KR101929328B1 - Manufacturing method of berries fermented liquid using lactic acid bacteria and berries fermented liquid manufactured by same method - Google Patents

Abstract

The present invention relates to a process for producing a fermented verrucous liquid using lactic acid bacteria and a fermented liquor fermented by the method. Specifically, the fermented liquid of berries produced according to the method of the present invention does not contain alcohol and thus has excellent quality.

Description

TECHNICAL FIELD The present invention relates to a method for producing a fermented liquor of fermented liquor using lactic acid bacteria and a method for producing fermented liquor fermented by using the fermented liquor fermented liquor,

The present invention relates to a process for producing a fermented verrucous liquid using lactic acid bacteria and a fermented liquor fermented by the method.

Gram-positive bacteria, lactic acid bacteria, are bacteria that break down carbohydrates such as glucose and lactose to produce lactic acid (lactic acid), and have no ability to decay proteins. Up to now, there have been about 300 ~ 400 kinds of lactic acid bacteria, of which 20 are used for fermented milk and fermentation industry. Lactic acid bacteria are widely distributed in the natural world and are a beneficial strain that has a direct or indirect relationship with human life.

The efficacy of lactic acid bacteria varies. Enter the intestinal tract and multiply in intestinal epithelial cells to prevent the colonization of harmful microorganisms and secrete antimicrobial substances to inhibit the growth of harmful microorganisms and to improve diarrhea and constipation as well as to improve immune activity, And so on. In addition, it is known that lactic acid bacteria are directly or indirectly added to foods to improve the storage stability of foods by lactic acid, which is a metabolite thereof, and improve the flavor and texture of foods.

Fermented foods, on the other hand, account for a large part of our diet and occupy a very important position in the nutrition and food culture of our diet. Examples of such fermented foods include soy sauce, soybean paste, kochujang, chonggukjang, kimchi, salted fish, and other marinated products such as fruit wine, takju, and vinegar. Recently, the public interest in health has been increased, and according to the expected effect of functional foods, plant fermentation liquid, which is one of the fermented foods, is attracting attention. The market for fermentation broth expanded rapidly from 6.9 billion won in 2005 to 38 billion won in 2011, and is expected to continue growing. Plant fermented liquids are expected to be diversified in the future due to the fact that the risk factors such as the smallness and hygiene and stability problems of the manufacturers of the plant coexist,

Most of the plant fermentation broth was prepared by mixing the plant with sugar and fermenting the fermentation broth for a certain period of time. The fermented broth was filtered through a fermentation broth. Antioxidant activity of plant fermentation broth (Lee J. J Food et al . , 25, 412, 2012), anti-obesity (Yang CY et al ., J. Animal Science and Technology , 53, p.75-81, 2011), anti-cancer (Kim MJ et al ., Korean J. Food Sci . , p. 432-437, 2011). Korean Patent Laid-Open No. 10-2016-0088271 discloses a plant fermentation broth and a method for producing the same, Korean Patent Laid-Open No. 10-2015-0080412 discloses a method for producing a fermentation broth having enhanced alcohol degradation ability and a method ≪ / RTI > However, there is still insufficient base for the scientific and standardization of fermentation broth. In addition, when producing a fermented liquid of a plant, alcohol is generated. When the content of alcohol is 1% or more in the main taxation law, it is defined as alcohol and it is difficult to sell it as a fermented food.

Accordingly, the present inventors have made efforts to produce a plant fermentation broth in which alcohol is not produced, and have succeeded in isolating and identifying Leuconostoc mesenteroides N8-2 strain which does not produce alcohol during the fermentation process, The present invention has been accomplished by confirming that Berry's fermentation broth does not contain alcohol even when the growth of lactic acid bacteria is vigorous, and that the quality is excellent.

It is an object of the present invention to provide a process for producing a fermentation broth of the present invention in which alcohol production is inhibited by using a strain of Leuconostoc mesenteroides N8-2.

Another object of the present invention is to provide a fermented liquid of berries produced by the above method.

In order to accomplish the above object, the present invention provides a method for producing berries, comprising the steps of: 1) preparing a juice of berries added with sugar; And 2) a step of inoculating lactic acid bacteria into the juice of the above step and fermenting the fermented milk, wherein the production of alcohol is inhibited.

In addition, the present invention provides a fermented liquid of berries produced by the above method.

The beryllium fermentation broth produced according to the method of the present invention does not contain alcohol and thus has excellent quality.

Fig. 1 is a photograph showing the growth of lactic acid bacteria in order to screen for lactic acid bacteria having resistance to the acid in the MRS solid medium.
FIG. 2 is a photograph showing the degree of growth of lactic acid bacterium in order to select lactic acid bacteria having acid resistance primarily in MRS solid medium.
FIG. 3 is a photograph showing the degree of growth of lactic acid bacteria in order to select lactic acid bacteria secondary to acid resistance in a MRS solid medium. FIG.
FIG. 4 is a photograph showing the degree of growth of lactic acid bacteria in order to select lactic acid bacteria having alcohol resistance in the MRS solid medium. FIG.
5 is a schematic view showing a process for producing a fermentation broth of Berry.
6 is a graph comparing the alcohol content of the fermented broth with fermentation period.

Hereinafter, the present invention will be described in detail.

The beryllium fermentation broth of the present invention comprises: 1) preparing a juice of berries to which sugar is added; And 2) a step of inoculating lactic acid bacteria into the juice of the above step and fermenting the fermented milk, wherein the production of alcohol is inhibited.

The lactic acid bacteria may be selected from the group consisting of Lactobacillus sp., Pediococcus sp., Leuconostoc sp., Bifidobacterium sp., And Streptococcus sp. ). ≪ / RTI > More specifically, the lactic acid bacteria is Lactobacillus know also required Ruth (Lactobicillus acidophilus), Lactobacillus Plastic tarum (Lactobacillus plantarum), Lactobacillus Charra Kasei (Lactobacillus paracasei), Lactobacillus ramno Versus (Lactobacillus rhamnosus), Lactobacillus Bulgaria kusu (Lactobacillus bulgaricus , Pediococcus, acidilactici), Phedi OKO kusu pen soil three mouse (Pediococcus pentosaceus), current Kono Stock mesen teroyi Death (Leuconostoc mesenteroides), flow sheets Stock Pocono Leeum (Leuconostoc citreum), Bifidobacterium lactis (Bifidobacterium lactis , Bifidobacterium bifidum , Bifidobacterium adolescentis , Streptococcus faecalis , Streptococcus lactis , or Streptococcus thermophilus ( Streptococcus faecalis , Streptococcus faecalis , Streptococcus lactis , Streptococcus thermophilus ). In one embodiment of the present invention, the lactic acid bacterium may be Leuconostoc mesenteroides, more particularly Leuconostomyces tendides N8-2 strain deposited with Accession No. KACC 92153P.

In a specific example of the present invention, the present inventors firstly selected lactic acid bacteria using morphological differences in the leukocytes and isolated and identified a total of 30 lactic acid bacteria through 16S rRNA gene sequence analysis (see Table 1).

In addition, the present inventors analyzed the resistance, acid resistance, alcohol resistance and alcohol-producing ability of the above strains, and eventually used the strain Rheoconostomyces teneroides N8-2 in the production of berries fermentation broth (Tables 1 to 6 and Figs. 4).

The sugar of step 1) is added in an amount of 10% (w / w) to 50% (w / w), specifically 20% (w / w) to 40% (w / w), more specifically 25% / w) to 35% (w / w). In one embodiment of the invention, the sugar may be added at 30% (w / w). If the sugar content is too low, alcohol may be formed, and if the sugar content is too high, it is too sweet. The sugar may generally include all sugar used in the production of a fermentation broth. In one embodiment of the invention, the sugar may be sugar.

The berries may be at least one selected from the group consisting of olive, blueberry, bramble, red currant, black currant, cranberry, raspberry, strawberry, aronia, ascorbic, gooseberry, acerola and elderberry. In one embodiment of the present invention, the berries may be audi, blueberries and macromolecules.

The juice may be prepared according to a conventional method of juicing. For example, it may be a rotary grinding juice, a pressed juice or a belt juice.

The fermentation of step 2) may be carried out at a temperature of 25 ° C to 35 ° C, specifically 27 ° C to 33 ° C, more specifically 28 ° C to 32 ° C. In one embodiment of the present invention, the fermentation temperature may be 30 < 0 > C. If the fermentation temperature is out of the above range, the activity of the lactic acid bacterium may be lowered and the fermentation of the beryllium juice may not occur. The fermentation period may be from 3 days to 10 days, more specifically from 4 days to 9 days, more specifically from 5 days to 8 days. If the incubation period is too short, the fermentation will not occur sufficiently. If the incubation period is longer than 2 weeks, alcohol may be generated during fermentation.

The method for producing a fermentation broth of the present invention may further comprise a step of sterilizing the fermentation broth before inoculation of the lactic acid bacteria. The sterilization may be carried out at a temperature of 55 ° C to 80 ° C, specifically 60 ° C to 75 ° C, more specifically 60 ° C to 70 ° C. In one embodiment of the invention, the fermentation temperature may be 65 < 0 > C. The fermentation time may be 15 minutes to 1 hour, specifically 20 minutes to 40 minutes, more specifically, 25 minutes to 35 minutes. In one embodiment of the present invention, the fermentation time may be 30 minutes.

In a specific example of the present invention, the inventors of the present invention prepared a fermented broth fermentation broth by inoculating a juice of each of Odie, Bokbunja and Blueberry as a raw material, inoculating Leuconostoc mesenteroides N8-2 strain, (See Figs. 1 to 5 and Tables 1 to 6).

The present invention also provides a fermented liquid of berries produced by the method of the present invention.

The beryllium fermentation broth can be prepared as described above. For example, a method for producing a fermentation broth according to the present invention comprises the steps of: 1) preparing a fermentation solution of berries to which sugar is added; And 2) inoculating the juice with lactic acid bacteria and fermenting the juice. In addition, the lactic acid bacterium may be Leuconostoc mesenteroides N8-2 strain. The method for producing a fermentation broth of the present invention may further comprise the step of sterilizing the fermented juice of berries prior to inoculation of the lactic acid bacteria.

In a specific example of the present invention, the present inventors confirmed that alcohol was not produced in the fermentation broth of fermented berries produced by inoculating Leuconostoc mesenteroides N8-2 strain (see FIG. 6 and Tables 7 to 10).

Hereinafter, the present invention will be described in detail by the following examples.

However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited thereto.

Selection of strains

<1-1> Isolation and Identification of Lactic Acid Bacteria

10 g of Nuruk (Chuncheon, Jincheon-gun, Jeonbuk, Wanju) was suspended in 90 ml of 0.85% NaCl solution, and 100 μl of the suspension was spread on MRS solid medium and cultured at 37 ° C for 48 hours. The morphological differences of the cultured microorganisms were used for primary selection, and 16S rRNA gene sequencing analysis was used to identify the isolates selected. For the 16S rRNA gene sequence analysis by PCR, the primers were a 27F primer (5'-AGAGTTTGATCCTGGCTCAG-3 ') having a nucleotide sequence of SEQ ID NO: 1 which is a universal PCR primer of 16S rRNA and a 1492R primer having a nucleotide sequence of SEQ ID NO: 2 (5'-GGTTACCTTGTTACGACTT-3 ') was used. The PCR was carried out 35 times with 1 cycle of initial denaturation for 5 minutes, denaturation at 94 ° C for 45 seconds, annealing at 55 ° C for 60 seconds, and elongation at 72 ° C for 60 seconds. The nucleotide sequence of the PCR product was analyzed with reference to Genocell Co.,

As a result, 30 kinds of lactic acid bacteria as shown in Table 1 were isolated and identified (Table 1).

Isolation of lactic acid bacteria division
(NO.) detach
(Isolates) Species M1 FMS02-84 Leuconostoc mesenteroidessubsp . mesenteroides
(Leuconostomesenteroidis subspecies Messeneroidesis) M2 K6-7 Leuconostoc Mesenteriodes (Ryukono Stock Meense Teroids) M3 N2-2 Pediococcus pentosaceus (Pediococcus pentosaceus ) M4 N4-5 Weissella cibaria ( Bissela sivaria ) M5 N7-3 Leuconostoc citreum &lt; / RTI &gt; M6 N8-2 Leuconostoc mesenteroides (Ryukono Stock Meense Teroids) M7 N8-3 Pediococcus acidilactici (Pediococcus axicilyl lactity ) M8 N13-4 Lactobacillus brevis (Lactobacillus brevis) M9 N13-5 Lactobacillus plantarum ( Lactobacillus plantarum ) M10 N15-4 Pediococcus acidilactici (Pediococcus axicilyl lactity ) M11 N17-5 Pediococcus pentosaceus (Pediococcus pentosaceus ) M12 N19-5 Lactobacillus curvatus ( Lactobacillus curvatus ) M13 N19-9 Lactobacillus plantarum ( Lactobacillus plantarum ) M14 N20-6 Lactobacillus sakei (Lactobacillus casei) M15 N20-7 Enterococcus durans ( enterococcus durance ) M16 N21-3 Lactobacillus plantarum ( Lactobacillus plantarum ) M17 N21-4 Lactobacillus brevis (Lactobacillus brevis) M18 N21-5 Pediococcus pentosaceus (Pediococcus pentosaceus ) M19 N22-6 Lactobacillus curvatus ( Lactobacillus curvatus ) M20 N25-7 Lactobacillus brevis (Lactobacillus brevis) M21 N27-4 Lactobacillus brevis (Lactobacillus brevis) M22 N34-5 Enterococcus faecium (Enterococcus fetium) M23 N35-3 Pediococcus acidilactici (Pediococcus axicilyl lactity ) M24 N36-3 Lactobacillus coryniformis ( Lactobacillus korinifomis ) M25 N41-12 Lactobacillus brevis (Lactobacillus brevis) M @ 6 N42-7 Pediococcus pentosaceus (Pediococcus pentosaceus ) M27 N43-11 Lactobacillus coryniformis ( Lactobacillus korinifomis ) M28 E2-4 Lactobacillus rhamnosus ( Lactobacillus lambosus ) M30 E3-10 Lactobacillus diolivorans ( Lactobacillus diolivorans ) M31 E9-8 Lactobacillus diolivorans ( Lactobacillus diolivorans )

<1-2> To my goodness  Selection of Lactic Acid Bacteria

Among the 30 kinds of lactic acid bacteria isolated and identified in Example <1-1>, lactic acid bacteria having resistance to the acidity were selected on a solid medium. Sugar was used as a sugar for strain selection.

Specifically, MRS agar (BD) medium was prepared using a 96 well plate at a concentration of 5, 10, 20, 30, 40, 50 or 60% (w / w) On the other hand, 30 kinds of lactic acid bacteria were prepared by culturing in MRS liquid medium for 48 hours. 200 [mu] L of the culture solution was dispensed into the MRS solid medium and incubated at 37 [deg.] C for 48 hours, and the growth of the strain was visually observed.

Comparison of resistance of lactic acid bacteria to sugar concentration in solid medium detach
(Isolates) Species FMS02-84 Leuconostoc mesenteroidessubsp . mesenteroides
(Leuconostomesenteroidis subspecies Messeneroidesis) K6-7 Leuconostoc Mesenteriodes (Ryukono Stock Meense Teroids) N2-2 Pediococcus pentosaceus (Pediococcus pentosaceus ) N4-5 Weissella cibaria ( Bissela sivaria ) N7-3 Leuconostoc citreum &lt; / RTI &gt; N8-2 Leuconostoc mesenteroides (Ryukono Stock Meense Teroids) N8-3 Pediococcus acidilactici (Pediococcus axicilyl lactity ) N13-4 Lactobacillus brevis (Lactobacillus brevis) N13-5 Lactobacillus plantarum ( Lactobacillus plantarum ) N15-4 Pediococcus acidilactici (Pediococcus axicilyl lactity ) N17-5 Pediococcus pentosaceus (Pediococcus pentosaceus ) N19-5 Lactobacillus curvatus ( Lactobacillus curvatus ) N19-9 Lactobacillus plantarum ( Lactobacillus plantarum ) N20-6 Lactobacillus sakei (Lactobacillus casei) N20-7 Enterococcus durans ( enterococcus durance ) N21-3 Lactobacillus plantarum ( Lactobacillus plantarum ) N21-4 Lactobacillus brevis (Lactobacillus brevis) N21-5 Pediococcus pentosaceus (Pediococcus pentosaceus ) N22-6 Lactobacillus curvatus ( Lactobacillus curvatus ) N25-7 Lactobacillus brevis (Lactobacillus brevis) N27-4 Lactobacillus brevis (Lactobacillus brevis) N34-5 Enterococcus faecium (Enterococcus fetium) N35-3 Pediococcus acidilactici (Pediococcus axicilyl lactity ) N36-3 Lactobacillus coryniformis ( Lactobacillus korinifomis ) N41-12 Lactobacillus brevis (Lactobacillus brevis) N42-7 Pediococcus pentosaceus (Pediococcus pentosaceus ) N43-11 Lactobacillus coryniformis ( Lactobacillus korinifomis ) E2-4 Lactobacillus rhamnosus ( Lactobacillus lambosus ) E3-10 Lactobacillus diolivorans ( Lactobacillus diolivorans ) E9-8 Lactobacillus diolivorans ( Lactobacillus diolivorans )

As a result, as shown in Table 2 and Fig. 1, Pediococcus pentosaceus N2-2. Bacillus cervaria N4-5, Leuconostocritium N7-3, Leuconostomyces teneroides N8-2, Pediococcus axilidyl lactis N8-3, Lactobacillus brevis N13-4, Pediococcus axicillus Lactobacillus N15-4, Pediococcus pentosaceus N17-5, Lactobacillus brevis N41-12, Pediococcus pentosaceus N42-7, Lactobacillus diolivolans E3-10 and Lactobacillus diolivolans E9 -8 strains were found to be resistant (Table 2 and Fig. 1).

<1-3> Acid resistance  Selection of Lactic Acid Bacteria

Lactic acid bacteria having an acid resistance were selected from the 12 resistant strains selected in the above Example <1-1>.

Specifically, the MRS agar (BD) medium was prepared by adjusting the pH to 4, 5, 6, 7 by diluting the NaOH solution using a 96-well plate. On the other hand, the 12 kinds of lactic acid bacteria selected in Example <1-2> were cultured in the MRS liquid medium for 48 hours, and 200 μl of the culture solution was dispensed to the plate. Each strain was repeated 7 times and spotted on the medium. The cultures were incubated at 37 ° C for 2 days, and six strains of lactic acid bacteria were selected by visual observation of the growth of the strains.

Comparison of acidity of lactic acid bacteria with acidity in solid medium Category (NO.)
Species
pH 7 6 5 4 N2-2 Pediococcus pentosaceus +++++ +++++ +++ + N4-5 Bissela Sivaria +++++ +++++ +++ + N7-3 Licorostocritium +++++ +++++ ++ ++ N8-2 Ryukono Stock Methenseeroides +++++ +++++ ++ ++ N8-3 Pedio Caucus +++++ +++++ ++ + N13-4 Lactobacillus brevis +++++ +++++ ++ ++ N15-4 Pedio Caucus +++++ +++++ ++ + N17-5 Pediococcus pentosaceus +++++ +++++ ++ + N41-12 Lactobacillus brevis +++++ +++++ ++++ +++ N42-7 Pediococcus pentosaceus +++++ +++++ +++ + E3-10 Lactobacillus diolriborans +++++ +++++ +++ + E9-8 Lactobacillus diolriborans +++++ +++++ +++ +

As a result, as shown in Table 3, Fig. 2 and Fig. 3, as strains having acid resistance, Ryukonosost Citrium N7-3, Ryukono Stokesensen Teroides N8-2, Lactobacillus brevis N13-4, Lactobacillus brevis N41-12, Pediococcus pentosaceus N42-7, and Lactobacillus diolivorans E9-8 were first selected.

Six selected strains were cultivated in pH 4 and pH 7 medium, and three kinds of lactic acid bacteria were selected by visual observation of the growth of the strain. Each strain was repeated 7 times and spotted on the medium.

As a result, Ryukono Stokes meenceroides N8-2, Lactobacillus brevis N41-12 and Pediococcus pentosaceus N42-7 strains were finally selected (Table 3, Fig. 2 and Fig. 3).

<1-4> My alcoholic  Selection of Lactic Acid Bacteria

The 12 types of lactic acid bacteria resistant to the tolerance selected in the above Example <1-2> were selected for the lactic acid bacteria having the alcohol resistance.

First, MRS agar (BD) medium was prepared by adding ethyl alcohol (Fisher) at a concentration of 2% (v / v), 4% (v / v) or 8% (v / v). Meanwhile, the 12 kinds of lactic acid bacteria selected in the above Example <1-2> were cultured in the MRS liquid medium for 48 hours, and then 200 μl was added to the solid medium. The inoculated strain was cultured at 37 ° C for 2 days, and the growth of the strain was visually observed. Each strain was repeated 7 times and spotted on the medium.

Comparison of Alcohol Content of Lactic Acid Bacteria with Alcohol Content in Solid Media Category (NO.)
Species
Alcohol(%) 0 2 4 8 N2-2 Pediococcus pentosaceus +++++ ++++ / +++ +++ / +++ ++ / + N4-5 Bissela Sivaria +++++ ++++ / +++ +++ / ++ + / + N7-3 Licorostocritium +++++ ++++ / +++ +++ / +++ ++ / + N8-2 Ryukono Stock Methenseeroides +++++ ++++ / +++ +++ / +++ ++ / + N8-3 Pedio Caucus +++++ +++ / +++ ++ / +++ ++ / + N13-4 Lactobacillus brevis +++++ +++ / ++++ +++ / +++ ++ / ++ N15-4 Pedio Caucus +++++ +++ / +++ ++ / +++ ++ / + N17-5 Pediococcus pentosaceus +++++ ++++ / +++ +++ / +++ ++ / + N41-12 Lactobacillus brevis +++++ +++ / +++ ++ / +++ + / + N42-7 Pediococcus pentosaceus +++++ ++++ / ++++ +++ / +++ ++ / + E3-10 Lactobacillus diolriborans +++++ ++++ / ++++ +++ / +++ ++ / ++ E9-8 Lactobacillus diolriborans +++++ ++++ / ++++ +++ / +++ ++ / ++

Based on /, the left shows the results of primary screening and the right shows secondary screening results.

As a result, as shown in Table 4 and Fig. 4, the lactic acid bacteria having the alcohol resistance were Pediococcus pentosaceus N2-2, Leuconostocisium N7-3, Leuconostomyceteroidesides N8-2, Bacillus brevis N41-12 and Pediococcus pentosaceus N42-7 (Table 4 and Fig. 4).

From the above, it was confirmed that Ryukono Stokes meenceroides N8-2, Lactobacillus brevis N41-12 and Pediococcus pentosaceus N42-7 were excellent in resistance to sugar, acid resistance and alcohol resistance.

<1-5> Determination of alcohol production by lactic acid bacteria in liquid medium

Three kinds of lactic acid bacteria, Lukonostomyces teneroides N8-2, Lactobacillus brevis N41-12, and Lactobacillus brevis, which are selected in Examples <1-2> to <1-4>, have acid resistance, Pediococcus pentosaceus strain N42-7 was used to confirm whether or not the strain produced alcohol in the fermentation process.

Specifically, the above three lactic acid bacteria were cultured in an MRS liquid medium for 48 hours, and then 200 μl of the culture liquid was inoculated into 10 ml of MRS liquid medium containing 30% (w / w) sugar. Sugar was used as sugar. After incubation at 37 ° C for 4 days, the content of alcohol contained in the culture was repeated three times and the average values were compared.

Comparison of Alcohol Content of Lactic Acid Bacteria in Liquid Medium Lactobacillus
Alcohol content (%) (v / v) One 2 3 Average N8-2 ND ND ND ND N41-12 2.3 2.6 2.2 2.37 N42-7 ND ND ND ND

ND (not ditect): If the alcohol content is 0

As a result, as shown in Table 5, about 2.4% (v / v) of alcohol was detected in the liquid medium inoculated with the Lactobacillus brevis N41-12 strain to confirm that the strain had alcohol-producing ability ). Therefore, further experiments were conducted using Leukonostomyces teneroides N8-2 and Pediococcus pentosaceus N42-7 strains, except for Lactobacillus brevis N41-12, which has an alcohol-producing ability.

<1-6> Confirmation of alcohol production by lactic acid bacteria depending on sterilization of fermentation medium

Using the two kinds of lactic acid bacteria selected in Example <1-5>, alcohol production was confirmed according to sterilization of the fermentation medium.

Specifically, 30% (w / w) sugar was added to the juice of Odi (Jeonbuk Gochang) to prepare a fermentation medium. Sugar was used as sugar. The prepared medium was not sterilized, or sterilized at 65 캜 for 30 minutes, at 75 캜 for 30 minutes, or at 75 캜 for 1 hour, and the two strains were inoculated in the sterilized medium. This was fermented at a temperature of 30 캜 to prepare a fermentation broth. On the 4th day after the fermentation start date, the alcohol content in the fermentation broth was repeated three times and the average values measured were compared.

Comparison of Alcohol Content of Lactic Acid Bacteria by Fermentation Conditions Lactobacillus
Sterilization condition
Alcohol content (%) (v / v) One 2 3 Average
N8-2

Sterilization ND ND ND ND 65 ° C, 30 minutes ND ND ND ND 75 ° C, 30 minutes ND ND ND ND 75 캜, 60 min ND ND ND ND
N42-7

Sterilization 15.2 14.6 15.1 15.0 65 ° C, 30 minutes 13.6 13.9 13.5 13.7 75 ° C, 30 minutes 13.8 13.6 13.2 13.5 75 캜, 60 min 14.0 14.0 13.8 13.9

As a result, as shown in Table 6, in the fermentation broth inoculated with strain Pediococcus pentosaceus N42-7, alcohol was generated irrespective of whether the fermentation medium was sterilized or not, whereas Leuconostomycetenoids N8-2 No alcohol was produced in the fermentation broth inoculated with the strain (Table 6). Therefore, a strain of Ryukono Stokes meenceroides N8-2 was selected as a final strain to produce a beryllium fermentation broth. The Ryukono Stokes meenceroides N8-2 strain was deposited with the Korean Agricultural Microorganism Resource Center (KACC) under accession number KACC 92153P.

Berry  Preparation of fermentation broth

&Lt; 2-1 > Preparation of an ODE fermentation broth

First, a group in which 30% (w / w) sugar was added to a group, and 30% (w / w) sugar was added to unseawed group and the fermented group was divided into two groups (Control group). Three groups of fermentation broths were prepared by adding a fermentation step to the one group of products. Further, the group of the obtained product was sterilized at a temperature of 65 캜 for 30 minutes, inoculated with the strain Ryukono Stokes meenceroides N8-2 (accession No. KACC 92153P), and then subjected to fermentation at a temperature of 30 캜 4 groups (experimental group), and 5 groups were prepared by the same conditions and methods as those of the group 4, except that sterilization was performed at 121 ° C for 15 minutes instead of the sterilization process (Table 7 and FIG. 5). Sugar was used as sugar for fermentation.

<2-2> Preparation of Bokbunja fermentation broth

First, the bokbunja were squeezed, and 30% (w / w) sugar added group was used as one group. The fermentation liquid prepared by fermenting 30% (w / w) 2 group (control group). Three groups of fermentation broths were prepared by adding a fermentation step to the one group of products. Further, the group of the obtained product was sterilized at a temperature of 65 캜 for 30 minutes, inoculated with the strain Ryukono Stokes meenceroides N8-2 (accession No. KACC 92153P), and then subjected to fermentation at a temperature of 30 캜 4 groups (experimental group), and 5 groups were prepared by the same conditions and methods as those of group 4, except that sterilization was performed at 121 ° C for 15 minutes instead of the sterilization process (Table 8 and FIG. 5). Sugar was used as sugar for fermentation.

<2-3> Blueberries  Preparation of fermentation broth

First, blueberries were squeezed, and a group to which 30% (w / w) sugar was added was used as one group. Fermentation was performed by adding 30% (w / w) sugar to unsweetened blueberries The fermentation broth was divided into 2 groups (control group), and the fermentation broth prepared by adding a fermentation step to 1 group was defined as 2 groups (control group). Three groups of fermentation broths were prepared by adding a fermentation step to the one group of products. Further, the group of the obtained product was sterilized at a temperature of 65 캜 for 30 minutes, inoculated with the strain Ryukono Stokes meenceroides N8-2 (accession No. KACC 92153P), and then subjected to fermentation at a temperature of 30 캜 4 groups (experimental group), and 5 groups were prepared by the same conditions and methods as in group 4, except that sterilization at 121 ° C for 15 minutes was added instead of the sterilization process (Table 9 and FIG. 5). Sugar was used as sugar for fermentation.

Berry  Analysis of quality characteristics of fermentation broth

<3-1> Analysis of quality characteristics according to manufacturing method

In order to compare the quality characteristics of the beryllium fermentation broth according to the manufacturing method, berries were fermented according to a conventional method. On the 7th day, the fermented broth obtained in Example 2 was sampled on day 0, and the sugar content, acidity, number of lactic acid bacteria and yeast number were measured.

Comparison of quality characteristics according to the manufacturing method of the fermented broth process
Sugar content
(° Brix) Acidity (%)
(Based on lactic acid) Lactobacillus
(log CFU / ml) leaven
(log CFU / ml) 1 group 46.0 ± 0.2 0.37 ± 0.01 - 5.1 2 groups 47.2 ± 0.7 0.6 ± 0.08 6.7 ± 0.04 7.8 3 groups 47.2 ± 0.1 0.44 + 0.02 6.0 ± 0.04 6.6 4 groups 46.6 ± 0.1 0.4 ± 0.01 6.1 ± 0.04 - 5 groups 47.1 ± 0.2 0.46 ± 0.01 6.1 ± 0.04 -

Comparison of quality characteristics according to the manufacturing method of bokbunja fermented broth process
Sugar content
(° Brix) Acidity (%)
(Based on lactic acid) Lactobacillus
(log CFU / ml) leaven
(log CFU / ml) 1 group 44.0 + - 0.0 1.0 ± 0.03 - 5.0 2 groups 45.5 ± 0.9 1.08 + 0.06 6.7 ± 0.02 7.7 3 groups 44.3 ± 0.2 1.1 ± 0.02 5.5 ± 0.04 7.4 4 groups 44.3 ± 0.1 0.98 + 0.03 5.7 ± 0.05 - 5 groups 44.6 ± 0.2 0.97 + 0.02 5.7 ± 0.02 -

Comparison of quality characteristics of blueberry fermented liquid according to the manufacturing method process
Sugar content
(° Brix) Acidity (%)
(Based on lactic acid) Lactobacillus
(log CFU / ml) leaven
(log CFU / ml) 1 group 44.1 ± 0.2 0.71 ± 0.01 - - 2 groups 51.0 ± 0.9 0.57 + 0.08 - 6.3 3 groups 44.6 ± 0.1 0.83 ± 0.01 - 5.0 4 groups 44.2 ± 0.0 0.78 ± 0.01 5.8 ± 0.01 - 5 groups 44.3 ± 0.0 0.77 ± 0.01 5.6 ± 0.02 -

As a result, as shown in Tables 7, 8 and 9, no yeast producing alcohol was detected in the fermentation liquid of the 4th and 5th groups, and the growth of the lactic acid bacteria inducing lactic acid fermentation was intense (Tables 7, 8 and 9 ).

<3-2> Analysis of quality characteristics according to fermentation period

In order to compare the quality characteristics of the fermentation broth with the fermentation period, berries were fermented according to a conventional method. On the 1st, 3rd, 5th, and 7th days, the fermentation broth prepared in Example 2 was collected and the alcohol content was measured. On the 7th day, samples were collected and the number of yeast Respectively. At this time, 2 groups (practice) were used as a control group and 4 groups (improvement) were used as experimental groups.

As a result, as shown in Table 10, no yeast producing alcohol was detected in the fermentation broth prepared by juicing and sterilizing beryllium fermentation broth and inoculating Leuconostoc mesenteroides N8-2 strain (Table 10). In addition, as shown in Fig. 6, no alcohol was produced in the fermented fermented broth (Fig. 6).

Comparison of yeast counts during fermentation period of berries fermented (log CFU / ml) division 2 Group (practice) 4 groups (improvement) Audi 7.8 0 Bokbunja 7.7 0 Blueberries 6.3 0

<110> Rural Development Administration <120> MANUFACTURING METHOD OF BERRIES FERMENTED LIQUID USING LACTIC          ACID BACTERIA AND BERRIES FERMENTED LIQUID MANUFACTURED BY SAME          METHOD <130> 2016P-10-042 <160> 2 <170> KoPatentin 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> 27F primer <400> 1 agagtttgat cctggctcag 20 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> 1492R primer <400> 2 ggttaccttg ttacgactt 19

Claims (11)

1) preparing a juice of berries added with sugar; And
2) a step of inoculating and fermenting Leuconostomyces tendens N8-2 strain deposited with the accession number KACC 92153P in the juice of step 1) above, and inhibiting the production of alcohol.
delete delete The process according to claim 1, wherein the sugar in step 1) is added in an amount of 10% (w / w) to 50% (w / w).
The method according to claim 1, wherein the berries are at least one selected from the group consisting of audi, blueberry, bramble, red currant, black currant, cranberry, raspberry, strawberry, aronia, ascorbic, gooseberry, acerola and elderberry A method for producing a fermentation broth.
The process according to claim 1, wherein the fermentation of step 2) is carried out at a temperature of 25 캜 to 35 캜.
The fermentation liquid according to claim 1, wherein the fermentation of step 2) is carried out for 3 to 10 days.
2. The method for producing a fermented vermicifluid according to claim 1, further comprising the step of sterilizing the juice of berries prior to inoculation of the strain Reoskonostomyces troidis N8-2 deposited with Accession No. KACC 92153P.
9. The process according to claim 8, wherein the sterilization is performed at a temperature of 55 to 80 캜.
The method for producing a fermented vermicifluate according to claim 8, wherein the sterilization is performed for 15 minutes to 1 hour.
A fermented verrucous liquid produced by the method of claim 1.

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