the conventionally known yeast KCTC3505 and KCTC13302 strains, The results are as follows.
SK-1: aquatic product kimchi prepared by adding Leuconostomercenteroid SK-1 strain
KCTC3505: Aquatic products prepared by adding a strain of Ryukono Stokescecente KCTC3505
KCTC13302: Aquatic products prepared by adding the strain KCTC13302
Lactic acid bacteria not added: Seafood Kimchi prepared without the strain of Leuconostoc sp.
FIG. 5 is a graph showing the results of a comparison between the aquatic product kimchi prepared by adding the yeast strain SK-1 of the present invention and the conventionally known yeast KCTC3505 and KCTC13302 strains, And the Ryukono stock contents of the aquatic products.
?: Aquatic products prepared by adding a strain of Ryukono stokescensentero SK-1
≪ / RTI >< tb >< tb >< SEP > KCTC3505 strain <
▲: aquatic product kimchi prepared by adding strain KCTC 13302
≪ RTI ID = 0.0 > x: < / RTI >

The present invention relates to a lactic acid bacterium isolated from Chinese cabbage kimchi added with aquatic products such as octopus and the like, leuconostoc mesenteroides ( Leuconostoc mesenteroides ) subsp. suionicum SK-1. < / RTI > The strains were separated from the cabbage kimchi prepared by adding octopus. However, the isolating range of the related strains is not limited thereto, and the strains may be separated from the cabbage kimchi prepared by adding octopus, octopus, octopus, abalone, Mesenteroides ( Leuconostoc mesenteroides ) subsp. The use of suionicum SK-1 is possible.

In the present invention, to produce aquatic kimchi having a cool taste and a unique flavor, Chinese cabbage kimchi added with octopus was isolated and the strain grown when the most delicious. Specifically, the lactic acid bacteria were isolated when the octopus kimchi was fermented at 4 ℃ and the pH was 4.55 and the acidity was 0.65%. Among the seven lactic acid bacteria isolated, the three lactic acid bacteria exhibiting excellent acid - Tran producibility was compared to identify the SK-1 strain having the best dextran production ability. As a result, it was confirmed that the strain belonged to Ryukono stokescenteide. suionicum SK-1. The strain was deposited at the Korean Culture Center of Microorganisms on Feb. 17, 2014 (Accession No .: KCCM11519P).

The present invention relates to the above-mentioned leucomoniast mesenteroid subsp. lt; RTI ID = 0.0 > SK-1 < / RTI > In general, the strain of succulent Lactobacillus mefenacetide is rapidly degraded under acidic conditions, especially at a pH of 4.0 or lower. However, the strain of succinostomercenterolide subsp. SUICIDUM SK-1 has an acid resistance at pH 3.2-5, especially at pH 3.5, and it shows strong acid resistance.

In addition, the present invention relates to a method for preparing a mixture of leuconostox mesenteroid subsp. suionicum SK-1 has excellent dextran production ability.

Among the conventionally known leuconostomercenteroids, leuconostox mesenteroide subsp. Compared to mesenteroides KCTC3505 and leuconostomercenteroid KCTC13302, dextran production yielded about 10% and 17% more dextran, respectively.

The leuconostox mesenteroide subsp. ≪ RTI ID = 0.0 > Aquaticum SK-1 or its culture can be added to produce aquatic product kimchi.

The present invention relates to a leuconostox mesenteroide subsp. culturing the strain of suicidium SK-1 in a PEM (Phenylethyl Alcohol Sucrose) medium, followed by primary pre-culture at 20 to 25 ° C for 20 to 24 hours; Inoculating 0.5-1% (v / v) of the primary culture broth into PES medium and then secondary culture at 20-25 ° C for 20-24 hours; It is preferable to inoculate the secondary culture broth with 0.5 ~ 1% (v / v) of the sterilized PES medium and then cultivate at 20 ~ 25 ° C for 20 ~ 24 hours.

The thus prepared luchonostomercenteroid subsp. The culture of the suikinum SK-1 strain was centrifuged at 4,000 rpm for 10 minutes to recover the cells of the lower layer and then diluted to 1 x 10 ^{7 to 8} CFU / mL in sterile physiological saline (0.85% NaCl) (W / v) based on the total weight of the kimchi seasoning. Most preferably, the total weight of the mixed kimchi sauce (red pepper powder, garlic, ginger, canned fish sauce, shrimp, glutinous rice paste, sugar, Aqueous kimchi can be prepared by adding 3% (w / v) of the suyonum SK-1 culture. If the culture solution of the strain is added in an amount exceeding 5% based on the total weight of the kimchi seasoning, the kimchi of the aquatic products may become overmatched and the sweet taste may become strong. When the content is less than 1%, the effect of the strain is insignificant.

As described above, leuconostox mesenteroide subsp. The acidity of kimchi prepared by adding suionicum SK-1 is slower than that of aquatic kimchi supplemented with conventional leucono-stokershenceotide strains, so that the optimum age (0.4 ~ 0.8% of acidity) of the aquatic kimchi is retained for a long time In the case of using the strain isolated in the present invention or a culture thereof when preparing aquatic product kimchi, the period of maintaining proper maturity is prolonged, so that a cool taste of the aquatic product kimchi can be maintained for a long time. In addition, the leuconostox mesenteroide subsp. The addition of the suyonum SK-1 strain and its culture broth in the preparation of aquatic product kimchi leads to fermentation as a dominant species during the fermentation of aquatic product kimchi and the production of a dextran-rich aquatic product kimchi is suppressed and maintains a cool taste for a long time .

Hereinafter, the present invention will be described in detail with reference to examples.

It should be understood, however, that the invention is not limited to the disclosed embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

Isolation of Lactic Acid Bacteria from Octopus Chinese Cabbage Kimchi

Octopus cabbage kimchi prepared by adding 5 to 10% (w / w) of octopus to the total weight of pickled cabbage was subjected to sensory evaluation while being aged at 4 ° C for 25 days, and the kimchi having the best flavor was analyzed with a lactic acid bacteria- Respectively. The sensory evaluation of octopus kimchi, which was aged 17 ~ 19 days, was evaluated as the best taste. The pH was 4.55 and the acidity was 0.65%. The kimchi juice of the octopus kimchi was diluted 10 times with 0.85% saline and 0.1 mL of PESA medium (Phenylethyl alcohol sucrose agar, 5 g of tryptone, 0.5 g of yeast extract, 20 g of sucrose, 2 g of ammonium sulfate, 0.244 g of magnesium sulfate heptahydrate, phosphate monobasic, 2.5 mL of phenylethyl alcohol, 15 g of DW / 1 L of agar) plate and inoculated onto a glass plate. Plates were then incubated for 5 days in a 20 < 0 > C incubator. Seven colonies of clear and large form were selected from among the colonies generated, and inoculated on PES agar plates, and cultured at 20 ° C for 3 days to isolate lactic acid bacteria colonies.

Of lactic acid bacteria isolated from octopus cabbage kimchi Acid resistance  Strain selection

To compare the acid resistance of seven kinds of lactic acid bacteria colonies isolated from octopus Chinese cabbage kimchi as in Example 1, Phenylethyl alcohol (PES) adjusted to pH 3.2, 3.5, 4.0, 4.5, 5.0 by using lactic acid 5 g of tryptone, 0.5 g of yeast extract, 20 g of sucrose, 2 g of ammonium sulfate, 0.244 g of magnesium sulfate heptahydrate, 1 g of potassium phosphate monobasic and 2.5 mL of DW / 1 L of phenylethyl alcohol were prepared, and 7 kinds of lactic acid bacteria colonies And incubated at 20 ° C for 72 hours to observe whether the cells were grown or not. The most resistant acid-resistant strains were selected in the pH range. The strains of colonies SK-1, SK-2 and SK-5 showed strong acid tolerance due to active growth at pH 3.5-5.0 (see Table 1).

Comparison of acidic activity of lactic acid bacteria isolated from octopus cabbage kimchi Colony number pH 3.5 4.0 4.5 5.0 SK-1 + + + + SK-2 + + + + SK-3 - + + + SK-4 - + + + SK-5 + + + + SK-6 - + + + SK-7 - + + +

Dextran Productive  Selection of excellent strains

In order to confirm the amount of dextran produced in acid resistant lactic acid bacteria SK-1, SK-2 and SK-5 selected from octopus Chinese cabbage kimchi as in Example 2, the above three lactic acid bacteria were added to 10 mL of PES medium Followed by subculture.

Then, the three lactic acid bacteria were inoculated in 100 mL of PES medium at a concentration of 1% (v / v), cultured at 20 ° C. for 5 days, and then the amount of dextran produced was measured to determine the amount of colony SK-1 The strains were finally selected. A method for measuring the amount of dextran production of a single isolated acid-resistant lactic acid bacteria strain was performed as follows; 50 mL of each culture was collected and centrifuged at 5,000 rpm for 10 minutes to obtain a clear supernatant. Ten times of ethanol (99.9%) was added to the above supernatant to recover the precipitated dextran, freeze-dried and measured the amount of dextran dried (see Table 2).

Comparison of dextran production of acid-resistant strains isolated from octopus kimchi Colony number Dextran production (%) SK-1 3.1 SK-2 1.6 SK-5 2.1

Identification of lactic acid bacteria SK-1 isolated from octopus kimchi

The morphological characteristics of SK-1 strain, which has acid resistance and high dextran production ability among seven lactic acid bacteria isolated from octopus cabbage kimchi, were analyzed. In order to analyze the morphological characteristics, SK-1 strain was cultured under optimal conditions and subjected to Gram staining. As a result, it was confirmed that the isolated lactic acid bacteria were gram-positive and had a bacterium shape. The results of catalase experiments showed negative responses, consistent with morphological features of the genus Leuconostoc sp.

Table 3 shows the results of using 49 types of carbohydrate using patterns of API-50CHL (BioMerieux SA, France) on SK-1 strain. The SK-1 strain may be selected from the group consisting of L-arabinose, ribose, D-xylose, D-mannose, mannitol, alpha-methyl-D- glucoside, N- acetylglucosamine, salicin, maltose, melibiose, trehalose, , D-turanose, gluconate, 2-keto-gluconate and 5-keto-gluconate, and it was found to be a strain belonging to Leuconostoc mesenteroides .

Investigation of Carbon Dioxide Utilization Pattern of Lactic Acid Bacteria SK-1 Isolated from Octopus Kimchi carbohydrate activation carbohydrate activation Glycerol - Esculin - Erythritol - Salincin + D-arabinose - Cellobiose - L-arabinose + Maltose + Ribos + Lactose - D-xylose + Melibiose + L-xylose - Sucrose - Adonitol - Trehalose + Beta-methyl-D-xyloside - Inulin - Galactose - Meredithos - D-glucose - D-raffinose + D-fructose - Starch - D-Mannos + Glycogen - L-sorbose - Xylitol - Rams North - Beta-genothiobiose - Dissytol - D-Turanos + Inositol - D-Rick sauce - Mannitol + D-tagatose - Sorbitol - L-Fucos - Alpha-methyl-D-mannoside - D-arabitol - Alpha-methyl-D-glucoside + L-arabitol - N-acetylglucosamine + Gluconate + Amigalline - 2-keto-gluconate + Arbutin - 5-keto-gluconate +

Meanwhile, the identity of the SK-1 strain isolated above was analyzed by 16S rRNA sequencing. In other words, the 16S rRNA base sequence was amplified by PCR using the purified DNA as a template according to a general method. The amplified DNA fragments were purified using a Big Dye Terminator Cycle Sequencing kit (Applied Biosystems, USA) and sequenced using a DNA sequencer (Applied Biosystems model 3730XL, USA). The determined nucleotide sequences were analyzed using EzTaxon server 2.1. As a result of 16S rRNA sequence analysis of SK-1 strain, Leuconostomercenteroid subsp. A strain having 99.8% identity to suionicum LMG 8159 ^{T was} named leuconostomercenteroid subsp. suyonum SK-1, and deposited it at the Korean Center for Microorganism Species Conservation (KCCM 11519P, Apr. 27, 2014).

The ratio of the lactic acid bacteria of the present invention to the conventional lactic acid bacteria Acid resistance  compare

The acid resistance of the present leukoconstrictor SK-1 isolated from octopus kimchi was compared with that of the previously known lactobacillus strains, Leuconostoxesterocyte KCTC3505 and Leuconostomercenteroid KCTC13302.

Each of the lactic acid bacteria was inoculated with 1 mL of the cell culture medium in 10 mL of PES medium adjusted to pH 3.2, 3.5, 4.0, 4.5, 5.0 with lactic acid. The cells were cultured at 20 ° C for 72 hours, and the presence or absence of growth was observed. The results are shown in Table 4. The leuconostomercenteroid SK-1 of the present invention is more resistant to acid than KCTC3505 and KCTC13302, which are strains belonging to the genus Leuconostochus, and thus the growth is active at pH 3.2 to 5.0.

Comparison of acid resistance between lactic acid bacteria of the present invention and conventional lactic acid bacteria pH The leuconostomesenteroid SK-1 of the present invention Leuconostomercenteroid KCTC3505 Ryukono Stock Messensteroid KCTC13302 5.0 + + + 4.5 + + + 4.0 + + + 3.5 + + + 3.2 + - -

The ratio of the lactic acid bacteria of the present invention to the conventional lactic acid bacteria Dextran Generation  compare

In order to compare the dextran production ability of leuconostomercenteroid SK-1 of the present invention with the conventionally known lactic acid bacterium, leucono stokescensentroid KCTC3505, which is known to have excellent dextran production ability, and yeast extract The ability to produce dextran was compared using leuconostomercenteroid KCTC13302. The lactic acid bacteria were inoculated in 1 mL of 100 mL of PES medium and cultured at 20 DEG C for 5 days. Thereafter, the amount of dextran produced in the culture solution was measured in the same manner as described in Example 3 above. As shown in FIG. 2, the lactic acid bacterium of the present invention had a dextran production amount as high as about 10% higher than that of Leuconostomyces teneroides KCTC3505, which is known to have excellent dextran production ability. 17% more dextran was produced. In the present invention, it has been confirmed that the leucono stokescenteide SK-1 isolated and identified has excellent dextran production ability as compared with the microorganism of the genus Leuconostoc.

Ryukono Stock Mesenteroid SK -1 of Seafood Kimchi  Addition culture

The culture broth of the strain for producing aquatic product kimchi using the strain Ryukono Stokeshensteeroide SK-1 of the present invention was prepared as follows.

The culture broth of Ryukono Stokesentenoid SK-1 strain was inoculated into the PES medium and then primary cultured at 20 ° C for 24 hours. The primary culture broth was inoculated 1% (v / v) in PES medium And then secondary culture was carried out at 20 ° C for 24 hours. The secondary culture broth was inoculated 1% in sterilized PES medium and cultured at 20 ° C for 24 hours.

The cultured medium of the thus-produced Ryukono Stokes meencellulosic SK-1 strain was centrifuged at 4,000 rpm for 10 minutes to recover the cells of the lower layer, and then sterilized with physiological saline (0.85% NaCl) at a concentration of 1 × 10 ^{7 to 8} CFU / mL was used.

Ryukono Stock Mesenteroid SK Production of Fish Kimchi Using Culture Medium

After removing the inside of aquatic products (octopus, squid, abalone, juku, octopus, etc.) and washing them thoroughly, water was removed and 3% (w / w) After storing the time, it was washed several times to remove the salty taste, and then it was boiled in the licorice for 3 to 5 minutes. The Chinese cabbage was dipped in 2% (w / w) weight of Chinese cabbage, and the 10% salted Chinese cabbage total weight And salted for more than 12 hours at about 18 ℃. When the salinity of the Chinese cabbage reached 1.8 ± 0.1%, it was washed twice in flowing water, drained into a wicker and dehydrated for 4 hours.

For the production of aquatic products kimchi, 78.3%, 2.35%, 2.35%, 2.35%, 1.17%, 1.57%, ginger (0.47%) and glutinous rice paste (3.38% (Octopus, squid, abalone, jukipo, octopus, etc.) treated with the above method were sliced into appropriate sizes, and the weight of the pickled cabbage Aquatic products were prepared so as to be about 10%. The cultured strain of Ryukono stokescensenteroid SK-1 strain prepared in the same manner as in Example 7 was measured to be 3% (w / v) based on the total weight of the kimchi sauce except for aquatic products, and added to the kimchi sauce and mixed. The aquatic products were prepared in the same manner as Kimchi spices mixed with the culture medium of Lukonovostomercenterol SK-1 strain. The final salinity of the aquatic kimchi prepared by this method was 2.2 ± 0.1%.

Ryukono Stock Mesenteroid SK - 1 strain  Fermentation Pattern of Added Kimchi Added

In order to compare the fermentation characteristics of the aquatic kimchi added with Leuconostokcesenteroid SK-1 strain isolated from octopus Chinese cabbage kimchi, the fermented kimchi was taken out at intervals of 7 days while being stored at 4 ° C. in a refrigerator at 4 ° C., Acidity, production of lactic acid and the number of bacteria in Leuconostoc were measured.

As a control, the same materials and compounding ratios were prepared, and a dipping fish kimchi (negative control) and luchonostomycesteroid KCTC3505 (positive control group 1) prepared in the same manner as in Example 7 were prepared without adding lactic acid bacteria such as Ryukono stock, ) And KCTC13302 (positive control 2) culture broth were prepared by the method of Example 8 above.

   (9-1) pH  And pH measurement

As in Example 8, pH and acidity of kimchi prepared by adding cultured medium of Ryukono Stokeshensteeroide SK-1 were measured and pH and acidity of each kimchi were compared. The sample was taken so that the ratio of marine products to Chinese cabbage was 1: 9 (g / g), minced and gauze was used as the kimchi solution. The pH of the solution was measured with a pH meter (Thermo Orion US / 320, Barrington, IL, USA) taking 20 mL of the Kimchi solution and neutralized to pH 8.1 using 0.1 N NaOH solution The amount of NaOH was converted into lactic acid content (%) and the acidity was measured.

As shown in Fig. 2, until the 14th day of fermentation, the aquatic kimchi of the negative control group to which no strain of Ryukono stock strain was added showed a higher pH value than the aquatic kimchi containing Ryukono SK-1 strain including positive control 1 and 2 . However, from the 21st day of fermentation, the pH of the aquaculture kimchi added with the culture medium of leucono stokescensenteroid SK-1 of the present invention was maintained at 4.56, The pH value of the aquatic kimchi of the negative control group to which KCTC13302 was added and to which no Kimchi and Liuchonobacter strains were added after 21 days of fermentation showed a tendency to decrease.

As shown in FIG. 3, the aquatic kimchi prepared using the leuko-stokescensenteroid SK-1 strain of the present invention reached a titratable acidity of 0.65% at 28 ° C. at 28 ° C. at 28 ° C., but the yeast strains KCTC3505 and KCTC13302 The kimchi of aquatic control 1 and 2, which were added, reached the optimum maturation pH of 0.65% at 21 and 17 days of ripening, respectively. In addition, although the acidity of the kimchi prepared by using the strain Ryukono Stokesentenoid SK-1 did not reach 0.8% even after 35 days of fermentation, the negative control, including fish kimchi not containing the strain of Ryukono-stock, The acidity of the kimchi prepared from the positive control group containing the stock methenseolide KCTC3505 and KCTC13302 reached 0.8% from the 28th day after storage and rapidly increased from then on.

   (9-2) Measurement of lactic acid production

The lactic acid production of the aquatic kimchi of the present invention prepared above and the aquatic kimchi used as a control group were measured and compared. The amount of lactic acid produced was measured according to the organic acid analysis method using HPLC (High performance liquid chromatography) as follows. Kimchi of aquatic products Kimchi of each sample was centrifuged at 13,000 rpm for 10 minutes, and the supernatant was filtered with a 0.22 μm syringe filter. The filtered sample was injected with 10 μL of sample using HPLC and flowed to 0.6 mL per minute using 0.008 MH ₂ SO ₄ mobile phase. The column used was Aminex HPX-87H (300 × 7.8 mm, 9 μm, Bio-rad, USA) and the column temperature was 35 ° C.

Similar to the results shown in Figs. 2 and 3, the negative control group fishery product kimchi added with no strain of Ryukono-stock strain at 4 캜 at the ripening-21st day, a positive control fish product kimchi and SK- 1 strain, the lactic acid production was somewhat lower than that of the aquatic kimchi. However, as shown in Fig. 4, the aquatic product kimchi prepared by using the leuko-stokescensenteroid strain SK-1 of the present invention showed 0.072% of lactic acid production on the 21st day of ripening at 4 DEG C, The yields were maintained constant at 0.076% and 0.079%, while the positive control group aquatic kimchi supplemented with marine aquatic kimchi, leuconostomercenteroid KCTC3505 and KCTC13302 strains as a negative control group, On the 28th day, lactic acid production was rapidly increased to 0.096%, 0.115% and 0.112%, respectively.

As a result, it can be concluded that the production of aquatic product kimchi by using the strain of Ryukono Stokes senoteloid SK-1 isolated from octopus Chinese cabbage kimchi, It was analyzed that long - term distribution and storage of aquatic kimchi could be possible by prolonging the period of maintaining the pH and acidity and decreasing lactic acid production.

   (9-3) Leuconostok sp. Bacterium  Measure

To measure the number of bacteria in Ryukono stock of each kimchi prepared in Example 8, each kimchi solution aged at 4 캜 for 35 days was diluted with 0.85% saline, and 0.1 mL of the kimchi dilution was added to a PESA medium plate And then plated on a glass plate. The agar plates were incubated in a constant temperature incubator at 20 ° C for 5 days, and the number of colonies formed was counted as each bacterium.

As shown in Fig. 5, the positive control group aquatic products kimchi supplemented with the leukoconstrictor SK-1 and leucono-stock strain of the present invention during the fermentation period of 35 days was a negative control group The number of Leuconostoc spp. Strains was significantly higher than that of aquatic products. In addition, since the number of bacteria in Ryukono-Stokes remains constant at a level of 10 ⁸ CFU / mL or more from the 14th day after ripening at 4 ° C, the kimchi prepared using Ryukono Stokes senoteloid SK-1 strain is maintained at a constant level It was confirmed that Leuconostomercenteroid SK-1 strains led to fermentation as a dominant species in the aquatic kimchi, and it was confirmed that the cool taste of the aquatic products kimchi could be maintained constantly, and that the yeast strains prepared using the positive control strains 1 and 2 The same tendency was also observed in the aquatic kimchi.

In the above example, Ryukono Stokes senoteloid SK-1 strain isolated from Chinese cabbage kimchi added with octopus is described. However, Ryukono stock produced from cabbage kimchi added with marine products such as octopus, squid, squid, abalone, It is obvious to those skilled in the art that the methacellulose SK-1 strain can be used separately, and such changes are within the scope of the present invention.

The drawings and the detailed description of the invention are merely illustrative of the invention and are used merely for the purpose of describing the invention and not for limiting the scope of the invention as set forth in the claims or the claims. Therefore, those skilled in the art will appreciate that various modifications and equivalent embodiments are possible without departing from the scope of the present invention. Accordingly, the true scope of the present invention should be determined by the technical idea of the appended claims.

Korea Microorganism Conservation Center (overseas) KCCM11519P 20140217

Claims (6)

Leuconostoc mesenteroides subsp. Kimchi, which has acid resistance at pH 3.2 ~ 5.0 isolated from cabbage kimchi prepared by adding octopus and has excellent dextran production ability. suionicum SK-1 strain (Accession No .: KCCM 11519P). delete A process for producing aquatic product kimchi characterized by adding leuconostomercenteroid SK-1 of claim 1 or a culture thereof. [Claim 4] The method according to claim 3, wherein the culture medium of the leukoconstrictor SK-1 strain is obtained by inoculating the strain into PES (Phenylethyl Alcohol Sucrose) medium and then pre-culturing the strain at 20 to 25 DEG C for 20 to 24 hours; Inoculating 0.5 to 1% (v / v) of the primary culture broth into PES medium, and then secondarily pre-culturing at 20 to 25 ° C for 20 to 24 hours; Culturing the microorganism for 20 to 24 hours at 20 to 25 DEG C after inoculating the second pre-culture broth with 0.5 to 1% (v / v) of the PES medium to be sterilized, . [4] The kimchi seasoning according to claim 3, wherein the cultured strain of leuconovirus methensertoid SK-1 is added to kimchi seasoning so that the culture medium is 1 to 5% (w / v) ≪ / RTI > The method of manufacturing a fish product according to claim 3, wherein the seaweed added to licorice is added so as to be 10% (w / w) based on the weight of the pickled Chinese cabbage.

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