KR101834231B1 - Vegetable Lactobacillus plantarum DSR KF15 having Activities on Antimicrobial And Antifungal for keeping freshness and Use Thereof - Google Patents

Abstract

The present invention relates to a plant lactobacillus Lactobacillus plantarum DSR KF15 (KCCM11772P) having antimicrobial and antifungal activity for maintaining freshness and its use. More specifically, strains isolating and identifying lactic acid bacteria from kimchi are not only antimicrobial properties that inhibit the growth of bacteria but also lactic acid bacteria that are excellent in anti-acidity, salt tolerance, infertility, and resistance to antibiotics as well as antifungal activity against yeast and fungi which are food harmful fungi. The strains of the present invention and the cultures thereof can be applied to foods as natural beverages to provide an effect of extending shelf life. It can also be provided as a pharmaceutical composition such as a dressing, a probiotic agent, a feed, an antimicrobial, etc., and a food or a functional food composition.

Description

Lactobacillus plantarum DSR KF15 having antimicrobial and antifungal activity for maintaining freshness and its use (Vegetable Lactobacillus plantarum DSR KF15 having activities of Antimicrobial and Antifungal for Antifungal and Antifungal Activity,

The present invention relates to a strain of Lactobacillus plantarum Lactobacillus DSR KF15 (strain No. KCCM11772P, Deposit date 2019.9.18) having antibacterial and antifungal activity for maintaining freshness and its use. More specifically, the lactic acid bacteria Lactobacillus plantarum DSR KF15 of the present invention is useful not only in antimicrobial activity for inhibiting the growth of bacteria but also in antibacterial activity against yeast and fungi, which are food harmful fungi, as well as lactic acid bacteria having excellent acid resistance, salt resistance, to be.

The strains of the present invention and the cultures thereof can be applied to foods as natural beverages to provide an effect of extending shelf life. It can also be provided as a pharmaceutical composition such as a dressing, a probiotic agent, a feed, an antimicrobial, etc., and a food or a functional food composition.

As consumers are increasing their income level and well-being, consumers are increasingly concerned about their health. Therefore, the need for the development of natural antimicrobial agents with excellent efficacy is increasing because of the increasing rejection of chemical preservatives. In addition, with regard to the development of natural antimicrobial agents, there is a growing interest in the antibacterial ability of cultured materials produced by microorganisms or microorganisms, and more importantly, strains with superior antibacterial activity among lactic acid bacteria, which are generally recognized as safe (GRAS) There is a growing interest in developing. However, in the case of lactic acid bacteria, it is difficult to apply it as a natural antimicrobial agent in a food with a narrow antimicrobial spectrum, and it is urgently required to develop a lactic acid bacterium having antibacterial activity against fungi such as yeast and mold as well as pathogenic bacteria. Lactobacillus strains known to be relatively safe among lactic acid bacteria are known to produce certain microorganisms in the intestinal tract of humans and animals and to enhance immunity, neutralization of harmful toxins, increase digestion and absorption of nutrients, and bioactivity. It is being taken in the form of a branch.

Pathogenic bacteria that cause food poisoning include Salmonella spp., Staphylococcus aureus, intestinal toxic Escherichia coli, Bacillus cereus, Yersinia enterocolitica and Listeria monocytogenes. Although synthetic or natural antibiotics are mainly used to prevent or treat infection with these pathogenic bacteria, they are strictly regulated due to problems such as tolerance and persistence, destroying intestinal beneficial microorganisms, and preventing the pathogenic bacteria infection caused by normal microorganisms . In addition, alkaloids, flavonoids, phytoalexins, antimicrobial peptides, organic acids and fatty acids are known as natural antibiotics, but they are not commercialized due to problems of antimicrobial activity, stability and formability. As an alternative to the problems of synthetic or natural antimicrobial agents, there are lactic acid bacteria, but lactic acid bacteria are excellent in acid resistance, biliary properties, intestinal viability, antibiotic resistance and pathogenic bacteria inhibiting ability to withstand various substances secreted from digestive organs The development of lactic acid bacteria is urgently required. If an antimicrobial agent using lactic acid bacteria that can solve these problems is developed, it is possible to increase the shelf life by inhibiting rancidity and it can be advantageously used for food production because it is safe for human body. In addition, antibiotic abuse can be suppressed due to inhibition of growth of pathogenic bacteria.

In the prior art related to the present invention, Korean Patent Registration No. 10-0536108 (acid-resistant leuconostomercenteroid isolated from kimchi and a method for producing a delicious kimchi using the same) was optimally aged at a low temperature and separated from the best-tasted kimchi Acidic Ryukonosu stock mesenteroide strains, and a method for producing kimchi which is improved in taste and functionality by adding the above strains and GABA having a blood pressure lowering and strain relieving function.

Korean Patent Registration No. 10-1485182 (New Lactobacillus plantarium isolated from kimchi which inhibits the growth of pathogenic bacteria) and its use has been shown to inhibit the growth of pathogenic bacterium and to be effective against Lactobacillus which is excellent in acid resistance, Pantalium DSR 330 and uses thereof. In particular, the Lactobacillus plantarum DSR 330 or the culture thereof according to the present invention can be usefully used for a dressing, a probiotic agent, a feed, a food additive, an antibacterial agent, a pharmaceutical composition and a fermented food.

Korean Patent Registration No. 10-1329644 discloses a method for producing lactic acid bacteria, which comprises fermenting lactic acid bacteria starter into a mixture of vegetable and water; And fermenting the fermented broth and fermenting the fermented broth to produce vegetables, and mixing the fermented broth with a pickling solution containing sugar and vinegar. The present invention relates to a method for preparing pickled vegetables, While giving a cool acidity and a soft acidity, but also excellent storage. Korean Patent Registration No. 10-1370942 (new Bacillus subtilis) discloses a new Bacillus subtilis strain CJMPB957 (KCCM11271P), a probiotic preparation containing the same, and a method for preventing or treating diseases of livestock will be.

Korean Patent Registration No. 10-1394348 (Lactobacillus plantarum DSR920 and its use for the treatment and prevention of metabolic diseases or inflammatory diseases) inhibits adipocyte hypertrophy by inhibiting the differentiation of adipocytes, while inflammatory cytokines The present invention relates to a novel Lactobacillus plantarum DSR920 which can be used for the treatment and prevention of obesity and ultimately for the prevention and treatment of metabolic syndrome diseases by improving the insulin resistance caused by chronic inflammatory reaction of adipose tissue, A food composition, and a feed composition. Korean Patent Registration No. 10-1369219 (Antibacterial Lactobacillus cells and Method for Producing the Antibacterial Lactic Acid Bacterium and Method for Producing the Antibacterial Lactic Acid Bacterium) Dried, or freeze-dried, followed by mixing the dried microbial cells with the fermentation filtrate. The present invention also relates to a method for producing the microbial cells. However, these prior arts have different technical constructions from those of the present invention.

The present invention relates to a method for isolating lactic acid bacteria having antifungal activity against yeast or fungi, which are food harmful fungi, in addition to antimicrobial activity against bacteria. It is also intended to extend the shelf - life by increasing the shelf - life by applying the strain and culture to food as a natural preservative. The present invention also provides a pharmaceutical composition, a food or a functional food composition which can be used for health promotion.

The present inventors first disclose the separation and identification of Kimchi lactic acid bacteria, which are resistant to pathogenic bacteria and fungal bacteria and have excellent acid resistance, salt resistance, bile resistance and antibiotic resistance, while isolating the antibacterial and antifungal lactic acid bacteria from kimchi. The strain of the present invention comprises a step of isolating a lactic acid bacterium from kimchi, a step of isolating a lactic acid bacterium that inhibits the growth of harmful microorganisms from the separated lactic acid bacteria, a step of testing the antibacterial and antifungal activities of the separated lactic acid bacteria, And separating and identifying the selected lactic acid bacteria. In order to confirm the mycological characteristics of the strain of the present invention, it includes a test for resistance to bubbles, an acid resistance test and an antibiotic test.

The lactic acid bacterium of the present invention can be used as a formulation, a probiotic, an antibacterial, a feed, a food additive, or a pharmaceutical composition for promoting the activity of human, livestock and fish.

An effective ingredient, excipient or carrier such as a culture of the Lactobacillus plantarum DSR KF15 strain of the present invention, a cell wall fraction, a cell wall fraction, a live cell, a dead cell, a filtrate from which a filtration or a centrifuged dry strain is removed, have. The content of the lactic acid bacteria in the composition may vary depending on the use of the composition and the formulation.

The pharmaceutical composition as described above may be used for the treatment of diseases such as Staphylococcus aureus, Escherichia coli, Bacillus cereus, Salmonella typhimurium, Salmonella cholera suis, Y. enterocolitica and Listeria monocytogenes, In order to prevent diseases and infectious diseases caused by L. monocytogenes or the like, an excipient or carrier may be added to the lactic acid bacteria and its active ingredient to form a composition. In addition, pharmaceutical compositions can be used for purulent diseases such as food poisoning, skin inflammation, otitis media, cystitis and the like.

The pharmaceutical composition, the composition for a dressing or a live bacteria, and the composition for an antimicrobial of the present invention can be mixed with a carrier and a flavoring commonly used in the art to form tablets, troche, capsules, elixir, syrup, Suspension, granule or the like. The carrier may be orally or parenterally administered using a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersant, a stabilizer, a suspending agent and the like.

The composition for feed can be produced by fermenting the lactic acid bacteria Lactobacillus plantarum DSR KF15 strain of the present invention and culturing the cells and the culture in the form of a pellet or a sill leaf as a fermented feed or a compound feed. Also, the Lactobacillus plantarum DSR KF15 strain and culture of the present invention can be used as a food additive for kimchi, beverage, baby food, dairy product, and starter for fermented food. In particular, the lactic acid bacteria Lactobacillus plantarum DSR KF15 strain of the present invention and the cultured product are mixed with kimchi seasonings such as red pepper powder, garlic, ginger, green onion, sugar and the like in the pickled cabbage, and then 0.1 to 5 wt% Kimchi can be made by adding Kimchi to Kimchi.

The culture medium of the Lactobacillus plantarum DSR KF15 strain of the present invention is a medium composed of a carbon source, a nitrogen source, a vitamin and a mineral. The culture can be carried out under usual microorganism culture conditions, It can be cultured for about an hour. The culture medium in the culture broth can be removed and centrifugation or filtration can be carried out to recover only the concentrated cells, and these steps can be carried out as required by those skilled in the art. The concentrated cells may be frozen or lyophilized according to a conventional method so as not to lose their activity.

The Lactobacillus plantarum DSR KF15 of the present invention is a lactic acid bacterium isolated from kimchi and exhibits an antimicrobial activity against fungi such as pathogenic microorganisms, fungi and yeast which can be grown on kimchi, and can produce a kimchi taste of a unique flavor by fermentation have. In addition, Lactobacillus plantarum DSR KF15 strain, culture medium and fungi substances are effective in the treatment and prevention of infectious diseases caused by biological preservatives and pathogenic bacteria by adding them to kimchi starters, foods, feeds and cosmetics

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a photograph of the antibacterial activity against Candida albicans of the present invention. FIG.

The strain of the present invention comprises a step of isolating a lactic acid bacterium from kimchi, a step of isolating a lactic acid bacterium that inhibits the growth of harmful microorganisms from the separated lactic acid bacteria, a step of testing the antibacterial and antifungal activities of the separated lactic acid bacteria, And separating and identifying the selected lactic acid bacteria. In order to confirm the mycological characteristics of the strain of the present invention, it includes a test for resistance to bubbles, an acid resistance test and an antibiotic test. Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

< Example  1>; Isolation of lactic acid bacteria from kimchi

20 kinds of kimchi prepared by a conventional kimchi preparation method were selected and the kimchi was used as a lactic acid bacteria-isolated kimchi sample. The kimchi samples were collected, diluted 10-fold with 0.85% saline, inoculated in 0.1 ml each MRS agar medium, and then plated on glass plates. Plates were then incubated for 1 day in a 37 &lt; 0 &gt; C incubator. Each of the resulting colonies was inoculated on MRS agar plates and cultured at 37 ° C for 1 day to isolate lactic acid bacteria colonies.

< Example 2 >; Isolation of Lactic Acid Bacteria that Prevent the Growth of Harmful Microorganisms

Among the hundreds of lactic acid bacteria isolated in Example 1, colonies were inoculated on MRS liquid medium, cultured at 37 ° C for 24 hours, and centrifuged to obtain a culture solution of lactic acid bacteria in order to select lactic acid bacteria inhibiting the growth of pathogenic bacteria.

As a harmful microorganism, yeast Candida albicans ( Candida albicans KCTC 17485), Fungus Fusarium oxysproum Three strains of KCTC11554, three pathogenic microorganisms, Bacillus cereus KCCM12142, Staphylococcus auerus KCCM 11335 and E. coli KCCM11234 were subcultured to the nutrient broth, The juveniles were inoculated 1% in the vagina, hardened on the plate, and dried for 20 minutes. In the YPD liquid medium, the third strain of transformed yeast was inoculated 1% in YPD agar medium, hardened on the plate, and dried for 20 minutes. The spores extracted above were inoculated into the PDA medium by 1%, hardened on the plate, and dried for 20 minutes.

Thereafter, a hole having a diameter of 9 mm was drilled in a cockbearer, and 100 μl of each of the lactic acid bacteria culture supernatants separated as described above was added dropwise. The diameter of inhibitory rings produced after incubation at 37 ℃ for 24 hours, yeast for 25-30 ℃ for 48 hours, fungus 25 ~ 30 ℃ for 48 hours or more were measured. The strain L1 (Lactobacillus plantarium DSR KF15) was selected which selected the most excellent strains among the lactic acid bacteria forming the inhibitory ring and ultimately suppressed the growth of the harmful microorganisms. The strains inhibiting the growth of selected harmful microorganisms were as shown in Table 1 below.

Indication strain Isolated lactic acid bacteria L1 L2 Candida albicans KCTC 17485 (yeast) 11 - Fusarium oxysproum KCTC11554 (mold) 12 - Bacillus cereus KCCM12142 (bacteria) 40 30 Staphylococcus auerus KCCM11335 (bacteria) 35 20 E. coli KCCM11234 (bacteria) 39 15

As a result, lactic acid bacterium L1 showed antimicrobial activity against yeast and mold as well as pathogenic harmful microorganisms, unlike other lactic acid bacteria.

< Example 3 >; Antimicrobial, Antifungal activity  Confirmation experiment

Yeast (YPD), fungus (PDA), and bacteria (NA) were used as the strain culture medium to confirm the antibacterial and antifungal activity of the selected lactic acid bacterium L1 and to compare the antibacterial activity with other lactic acid bacteria. Yeast, fungi and bacteria used as the test strains were as shown in Table 2 below.

division Strain name
mold Fusarium oxysproum KCTC11554 Fusarium oxysproum KCTC16909 Aspergillus flavus KCTC16682 Aspergillus niger KCTC16683 leaven Candida albicans KCTC 17485 Pichia membranifaciens

Pathogenic microorganism Bacillus cereus KCCM11774 Staphylococcus auerus KCCM11335 E. coli KCCM11234 Listeria monocytogenes KCCM40307 Salmonella typhimurium KCTC2057 Spttreococcus mutans KCTC3065

< Pathogenic bacteria  Antibacterial activity Test>

For the test of the antibacterial activity of L1, the cells were subcultured in a MRS liquid medium and Nutrient agar was added with pathogenic bacterium Bacillus cereus KCCM11774), Staphylococcus aureus (Staplyococcus aureus KCCM 11335), L. monocytogenes to Ness (Listeria monocytogenes KCCM40307), E. coli KCCM11234, Salmonella typhimurium KCTC2057) and Streptococcus mutans KCTC3065 were each inoculated at a rate of 1%, and then hardened on a plate, followed by drying for 20 minutes. 100 ㎕ of culture supernatant of L1, which had been punched through a hole having a diameter of 9 mm and then concentrated 5 times, was added thereto, followed by incubation at 37 캜 for 24 hours to measure the size of the inhibitory ring.

< Anti-yeast  Test>

The yeast Candida prepared for the anti-yeast test of L1 albicans KCTC 17485, Pichia The membranifaciens were cultured in the YPD liquid medium in a third subculture, 1% inoculated on YPD agar medium, inoculated on a plate, and dried for 20 minutes. 100 占 퐇 of culture supernatant of LAB1, which had been purged with a 9-mm diameter hole with a cockbearer, was cultured at 25 占 폚 for 48 hours, and the size of the inhibitory circle was measured.

< Antifungal  Test>

For the antifungal test of L1, the fungus Fusarium oxysproum KCTC11554 prepared in the PDA medium, Fusarium oxysproum KCTC16909, Aspergillus flavus KCTC16682, Aspergillus niger KCTC16683, and the spores are prepared as described above. The extracted spores were inoculated 1%, hardened on the plate, and dried for 20 minutes. Then, 100 μl of culture supernatant of L1, which had been punched through a hole with a diameter of 9 mm and concentrated 5 times, was added thereto and cultured at 25 ° C for 48 hours or more to measure the size of the inhibitory ring. The results of the antimicrobial activity test are shown in Table 3 below.

division L1 LF CCM12116 LF KCT1048 Fusarium oxysproum KCTC11554 15 0 0 Fusarium oxysproum KCTC16909 15 0 0 Aspergillus flavus KCTC16682 13 0 0 Aspergillus niger KCTC16683 12 0 0 Candida albicans KCTC 17485 10 0 0 Pichia membranifaciens 10 0 0 Bacillus cereus KCCM11774 40 30 38 Staphylococcus auerus KCCM11335 38 35 35 E. coli KCCM11234 39 33 38 Listeria monocytogenes KCCM40307 35 30 35 Salmonella typhimurium KCTC2057 33 12 35 Streptococcus mutans KCTC3065 35 18 35

The selected lactic acid bacterium L1 showed antibacterial activity against yeast and mold compared with the same kind of lactic acid bacterium, and confirmed that the antimicrobial spectrum was wide.

< Test Example  1>; My bile  Test

In order to confirm the bile resistance of the lactic acid bacteria isolated from Kimchi as described above, L1 isolated from Examples 1 to 3, Lactobacillus plantarium KCCM12116 and Lactobacillus plantaitium KCTC1048 were dissolved in 0.3%, 0.5 %, 1%, 2%, and 3%, respectively. Then, the growth of the strain at the artificial bile concentration was observed while culturing at 37 캜 for 2 hours. The results of the test for biliary properties were as shown in Table 4 below.

Survival rate (%) / Oxgall concentration (%) 0.3 0.5 One 2 3 L 1 100 100 100 100 98 Lactobacillus plantarium KCCM12116 95 85 50 48 40 Lactobacillus platarium KCTC1048 100 90 70 60 55

From the above results, L1 has a survival rate of 100% even at the Oxgall concentration of 0.3-1% and a survival rate of 98% even at the Oxgall concentration of 3%.

< Test Example  2>; Acid resistance  Test

Lactobacillus plantarium KCCM12116 and Lactobacillus plantarium KCTC1048 isolated in Examples 1 to 3 were mixed with pH 2.5, 3.0, 3.5, 4.0, 4.5, and 5 using hydrochloric acid to confirm the acid resistance of the lactic acid bacteria isolated from Kimchi. Lt; RTI ID = 0.0 &gt; ml &lt; / RTI &gt; Then, the viable cells were counted while incubating at 37 DEG C for 2 hours. The acid resistance test results are shown in Table 5 below.

Survival (%) / pH 2.5 3.0 3.5 4.0 L 1 88 93 96 100 Lactobacillus plantarium KCCM12116 56 66 80 95 Lactobacillus platarium KCTC1048 55 75 90 100

From the above results, it was shown that L1 was active at pH 2.5 ~ 5, survived more than 85% even at pH 2.5, and showed survival rate of about 90% or more at pH 3, showing strong acid resistance.

< Test Example  3>; My antibiotics  Test

In order to confirm resistance of the lactic acid bacteria isolated from kimchi to the antibiotics, L1 lactic acid bacteria isolated in Examples 1 and 2 were inoculated into MRS solid medium, and a filter (diameter 6 mm) containing antibiotics was placed on the plate and cultured for 24 hours to obtain antibiotics The size of the inhibitory ring formed by the test was measured. The measurement results are shown in Table 6 below and the unit is mm (millimeters).

Antibiotic L1 LF KCCM12116 Tetracycline (30 占 퐇) 15 21 Erythromycin (15 μl) 16 22 Ampicillin (10 μl) 13 20 Penicillin (10 IU / IE / UI) 18 19 Separocin (30 μl) 20 22

From the above results, L1 is resistant to various antibiotics, so that its activity can be maintained and used effectively in antibiotics, formulations, probiotics, feeds, food additives, antimicrobials, pharmaceutical compositions and fermented foods Could know.

< Experimental Example >; Identification of selected lactic acid bacteria

One) Burgess  Classification Bacteriology Manual

After isolating the lactic acid bacteria selected in Examples 1 to 3 into single colonies, morphological and biochemical characteristics were examined according to Bergy's manual of systematic bacteriology, and Gram staining was performed. The results are shown in Table 7 below.

characteristic Lactobacillus platarium DSR KF15 Gram + Colony form Rod Colony color yellow Motility - Catalase (Catalase) - Fermentation form homo Spore - Growth at 16 ℃ + Growth in 4% NaCl +

It was confirmed that the strain isolated as described above is a Gram-positive strain and has a bacterium form.

2) Analysis using API system

Thereafter, the strain was identified as an API system (La Balme-les-Grottes, France). First, the sterilized platinum microbial colonies were each taken and suspended in 2 ml of sterile distilled water. Again, the bacterial solution was suspended in 5 ml of sterilized distilled water to a concentration of MccFaland Standard Solution No. 2 supplied by API 50CH kit (BioMerieux, France). The supernatant was added to the liquid medium of the API 50CH kit and homogenized, and 200 μl of each of the 50 tubes of the API 50CH kit was inoculated. The tube was overlaid with mineral oil and then cultured at 30 DEG C for 48 hours. The culture solution was analyzed by an API system to identify 49 types of carbohydrate fermentation patterns, and the results were entered into an ATB identification computer system.

As a result, the DSR KF 15 lactic acid bacteria selected in Examples 1 to 3 were found to be strains belonging to the genus of Lactobacillus having the carbohydrate fermentation pattern shown in Table 8 below.

Control group - D-Manno + Salincin + Kentio Vioz + Glycerol - L-Solbos - Cellobiz + D-Turanz - Erythritol - Rannoz + Malts + D-rings - D-Ariibinose - Dissytol - Laktos + D-Tagatoz - L-arabinose + Inositol - Melibiose + D-fucose - Riboze + Mannitol + Saka Rose + L-fucose - D-xylose - Sorbitol + Trehalose + D-arabitol - L-xylose - D-mannoside - Inulin + L-arabitol - Adonitol - D-glucoside - Melrietto + Gluconate - Xyloside - Glucosamine + D-Rapinoz + 2-Setto-gluconate + Galactos + Amigalline + Amidon - 5-Seto-gluconate - D-glucose + Arbutin + Glycogen - - -

Lactic acid bacteria were found to be strains belonging to the genus Lactobacillus having the above carbohydrate fermentation pattern.

3) 16S rDNA  Sequencing

The base sequence of 16S rDNA of the selected lactic acid bacteria was analyzed. As a result, the present inventors named L1 lactic acid bacteria as "Lactobacillus plantarum DSR KF15 ", and deposited it in Korean Culture Center of Microorganisms as Deposit No. KCCM11772P on September 15, 2015. The results of this nucleotide sequence analysis are as shown in the following identification item [Sequence Listing Free Text].

< Application example >; Manufacture of kimchi using lactic acid bacteria

3 parts by weight of red pepper powder, 2.4 parts by weight of garlic, 0.6 parts by weight of ginger, 2.4 parts by weight of green tea, 10.9 parts by weight of iris and 0.6 part by weight of sugar were mixed with 100 parts by weight of Chinese cabbage having salinities of 2.1-2.3% Then, 1% by weight of the lactic acid bacterium of the present invention was added as a starter to the total weight of the kimchi, and no starter was added to the control group. Kimchi was added to pickled Chinese cabbage, and kimchi was immersed in the kimchi. Then, it was aged at 5 ℃ for 1, 2, and 4 weeks. The acidity of kimchi fermented for 2 weeks was 0.6%, and the addition was 0.7% (control). The acidity was lower than that of the control kimchi, so the acidity was gradually increased at the latter half of kimchi fermentation to improve the quality of kimchi Can be achieved. The acidity of Kimchi and the number of lactic acid bacteria were measured and the results are shown in Table 9 below.

division Invention Control group Acidity (%) 0.6 0.7 Number of lactic acid bacteria (log cfu / ml) 9.55 8.51

The lactic acid bacteria of the present invention were found to be 9.55 (log cfu / ml) and no addition (control group) of 8.51 (log cfu / ml) as the starter of the present invention, It can be seen that it is excellent as a starter.

< Test Example >; Sensory test

Five points scale method (5 points: excellent, 4 points: excellent, 3 points: excellent, 3 points: good, good, good) were evaluated for the texture, aging, overall taste, smell, ; Average, 2 points or less; insufficient), and is shown in Table 10 below. The panel was selected for students (20s, 30s, 40s, and 8 boys and girls) who were proficient in food, college students, workers and housewives. The results of the sensory test are shown in Table 10 below.

From the above results, the longer the ripening period, the lower the overall quality. However, it can be seen that the kimchi using the lactic acid bacteria of the present invention as a starter has a delayed acidity rate and a prolonged good taste period due to the proliferation of lactic acid bacteria, as compared with the control group.

< Manufacturing example  1> Feed composition

2.0 parts by weight of Lactobacillus plantarum DSR KF15 lactic acid bacterium of the present invention, 1.0 part by weight of glucose, 1.0 part by weight of peptone, 0.2 part by weight of yeast extract, 0.05 part by weight of phosphorus acid, 0.05 part by weight of magnesium sulfate, And incubated aerobically for 2 days at 37 ° C. The culture medium of each strain and the excipient degreasing solution were simply mixed at a weight ratio of 1: 1, followed by drying at 40 ° C or lower, and the dried mixture was pulverized to prepare a feed composition. At this time, the prepared feed composition contained the lactic acid bacteria of the present invention at a concentration of 1 × 10 ⁹ cfu / g or more.

< Manufacturing example  2> Probiotic agent composition

2.0 parts by weight of Lactobacillus plantarum DSR KF15 lactic acid bacterium of the present invention, 1.0 part by weight of glucose, 1.0 part by weight of peptone, 0.2 part by weight of yeast extract, 0.05 part by weight of phosphoric acid, 0.05 part by weight of magnesium sulfate, And incubated aerobically for 2 days at &lt; RTI ID = 0.0 &gt; 37 C. &lt; / RTI &gt; The culture broth of each of the above strains was inoculated at 10% by weight in a solid culture medium in which a defatted steel, soybean meal, corn and molasses were mixed at a weight ratio of 3: 1: 1: 1, Lt; / RTI &gt; The fermentation broth composition was prepared by anaerobically fermenting the fermentation broth with stirring at 40 ° C for 3 days. The broth composition prepared herein contained 1 × 10 ⁹ cfu / g or more of the lactic acid bacteria of the present invention.

< Manufacturing example  3> Composition for food additive

2.0 parts by weight of Lactobacillus plantarum DSR KF15 lactic acid bacteria of the present invention, 1.0 part by weight of glucose, 1.0 part by weight of peptone, 0.2 part by weight of yeast extract, 0.05 part by weight of phosphorus acid and 0.05 part by weight of magnesium sulfate, And cultured aerobically for 2 days at 37 ° C. 60% by weight of the culture solution was added to a mixture of milk powder or skimmed milk and starch or lactose at a weight ratio of 1: 1, and the resulting mixture was rapidly frozen at -70 캜 or below, followed by lyophilization at -60 캜 for 36 hours, A composition for additives was prepared. At this time, the temperature of the hot plate for increasing the efficiency of moisture evaporation during lyophilization was set to 30 ° C, and the prepared food additive composition contained 1 × 10 ⁹ cfu / g or more of the lactic acid bacteria of the present invention.

< Manufacturing example  4> Pharmaceutical composition

Each of the bacterial composition powders obtained in Preparation Examples 1, 2 and 3 was molded into tablets, pills and capsules according to a conventional method to prepare a pharmaceutical composition. The composition prepared herein had a composition of 1 × 10 ⁹ cfu / g or more.

The strains of the present invention and the cultures thereof can be applied to foods as natural beverages to provide an effect of extending shelf life. Also, since it is provided as a pharmaceutical composition and a food composition such as a dressing, a probiotic agent, a feed, and an antibacterial, it is industrially applicable.

Korea Microorganism Conservation Center (overseas) KCCM11772P 20150918

<110> Daesang F & F <120> Vegetable Lactobacillus plantarum DSR KF15 having Activities on          Antimicrobial and Antifungal for keeping freshness and Use          Thereof <130> 10273 <160> 1 <170> KoPatentin 3.0 <210> 1 <211> 1502 <212> DNA <213> Lactobacillus plantarum <400> 1 gtggcggcgt gctatacatg cagtcgacga actctggtat tgattggtgc ttgcatcatg 60 atttacattt gagtgagtgg cgaactggtg agtaacacgt gggaaacctg cccagaagcg 120 ggggataaca cctggaaaca gatgctaata ccgcataaca acttggaccg catggtccga 180 gtttgaaaga cctggaaaca gatgctaata ccgcataaca acttggaccg catggtccga 240 tggggtaacg gctcaccatg gcaatgatac gtagccgacc tgagagggta atcggccaca 300 ttgggactga gacacggccc aaactcctac gggaggcagc agtagggaat cttccacaat 360 ggacgaaagt ctgatggagc aacgccgcgt gagtgaagaa gggtttcggc tcgtaaaact 420 ctgttgttaa agaagaacat atctgagagt aactgttcag gtattgacgg tatttaacca 480 gaaagccacg gctaactacg tgccagcagc cgcggtaata cgtaggtggc aagcgttgtc 540 cggatttatt gggcgtaaag cgagcgcagg cggtttttta agtctgatgt gaaagccttc 600 ggctcaaccg aagaagtgca tcggaaactg ggaaacttga gtgcagaaga ggacagtgga 660 actccatgtg tagcggtgaa atgcgtagat atatggaaga acaccagtgg cgaaggcggc 720 tgtctggtct gtaactgacg ctgaggctcg aaagtatggg tagcaaacag gattagatac 780 cctggtagtc cataccgtaa acgatgaatg ctaagtgttg gagggtttcc gcccttcagt 840 gctgcagcta acgcattaag cattccgcct ggggagtacg gccgcaaggc tgaaactcaa 900 aggaattgac gggggcccgc acaagcggtg gagcatgtgg tttaattcga agctacgcga 960 agaaccttac caggtcttga catactatgc aaatctaaga gattagacgt tcccttcggg 1020 gacatggata caggtggtgc atggttgtcg tcagctcgtg tcgtgagatg ttgggttaag 1080 tcccgcaacg agcgcaaccc ttattatcag ttgccagcat taagttgggc actctggtga 1140 gactgccggt gacaaaccgg gaaaaggtgg ggatgacgtc aaatcatcat gccccttatg 1200 acctgggcta cacacgtgct acaatggatg gtacaacgag ttgcgaactc gcgagagtaa 1260 gctaatctct taaagccatt ctcagttcgg attgtaggct gcaactcgcc tacatgaagt 1320 cggaatcgct agtaatcgcg gatcagcatg ccgcggtgaa tacgttcccg ggccttgtac 1380 acaccgcccg tcacaccatg agagtttgta acacccaaag tcggtggggt aaccttttag 1440 gaaccagccg cctaaggtgg gacacatgat tagggtgaag tcgtaacaag gttaacccgt 1500 aa 1502

Claims (8)

As a lactic acid bacteria Lactobacillus plantarum DSR KF15 (KCCM11772P) strain having antibacterial and antifungal activity for fresh keeping,
Wherein the fungus is selected from the group consisting of Fusarium oxysporum , Aspergillus niger Wherein the strain is one or more selected from the group consisting of Aspergillus niger and Pichia membranifaciens . delete The method of claim 1, wherein the strain is selected from the group consisting of vegetable lactic acid bacteria Lactobacillus plantarum DSR KF, which inhibits the growth of antibacterial, antifungal and pathogenic bacteria and has acid resistance at pH 2.5 to 4, biliary and antibiotic resistance at 0.3 to 3% 15 (KCCM11772P). A composition for antifungal and antifungal use characterized by containing the plant Lactobacillus plantarum Lactobacillus plantarum DSR KF15 (KCCM11772P) strain or the culture thereof according to claim 1 to inhibit the growth of harmful microorganisms. delete A food composition comprising the plant lactic acid bacterium Lactobacillus plantarum DSR KF15 (KCCM11772P) strain of claim 1 or a culture thereof. A prophylactic composition comprising the plant Lactobacillus plantarum Lactobacillus plantarum DSR KF15 (KCCM11772P) strain of claim 1 or a culture thereof. 11. A dressing composition comprising the plant lactobacillus Lactobacillus plantarum DSR KF15 (KCCM11772P) strain of claim 1 or a culture thereof.

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