KR100571050B1 - Method for preparing fermented food using strain enhanced productivity of bacteriocin - Google Patents

Abstract

 The present invention relates to a method for producing fermented foods using bacteriocin-producing fortified strains. In particular, the present invention relates to a method for enhancing bacteriocin production from bacteriocin-producing strains by non-genetic manipulation by identifying bacteriocin-producing substances in susceptible strains specific for bacteriocin-producing strains. It can be used as a seed of. Therefore, it is possible to maintain the taste and quality of fermented foods uniformly and to prevent various kinds of microbial contamination, thereby obtaining the effect of extending the storage period.

Bacteriocin Production Enhancement, Fermentation System Using Fortified Strains, Kimchi Lactobacillus, Bacteriocin Producing Strains, Susceptible Strains, Bacteriocin Producing Substances, Species, Fermentation

Description

METHODS FOR PREPARING FERMENTED FOOD USING STRAIN ENHANCED PRODUCTIVITY OF BACTERIOCIN

1 is an electrophoresis picture of bacteriocin inducer isolated from Lactobacillus platarum KFRI 464.

[TECHNICAL FIELD OF THE INVENTION]

The present invention relates to a method for producing fermented foods using bacteriocin-producing strains, and more specifically, bacteriocin-producing strains in non-sterile open fermented foods, bacteriocin-producing susceptible strains containing bacteriocin-producing substances, and methods for enhancing bacteriocin production using the same. And its application to fermented foods.

[Private Technology]

Kimchi is one of the representative fermented foods that can be manufactured by anyone at home without special knowledge. However, Kimchi has not established a system that can quantify or qualitatively ferment kimchi compared to its long history. This is in contrast to the early development and introduction of artificial sterilization and spawning technology in the fermentation system since dairy products fermented products, which are representative fermented foods in the West, are easy to breed harmful microorganisms such as E. coli.

In Kimchi's commercialization, quality uniformity, standardization, and long-term integrity remain unresolved. The biggest reason is that kimchi is not 'separated' unlike fermented dairy products. That is, raw and subsidiary materials of kimchi cannot be sterilized by heat, so the initial number of bacteria and the number of species transferred from kimchi material are not constant, so fermentation systems such as fermented dairy products cannot be applied. In the previous research, it was confirmed that the kimchi fermentation system, which does not produce raw material sterilization even after developing excellent spawn, should be inoculated with a relatively large amount of spawn at the initial stage. have. Therefore, we try to reduce the rancidity of kimchi and prolong the eating period by physicochemical methods such as pasteurization, irradiation, preservative treatment, addition of buffers or addition of salt mixtures for the purpose of prolonging the fermentation period. It has not been put to practical use due to such problems.

In addition, in the case of fermented foods, a product showing a unique flavor is produced according to the difference of fermentation, and it is common to grade products according to the degree of fermentation. On the other hand, in the case of kimchi, the diversification of kimchi products is limited only to the diversification of main ingredients such as Chinese cabbage, radish, cucumber, mustard, and perilla, and the variety of seasonings. No system has been established that can diversify the flavors by varying the types or combinations thereof. However, the development of various kimchi using these spawn is a challenge at present, and may be a good opportunity to increase the added value of traditional kimchi in the future.

Kimchi quality is determined by the transition pattern and the degree of development of the microbial population during the ripening process. The lactic acid bacteria that are directly involved in kimchi fermentation include Pediococcus, Lactobacillus, Lactococcus, and Luconostoc, and about 200 microorganisms was separated from (So, MH, Kim, YB Identification of psychrotrophic lactic acid bacteria isolated from Kimchi Korean J. Food Sci Technol 27 (4):... 495-505 (1995), Kang, SM, Yung, WS, Kim , YC, Joung, EY and Han, YG Strain improvement of Leuconostoc mesenteroides for Kimchi fermentation and effect of starter.Korean J. Appl.Microbiol.Biotechnol. 23 (4): 461-471 (1995), Lee, CW, Ko, CY and Ha, DM Microfloral of the lactic acid bacteria during Kimchi fermentation and identification of the isolates.Korean J. Appl.Microbiol.Biotechnol. 20 (10): 102-109 (1992))

In the early stage of fermentation of kimchi, heterofermental lactic acid bacteria, such as luconosestock mesenteroides, begin to multiply, and the growth of heterologous lactic acid bacteria peaks during the ripening season when kimchi is best eaten. This indicates that the taste of kimchi and the growth of heterologous lactic acid bacteria are closely related. After the middle of fermentation, the pH is lowered to 4.0 or less, and lactobacillus plantarum, a highly acidic homogeneous lactic acid bacterium, grows rapidly and produces a large amount of acid, causing the rancidity of kimchi.

On the other hand, the artificial control of Kimchi fermentation using microorganisms is still in its infancy because of the characteristics of Kimchi fermented food. Currently, bacteriocin producing strains that can be used as starting strains are screened, and strains are mainly selected from sauerkraut or dairy fermentation related strains. Enterococcus hirae is selected from kimchi, but it is a bacterium that controls the harmful environment of kimchi, rather than contributing to improve the taste of kimchi, and this bacterium is GRAS (Genenally Recognized As Safe). It is not an officially approved species.

 Therefore, as a starting point for the development of the Kimchi fermentation system, the development of strains that produce bacteriocins with excellent fermentation properties and antibacterial activity against various bacteria should be prioritized, and it is important to establish an optimal fermentation system using them. Do.

It is an object of the present invention to provide a method for enhancing bacteriocin production of a bacteriocin producing strain.

In addition, an object of the present invention is to provide a bacteriocin-producing enhanced strain of bacteriocin-producing strains.

It is another object of the present invention to provide a bacteriocin-producing fortified strain that can be used as a seed for non-sterile open fermented food.

It is another object of the present invention to provide a fermentation system using bacteriocin producing strains and susceptible strains.

It is another object of the present invention to provide a susceptible strain that specifically increases the bacteriocin production of the bacteriocin producing strain.

In addition, an object of the present invention is to provide a kimchi excellent in flavor and suppressed growth of harmful bacteria.

In order to achieve the above object, the present invention provides a method for enhancing bacteriocin production, comprising the step of adding a susceptible strain or a fraction thereof to the bacteriocin producing strain, the susceptible strain is inhibited growth by the bacteriocin producing lactic acid bacteria and specific to the bacteriocin producing lactic acid bacteria It provides a method for enhancing bacteriocin production by acting as a strain to enhance bacteriocin production.

The present invention also provides a method for producing bacteriocin-producing fortified strains comprising culturing the bacteriocin-producing strain in the presence of a susceptible strain or a fraction thereof, wherein the susceptible strain is inhibited in growth by bacteriocin-producing lactic acid bacteria and produces bacteriocin. It provides a method for producing bacteriocin-producing fortified strains that specifically acts on lactic acid bacteria to enhance bacteriocin production.

In another aspect, the present invention provides a bacteriocin-producing enhanced strain produced by the bacteriocin-producing enhanced strain manufacturing method.

In another aspect, the present invention provides a seed food for fermented food containing bacteriocin-producing fortified strains.

In addition, the present invention is fermentation comprising the fermented food seedlings added to 0.0001 to 0.01 parts by weight (wet part by weight) based on 100 weight of the food substrate and put it at 10 to 30 ℃ fermented by the seedlings grown as dominant species Provide a system.

In another aspect, the present invention provides a fermented food prepared by adding the seed for the fermented food.

The present invention also provides bacteriocin production inducer, which induces bacteriocin production of Luconosstock Citrium GJ7, which comprises a N-terminal amino acid sequence of SEQ ID NO: 1 and has a molecular weight of about 6,500 Da.

Hereinafter, the present invention will be described in detail.

The present invention relates to a method for enhancing bacteriocin production using a susceptible strain, a method for producing bacteriocin-producing fortified strains, and a seeding in fermented food of bacteriocin-producing fortified strains.

The susceptible strains referred to in the present invention are strains that are bacteriostatic or killed by bacteriocin producing strains, and are preferably strains that induce the production of bacteriocins from specific bacteriocin producing strains. The target microorganism of each susceptible strain is specifically present. Susceptible strains can be used to maximize the production of bacteriocin without the use of conventional genetic engineering to improve the bacteriocin production rate of the bacteriocin producing strain, and provide a non-genetic method that does not fall under the regulations for genetically modified organisms (GMOs). Make it possible.

The bacteriocin production inducing action of the susceptible strain is due to a specific material present in the susceptible strain, which is described herein as a bacteriocin inducer. Bacteriocin inducers of susceptible strains have different intracellular positions depending on the type of susceptible strain, and are preferably present in intracellular fractions or cell debris. The intracellular fraction can be prepared from the supernatant of the crushed cells, the cell lysates can be prepared from pellets of the crushed cells.

In one embodiment of the present invention, bacteriocin inducer was isolated and purified from Lactobacillus platarum KFRI 464. The bacteriocin inducer can induce the production of bacteriocins of Luconosstock Citrium GJ7, Luconosstock Paramescenteroides GJ77 and Lactobacillus Platarum GJ78, and comprises an N-terminal amino acid sequence of SEQ ID NO: 1, about 6,500 It is a protein having a molecular weight of Da. The bacteriocin production inducer is thermostable at 4 to 121 ° C. and also exhibits pH stability at pH 2.5 to 9.5.

Bacteriocin-producing strains referred to in the present invention are usually all kinds of strains producing bacteriocin, and are preferably lactic acid bacteria producing bacteriocins. Examples of bacteriocin-producing lactic acid bacteria include Luconostoke Citrium GJ7, Lukonost Paramesenteroides GJ77, Luconostok Paramescenteroides GJ74, and Lactobacillus Platarum GJ78, and most preferably Luconosstock Citrium GJ7. to be.

Among the susceptible strains specific for each of the four bacteriocin-producing lactic acid bacteria, the susceptible strain containing the bacteriocin inducing substance is as follows:

The susceptible strains for Luconostock Citrium GJ7 are Lactobacillus plantarum KFRI 464, Lactobacillus delbrueckee KFRI 347, Lactobacillus exidophilus KFRI 150, and Luconostock mesenteroides KCTC 1628, respectively.

The susceptible strains for Luconostock Paramescenteroides GJ77 are Lactobacillus plantarum KFRI 464, Lactobacillus delbrueckee KFRI 347, Luconostock mesenteroides KCTC 1628, Lactobacillus plantarum KFRI 236 and Lukonostok, respectively. Mesenteroides KFRI 218,

The susceptible strains for Luconostock Paramescenteroides GJ74 are Lactobacillus delbrueckee KFRI 347, Luconostock mesenteroides KFRI 218, Lactobacillus exidophilus KFRI 150, and Luconostock mesenteroides KCTC 1628, respectively.

Susceptible strains for Lactobacillus platarum GJ78 are Lactobacillus plantarum KFRI 464, Lactobacillus exidophilus KFRI 150 and Lukonostoke mesenteroides KCTC 1628, respectively.

Both bacteriocin producing strains and susceptible strains of the present invention are dietary lactic acid bacteria officially approved as GRAS (Genenally Recognized As Safe).

The present invention provides a method for enhancing bacteriocin production using the susceptible strain. Bacteriocin production enhancement method of the present invention is to culture the bacteriocin production strain in the presence of a susceptible strain or fractions thereof. At this time, the susceptible strain is naturally used to have a bacteriocin-inducing action depending on the type of bacteriocin producing lactic acid bacteria. The susceptible strain may be live or dead, and the fraction may be selected from the group consisting of intracellular fractions, cell debris and bacteriocin inducers purified therefrom.

The culturing may be carried out by adding a sensitive strain or a fraction thereof to the bacteriocin producing strain in a ratio of 1: 0.1 to 10, and anaerobic culture at 20 to 30 ° C. for 16 to 48 hours. At this time, the bacteriocin producing strain is inoculated at 0.1 to 10% by volume in the medium. At this time, the medium to be used is a conventional culture medium, and it is preferable to use an appropriate medium depending on the type of bacteriocin producing strain and the method of use. The mixing ratio and condition range during the culturing are to enhance the bacteriocin productivity of the bacteriocin producing strain, and it may be difficult to optimize the bacteriocin productivity if it is out of the range.

In carrying out the method of enhancing bacteriocin production of the present invention, when the susceptible strain is used as a live bacterium, the susceptible strain is killed by the bacteriocin producing strain and at the same time enhances the bacteriocin production of the bacteriocin producing strain. Therefore, only the bacteriocin-producing strains with enhanced bacteriocin production survive after the culture, and the culture medium may be centrifuged or filtered to obtain strains with enhanced bacteriocin production.

In the present invention, bacteriocin-producing strains in which bacteriocin production is enhanced by a susceptible strain or a fraction thereof are described as bacteriocin-producing enhanced strains. That is, the bacteriocin-producing enhanced strain of the present invention is a bacteriocin-producing strain cultured in the presence of a susceptible strain or a fraction thereof, and the bacteriocin-producing enhanced strain may be obtained by drying the bacteriocin-producing strain.

Bacteriocin-producing fortified strains of the present invention can be used as a seed for fermented foods, and particularly as a seed for non-sterile open fermentation systems. Non-sterile open fermentation is a type of food fermentation mainly used in the Asian region. Aseptic fermentation environment is achieved by completely enclosing fermentation vessels that do not sterilize raw materials during fermentation and prevent the introduction of bacteria from the outside during fermentation. It means not to use, the bacteriocin-producing fortified strain of the present invention grows as a dominant species in the fermentation system effectively inhibits the propagation of various bacteria, so it is very suitable for non-sterile open fermentation system.

In addition, bacteriocin-producing fortified strains of the present invention are provided as a seed for fermented food. The spawn for the fermented food may further comprise a susceptible strain or fractions thereof. The mixing ratio is 1: 0.1 to 10 by weight ratio of the susceptible strain or fractions thereof to the bacteriocin-producing enhanced lactic acid bacteria. When the mixing ratio is out of the range, it may be difficult to optimize the bacteriocin productivity of the bacteriocin-producing strains. The spawn for the fermented food maintains the taste and quality of the fermented food uniformly, and it is hygienic and can extend the shelf life at the same time by blocking various bacterial contamination.

In another aspect, the present invention provides a fermentation system using a seed for fermented food. The fermentation system is a non-sterile open type fermentation is added to the 0.0001 to 0.01 parts by weight (wet parts by weight) based on 100 weight of the food substrate and put at 10 to 30 ℃ fermentation. A fermentation system using bacteriocin-producing strains inhibits the growth of unsterilized food ingredients or other fungi, such as Escherichia coli and yeast introduced during the open fermentation process, and grows only bacteriocin-producing lactic acid bacteria as dominant species. Only fermentation is performed.

The present invention also provides a fermented food fermented with a seed for fermented food. Fermented foods may be prepared by adding the above seed to the food substrate and aged, wherein the amount of the seed is appropriately adjusted according to the type of fermented food and the type of seed. Preferably, it is inoculated at 0.0001 to 0.01 part by weight (wet part by weight) based on 100 parts by weight of the food main ingredient. Fermented foods that can be prepared include kimchi, salted fish, soybean paste or soy sauce, and other non-sterile open fermented foods can be used.

In particular, kimchi is used without sterilizing the main ingredients, and it is manufactured by mixing various kinds of subsidiary materials (spices), and it is difficult for artificial fermentation control and spawning, but in the present invention, various germs other than the spawn are controlled by using the spawn. Along with the effect, it facilitates flavor maintenance and long-term storage. Kimchi can be prepared by adding the seed. Kimchi main ingredients vary depending on the type of kimchi, and means the ingredients other than those classified as seasonings. Specifically, there are cabbage, radish, cucumber, radish, leek and leek. As a method of adding spawn during kimchi production, it is preferable to add spawn to kimchi seasoning atmosphere. Kimchi prepared by the above method is cooked for 2 to 3 days at room temperature, and then stored at 4 to 10 ℃.

In one embodiment of the present invention, Chinese cabbage kimchi was prepared by adding only Luconostock Citrium GJ7 or by adding Luconostock Citrium GJ7 fortified strain, and confirming the type variation and flavor of the fungi in kimchi samples by the kimchi ripening day. It was. In both cases, the experimental group maintained constant Lukonostok citreum GJ7 at 10 ^{9-10 10} cfu / mL even after 14 days, 30 days and 60 days, and no growth of bacteria was observed. Showed a significant difference. In addition, in the sensory test, the control group had a strong sour taste and seasoning taste, but the experimental group had a slightly sweet taste and a bitter taste and was soft overall, showing a remarkably good flavor.

In another aspect, the present invention provides a method for producing a fermented food comprising adding the seed spawn to the food main ingredient to produce a fermented food. For example, in the case of Chinese cabbage kimchi, seasoning is used in a weight ratio of 1: 0.1 to 0.2 for cabbage marinated in about 1.5% salinity, and spawn is used in a weight ratio of 1: 0.0001 to 0.001 for seasoning. The seasoning may vary depending on taste, but may include garlic, pepper, ginger, sugar, fish sauce or other additives. Additives include MSG, sauerkraut, chives, radishes, pears, carrots, and the like. As for the composition ratio in seasoning, it is good to adopt a normal ratio.

 Fermented food production method of the present invention by maximizing the production of bacteriocin without going through the conventional genetic manipulation to improve the bacteriocin production rate of bacteriocin production strain, it is very effective in the production of non-sterile open fermented food.

Hereinafter, examples of the present invention will be described. The following examples are only for illustrating the present invention and the present invention is not limited to the following examples.

Luconosestock Citrium GJ7 is a lactic acid bacterium isolated from Kimchi in optimum ripening state. It is a gram-positive and spherical form of Lactobacillus, deposited on July 15, 2003 with the Korea Microorganism Conservation Center and assigned accession number KFCC-11322. Granted. Luconosestock Citrium GJ7 is grown in a conventional lactic acid bacteria culture medium and incubated under anaerobic conditions at a culture temperature of 30 ± 5 ℃.

In addition, the GJ series used in the present invention are isolates and identified strains of the laboratory, the KFRI series was used to purchase strains sold by the Korea Food Research Institute, and the KCTC series were strains sold by the Korea Institute of Bioscience and Biotechnology. It was purchased and used.

Example 1 Antibacterial Activity of Bacteriocin Producing Lactic Acid Bacteria Against Other Lactic Acid Bacteria

Lactobacillus (Lactobacillus plantarum KFRI 464, susceptible to bacteriocin-producing strains, Luconosstock Citrium GJ7, Luconosstock Paramescenteroides GJ77, Luconosstock Paramescenteroides GJ74, and Lactobacillus platarum GJ78, respectively) Lactobacillus delbrueki KFRI 347, Lactobacillus plantarum KFRI 236, Luconostock mesenteroides KFRI 218, Lactobacillus exidophilus KFRI 150 and Luconostock mesenteroides KCTC 1628) Table 1 shows.

Bacteriocin Producing Strains Bacteriocin Susceptible Strains Growth inhibition ring diameter (cm) Luconostock Citrium GJ7 Lactobacillus plantarum KFRI 464 1.2 Lactobacillus delbrueki KFRI 347 1.4 Lactobacillus plantarum KFRI 236 1.2 Luconostock Mesenteroides KFRI 218 1.3 Lactobacillus exidophilus KFRI 150 2.0 Luconosstock Mesenteroides KCTC 1628 1.3 Luconostock Paramescenteroides GJ77 Lactobacillus plantarum KFRI 464 1.8 Lactobacillus delbrueki KFRI 347 2.0 Lactobacillus plantarum KFRI 236 1.3 Luconostock Mesenteroides KFRI 218 2.0 Lactobacillus exidophilus KFRI 150 2.2 Luconosstock Mesenteroides KCTC 1628 1.9 Lucono Stock Paramesenteroides GJ74 Lactobacillus plantarum KFRI 464 1.9 Lactobacillus delbrueki KFRI 347 2.0 Lactobacillus plantarum KFRI 236 1.9 Luconostock Mesenteroides KFRI 218 2.2 Lactobacillus exidophilus KFRI 150 2.6 Luconosstock Mesenteroides KCTC 1628 2.1 Lactobacillus Platarum GJ78 Lactobacillus plantarum KFRI 464 1.5 Lactobacillus delbrueki KFRI 347 2.0 Lactobacillus plantarum KFRI 236 2.0 Luconostock Mesenteroides KFRI 218 2.3 Lactobacillus exidophilus KFRI 150 2.3 Luconosstock Mesenteroides KCTC 1628 2.0

The direct method of direct application of the cells (Kim, SI, Kim, IC and Chang, HC Isolation and identification of antimicrobial agent producing microorganisms and sensitive strain from soil.J. Korean Soc.Food Sci.Nutr. 28 (3): 526-53 (1999)), and the sterile culture solution was added to a paper disk to use agar diffusion (Tagg, JR and Mcgiven, AR Assay system for bacteriocin. Appl. Microbiol. 21: 943 (1971)). The antibacterial activity of each bacteriocin producing strain was different depending on the susceptible strain or the bacteriocin producing strain.

Example 2: Antimicrobial Activity Enhancement of Bacteriocin Producing Strains

2-1. Cell fractionation for location of bacteriocin-producing substances

end. Preparation of each fraction of susceptible strains

 Lactobacillus plantarum KFRI 464, Lactobacillus delbruecki KFRI 347, Lactobacillus plantarum KFRI 236, Luconostock mesenteroides KFRI 218, Lactobacillus exidophilus KFRI 150 and Luconostock mesenteroides KCTC 1628 Thus, a total of six species were used as susceptible strains for fractionation of bacteriocin derivatives.

Extracellular Extract

: 100 ml of liquid medium was inoculated with susceptible strains, incubated at 30 ° C. for 24 hours, followed by centrifugation (9,950 × g, 4 ° C., 15 min) to separate the supernatant.

Intracellular extract

: 100 ml of liquid medium was inoculated with susceptible strains, incubated at 30 ° C. for 24 hours, followed by centrifugation (9,950 × g, 4 ° C., 15 min) to obtain pellets, and 20 ml of 50 mM Tris-HCl (pH 8.3) was added thereto. Ultrasonic pulverization (Ampi: 60, Pulse: 2 sec, Time: 10 min, Sonics. VC 130), the pulverization was centrifuged (9,950 xg, 4 ℃, 25 min), then the supernatant was filtered to separate the filtrate.

Cell debris

: 100 ml of liquid medium was inoculated with susceptible strains, incubated at 30 ° C. for 24 hours, followed by centrifugation (9,950 × g, 4 ° C., 15 min) to obtain pellets, and 20 ml of 50 mM Tris-HCl (pH 8.3) was added thereto. Ultrasonic grinding (Ampi: 60, Pulse: 2 sec, Time: 10 min, Sonics. VC 130) was carried out, and the pellet was centrifuged (9,950 xg, 4 ° C, 25 min) to separate pellets. The pellet was used by dissolving in 20 ml MRS liquid medium.

I. Confirmation of antimicrobial activity effect of each fraction

Luconosstock Citrium GJ7, Luconosstock Paramescenteroides GJ77, Luconosstock Paramescenteroides GJ74, and Lactobacillus Platarum GJ78, respectively, were inoculated in 100 ml MRS liquid medium at 1%, each of the six susceptible strains. Extracellular fraction, intracellular fraction and cell debris were added at 2.5% and incubated at 30 ° C. for 4 hours. After incubation, the culture filtrate was obtained and the antibacterial activity was confirmed by the agar diffusion method.

2-2. Induction of Bacteriocin Production by Susceptible Strain Cell Fraction

The effect of increasing bacteriocin production of the bacteriocin producing strains by the extracellular fraction, the intracellular fraction, and the cell debris of each susceptible strain was confirmed.

In this experiment, a total of 6 bacteriocin-sensitive strains (Lactobacillus plantarum KFRI 464, Lactobacillus delbrueckee KFRI 347, Lactobacillus plantarum KFRI 236, Luconostoke mesenteroides KFRI 218, Lactobacillus exidophilus KFRI 150 and Lukonostoke mesenteroides KCTC 1628) were used. Among the fractions of each susceptible strain, the most potent bacteriocin-inducing activity was shown in bold and italic. Susceptible strains not listed in bold and italy in any fraction indicate no bacteriocin inducers present.

Extracellular fractions did not increase bacteriocin production, and intracellular fractions and cell debris significantly increased bacteriocin production. Particularly, the fractions that increase bacteriocin production were identified differently according to susceptible strains, and some susceptible strains had an effect of increasing bacteriocin production in both intracellular fractions and cell debris, and some susceptible strains showed the above activity in only one fraction. . Detailed results are shown in Table 2 below.

Bacteriocin Producing Strains Susceptible strains Fraction of Susceptible Strains Control Cell debris Intracellular fractionation Luconostock Citrium GJ7 Lactobacillus plantarum KFRI 464 1.2 2.0 1.5 Lactobacillus delbrueki KFRI 347 1.4 1.7 1.4 Lactobacillus plantarum KFRI 236 1.2 1.2 1.2 Luconostock Mesenteroides KFRI 218 1.3 1.3 1.3 Lactobacillus exidophilus KFRI 150 2.0 2.0 2.8 Luconosstock Mesenteroides KCTC 1628 1.3 2.0 1.3 Luconostock Paramescenteroides GJ77 Lactobacillus plantarum KFRI 464 1.8 2.1 2.2 Lactobacillus delbrueki KFRI 347 1.7 1.7 2.2 Lactobacillus plantarum KFRI 236 1.3 2.0 2.0 Luconostock Mesenteroides KFRI 218 2.0 2.3 2.2 Lactobacillus exidophilus KFRI 150 2.8 2.8 2.8 Luconosstock Mesenteroides KCTC 1628 1.9 1.9 2.2 Lucono Stock Paramesenteroides GJ74 Lactobacillus plantarum KFRI 464 2.3 2.3 2.3 Lactobacillus delbrueki KFRI 347 2.0 2.2 2.5 Lactobacillus plantarum KFRI 236 1.2 1.2 1.2 Luconostock Mesenteroides KFRI 218 2.2 2.2 2.5 Lactobacillus exidophilus KFRI 150 2.6 2.7 2.9 Luconosstock Mesenteroides KCTC 1628 2.1 2.5 2.3 Lactobacillus Platarum GJ78 Lactobacillus plantarum KFRI 464 1.5 2.2 1.7 Lactobacillus delbrueki KFRI 347 1.7 1.7 1.7 Lactobacillus plantarum KFRI 236 2.0 2.0 2.0 Luconostock Mesenteroides KFRI 218 2.2 2.2 2.2 Lactobacillus exidophilus KFRI 150 2.3 2.3 2.5 Luconosstock Mesenteroides KCTC 1628 2 2.4 2.2

In particular, the bacteriocin inducer for Luconosstock Citrium GJ7 is present in cell debris in susceptible strains Lactobacillus plantarum KFRI 464, Lactobacillus delbrueckee KFRI 347 and Luconosestock mesenteroides KCTC 1628, but Lactobacillus exis Dophilus KFRI 150 was present in the intracellular fraction.

Bacteriocin inducers for Luconostok Paramesenteroides GJ77 include intracellular fractions in susceptible strains Lactobacillus plantarum KFRI 464, Lactobacillus delbruecki KFRI 347 and Lukonostok mesenteroides KCTC 1628 in the intracellular fraction. Terroides KFRI 218 was present in cell lysates and Lactobacillus plantarum KFRI 236 had inducers in both intracellular fractions and cell lysates.

Bacteriocin inducers for Luconosestock Paramesenteroides GJ74 are found in the intracellular fractions of Lactobacillus delbrueckee KFRI 347, Luconostoke mesenteroides KFRI 218 and Lactobacillus exidophilus KFRI 150, and Luconosstock mesenteroide Des KCTC 1628 cell lysate was present.

Bacteriocin inducers for Lactobacillus platarum GJ78 were present in cell lysates for Lactobacillus plantarum KFRI 464 and Lukonostoke mesenteroides KCTC 1628, and Lactobacillus exidophilus KFRI 150 was present in intracellular fractions.

In addition, Lactobacillus plantarum KFRI 464 and Lactobacillus plantarum KFRI 236 are both Lactobacillus plantarum and susceptible to Luconosstock Citrium GJ7, while KFRI 464 is derived from Luconostock Citrium GJ7. Induced bacteriocin production, but KFRI 236 was found to have no inducing effect. In other words, susceptible strains containing inducing factors that can induce bacteriocin production for specific bacteriocin-producing strains are not determined by the genes and species of the microorganism. It can be seen that is very specific.

2-3. Co-culture

When bacteriocin inducers were present in the cell lysate, it was confirmed whether co-culture of bacteriocin-producing strains and susceptible strains could increase bacteriocin production.

Bacteriocin-producing strains and susceptible strains were inoculated into MRS medium by 1%, respectively, and co-cultured at 30 ± 5 ° C. for 24 hours, followed by cell counting. In addition, in order to confirm whether the antimicrobial activity is added by the result of lactic acid produced in lactic acid bacteria, only susceptible strains were cultured and their antimicrobial activity was also confirmed.

Both antimicrobial activity was increased by mixed cultures of 1% or more susceptible strains, but the dominant species were identified as bacteriocin producing strains. The antimicrobial activity was determined by bacteriocin producing strains. It can be seen that.

In addition, when co-culture of 1% bacteriocin-producing strains and 10% susceptible strains, the dominant strains in the medium were bacteriocin-producing strains inoculated at 1%, not 10% susceptible strains. This suggests that the more susceptible strains are present on the medium containing the bacteriocin producing strains, the more bacteriocin producing strains produce more bacteriocins to control the medium environment. Thus, the susceptible strain stimulates the bacteriocin production against the bacteriocin producing strain, and at the same time, growth is inhibited by the produced bacteriocin, so that the bacteriocin producing strain is maintained as a dominant species in the culture medium when mixed.

Therefore, the inducer present in the cell wall of the susceptible strain is known to stimulate the bacteriocin producing strain when mixed with the bacteriocin producing strain, thereby promoting bacteriocin production. In other words, the bacteriocin-producing strain recognizes a specific substance secreted by susceptible strains and assumes that it is an inducer and secretes a much stronger or higher amount of antimicrobial substances depending on the instinct physiology of the microorganism that predominantly controls the environment. Makes it possible.

In addition, Lactobacillus plantarum, one of the susceptible strains, is a normal lactic acid bacterium type, and can produce a large amount of lactic acid and exhibit antibacterial activity. To confirm this, the mixed culture of Luconostock Paramescenteroides GJ74 and Lactobacillus delbrueckee KFRI 347 , or the mixed culture of Luconostock Citreum GJ7 and Lactobacillus plantarum KFRI 464, after incubation, pH, Cell growth degree (OD after culture) and diameter of the inhibitory ring were observed in accordance with the mode observed in the bacteriocin producing strain, and the bacteriocin producing strain was maintained at about 100% dominant species in the medium after the culture. This means that even though mixed strains (susceptible bacteria that produce lactic acid and bacteriocin-producing strains) exist under the same medium environment, the bacteriocin-producing strains can completely control the medium environment. It indicates that it can be applied to the production of sterilized fermented food.

Pear PH after incubation O.D. after incubation. Inhibitory ring diameter (cm) Fungal population after incubation GJ74 4.44 8.386 2.0 GJ74 100% GJ74 + Lb. del 1% 4.44 8.456 2.1 GJ74 100% GJ74 + Lb. del 2% 4.45 8.491 2.1 GJ74 100% GJ74 + Lb. del 5% 4.47 8.498 2.3 GJ74 100% GJ74 + Lb. del 10% 4.48 8.435 2.4 GJ74 99.9% Lb. del 1% 4.88 4.893 1.3 Lb. del 100% Lb. del 2% 5.03 4.865 1.5 Lb. del 100% Lb. del 5% 5.04 5.054 1.5 Lb. del 100% Lb. del 10% 5.03 4.774 1.5 Lb. del 100% GJ7 5.02 2.52 1.2 GJ7 100% GJ7 + Lb. plant 1% 4.89 2.624 1.5 GJ7 100% GJ7 + Lb. plant 2% 4.86 3.032 1.7 GJ7 97.6% GJ7 + Lb. plant 5% 4.78 3.268 1.7 GJ7 96.3% GJ7 + Lb. plant 10% 4.50 4.232 1.8 GJ7 91.96% Lb. plant 1% 4.54 8.26 2.0 Lb. plant 100% Lb. plant 2% 4.49 8.428 2.1 Lb. plant 100% Lb. plant 5% 4.50 8.351 2.3 Lb. plant 100% Lb. plant 10% 4.50 8.197 2.4 Lb. plant 100% * GJ74: Luconostock Paramesenteroides GJ74 Lb. del : Lactobacillus del bruecki KFRI 347 GJ7: Luconostock Citrium GJ7 Lb. plant : Lactobacillus plantarum KFRI 464

Example 3: Enhancement of Antimicrobial Activity of Luconotstock Citrium GJ7

3-1. Co-culture

Changes in antimicrobial activity were confirmed by co-culture with Lactobacillus delbrueckee KFRI 347, Lukonostoke mesenteroides KFRI 218, and Lactobacillus plantarum KFRI 464, which are susceptible strains of Luconostock Citrium GJ7.

Luconosstock Citrium GJ7 was inoculated at 1% in 100 ml of MRS, and after incubation for 24 hours by inoculation of 1% of susceptible strains, the culture filtrate was obtained and the antibacterial activity was confirmed. The results are shown in Table 4 below.

Co-culture Antimicrobial activity Luconostock Citrium GJ7 + Luconostock Citrium GJ7 + Lactobacillus plantarum KFRI 464 ++++ Luconostock Citrium GJ7 + Lactobacillus del brewerie KFRI 347 +++ Luconosstock Citrium GJ7 + Luconosstock Mesenteroides KFRI 218 + Luconostock Citrium GJ7 + Lactobacillus plantarum KFRI 236 + Luconostock Citrium GJ7 + Lactobacillus exidophilus KFRI 150 + Luconostock Citrium GJ7 + Luconostock Mesenteroides KFRI 1628 +++   * Antimicrobial activity is measured by the size of growth inhibition ring ++++: 1.9-2.0 cm, +++: 1.7-1.8 cm, ++: 1.4-1.6 cm, +: 1.1-1.3 cm,-: none

In Table 4, it was confirmed that the antimicrobial activity is significantly increased when co-culture of susceptible strains to Lukonosciteum GJ7. However, the susceptible strains Lukonosstock Mesenteroides KFRI 218, Lactobacillus plantarum KFRI 236, and Lactobacillus exidophilus KFRI 150 were found to have no effect on the antimicrobial activity.

Looking at the results of Table 4 together with the results according to the location of the inducer in the susceptible strains of Table 2, in Table 2, the bacteriocin inducer is not present in Luconostok Mesenteroides KFRI 218 and Lactobacillus plantarum KFRI 236 In case of Lactobacillus exidophilus KFRI 150, although bacteriocin inducers are present but intracellularly, co-culture of Table 4 does not appear to enhance bacteriocin production. In Table 2, the presence of an inducer in the cell lysate (Lactobacillus plantarum KFRI 464, Lactobacillus delbrueckee KFRI 347, Luconostock mesenteroides KFRI 1628, Lactobacillus exidophilus KFRI 150) are all susceptible. Coculture of strains and bacteriocin producing strains may increase bacteriocin production.

That is, not all susceptible strains have bacteriocin induction effects. Intracellular distribution of inducers between susceptible strains (Lactobacillus plantarum KFRI 464, Lactobacillus delbrueckee KFRI 347, Luconostock mesenteroides KFRI 1628, Lactobacillus exidophilus KFRI 150). Depending on the location, the co-culture alone induces an effect (Lactobacillus plantarum KFRI 464, Lactobacillus delbrueckee KFRI 347, Luconostock mesenteroides KFRI 1628) and susceptible bacterial cell fractions during the culture of the bacteriocin production strain. Only when the bacteriocin production enhancer may appear.

Example 4 Properties of Bacteriocin Production Inducers

Intracellular fractions were prepared from Lactobacillus platarum KFRI 464, which induces bacteriocin production-promoting effects against Luconosstock Citrium GJ7, and their pH stability and thermal stability were confirmed.

4-1. pH stability

Intracellular fractions were pH 2.5 (50 mM Glycine-HCl), pH 4.5 (50 mM Sodium acetate), pH 6.0 (50 mM Sodium citrate), pH 7.0 (50 mM Phosphate), pH 8.0 (50 mM Tris-HCl) and It was dissolved in each buffer solution of pH 9.5 (50 mM Glycine-NaOH), left at 4 ° C. for 12 hours, and added to the leuconstock citreum GJ7 culture process for 24 hours, followed by antimicrobial activity. The results are shown in Table 5 below.

Control Intracellular fraction / pH Luconostock Citrium GJ7 Luconosestock Citrium GJ7 + Intracellular Fraction 2.5 4.5 6.0 7.0 9.5 + ++ ++ ++ ++ ++ ++ * Antimicrobial activity is measured by the size of growth inhibition ring ++: 1.4 ~ 1.6 ㎝, +: 1.1 ~ 1.3 ㎝,-: none

The intracellular fraction was found to still increase the bacteriocin production of the bacteriocin producing strains when treated under conditions of pH 2.5 to 9.5, indicating that the bacteriocin production inducers are pH stable.

4-2. Thermal stability

The intracellular fractions were heat treated at 4, 30, 50 and 70 ° C. for 3 hours each. In addition, heat treatment was performed at 100 and 121 ° C. for 30 minutes, respectively. The heat-treated intracellular fractions were co-cultured with Luconosestock Citrium GJ7 to confirm their effect on antimicrobial activity. The results are shown in Table 6 below.

Control Antimicrobial activity in intracellular fractionation, temperature ℃ (hours) Luconostock Citrium GJ7 Luconosestock Citrium GJ7 + Intracellular Fraction 4 (3h) 37 (3h) 50 (3h) 70 (3 h) 100 (30 minutes) 121 (30 minutes) + ++ ++ ++ ++ ++ +++ +++ * Antimicrobial activity is measured by the size of growth inhibition ring ++: 1.4 ~ 1.6 ㎝, +: 1.1 ~ 1.3 ㎝,-: none

The bacteriocin inducer maintained its induction activity even in the range of 4 to 121 ° C, and increased induction activity at 100 ° C and 121 ° C. This leads to the modification of the tertiary structure of protein molecules by high heat, and this modified form is rather high induction activity.

4-3. Enzyme treatment

In order to confirm whether the bacteriocin inducer is a protein, the proteolytic enzyme treatment was performed to investigate bacteriocin production.

Intracellular fractions were reacted with proteinase K, trypsin and α-chymotrypsin at 37 ° C. for 5 hours at a final concentration of 2 mg / ml, and protease inhibitor AEBSF (4- (2-aminoethyl) -benzenesulfonyl fluoride) was 100. Addition at mM concentration was performed for 12 hours at 37 ℃. Each treatment was then added during culturing Lukonotstrum GJ7 to confirm the antimicrobial activity change. The results are shown in Table 7 below.

Culture filtrate Enzyme treatment Antimicrobial activity Control Luconostock Citrium GJ7 No treatment + Luconosestock Citrium GJ7 + Intracellular Fraction No treatment ++ Experimental group Luconosestock Citrium GJ7 + Intracellular Fraction Proteinase K + Trypsin + α-chymotrypsin + * Antimicrobial activity is measured by the size of growth inhibition ring ++: 1.4 ~ 1.6 ㎝, +: 1.1 ~ 1.3 ㎝,-: none

The intracellular fraction was confirmed that the proteolytic enzyme treatment did not induce bacteriocin production, it can be seen that the bacteriocin production inducer is a protein.

4-3. Molecular Weight and N-terminal Amino Acid Sequence

A crude inducing factor was prepared in intracellular extracts from Lactobacillus platarum KFRI 464, which induces bacteriocin production-promoting effects on Luconosstock Citrium GJ7. The inducer was first purified using a DEAE-sephacel column, followed by secondary purification through HPLC (μBondapack C18 column, 3.9 × 300 mm, Waters, Co.) to separate the protein fraction with inducible activity. It was. Purified inducers were tricine-polyacrylamide gel eletrophoresis.

Figure 1 is an electrophoresis picture of bacteriocin derivatives isolated from Lactobacillus platarum KFRI 464, the inducer was identified as a protein having a molecular weight of about 6,500 Da. In Figure 1 lane 1 is the inducer and lane 2 is the polypeptide standard size marker (Bio-Rad, USA).

The N-terminal amino acid sequence of the pure purified inducer was analyzed and 16 amino acid sequences starting from the N-terminus starting with methionine were identified. The sequence is set forth in SEQ ID NO: 1.

Example 5: Construction of Kimchi Fermentation System

5-1. Preparation of Kimchi Added with Lucono Stock Citrium GJ7

end. in vitro

Kimchi of the same compounding ratio as commonly used kimchi composition was ground in a mixer, squeezed with sterile gauze, and then filtered with 0.45 μm filter paper of Millipore, to prepare a kimchi liquid medium. The kimchi liquid medium was inoculated with 1% by volume of Lucono Stock Citrium GJ7, and the number of bacteria in the medium was measured while anaerobic culture at 30 ° C. for 2 days. CaCO ₃ , MRS and LB plate medium were used as the bacterial counting medium, and colonies formed for each medium were randomly selected and their morphology was observed under a microscope. The results are shown in Table 8 below.

Reputation Control Luconostock Citrium GJ7 30 ℃, 0h culture 30 ℃, 48h culture 30 ℃, 0h culture 30 ℃, 48h culture CaCO ₃ Too many germs (counting specific strains) Very many bacteria (yeast, various kinds of lactic acid bacteria and various bacteria, inability to count specific strains) Too many germs (counting specific strains) Single species: GJ7 (1.8 × 10 ¹⁰ ) MRS Too many germs (counting specific strains) Very many bacteria (yeast, various kinds of lactic acid bacteria and various bacteria, inability to count specific strains) Too many germs (counting specific strains) Single species: GJ7 (3.0 × 10 ¹⁰ ) LB Too many germs (counting specific strains) Very many germs (various flora) Too many germs (counting specific strains) Single species: GJ7 (2.8 × 10 ¹⁰ )

From Table 8, the kimchi liquid medium contains a variety of microorganisms, but when inoculated with the ruconotstock citrium GJ7 of the present invention 48 hours after the cultivation other kinds of microorganisms are not observed and Lukonostok citrium GJ7 Only dominant species were found to exist. In other words, it was found that Lukonot stock citreum GJ7 exists as a dominant species by inhibiting the growth of other microorganisms in kimchi liquid medium.

I. in vivo

After the kimchi was cooked by inoculating Luconot Stock Citrium GJ7 under the same conditions as the actual soaking kimchi, the changes in the flora were observed.

Kimchi seasoning 300-580 g (garlic: 27-52 g, red pepper: 55-105 g, ginger: 14-26 g, sugar: 18-34 g, MSG) for 2,700-3,000 g of pickled cabbage (about 1.5% salinity) : 8-16 g, fish sauce: 95-185 g, savory rice: 2-4 g, green onion: 27-53 g. Radish: 20-39 g, pear: 20-39 g, carrot: 14-27 g) 100 mg (wet weight, 10 ⁹ to 10 ¹⁰ cfu / ml) of Luconot Stock Citrium GJ7 cells was added to the seasoning. Luconosstock Citrium GJ7 cells were prepared by centrifugation at 9,950 g and 4 ° C. for 5 minutes at 30 ° C. for 24 hours in MRS medium, and pellets were washed twice with sterile water.

The prepared kimchi was stored at room temperature (20-28 ° C.) for 2 days and then refrigerated. Part of the kimchi sample was taken while storing the internal organs to check the bacterial count and the total flora. Table 9 shows the total bacterial counts and bacterial counts in kimchi after 14 days of refrigeration. In Table 9, the experimental group to which Lukono Stock Citrium GJ7 was added during Kimchi was kept constant at 10 ⁹ cfu / ml after 14 days.

Reputation Control group (number of colonies of bacteria) Experimental group CaCO ₃ Very high bacteria (cannot be measured for a specific strain) 3.3 × 10 ⁹ cfu / mL (GJ7) MRS      Very high bacteria (cannot be measured for a specific strain) 4.1 × 10 ⁹ cfu / mL (GJ7) LB      Too many germs (cannot measure specific strains) 2.5 × 10 ⁹ cfu / mL (GJ7) Sensory evaluation It has a sour taste and a lot of seasoning taste. Both color and taste are poor quality compared to the experimental group Slightly sweet and tangy. The sour taste is weak, so the taste is soft pH 4.09 4.28

In addition, the kimchi taste of the experimental group showed a soft sweetness and refreshing taste while refreshing and cool. The above results confirm the same result by repeating the same experiment five times.

5-2. Kimchi production using bacteriocin inducer

end. Preparation of Fortified Strains

Luconosstock Citrium GJ7 was inoculated at 1% in MRS medium, and the intracellular fraction of Lactobacillus plantarum KFRI 464 was added at 2.5% and then incubated at 30 ° C for 24 hours. The culture solution was centrifuged (9,950 xg, 4 ℃, 15 min) to separate the cells and the supernatant, the cells were washed twice in sterile water and used as a fortified kimchi production strain. When the fortified strains are not used immediately, do not suspend the cells by centrifugation and store them at -70 ° C, and thaw them if necessary.

I. Preparation of Kimchi Using Fortified Strains

Kimchi was prepared in the same manner as in vivo, and 100 mg (wet weight) of fortified strain was mixed with kimchi seasoning. At this time, intracellular fractions were prepared from Lactobacillus plantarum KFRI 464 cells of 10 to 1000 mg (wet weight) and mixed with the fortified strains.

Table 10 shows the total bacterial count and the number of bacteria in kimchi after 60 days of refrigerated storage after cooking for two days at room temperature (20-28 ℃) after the production of kimchi using fortified strains. After 60 days of ripening at room temperature, the reduction of inoculation bacteria and the growth of other bacteria were not observed due to the transition of Kimchi fermented lactic acid bacteria, and the taste of kimchi ripening was still observed without excessive soaking. Therefore, by using the Lukonot Stock Citrium GJ7 reinforced strain of the present invention in the production of kimchi, it is possible to improve the spawning of kimchi, improvement of shelf life and flavor of kimchi.

Reputation Control group (number of colonies of bacteria) Experimental group CaCO ₃ Very many bacteria (large colonies, bacillus: 2 × 10 ⁹ cfu / ml medium colonies, cocci: 8.3 × 10 ¹⁰ cfu / ml yeast: 3 × 10 ⁸ cfu / ml) 8.6 × 10 ⁹ cfu / mL (GJ7) MRS Very high bacterial count (large colony, bacillus: 8 × 10 ⁹ cfu / ml medium colony, cocci: 2 × 10 ⁹ cfu / ml) 5.9 × 10 ⁹ cfu / mL (GJ7) LB Very large bacteria (large colonies, bacillus: 2 × 10 ⁹ cfu / ml medium colonies, cocci: 8.3 × 10 ¹⁰ cfu / ml small colonies, simple bacilli: 5.2 × 10 ¹⁰ cfu / ml) 3.7 × 10 ¹⁰ cfu / mL (GJ7) Sensory evaluation A lot of sour taste, seasoning taste and light taste Cool with tapping taste, weak acidity and sweet taste pH 4.06 4.19

 All. Comparison of microbial flora in kimchi and general kimchi using fortified strains

Samples of Kimchi prepared in B. and two kinds of Kimchi (Jongga House Kimchi, Pulmuone Kimchi) and commercial household Kimchi (Gwangju Metropolitan City), which are currently on the market, were searched for kimchi samples after 23-30 days. The strain and the salinity, pH were observed, and the results are shown in Table 11 below.

In Kimchi using Lukonot Stock Citrium GJ7-reinforced strains, only GJ7 was detected as in Table 9 and Table 10.However, other commercial bacteria, yeasts, molds, etc. About species were detected. Like this, the kimchi we usually eat has a myriad of bacteria in addition to beneficial lactic acid bacteria. For this reason, when the international standard for kimchi was enacted by the 24th International Code of Food Standards (Codex) on July 6, 2001 (Geneva, Switzerland, 5 pm local time), it was reduced to Control of microorganisms is not possible with current technology, and microbiological specifications have not been included in the list. Therefore, when the fortified strains of the present invention are applied to fermented foods, not only can control germs effectively, but also microbiological standardization can be achieved.

Kimchi Search strain: Number of bacteria pH Salinity GJ7 fortified kimchi (30 days) GJ7: 9.7 × 10 ⁹ 4.20 1.5 Chongga Kimchi (30 days) 5 kinds ① Lactobacillus (1 type): 7.5 × 10 ⁹ ② Bacteria (2 types): 4.5 × 10 ⁸ ③ Yeast (1 type): 7 × 10 ¹⁰ ④ Mold (1 type): 2 × 10 ¹⁰ 4.06 1.6 General Home Kimchi (23 Days) 6 species ① Lactobacillus (2 species): 4 × 10 ⁸ ② Bacteria (2 species): 5 × 10 ⁸ ③ Yeast (1 species): 5 × 10 ⁹ ④ Mold (1 species): 2 × 10 ⁷ 4.10 1.5 Pulmuone (26 days) 7 species ① Lactobacillus (2 species): 5.5 × 10 ⁷ ② Bacteria (2 species): 5 × 10 ⁸ ③ Yeast (1 species): 9 × 10 ⁶ ④ Mold (1 species): 1 × 10 ⁷ 4.19 1.7

As described above, the present invention provides a method for enhancing bacteriocin production from a bacteriocin producing strain through a non-genetic manipulation method, and provides a bacteriocin-enhanced strain as a seed of an unsterilized open fermented food. Due to the present invention, it is possible to make specific microorganisms in non-sterile open fermented foods such as kimchi, salted fish, and soy sauce, and thus, to improve the hygienic effect and the shelf life by preventing the uniformity of taste and quality of the fermented foods and contamination of various bacteria. The effect can be obtained.

<110> CHANG, HAE CHOON <120> METHOD FOR PREPARING FERMENTED FOOD USING STRAIN ENHANCED          PRODUCTIVITY OF BACTERIOCIN <130> dpp20032204kr <160> 1 <170> KopatentIn 1.71 <210> 1 <211> 16 <212> PRT <213> Lactobacillus plantarum KFRI 464 <400> 1 Met Val Lys Ile Arg Leu Lys Arg Met Gln Ser Lys Lys Asn Pro Phe   1 5 10 15

Claims (24)

A method for enhancing bacteriocin production, comprising adding a susceptible strain or a fraction thereof to a bacteriocin producing strain, The susceptible strain is inhibited growth by bacteriocin-producing lactic acid bacteria and is a strain that acts specifically to the bacteriocin-producing lactic acid bacteria to enhance bacteriocin production. According to claim 1, wherein the bacteriocin producing strain is a bacteriocin producing lactic acid bacteria; The susceptible strain is live or dead; And The fraction of the susceptible strain is an augmentation method is selected from the group consisting of intracellular fractions, cell debris and bacteriocin inducers purified therefrom. The method of claim 1, Adding a sensitive strain or a fraction thereof to the bacteriocin producing strain is culturing by adding a sensitive strain or a fraction thereof to the bacteriocin producing lactic acid bacteria at a ratio of 1: 0.1 to 10; And Said step is an augmentation method comprising anaerobic culture at 20 to 30 ℃ for 16 to 48 hours. A method for producing a bacteriocin-producing enhanced strain comprising culturing a bacteriocin-producing strain in the presence of a susceptible strain or a fraction thereof, thereby obtaining a bacteriocin-producing enhanced strain, The susceptible strain is inhibited in growth by bacteriocin-producing lactic acid bacteria and acts specifically to the bacteriocin-producing lactic acid bacteria to enhance the production of bacteriocin. The method of claim 4, wherein The bacteriocin producing strain is a bacteriocin producing lactic acid bacterium; The susceptible strain is live or dead; And The fraction of the susceptible strain is selected from the group consisting of intracellular fractions, cell debris and bacteriocin inducers purified therefrom; The preparation method includes culturing the bacteriocin producing strain by adding a sensitive strain or a fraction thereof in a ratio of 1: 0.1 to 10; And The manufacturing method comprises the anaerobic culture for 16 to 48 hours at 20 to 30 ℃. The method according to claim 4, wherein the bacteriocin producing strain is leukonstock citrium GJ7 (KFCC-11322). A seed composition for fermented foods containing bacteriocin-producing fortified strains of Luconosstock Citrium GJ7, wherein bacteriocin production is enhanced by the method of claim 6. The method of claim 7, wherein The spawn for the fermented food further comprises a susceptible strain or fractions thereof; And The spawn for the fermented food further comprises a sensitive strain or fractions thereof in a bacteriocin-producing fortified strain 1: 0.1 to 10 by weight, The susceptible strain is a strain composition selected from the group consisting of Lactobacillus plantarum KFRI 464, Lactobacillus delbrueckee KFRI 347, Lactobacillus exidophilus KFRI 150 and Lukonostoke mesenteroides KCTC 1628. The method of claim 8, The susceptible strain is live or dead; And The fraction of the susceptible strain is a seed composition which is selected from the group consisting of intracellular fractions, cell debris and bacteriocin inducers purified therefrom. delete delete delete delete The spawn composition according to any one of claims 7 to 9 is added at 0.0001 to 0.01 part by weight (wet part by weight) based on 100 parts by weight of the food base, and placed at 10 to 30 ° C. to grow as a dominant species. Fermentation system comprising fermentation. A fermented food prepared by adding the seed composition according to any one of claims 7 to 9. The fermented food of claim 15, wherein the fermented food is kimchi. delete delete Induces bacteriocin production of leukonstock citrium GJ7; Purified from Lactobacillus platarum KFRI 464; The N-terminal amino acid sequence of SEQ ID NO: 1; The molecular weight is 6,500 Da; Thermally stable at 4 to 121 ° C; having stability at pH 2.5 to 9.5, Bacteriocin Production Inducing Protein. delete Enhances bacteriocin production of leunostock citrium GJ7; And Susceptible strains against Luconosstock Citreum GJ7 selected from the group consisting of Lactobacillus plantarum KFRI 464, Lactobacillus delbruecki KFRI 347, Lactobacillus exidophilus KFRI 150 and Lukonostoke mesenteroides KCTC 1628. . delete delete delete

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