KR20150125836A - Lactobacillus plantarum js1 and synbiotics functional makgeolli prepared therewith - Google Patents

Abstract

The present invention relates to a novel Lactobacillus plantarum strain used in producing makgeolli, makgeolli produced by using the same, and a method for producing makgeolli by using the same. A novel Lactobacillus plantarum JS1 strain according to the present invention is an appropriate strain that is suitable in improving synbiotics functions of makgeolli, and has excellent probiotics properties compared to conventional makgeolli. Makgeolli produced by using the same has excellent functional flavors. Furthermore, a method for producing makgeolli using a novel lactobacillus plantarum JS1 strain according to the present invention has improved synbiotics functions, compared to conventional makgeolli, so added value in the industry of makgeolli can be improved.

Description

Lactobacillus plantarum JS1 and SYNBIOTICS FUNCTIONAL MAKGEOLLI PREPARED THEREWITH This invention relates to Lactobacillus plantarum JS1,

The present invention relates to a novel lactic acid bacterial strain used in the production of makgeolli, makgeolli prepared using the same, and a method for producing functional makgeolli using the same.

 Makgeolli is a traditional wine made from cereals and yeast, and is made by bottle fermentation. It is filtered out after fermentation and its appearance is opaque, and the sweetness, sour, sincerity, gummy and sour taste are harmoniously balanced with alcohol content of 6% ~ 8% to be. In particular, it is known that Makkolli has higher health functionalities than suspended or filtered sake after fermentation because the suspension of the fermented product, ie, the fermented product as a raw material, is contained in the final product in an unfiltered state. The suspended solids of rice wine include starch polysaccharides, oligosaccharides, organic acids, peptides and various useful physiologically active substances derived from raw materials and fermented products including yeast and lactic acid bacteria (lactic acid bacteria), which are microbial cells. Specifically, Makgeolli is a natural food fermented naturally by pure microorganisms. As a health food, it is a healthy food. It contains more than 10 kinds of essential amino acids which are involved in metabolism in the human body, It is known to be good for skin cosmetics because it contains all the organic acid which stops the thirst by making a fresh taste, acetylcholine, protein which helps the liver function, vitamin B, inositol, choline etc. .

Probiotics Using Lactic Acid Bacteria Functional improvement has already been proven in many documents and studies, and the field of foods utilizing them has been increasing. Recently, Synbiotics, which combines probiotics and prebiotics in the intestines, is attracting attention. Accordingly, various forms of foods such as fermented milk, cheese, health drinks, sauces, and health functional foods have been manufactured, and the market for synbiotics has been greatly growing. Lactic acid bacteria is a representative synbiotics strain that functions as a probiotic function in itself, and simultaneously functions as a synbiotics by restricting intestinal microorganisms, which are the roles of prebiotics in the large intestine, and improving the growth of useful microorganisms. Representative foods that can take such lactic acid bacteria as food are various such as soy sauce, salted fish, kimchi, and fermented milk. Taking advantage of the fact that it is possible to use the improved probiotics already known to be beneficial as synbiotics food, dairy foods that contain many probiotics are greatly growing the new biotics market. In recent years, the market for synbiotics has been expanding its range of beverages and health functional foods, and makgeolli, which contains lactic acid bacteria and a suspension material derived from raw materials at the same time, is attracting attention as a product capable of exhibiting a strong new biotics function.

In this regard, Korea Patent Laid-Open Publication No. 2006-0078894 discloses a novel Lactobacillus K yarn with the alcohol, the acid resistance and low-temperature characteristics sex (Lactobacillus sakei ) and a process for producing the fermented soybean using the fermented soybean , and more particularly to a starter for fermentation of a fruit wine.

It is an object of the present invention to select optimal lactic acid bacteria for improving the synbiotic function of makkolli, and further to develop an optimum fermentation process using the same.

The inventors of the present invention have conducted intensive research to achieve the above object. As a result, the inventors of the present invention have found that the lactobacillus plantarum strain has excellent fermentation characteristics for producing makgeolli, and that the makgeolli, And thus the present invention has been completed.

The novel lactobacillus Lactobacillus strain Lactobacillus strain JS1 of the present invention is a strain suitable for enhancing the functionality of the new biotics of makgeolli. The strain has superior probiotic characteristics as compared with the conventional makgeolli, and the makgeolli prepared using the same has excellent palatability Do. In addition, it is expected that the method of producing makgeolli using the novel Lactobacillus plantarum JS1 strain of the present invention can further increase the value of the makgeolli industry by further improving the synbiotics function as compared with the conventional makgeolli.

1 is a flowchart illustrating a method of manufacturing a synbiotic makgeolli according to an embodiment of the present invention.
Fig. 2 shows the change in the amount of the bacterial cells according to the immersion setting by inoculating Lactobacillus planta JS1.
Figure 3 shows the growth of yeast during fermentation of rice wine with Lactobacillus plantarum JS1 inoculated with yeast.
Figure 4 compares the reference strain of the same species as the Lactobacillus flutarium of the present invention with the 16S rRNA sequence (using 518F, 800R primers).

Hereinafter, the present invention will be described in detail. However, it should be understood that the present invention may be embodied in many other specific forms without departing from the spirit or essential characteristics thereof.

In one aspect, the present invention relates to a novel strain of lactobacillus Lactobacillus plantarum JS1 (Accession No. KFCC11566P).

The inventors of the present invention have found that the lactic acid bacteria isolated from various rice wine are resistant to pH tolerance and bile salt resistance for the production of synbiotics makgeolli and have excellent sensory tastes of makgeolli prepared by using the same to improve the synbiotic function of makgeolli The selected lactic acid bacteria were identified and identified as Lactobacillus plantarum. The Lactobacillus plantarum strain according to the present invention is a wild lactic acid bacterium isolated from a makgeolli, and has excellent fermentation characteristics for producing makgeolli, and the makgeolli prepared using the lactic acid has excellent properties. As a result of the analysis on the genetic characteristics, it was confirmed that it is a new strain different from the known strain of the same species and the industrialized strain. This strain was named as JS1 strain and deposited on November 5th, 2013 with the deposit number KFCC11566P at the Korea Microorganism Conservation Center.

There are many prebiotics available for probiotics in makgeolli, one of which is beta-glucan, a cell wall component of yeast. In this connection, the fact that the lactic acid bacteria selected in the present invention do not inhibit the growth of yeast means that normal rice wine fermentation is induced and yeast is not reduced as a prebiotic.

In addition, lactic acid is added to control the pH of the yeast before fermentation to produce a typical rice wine. In the present invention, lactic acid is produced by directly inoculating a live strain producing lactic acid instead of adding lactic acid. And the inoculum itself acts as a probiotic, which is one of the characteristics of the present invention.

In another aspect, the present invention relates to a rice wine having improved synbiotics functionality prepared using the Lactobacillus plantarum JS1.

In another aspect, the present invention relates to a method for producing makgeolli having improved synbiotics functionality using the Lactobacillus plantarum JS1.

A typical method for producing makgeolli is a method comprising the steps of: an immigration step of mixing and culturing white germ with the added cereal grains; a first fermentation step of fermenting the yeast culture solution and water by mixing with the yeast culture liquid; a step of mixing rice and water added to the first fermentation broth A secondary fermentation step for fermentation by filtration, a filtration step and the like.

On the other hand, the method of the present invention for producing a macheled rice comprises the steps of: a) mixing and cultivating white germ with baked cereal to prepare an inlet; b) mixing the yeast culture broth, Lactobacillus planta JS1 culture broth, and water to enter the broth and fermenting to prepare a first fermentation broth; And c) fermenting the primary fermentation broth by mixing starch-containing main raw material and water to prepare a secondary fermentation broth. And d) a filtration step after step c).

That is, it is a key technical feature of the present invention that Lactobacillus plantarum JS1 strain selected in the present invention is applied in the first fermentation step.

FIG. 1 is a flowchart illustrating a method of manufacturing a synbiotic makgeolli according to an embodiment of the present invention. Referring to FIG. 1, an embodiment of a method of manufacturing a makgeolli according to the present invention will be described step by step.

In order to prepare rice wine using the Lactobacillus plantarum JS1 strain according to the present invention, the entry is first made. Entry can be made by conventional methods by inoculating white rice bran into cereals such as rice or barley. For example, inoculation can be made by inoculating white rice germ in a rice grate and propagating white rice germs in a thermo-hygrostat for 48 hours while maintaining the temperature at 33 ° C and a humidity of 95%.

An aspherical microorganism, preferably Aspergillus luchuensis , is used as the white germ, but is not limited thereto.

The yeast strain of the first fermentation stage can be cultured in a large amount by a conventional culture method of Saccharomyces sp. As the culture medium, a medium composed of carbon source, nitrogen source, vitamins and minerals can be used. For example, a yeast malt extract agar medium, a potato dextrose agar (PDA) medium, or an immobilized glycoside can be used as a medium. The immigration saccharified liquid was added to 300 parts by weight of water with respect to 100 parts by weight of the immigration using the inlet prepared in the above step, stirred and saccharified at 60 to 65 ° C for 3 hours to 5 hours, and then stirred at 4,500 rpm to 6,000 rpm for 20 minutes Followed by centrifugation for 30 minutes, followed by filtration, sterilization, and dilution to a degree of 10 ° Brix to 14 ° Brix. The culture can be carried out using the culture medium under aerobic conditions at a temperature of 25 ° C to 30 ° C for 36 hours to 72 hours and then stored at 4 ° C.

Water is added to the impregnation prepared in the above step and mixed, and the temperature is adjusted to 25 ° C. It is preferable that 120 parts by weight to 140 parts by weight of water is added to 100 parts by weight of the incoming water.

In one embodiment, the inoculation amount of the yeast in step b) may be an inoculum amount used in the production of general rice wine. It was confirmed through the preliminary experiment of the present invention that makgeolli was prepared at the same level as the yeast inoculation amount of Lactobacillus plastataum JS1, and it was confirmed that the makgeolli having a very strong acidity was produced. Through repeated experiments, Lactobacillus plutarium JS1 It was confirmed that the range of 15% to 20% is preferable in terms of the symbol.

That is, the Lactobacillus plantarum JS1 culture medium is preferably 15% to 20% by weight based on the yeast culture. The Lactobacillus plantarum JS1 culture medium is most preferably 20% by weight based on the yeast culture medium.

In a preferred embodiment, 0.06% of the yeast is inoculated with respect to the rice weight, and the lactobacillus is cultured in the MRS liquid medium and the recovered pellet is recovered by 0.012% Followed by fermentation at 25 ° C for 2 days to 3 days.

The starch-containing raw material may be at least one selected from the group consisting of rice, barley, corn, sorghum, wheat, potatoes and sweet potatoes, but is not limited thereto. The starch-containing main raw material is most preferably rice which has not been added. By using the raw material without additives, it is possible to achieve an effect of reducing the manufacturing cost as well as an excellent synbiotics effect.

The primary fermentation broth prepared in the above step and the starch-containing raw material are mixed and fermented to prepare a soup base.

In a preferred embodiment, the primary fermentation broth prepared in the above step may be mixed with starch-containing main raw material and water, and then the secondary fermentation may be carried out at 25 ° C for 5 days to 6 days.

That is, in a preferred embodiment, the primary fermentation of step b) is carried out at 25 ° C for 2 to 3 days, and the secondary fermentation of c) is carried out at 25 ° C for 5 to 6 days.

In the filtration step, the obtained fermentation broth can be filtered with a 120 mesh sieve.

The present invention will be described in more detail by way of the following examples. However, the following examples are for the purpose of illustrating the present invention and are not intended to limit the scope of the present invention.

[Example]

1st round selection

In this study, 10 kinds of Makkolli were purchased and the wild lactic acid bacteria were isolated and identified. As a result, Lactobacillus casei (Lactobacillus casei , Lactobacillus paracasei , Lactobacillus < RTI ID = 0.0 > plantarum), Lactobacillus brevis (Lactobacillus brevis and Pediococcus acidilactici were isolated from the strains.

Makgeolli manufacturing

The makgeolli was prepared for the makgeolli inoculated with 5 isolates.

For the manufacture of the immigration, the rice was immersed for 3 hours, drained and then added. In 10 kg of the expanded rice, 2 g of Aspergillus lucens spore formation was inoculated and cultured for 48 hours at 33 캜 and 95% humidity.

For the primary fermentation, 513 g of the above-prepared entrant and 718 g of water were mixed, and then 0.06% by weight of yeast was inoculated with rice, and lactic acid bacteria were added at a rate of 0.012% by weight of rice. The mixed solution was fermented at 25 캜 for 2 days to prepare a primary fermentation broth.

For the second fermentation, rice without added rice was prepared. In order to remove foreign matter or contamination of rice, it was thoroughly washed with water and immersed in water at room temperature for 3 hours for sufficient absorption of water and then ground. 1,743 g of water and 1,026 g of ground rice were added to the primary fermentation broth and fermented at 25 DEG C for 5 days to produce synbiotics makgeolli. The fermented makgeolli was filtered with a 120 mesh sieve.

Fermentation characteristics

The fermentation characteristics for the makgeolli were analyzed and the results are shown in Table 1 below.

During fermentation, lactic acid bacteria were inoculated with Lactobacillus paracasei and Pediococcus aceyltactii at a pH value of 4.0 or higher. However, Lactobacillus brevis, Lactobacillus plantarum and Lactobacillus casei were inoculated One rice wine showed a low pH value of 3.5.

On the other hand, total acidity was significantly higher in the order of Lactobacillus brevis, Lactobacillus casei and Lactobacillus plantarum in the order of lactic acid bacteria, lactic acid bacteria and lactobacillus paracase.

Although there was some difference in each lactic acid bacterial inoculation group, it was confirmed that the rice wine inoculated with 5 kinds of lactic acid bacteria isolated showed normal pH, total acidity and alcohol production rate during the fermentation process and showed a fermentation range of normal fermentation range Respectively.

Sensory evaluation

The likelihood of the makgeolli prepared with the above 5 strains was evaluated and shown in Table 2 below.

(9 points: very good, 7 points: good, 5 points: normal, 3 points: not good, 1 point: very poor). The evaluators were repeatedly evaluated by 9 lecturers on two occasions, and a brief description of the evaluation was given. The results of the evaluation showed that the taste of makgeolli made with Lactobacillus casei and Lactobacillus plantarum was the highest.

Therefore, Lactobacillus casei and Lactobacillus plantarum were firstly selected by comparing the lactic acid bacteria characteristics and sensory evaluation results for the production of synbiotics makgeolli.

As a result of the analysis on the genetic characteristics, it was confirmed that the two strains selected first were different from the known strains of the same species and the strains already used industrially. Lactobacillus planta was named JS1 strain (see Fig. 4 and the sequence listing), and Lactobacillus casei was named JS13 strain.

Second selection

The acidity and bile acid tolerance of the first selected lactobacillus Lactobacillus plantarum JS1 and Lactobacillus casei JS13 were investigated. KCTC3104 (Lactobacillus plantarum) and KCTC3545 (Lactobacillus helveticus), which were distributed by KCTC (Korea Collection for Type Culture), were investigated in order to compare the resistance of selected lactic acid bacteria. Table 3 shows the results of an experiment on the pH tolerance of lactic acid bacteria, and Table 4 shows the results of experiments on the bile acid resistance of lactic acid bacteria.

Based on the above results, Lactobacillus plastarium JS1, a strain with low pH and strong resistance to bile, was finally selected.

Of the selected strain Synbiotics  Enhancements

Fig. 2 shows the results of observing changes in the amount of Lactobacillus planta JS1 cells (live cells) according to the immersion setting by inoculating the finally selected Lactobacillus planta Room JS1.

In case of makgeolli (Nuruk / steamed rice in Fig. 2), fermented at 23 ℃ and 25 ℃, the amount of bacteria was fermented at 10 ⁶ CFU / mL on the 3rd day of fermentation and 10 ¹⁰ CFU / mL. On the 7th day of fermentation, it decreased to 10 ⁸ CFU / mL. On the other hand, mackerel fermented at 28 ℃ showed relatively small amount of bacterial cells, but gradually increased to 10 ⁸ CFU / mL on the 5th day after fermentation.

Fermented at 23 ° C and 28 ° C, fermented at 10 ⁷ CFU / ml on day 3 of fermentation, and 10 ¹⁰ CFU / whereas it increased to a mL down to the end of fermentation, and decreased when sikyeoteul fermentation at 25 ℃ then increased up to 5 primary fermentation ¹⁰ 10 CFU / mL to about 10 ¹¹ CFU / mL in the primary fermentation 7 10 ⁸ CFU / mL.

In the case of makgeolli fermented with rice added to the market (Kogi / steamed rice in Fig. 2), it was increased from 10 ⁷ CFU / mL to 10 ⁸ CFU / mL at the beginning of fermentation at 23 ° C., When fermented at 25 ℃, it increased rapidly from 10 ⁷ CFU / mL to 10 ⁸ CFU / mL and rapidly decreased to 10 ⁶ CFU / mL at the end of fermentation.

In the case of makgeolli fermented with rice that has not been added to the market (Kogi / non-steamed rice in Fig. 2), it was relatively low as 10 ⁵ CFU / mL to 10 ⁶ CFU / mL at the beginning of fermentation, to ¹⁰ 10 CFU / mL to about 10 ¹¹ CFU / mL it was increased rapidly. Particularly, when fermented at 25 ° C, the amount of the lactobacillus plantarum JS1 cell was maintained at 10 ⁸ CFU / mL to 10 ⁹ CFU / mL until the end of fermentation.

In addition, the growth of yeast during the fermentation process was examined in order to examine whether or not the rice wine inoculated with the yeast in the early lactobacillus plantarum JS1 fermented was fermented.

The fermented rice wine (Nuruk / steamed rice and Kogi / steamed rice in Fig. 3) fermented at 28 ° C using yeast and entrainment as the fermentation agent showed a tendency that the yeast growth tended to decrease during the fermentation process. Was maintained normally.

 Therefore, the fermentation process can be carried out at a temperature of 25 ° C under the condition that the final lactobacillus plantarum JS1 cell mass is high, that is, the fermentation medium is used for entry into the fermentation broth without any inhibition of yeast growth during fermentation, The optimum process for enhancing the function of the new biotics of Makkolli was judged.

Korea Microorganism Conservation Center KFCC11566P 20131105

<110> KOREA FOOD RESEARCH INSTITUTE <120> LACTOBACILLUS PLANTARUM JS1 AND SYNBIOTICS FUNCTIONAL MAKGEOLLI          PREPARED THEREWITH <130> IPDB-52831 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 2096 <212> DNA <213> 16S rRNA of Lactobacillus plantarum JS1 (KFCC11566P) <400> 1 gaaggtttca cgctccatgt gaagtgactc aggagttagt cccatctcgc gtgtgaaacg 60 cgccctatct ctattgggcg cgacagagag atatcttcta catatagaag aatgtttgtg 120 tggaaccccg cccgcagaaa aaagatctcg ctgctgttaa tacatcttct tcaaaaagag 180 atatcttcgc gaaagactcc gcttcgggtt tcacaaaccg caacgtttta gccttacgat 240 cacctatcat cgtcccacta gcgctgtcta aaaggttacc ccaccgactt tgggtgtaca 300 aactctcatg gtgtgacggg cggtgtgtac aagggcccgg gaacgtattc ccacccgcgg 360 catgtgatcc gcgattacta gcgattccga cttcatgtag gcgagttgca gcctacaatc 420 cgaactgaga atggctttaa gagattagct tactctcgcg agttcgcaac tcgttgtacc 480 atccattgta gcacgtgtgt agcccaggtc ataaggggca tgatgatttg acgtcatccc 540 caccttcctc cggtttgtca ccggcagtct caccagagtg cccaacttaa tgctggcaac 600 tgataataag ggttgcgctc gttgcgggac ttaacccaac atctcacgac acgagctgac 660 gacaaccatg caccacctgt atccatgtcc ccgaagggaa cgtctaatct cttagatttg 720 catagtatgt caagacctgg taaggttctt cgcgtagctt cgaattaaac cacatgctcc 780 accgcttgtg cgggcccccg tcaattcctt tgagtttcag ccttgcggcc gtactcccca 840 ggcggaatgc ttaatgcgtt agctgcagca ctgaagggcg gaaaccctcc aacacttagc 900 attcatcgtt tacggtatgg actaccaggg tatctaatcc tgtttgctac ccatactttc 960 gagcctcagc gtcagttaca gaccagacag ccgccttcgc cactggtgtt cttccatata 1020 tctacgcatt tcaccgctac acatggagtt ccactgtcct cttctgcact caagtttccc 1080 agtttccgat gcacttcttc ggttgagccg aaggctttca catcagactt aaaaaaccgc 1140 ctgcgctcgc tttacgccca ataaatccgg acaacgctac ccacctacgt attaccgcgg 1200 ctgctggcac gtagttagcc gtggctttct ggttaaatac cgtcaatacc tgaacagtta 1260 ctctcagata tgttcttctt taacaacaga gttttacgag ccgaaaccct tcttcactca 1320 cgcggcgttg ctccatcaga ctttcgtcca ttgtggaaga ttccctactg ctgcctcccg 1380 taggagtttg ggccgtgtct cagtcccaat gtggccgatt accctctcag gtcggctacg 1440 tatcattgcc atggtaagcc gttaccccac catctagcta atacgccgcg ggaccatcca 1500 aaagtgatag ccgaagccat ctttcaaact cggaccatgc ggtccaagtt gttatgcggt 1560 attagcatct gtttccaggt gttatccccc gcttctgggc aggtttccca cgtgttactc 1620 accagttcgc cactcactca aatgtaaatc atgatgcaag caccaatcaa taccagagtt 1680 cgttcgactt gcatgtatta ggcacgccgc cagcgtcgtc ctgacgtgaa aagccctctc 1740 tctcttatgc ccgccagaaa cggagaagag ccttgtggtt ccttggacta caccacccct 1800 gctgtataca gtaagaagta tataagtcac atcggggagg tacaagcaga atagaagtcg 1860 acgagtaggt gataaggaca caatatggga atgtgtgtca ttaatattta ccgcgcggga 1920 gacgtgttgg gctgtgcttt atggtaaaat acagacaaac ctacactgtc acgcaaaatg 1980 ttcgtgtgag aaatgattca gagcgaactg gtagtacgga cggtcgcaca tacaacttct 2040 atggaaatcg gtaggctggc ctaagactgg ccgtacacga agtgaaatcc tagtcg 2096

Claims (7)

CLAIMS What is claimed is: 1. A pharmaceutical composition comprising Lactobacillus plantarum JS1 plantarum JS1, accession no. KFCC11566P):
a) mixing and culturing white germ with the added cereal to produce an entry;
b) mixing the yeast culture broth, Lactobacillus planta JS1 culture broth, and water to enter the broth and fermenting to prepare a first fermentation broth; And
c) mixing the primary fermentation broth with starch-containing raw material and water and fermenting to produce a second fermentation broth. The method according to claim 1,
Wherein the starch-containing main ingredient is at least one selected from the group consisting of rice, barley, corn, sorghum, wheat, potatoes and sweet potatoes which have not been increased. The method according to claim 1,
wherein the Lactobacillus plantarum JS1 culture medium is 15 wt% to 20 wt% based on the yeast culture in step b). The method according to claim 1,
characterized in that the primary fermentation of step b) is carried out at 25 DEG C for 2 to 3 days, and the secondary fermentation of step c) is carried out at 25 DEG C for 5 to 6 days. A makgeolli produced by the method of any one of claims 1 to 4. Lactobacillus plantarum JS1 ( Lactobacillus plantarum JS1, Accession No. KFCC11566P). Lactobacillus plantarum JS1 ( Lactobacillus plantarum JS1, accession number KFCC11566P).

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