KR101063624B1 - Schizandra fermented product, fermentation method thereof and food composition comprising the fermented product - Google Patents

Abstract

The present invention relates to a Schizandra fermentation product, a fermentation method thereof, and a food composition including the fermentation product, and more particularly, Enterococcus sp. Fermented Schisandra chinensis with TO-94, Schisandra chinensis fermented product with increased contents of fructose, glucose, lactic acid, L-tartaric acid, vitamins, polyphenols and anthocyanins, fermentation method thereof, and a food composition comprising the fermentation product will be.

Description

Fermentation method of Schisandra chinensis, its fermentation method and food composition containing the fermentation {Ferments of Schisandra chinensis, the method for fermenting etc and the food composition containing the same}

The present invention relates to a Schizandra fermentation product, a fermentation method thereof, and a food composition including the fermentation product, and more particularly, Enterococcus sp. Fermented Schisandra chinensis with TO-94, Schisandra chinensis fermented product with increased contents of fructose, glucose, lactic acid, L-tartaric acid, vitamins, polyphenols and anthocyanins, fermentation method thereof, and a food composition comprising the fermentation product will be.

Schisandra chinensis is a deciduous vine plant belonging to the Schisandra chinensis family, and various physiological functions such as blood pressure lowering, anticancer, anti-aging, immunomodulation and antimicrobial activity have been reported (Ames BN, et al., Proc. Natl. Acad. Sci. USA, 1993, 90: 7915-7922; Choi JS, et al ., Phytochemistry, 1995, 40: 997-999; Devy C, et al ., Biochem. Parmacol., 1990, 39: 399-405; Kuramoto T, Up to date Food Processing , 1992, 27 (3): 22-23; Hae J, Dong Eu Bo Gam , Nam San Dan , Seoul ., 1990, p. 140-142; Takashi N, et al ., FEBS Lett ., 2002, 514: 315-322; And Kamei R, et al ., Life Sci . , 2003, 73: 2091-2099), anthocyanin, a red pigment unique to Schisandra chinensis, is known to have antioxidant activity. The five-flavored Schisandra chinensis is attracting attention as a highly commercial raw material in line with the trend of the global beverage market, where various functionalities and natural colored beverages are increasing (Lee WY, et. al ., Korean J. Food Preserv . , 2006, 13: 252-258). However, in order to stabilize the properties of Schizandra chinensis, the process of extracting water at low temperature inside and outside the temperature of 60 ℃ is less effective because of its low yield and activity (Lee WY, et. al ., Korean J. Food Preserv. , 2006, 13: 252-258; And Kim HK, et al ., Korean J. Food Culture , 2004, 19: 484-490). In addition, the sugar content contained in the fruit itself is only about 4.05%, so it is not suitable for fermentation because it is insufficiently used as a microbial nutrient source (Hyun HK, et. al ., Korean J. Plant Res . , 2002, 15: 1-7), with an initial pH of 2.5 and low acidity, which is not suitable for the growth of fermented microorganisms.

Recently, various studies have been attempted to increase the weight of nutrients from low nutrition materials using lactic acid bacteria that can hydrolyze starch raw materials, thereby developing high energy supplement foods. Lactic acid bacteria are microorganisms that can grow under low acidity conditions and are fermented by lactic acid to prevent food spoilage and secrete antimicrobial substances such as bacteriocin to inhibit the growth of food poisoning bacteria and gut rot bacteria (Park YS, et. al ., J. Korean Soc . Agric . Chem . Biotechnol ., 2003, 461: 367-375). For example, Songre-Ouattar et al. (Songre-Ouattara LT, et. al ., International Journal of Food Microbiology , 2009, 130 (3): 258-264) Lactobacillus Peanut Sludge Starch Partial degradation using plantarum A6 reported an increase in the weight of the nutrients. Other corn (Nakamura LK, International Journal of Systematic Bacteriology , 1998, 31: 56-63), potato (Chatterjee M, et al., Biotechnology Letters , 1997, 19: 873-874) and Cassava; Giraud E, et al., Applied Microbiology and Lactobacillus fermenting amylaceous raw material ( Biotechnology , 1991, 36: 379-383) amylovorus And Lactobacillus plantarum have been reported. Also Thi is Lactobacillus rice / bean Fermentation with plantarum has led to the development of nursing supplements (Nguyen TT, et. al ., Food Chemistry 100. , 2007, p. 623-631). However, in the case of Schisandra chinensis, it is difficult to ferment by lactic acid due to lack of sugar content. Commercial enzymes may be used to increase the carbohydrate content of Schisandra chinensis, but in this case, the purchase of enzymes that are active under low pH conditions of Schisandra chinensis will not be smooth, as well as increased production costs and additional equipment for further processing. There is no practical use.

Accordingly, the present inventors can isolate and identify specific fermentation bacteria that can ferment Schizandra chinensis in order to increase the effectiveness of Schizandra chinensis and fermented Schizandra chinensis fermented products having low acidity and high fiber content. The present invention was completed.

Accordingly, it is an object of the present invention to provide a Schizandra fermented product, a fermentation method thereof, and a food composition comprising the same, using novel fermentation bacteria.

In order to achieve the above object, in the present invention, Enterococcus sp. Provide Schizandra fermentation fermented with TO-94.

Also in the present invention, Enterococcus sp. Of Accession Number KFCC11477P. Provided is a method of fermenting Schisandra chinensis with TO-94.

The present invention also provides a food composition containing the Schisandra chinensis.

Schisandra chinensis of Enterococcus sp. Fermented products prepared by fermentation with TO-94 contained more fructose, glucose, lactic acid, L-tartaric acid and vitamins than non-fermented Schisandra chinensis, and sucrose, maltose, oxalic acid, and formic acid. ) And malic acid content decreased. In addition, the fermented product increased the content of polyphenols and anthocyanins by 3.8 times and 1.2 times compared to Schizandra chinensis, respectively.

1 is Enterococcus sp . 16S rDNA sequence and Phylogenetic tree of TO-94 are shown.
2 Entrococcus paecium IFO13712 and Enterococcus sp . The fatty acid composition of TO-94 total membrane lipid is shown.
3 shows Enterococcus sp. TO-94 was inoculated in MRS medium and Schisandra chinensis, respectively, and cultured at 37 ° C., and it was a graph comparing the growth of bacteria at 4 hour intervals (■: Schisandra chinensis broth and ●: MRS medium).
4 is Enterococcus sp. It is a graph which measured the change of the content of the free sugar and the total sugar of the fermentation product obtained by the process of Schisandra chinensis by TO-94 [■: maltose, ▲: sucrose, ●: glucose, *: fructose, and ◇: total sugar content].

Hereinafter, the present invention will be described in more detail.

The present invention Enterococcus sp. The present invention relates to a Schisandra fermented product fermented with TO-94 and a fermentation method thereof. Enterococcus sp. TO-94 was obtained from isolates of lactic acid bacteria with fibrous degradability from infant feces less than 2 months old, and then tested for their suitability for fermentation of Schizandra chinensis. Deposited with the microbial preservation center was given accession number KFCC11477P.

The fermentation method of Schisandra chinensis according to the present invention comprises the following steps:

1) crushing using a homogenizer after adding the same amount (v / v) of water to Schizandra;

2) heat sterilizing the lysate of 1) at 70 to 75 ° C. for 5 to 10 minutes;

3) Enterococcus sp. Of accession number KFCC11477P. Preparing a dilution (6.0-8.0 CFU / ml) diluted with TO-94 in physiological saline; And

4) mixing the dilution of 3) to the sterile lysate of 2) and incubated at 37 ° C. for 72 hours.

In another aspect, the present invention provides a food composition containing 10 to 100% by weight of the Schizandra fermentation product based on the total weight of the composition.

The food composition according to the present invention may be prepared as various cooked foods such as beverages, baking, drinks, oils, ice cream, sweets, rice cakes, snacks, baby foods, alcoholic beverages, seasonings, retorts or canned foods, but is not limited thereto. .

Hereinafter, the present invention will be described more specifically with reference to examples and test examples. These examples are provided only for understanding the contents of the present invention, and the scope of the present invention is not limited to these examples, and modifications, substitutions, and insertions commonly known in the art may be performed. This is also included in the scope of the present invention. In Examples and Test Examples of the present invention was statistically processed by the one-way analysis of variance (ANOVA) method. Statistical results were expressed as mean ± standard deviation of the three analysis results.

Example 1 Enterococcus sp. Isolation and Identification of TO-94

<Isolation and Identification of Lactic Acid Bacteria with Fibrous Resolution>

A total of 17 feces of infants under 2 months of age were collected to isolate lactic acid bacteria having fibrinolytic capacity for Schisandra fermentation. Collected infant feces were diluted to an appropriate magnification (10 ^{−8 /} ml) after filtration. The diluted solution was plated on MRS medium (Lactobacilli MRS broth; Difco Co., France) and incubated at 37 ° C. for 24 hours. Colonies formed after the culture were first picked strains forming a transparent ring by tooth picking on MRS medium to which 2% CaCO ₃ was added. Among the 1,150 colonies of the first screened colonies, 10 g / L carboxymethyl cellulose was excluded, except for dextrose and ammomium citrate, in order to select strains with high carboxymethyl cellulose (CMC) resolution. cellulose) was selectively inoculated with MRS agar medium for primary selection strain and incubated at 37 ° C for 24 hours. After incubation, the strain was stained with 1% Congo red solution and washed with 1% NaCl solution to form a final strain of yellow transparent rings.

Morphological Characterization

The size and shape of the isolates were determined by Gerhatdt et al. (Gerhardt P, et. al ., American Society for Gram staining (BD Co., USA) was performed according to the method of Microbiology , Washington, 1984, and morphological characteristics were observed with an optical microscope (Nicon, FK-IIA, Japan). At this time, the shape, size and color of colonies grown in glucose nutrient agar medium, skim milk medium and LB (Luria-Bertani) medium were observed, and the semi-fluid solid medium (tryptose 1.0%, Mobility was examined by incubating in 0.5% sodium chloride and 0.5% agar. The isolates were round in colony form in acidic plate media (pH 3.0), bright milky white and convex and smooth. In addition, the growth was good and anaerobic in acidic liquid medium, and the cells showed precipitation.

Physiological Characterization

To investigate the physiological characteristics of the isolates, the microbial experiment manual (Cappuccino JG, et al ., Benjamin and Cummings , California , 1975), casein resolution, starch resolution, gelatin liquefaction, sugar fermentation, indole production, nitrate reduction, catalase and Oxidase production capacity was investigated.

In addition, the fibrous resolution of the isolate was determined by measuring the activity of CMCase and pectinase. The activity of CMCase was quantified by measuring the reducing sugars liberated from CMC by DNS method (Miller GL, Anal . Chem . , 1959, 31: 426-428). For the standard measurement of this experiment, 500 μl of CMC (1.0%, w / v) dissolved in distilled water, 250 μl of 200 mM Na-citrate buffer (pH 5.5) and enzyme solution were mixed to the final 1 ml, and then 55 The reaction was carried out at 1 ° C. for 1 hour 30 minutes. 3.0 ml of DNS reagent and 0.1 ml of glucose solution (0.1 mg / ml) were added to stop the reaction, boil for 5 minutes, allow to cool and measure absorbance at 550 nm. The amount of reducing sugar produced was determined from a standard curve prepared using a glucose standard solution. The activity of 1.0 unit of enzyme was defined as the amount of enzyme that produced a reducing sugar corresponding to 1.0 μmol of glucose in 1 minute under 55 ° C. using CMC as a substrate.

Physiological characteristics of the isolates are shown in Table 1 below:

Growth temperature 15 to 45 ℃ Growth pH 2.0 ~ 9.0 NaCl resistance ≤ 10% Catalase - Oxidase - Lecithinase + Lipase - Phenylalanine Deamination - Hydrolysis Starch + casein + cellulose + Esculin + Indole production + Levan formation from sucrose + NH ₃ production from argin + NH ₃ production from peptone + Gelatin liquefaction + Methyl red test - Vogs-Proskauer reaction + Nitrate reduction + Action in milk Coagulation + Peptonization + Oxidation-fermentation test Fermentation Growth in nutrient plates +

In Table 1, the growth pH range of the isolate strain was 2.0 to 9.0, and the growth temperature range was 15 to 45 ℃. In addition, it was possible to grow up to 10% salt concentration, hydrolyzed catalase negative, oxidase negative, starch and casein, and cellulose hydrolysability was positive. In addition, it had the ability to produce indole, NH ₃ was produced from arginine and peptone, and growth was also confirmed in neutral nutrient medium. L-rhamose was not used in the sugar fermentation experiment, but glucose, sucrose and raffinose were well used. Acid was produced from some sugars, including glucose, but no gas was produced in all sugars.

Biological Characterization

The biological properties of the isolates are described by Bochner et al. (Bochner BR, et. al ., Genome research .. 2001, 11: 1246-1255; Bochner BR, "Breath prints" at the microbial level, 1989, ASM News . 55: 536-239; And Bochner BR, Nature , 1989, 339: 157-158) and Lee et al. (Lee JS, et. al ., Int . J. Syst . Bacteriol ., 2002, 52: 2110-2123). The isolates were precultured in a nutritionally limited medium (Resasoner DJ, et al., Appl . Environ. Microbiol ., 1985, 49: 1-7) for 24 hours and then suspended in sterile saline to an appropriate concentration, and various carbon and nitrogen sources. , a busy phosphate source (phosphate source), hwangwon (sulfur source), independent supplement (supplement autotrophic) and inhibitors (inhibitory compound) per 150 μL in the ^TM GP Biolog microplate containing the cells were cultured for the next 4-24 hours . During this period, the cell's respiratory activity causes various sources to oxidize, and the tetrazolium dye contained in the plate forms purple. Enterococcus sp. Using a biolog data base. As a result of comparing the similarity between TO-94 and standard strains, Enterococcus sp. TO-94 is Lactococcus lactis with about 43.3% and Streptococcus The thermophilus showed a similarity of about 20.7%, which turned out to be a new microorganism different from the existing lactic acid bacteria.

<Chemical Characteristic Analysis>

Chemical characteristics of the isolates were determined by Komagata et al. (Komagata K, et. al ., Method in Microbiology . Acadeic press., 1987, 19: 161-207) (Method in Microbiology) and Miller (Miller GL, Anal . Chem . , 1959, 31: 426-428) were analyzed according to the microbial identification system (MIS). As analytical methods, fatty acid type, DNA base composition (G + C molar content) and DNA sequence in the cell wall were examined.

Gas chromatography (GC, HP 6890) was used to confirm the total cell fatty acid composition of the isolate strain. As a standard fatty acid, a standard fatty acid (calibration standard kit) provided by HP (Hewlett-Packard) was used, and the culture of the strain was incubated for 24 hours at 30 ℃ using agar (Sabouraud's agar; Difco, USA) medium. The cultured cells were then taken to a test tube, and then 1 mL of saponification reagent (45 g of sodium hydroxide, 150 mL of methanol, and 150 mL of distilled water) was added thereto, followed by cooling for 30 minutes at 100 ° C., followed by cooling in running water.

2 mL of a mixture of 325 mL of 6 N HCl and 275 mL of methanol was added to the saponified cells, followed by methylation at 80 ° C. for 10 minutes, and then 1.25 mL of a mixture of 200 mL of hexane and 200 mL of methyl-t-butylether was added thereto. After shaking slowly at room temperature for 10 minutes, the fatty acid was extracted by removing the lower layer solution. Washed with 5.4% NaCl and used as analytical sample. Analytical samples were analyzed using GC and then Sherlock program, a strain identification program of MIDI.

16S rDNA gene analysis was performed using a PCR purification kit (Wizard Genomic DNA Prep Kit, Promega). At this time, the reaction conditions were denatured at 94 ° C. for 1 minute, annealed at 60 ° C. for 1 minute 30 seconds, and then repeated a total of 30 cycles of expansion at 72 ° C. for 1 minute. The primers used were 27F (5'-AGAGTTTGATCMTGGC TCAG-3 '; SEQ ID NO: 1) and 1492r (5'-TACGGYTACCTTGTTACGACTT-3'; SEQ ID NO: 2). The amplified fragment (16S rDNA region) was inserted into pGEM T easy vector (Promega) and transformed by host molecular cloning method using E. coli DH5α as a host cell, followed by DNA purification system (Wizard Plus The plasmid was purified (Mini-preparation) using the SV miniprep DNA purification system and sequenced using an automated DNA sequencing device (ABI 3700, Applied Biosystems Inc.). Using Clustal X (ver. 1.8) software, the sequences were aligned with reference to the rRNA secondary structure, and then similarity (%) values were obtained. In addition, the gene sequence was compared with Genebank through “Advanced Blast search,” and treeview (ver. 1.66) was used to calculate the tree tree (phylogenetic tree and dendrogram) by neighbor-joining analysis (Lee JS, et. al ., Int . J. Syst . Bacteriol ., 2002, 52: 2110-2123) to create a phylogenetic tree (phylogenetic tree) and the results are shown in FIG. The scale bar of the phylogenetic tree means 0.1 substitutions per site.

16S rDNA gene alignment was performed using Clustal X (ver. 1.8) software to determine the 1,674 bp sequence of SEQ ID NO: 3 and the results are shown in FIG. 1. In addition, as a result of examining the cell fatty acid component of the isolated strain in FIG. 1, the main components were 16: 0 and 19: 0 cyclopropane w8c, respectively, showing 20.29% and 24.37%, respectively. Specifically, the contents of 19: 0 cyclopropane w8c and 17: 0 cyclopropane were high. Which is Leuconostoc The genus Lactobacillus has an average of 11.29% and 0.38% (Giraud E, et. al ., Applied Microbiology and Biotechnology , 1991, 36: 379-383) Enterococcus In lactic acid is 17: 0 without cyclopropane 19: 0 cyclopropane w8c only have a very small amount (2.5%) and (Grazia FM, et al, nov Inter J. Sys Evol Micro, 2004, 54:...... 1717- 1721), Streptococcus Lactobacillus spp. And genus Lactobacillus are reported to contain 39.8% and 10.7% (Goldberg I, et. al ., Appl. Environ . Microbio ., 1977, 33: 489-496) did not find a match. Leucomostoc compared to references pseudomesenteroides showed a similarity of about 33.6% and Lactobacillus fermentum is about 22.9%, Tetragenococcus It was about 21.5% with halophila and showed very low similarity, but Entrococcus faecium It was confirmed that the bacteria belong to the genus Entrococcus by showing a similarity of 95.7% with the standard strain. However, the strain isolated from the present invention and Entrococcus faecium Comparison of IFO13712 (Youko S, et al ., Microbiology, 2003, 149: 2901-2908) (FIG. 2), showing so many differences that it is impossible to compare between the two strains, which is likely a new microorganism that has not been reported so far.

Therefore, the strain isolated in the present invention is judged to be a new species different from the lactic acid bacteria found so far " Enterococcus sp. TO-94 ", and was deposited on January 12, 2010 to the Korea Microbiological Conservation Center affiliated with the Korean spawn association, and received the accession number KFCC11477P.

Example 2 Culture of Lactic Acid Bacteria

Enterococcus sp. Isolated from Example 1 above. TO-94 was used in the experiments while stored at -80 ° C in 40% glycol solution. The growth of bacteria was incubated at 10% in MRS medium supplemented with 1% CMC and incubated at 37 ° C for 24 hours. Growth of bacteria at 600 nm was carried out using Spectronic 401 spectrophotometer (Milton Roy, paris, France) (A600 vs. cfu). / ml) was measured. After incubation, the cells were collected by centrifugation at 14,000 g × 10 minutes. The cells were washed in physiological sterile solution (0.9%, w / v saline) and recentrifuged under the same conditions to be used as inoculum of Schizandra fermentation. The growth of lactic acid bacteria during fermentation was measured by viable count after step dilution.

Example 3 Fermented Schisandra chinensis by Lactic Acid Bacteria

Enterococcus sp. Fermentation of Schisandra chinensis by TO-94 was carried out as follows.

In the present invention, the Schizandra chinensis purchased from Chonbuk Jangsu Nonghyup was used while frozen at -20 ℃. The same amount (W / V) of purified water was added to the frozen Schisandra chinensis, and then crushed using a homogenizer, taking care not to crush the seeds.

To inoculate the fermentor (starter) for lactic fermentation, the shredded Schizandra solution was heat sterilized at 75 ° C. for 10 minutes to remove contaminants present in Schizandra pericarp. Enterococcus sp. TO-94 was diluted in physiological saline and inoculated with 8.0 cfu / ml and incubated at 37 ° C. for 72 hours. After incubation, the cells, seeds and unfermented by-products were removed by filtration (whatman No. 5) and the clarification supernatant was used as a fermentation product. At this time, the amount of fermentation broth was 160 g / Schisandra chinensis and 100 g, the yield was 160%.

Test Example 1 Measurement of pH and Acidity

< Enterococcus sp. Growth Curve of TO-94>

In general, lactic acid bacteria are microorganisms that ferment sugars to produce lactic acid, and are microorganisms that impart various functionalities such as inhibiting food spoilage and food poisoning causing bacteria. In the present invention, Lactic acid bacteria Enterococcus sp. TO-94 was inoculated in MRS medium and Schisandra chinensis broth, respectively, and cultured at 37 ° C., and the growth of bacteria was compared at 4 hours, and the results are shown in FIG. 3. ].

In the results of FIG. 3, the logarithmic growth phase started from 4 hours of inoculation at the time of growth in MRS medium and reached the stop phase 12 hours later. On the other hand, when inoculated with Schizandra chinensis solution, the induction period lasted up to 8 hours, and the logarithmic growth phase also lasted up to 20 hours. This means that the content of sugar used for fermentation is 4.05% (Hyun HK, et. al ., Korean J. plant Res ., 2002, 15: 1-7) The fruit itself contains little fermentable sugar and there are few ingredients that can be used as a nutrient source. It seems to increase.

<pH measurement>

PH was measured every 4 hours during the fermentation process of Example 3 by using a pH meter (sp-701, Suntex Inc. Korea), the acidity was 20 mL of the fermentation broth by the AOAC method (#) until the pH reached 8.3 After titration with 0.1 N NaOH solution, 0.1 N NaOH requirement was converted to lactic acid content (%). No significant changes in pH and acidity were observed.

Organic acid measurement

In addition, the measurement of organic acids in Schisandra chinensis fertilizer was carried out in ACN / DW (1: 1) 10 in a C18 cartridge according to the 2008 Health Functional Food Code (Lee JS, et al., J. Microbiology , 1996, 34: 225-228). 10 ml of the fermentation broth was added to remove 4-5 ml of the initial eluate, and then measured using a Shiseido HPLC system (Shiseido, Tokyo, Japan). Analysis conditions of the organic acid are shown in Table 2 below.

column Prevail Organic acid, 5 μm (4.6 × 250 mm) Temperature 40 ℃ Injection volume 10 μL Mobile phase 25 mM Kh2PO4 pH 2.5 Flow rate 1 ml / min Detector PDA detector (210 nm)

Enterococcus sp. The results of measuring the change in the content of organic acid during the fermentation process of Schizandra chinensis by TO-94 are shown in Table 3 below.

Organic acid Content (mg / g) 0 hours later After 20 hours Oxalic acid 0.18 0.03 L-tartaric acid Not detected 1.83 Formic acid 2.20 Not detected L-malic acid 18.91 14.17 L-lactic acid Not detected 9.84 Acetic acid 0.38 2.08 Citric acid 37.41 36.35 Suche mountain Not detected Not detected

As can be seen from the results of Table 3, changes in various organic acids were measured during the fermentation process. The main product of lactic acid increased from 0 to 9.84 mg / g, and specifically the acetic acid and L-tartaric acid contents also increased from 0.38 to 2.08 mg / g and from 0 to 1.83 mg / g. On the other hand, the content of oxalic acid, formic acid and malic acid was reduced. In addition, citric acid did not show a change. The results showed that the content was changed without a big change in the overall acidity.

Test Example 2 Measurement of Sugar and Free Sugar

The fermentation broth was taken every 4 hours in the fermentation process of Example 3, and the sugar content was measured using a sugar meter (Master-M, Atage, Tokyo, Japan), and the free sugar was filtered through a membrane filter (0.45 μM) and then Shiseido HPLC system (Shiseido). ; Tokyo, Japan). Analysis conditions of the free sugars are shown in Table 4 below.

column Asahipack NH2P-50 4E (4.6 × 250 mm) Temperature 35 ℃ Injection volume 20 μL Mobile phase 78% ACN Flow rate 1 ml / min Detector RID

Enterococcus sp. The result of measuring the change of free sugar content and total sugar content during the fermentation of Schizandra chinensis by TO-94 is shown in FIG. 4. [■: maltose, ▲: sucrose, ●: glucose, *: fructose, and ◇: total Sugar content].

In the results of FIG. 4, the content of fructose and glucose increased to 5.83 and 4.30 mg / g after 20 hours of fermentation, while the sucrose and maltose of disaccharides increased to 6.0 and 3.0 mg / g after 8 hours of fermentation. After 20 hours of fermentation they were reduced to 0.35 and 0.90 mg / g, respectively. This was initially done with Enterococcus sp. Due to the degradation of non-fermentable sugars by TO-94, an increase in disaccharides was observed, which is believed to have been used as a nutrient source for bacterial growth. In addition, the total sugar content increased about 5 times from 1.8 Brix to 10.2 Brix. This is expected to play a big role in the development of Schisandra fermented food, which is difficult to develop fermented food due to lack of carbon source.

Test Example 3 Measurement of Vitamin

Vitamins of Schisandra chinensis were measured using a Shiseido HPLC system (Shiseido, Tokyo, Japan) in accordance with the Healthy Functional Food Code Trace Component Test Method. Analytical conditions of vitamins are shown in Table 5 below.

column Capsule pack (Capcell pak) C18 UG 120 (3.0 * 250mm) Temperature 40 ℃ Injection volume 10 μL Mobile phase Vitamin B ₁ , B ₆ , Nicine, Folic Acid A: DW 957 ml + MeOH 25 ml + PicB7 20 ml
B: 500 ml DW + 500 ml MeOH + 20 ml PicB7 Vitamin B ₂ A: 10 ml NaH ₂ PO ₄ in DW (1 L) (pH 5.5)
B: MeOH A: B = 78: 22 Pantothenic Acid (Vitamin B ₅ ), Vitamin C 20 ml NaH ₂ PO ₄ (pH 2.1): ACN = 98: 2 Cyanocobalamin Biotin
(Cyanocobalamin Biotine) A: 5 mM KH ₂ PO ₄ + 0.1% phosphoric acid
B: 5 mM KH ₂ PO ₄ / MeOH = 80/20
A: B = 500: 10 Vitamin K MeOH Flow rate 1 ml / min Detector PDA detectors (210 nm, 248 nm) and UV detectors (200 nm, 270 nm)

Enterococcus sp. Table 6 shows the results of measuring the vitamin content change during the fermentation process of Schizandra chinensis by TO-94.

Vitamin types Content (mg / 100g) 0 hours later After 20 hours B ₁ 0.19 0.52 B ₂ 0.024 0.08 B ₅ 3.5 Not detected B ₆ Not detected Not detected B ₇ Not detected Not detected Nisin 0.83 1.44 Folic acid 502,05 901.26 B ₁₂ Not detected Not detected C 12.65 38.0 K ₁ Not detected Not detected β-carotene Not detected Not detected

In the results of Table 6, vitamin B ₁ , B ₂ , niacin, folic acid and C predominantly and all the vitamins increased about 1.5 to 3 times compared to before fermentation. The content before fermentation is ground juice and is considered to be related to the efficiency of dissolution. Enterococcus sp. During the fermentation of Schizandra chinensis by TO-94, it is believed that the content increased due to the increase of elution efficiency due to the decomposition of the fibers constituting the cell wall.

Schisandra chinensis mainly consists of hot water, water and alcohol extraction (Kim SI,et al ., The Korean journal of food and nutrition, 2009, 22: 41-47; Choi EO,et al ., Korean journal of biotechnology and bioengineering, 2008, 23: 271-275; Kwon HJ,et al ., Korean journal of food preservation, 2008, 15: 587-592; Kim HK,et al ., Journal of the Korean society of food culture, 2004, 19: 484-490; And Kim SI,et al ., Korean J Food and nutr., 2009, 22: 41-47) Examples of vitamin content are difficult to find. It is believed that this is due to the loss of solubility and the loss of the vitamin by heating and solvent. Therefore, the fermentation method by lactic acid bacteria having a fiber protection ability is considered to be useful for the development of nutritionally improved Schizandra chinensis.

Test Example 4 Changes in Polyphenol and Anthocyanin Contents during Fermentation

<Measurement of Total Phenolic Compound>

To 5 mL of the 10-fold dilution of the Schizandra fermented product of Example 3, 5 mL of Folin-Ciocalteu reagent was added and mixed. After 3 minutes, 5 mL of 10% Na ₂ CO ₃ solution (w / v) was added thereto, followed by shaking. Then colorimetric was determined at 700 nm. The total phenolic compound content was measured from a calibration curve using tannic acid as a standard.

<Anthocyanin measurement>

10 mL of the Schisandra fermentation of Example 3 was mixed in 40 mL of 0.1% HCl-80% MeOH solution (v / v) and shaken for 24 hours. The extracted pigment was subjected to centrifugation at 3,000 g x 10 minutes, and the supernatant obtained by 0.1% HCl-80% MeOH solution (v / v) was applied to 100 mL and colorimetrically determined at 528 nm. Anthocyanin content was measured from a calibration curve prepared with cyanidine as a standard.

Enterococcus sp. The results of measuring the change in the content of polyphenols and anthocyanins during the fermentation of Schizandra chinensis by TO-94 are shown in Table 7 below.

Organic acid content 0 hours later After 20 hours Polyphenol (%) 0.211 0.823 Anthocyanin (μg / mL) 45.23 51.40

In the results of Table 7, the polyphenol increased from about 0.211% at the beginning of fermentation to about 0.823% after 20 hours, and about 3.8 times from anthocyanin to about 1.2 times from 45.23 μg / ml to 51.40 μg / ml.

Korea Microbial Conservation Center (Domestic) KFCC11477 20100112

Attach an electronic file to a sequence list

Claims (3)

Enterococcus sp. Of Accession No. KFCC11477P. Schisandra fermented with TO-94. 1) crushing using a homogenizer after adding the same amount (v / v) of water to Schizandra;
2) heat sterilizing the lysate of 1) at 70 to 75 ° C. for 5 to 10 minutes;
3) Enterococcus sp. Of accession number KFCC11477P. Preparing a diluted solution of TO-94 diluted in physiological saline; And
4) mixing the dilution of 3) to the sterile lysate of 2) and incubating at 37 ° C. for 72 hours;
Fermentation of Schizandra chinensis through. A food composition containing the Schisandra chinensis according to claim 1.

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