KR20090039941A - Function-related fermentation raw materials using the composition which mixed chlorella in rubus coreanus and method to produce thereof - Google Patents

Abstract

A method for preparing functional fermented material using Korean raspberry and chlorella mixture is provided to allow functional food products to be manufactured using the functional fermented materials, thereby enabling a consumer to easily obtain medicinal effects of chlorella. A method for preparing functional fermented material comprises the following steps of: inoculating fermented microorganisms to a mixture of 95~50wt% of Korean raspberry and 5~50wt% of chlorella; and aging the inoculated mixture at 30~40°C for 3~7 days. The Korean raspberry represents Korean raspberry itself, frozen Korean raspberry or Korean raspberry juice. The fermented microorganisms represent lactic acid bacteria, Bacillus subtilis or a mixed strain thereof. The prepared functional fermented material has a formulation of powder, liquid or viscous liquid.

Description

Functional-related fermentation raw materials using the composition which mixed chlorella in Rubus coreanus and Method to produce Technical}

The present invention relates to a functional fermentation raw material using a bokbunja and chlorella mixture, and to a method for producing the same, and more specifically, to a fermented product to be fermented for a predetermined time by inoculating fermentation microorganisms in the bokbunja in an appropriate amount, The present invention relates to a functional fermentation raw material using a bokbunja and chlorella mixture which is processed into a liquid phase and a method for producing the same.

Bokbunja is an old document named Myeong-seok-ri, which is called "Agamgam Pyeong-ju Jukyeong-Gyeonggi-Bubok Owolchae". Since it is written, the herbal literature has been used to treat symptoms such as Yangyang, Yujeong, Hanseol, Urinary, Excessive urination, and infertility. It is written. What is Bokbunja? If you type lobe honor based on a recent yeongumul of such a report, a comprehensive reference of the Enlightenment, such as form and color, maturity period of the fruit was historical research to say bokbunja berries Rubus immature fruit of coreanus Miq. Belong to the rose family Rosaceae Strawberry in Rubus . Fruits of the same genus that have a flexible relationship with bokbunja are used or mixed with bokbunja depending on the region, and wild cog plants have 50 kinds of plants, 27 kinds of Japan, 22 kinds of Korea, and fruit of plants of Rubus genus in USA and Europe. Collected as raspberry, more than 400 species of this genus have been investigated.

The plant used as bokbunja has been called 樹 掌 覆盆 葉 覆盆子Rubus since the Qing Dynasty. chingii Hu., and R. palmatus Thunb., R. idaeus L., R. corchorifolius L., R. ieaeopsis Forke, R. foliolosus D. Don, R. lasiostylus Forke, R. crataegilifolius Bunge , R. idaeus var. concolor is used.

Bokbunja ( Rubus coreanus Miq.) has been used in Korea as a medicinal or old plant, and started to grow in the vicinity of Gochang Sununsa since the 1960s, and to support farmland income crops in 1989 and Gochang-gun subsidies in 1991. In 1995, the cultivation of Bokbunja cultivation technology at the Gochang-gun Agricultural Technology Center led to the establishment of Gochang-gun as a special crop for farm household income.

The national production and cultivation area of Bokbun was 1,300 MT / 703.21 ha in 2003, 2,750 MT / 654 ha in 2004, and 6,033 MT / 1,086 ha in 2005. In 2006, the cultivated area was approximately twice that of the previous year. Is estimated to be about 1.6 times 9,800 MT / 2,188 ha.

 If you look at the types of processed foods using Bokbunja, you can see Bokbunja Juice, Fermented Wine, Tea, Soup, Jelly, Gochujang, Cheonggukjang, etc., to solve the problems caused by the overproduction of Bokbunja. There is a need for research on ways to increase functionality with searched and already active ingredients.

Accordingly, an object of the present invention is to provide a functional fermentation raw material and a method for producing the same, in which the functionality is increased in the bokbunja by searching for a new functional substance and adding an active ingredient which has already been investigated.

Therefore, the present inventors have a high protein content microorganisms, and promote the growth of microorganisms, animal growth promoting effect, action as a physiologically active substance, plant hormone effect, netting action and chlorella growth factor (CGF) and medical treatment Effects include hepatic scar prevention, increased resistance to dysentery bacteria, the treatment of digestive ulcers, the promotion of intestinal fluid movement, the prevention of leukocyte reduction, the healing of wounds, the antiviral effect, the lowering of total cholesterol, the healing effect of milk allergy and the blood of children. Chlorella, a microalgae that has reported effects on protein composition, electrolyte and albumin content in the form of nutrients, is rich in nutrients. When fermented with mixed bokbunja fermentation to improve the functionality of the bokbunja And it was found that there is a supplemental effect (supplementary effect) of protein among the nutrients of bokbunja completed the present invention.

In order to solve the above problems, the present invention is inoculated with a mixture of 95-50% by weight of bokbunja and 5-50% by weight of chlorella to inoculate fermentation microorganisms, which are two strains of lactic acid bacteria, Bacillus subtilis or Bacillus subtilis and Lactobacillus at a temperature of 30-40 ° C. It provides a method for producing a functional fermentation raw material using a bokbunja and chlorella mixture, characterized in that it comprises a process for formulating the fermentation broth obtained in a liquid or powder form after fermentation for 3 to 7 days under the conditions.

In addition, the present invention provides a method for producing a functional fermentation raw material using bokbunja and chlorella mixture, characterized in that the bokbunja is natural bokbunja, frozen bokbunja or bokbunja juice.

In addition, the present invention, the lactic acid bacteria is Lactobacillus casei, Lactobacillus plantarium, Lactobacillus lactis, Lactobacillus asidophilus, Lactobacillus brevis, Lactobacillus vulgaris, Lactobacillus rhamnosus, Pediococcus pentosa A mixture of bokbunja and chlorella, characterized in that it is one or two or more mixed strains selected from the group consisting of Seutus, Streptococcus thermophilus, Bifidobacterium butineri, Bifidobacterium infantis, and Bifidobacterium longum. It provides a method for producing a functional fermentation raw material using.

In addition, the present invention is the Bacillus subtilis, Bacillus licheniformis, Bacillus natto, Bacillus sirius, Bacillus Spices (cam) and Bacillus Sushi (Ewha) is one or two or more mixed strains selected from the group consisting of It provides a method for producing a functional fermentation raw material using a bokbunja and chlorella mixture, characterized in that.

In addition, the present invention is a mixed strain of lactic acid bacteria and Bacillus subtilis mixed strains of Lactobacillus casei and Bacillus subtilis, mixed strains of Lactobacillus casei and Bacillus sp. (Cam), Lactobacillus casei and Bacillus sp. Group consisting of mixed strains of Lactobacillus plantarium and Bacillus subtilis, mixed strains of Lactobacillus plantarium and Bacillus sp. (Cam) or mixed strains of Lactobacillus plantarium and Bacillus sp. It provides a method for producing a functional fermentation raw material using a mixture of bokbunja and chlorella, characterized in that one species selected from.

In addition, the present invention is the fermentation microorganisms are cultured until the absorbance (Abs.) Measured at 620nm by the culture of the fermentation microbial bokbunja characterized in that inoculated in the bokbunja and chlorella mixtures It provides a method for producing a functional fermentation raw material using a chlorella mixture.

In addition, the present invention provides a method for producing a functional fermentation raw material using the bokbunja and chlorella mixture, characterized in that the inoculation amount of the fermentation microorganism is inoculated to be 2% (V / V) compared to the bokbunja and chlorella mixture.

In addition, the present invention is inoculated so that the fermentation microorganisms are two kinds of mixed strains, each of which is mixed with each other in a ratio of 1 to 1 (V / V) compared to the bokbunja and chlorella mixture It provides a method for producing a functional fermentation raw material using a bokbunja and chlorella mixture, characterized in that.

In addition, the present invention after the fermentation of the bokbunja and chlorella mixture pH range is 3 to 4, the acidity range is 1.9 to 2.7, the sugar range is 4.7 ~ 11.7 Brix functional fermentation raw material using the bokbunja and chlorella mixture, characterized in that It provides a method of manufacturing.

In another aspect, the present invention provides a method for producing a functional fermentation raw material using the bokbunja and chlorella mixture, characterized in that the bokbunja juice to make the bokbunja liquid by mixing the bokbunja and mixed chlorella in the bokbunja liquid.

In addition, the present invention provides a functional fermentation raw material using the bokbunja and chlorella mixture prepared by any one of the above-described manufacturing method.

In addition, the present invention provides a functional fermentation raw material using the bokbunja and chlorella mixture, characterized in that the form of the functional fermentation powder, liquid or mucus.

Fermentation raw materials and a method for producing the same using a mixture of chlorella mixed with bokbunja or bokbunja juice according to the present invention has the following effects.

1. The main consumption type of Bokbunja is the main form of fruit juice and wine, and it is limited to other kinds of soybeans and cereal products, such as jangjang, cracked rice cake, rice cake, and noodles, and it is a fermented raw material according to the fermentation raw material manufacturing method according to the present invention. New functional foods can be developed using.

2. By establishing a process for fermenting bokbunja and chlorella mixtures and selecting good microorganisms, new product can be developed by using selected strains when mixing fermented bokbunja and other raw materials to develop fermented food.

3. It is possible to utilize technology in developing high value-added products by utilizing resources with high protein content.

4. The evaluation method of bioactive function of mixed fermented bokbunja and chlorella can be applied to other fermented foods.

5. Fermentation of fruits and vegetables can also be applied to the development of new products by applying the fermentation technology developed in this study.

6. The technique can be applied to the fermentation of fruits of plants that have a flexible relationship with bokbunja.

7. Utilizing the fermentation technology (temperature control, pH control, microbial strain control, fermentation time) accumulated by fermenting Bokbunja and Chlorella mixtures, the new health functional foods will be improved by improving the fermentation management ability using other materials that replace Chlorella. Can be used to develop

8. It is possible to use fermentation technology developed by this research as a new well-being food development model to develop new products by combining bokbunja with new functional materials.

9. It can be used to evaluate the biological activity of new products developed by other technologies by introducing the biological activity evaluation technology of this research and development product.

10. It is effective in contributing to the development of the food industry by developing and producing new types of fermented foods with enhanced functionality by combining with other functional food materials other than Bokbunja.

Hereinafter, the present invention will be described in more detail with reference to the accompanying drawings.

1 is a graph showing the change in moisture content according to the mixing ratio of frozen bokbunja and bokbunja juice and chlorella, Figure 2 shows the culture results of lactic acid bacteria and Bacillus subtilis for the change in water content according to the mixing ratio of frozen bokbunja and bokbunja juice and chlorella. Figure 3a to 3j is a graph showing the acid production amount of lactic acid bacteria according to the chlorella addition level of lactic acid bacteria used in the present invention, Figures 4a to 4h of the lactic acid bacteria according to the chlorella addition level of Bacillus subtilis used in the present invention 5A to 5K are graphs showing the amount of acid production, and Fig. 5A to 5K are graphs showing the acid production amount change of the selection strain according to the level of chlorella added to bokbunja according to one embodiment of the present invention, and Figs. 6A to 6F illustrate one embodiment of the present invention. It is a graph showing the change of lactic acid production amount according to temperature of the bokbunja and chlorella mixture according to the example, Figure 7a to 7d is an embodiment of the present invention As a graph showing a change in pH by production of lactic acid mixture according to bokbunja and Chlorella, it will be described in detail the present invention with reference to these figures.

The general components of bokbunja and chlorella used in the present invention are shown in Table 1, and analyzed according to the Food Code Analysis Method (Food and Drug Administration, 2005).

The general components of Bokbunja of Table 1 are crude protein 1.13%, carbohydrate 8.0%, crude fat 3.30%, moisture 83.5%, ash 4.0%, the general components of chlorella are crude protein 49.49%, carbohydrates 29.1%, crude fat 7.90%, moisture 7.4%, Since the level of ash was 8.60%, it was thought that chlorella was added to the bokbunja.

On the basis of these facts, it is thought that supplementing the nutrients lacked in bokbunja could be expected when using bokbunja as a main ingredient and chlorella as an ingredient. It was considered possible to develop.

The present invention is to prepare a bokbunja and chlorella mixture by mixing chlorella in appropriate amounts based on the general ingredients of bokbunja and chlorella, and inoculate the fermentation microorganisms, which are lactic acid bacteria, Bacillus subtilis or two mixed strains of Bacillus and Lactobacillus. By fermentation 3 to 7 days under the conditions of the temperature of 30 ~ 40 ℃ to prepare a fermented product, by processing the fermented product in the form of powder or liquid to produce a functional fermented material.

According to the present invention, the pH suitable for the lactic acid production of the bokbunja and chlorella mixture after inoculation of the fermentation microorganism is preferably 4 to 6, and after mixing the bokbunja and chlorella, the pH is measured by a pH meter and then adjusted by a conventional pH adjustment method. Can be adjusted.

The mixed amount of bokbunja and chlorella is mixed with 95-50% by weight of bokbunja and 5-50% by weight of chlorella. More preferably, it is more preferable to mix 95-65 weight% of bokbunja and 5-35 weight% of chlorella. More preferably, 70 to 65% by weight of bokbunja and 30 to 35% by weight are preferably mixed. If the mixing amount of the chlorella exceeds 50%, there is a disadvantage in that the content of the natural odor and ingredients of the chlorella is increased to reduce palatability, and when the addition of less than 5%, the nutrition to be obtained in the present invention. There is a disadvantage that can not expect the synergistic effect of the components. The bokbunja uses a natural bokbunja, frozen bokbunja or bokbunja juice, preferably lyophilized bokbunja. The reason is that the hot air drying has the advantage of reducing the production cost, but since the color of the product is dark green in appearance after the mixture fermentation has the disadvantage that it looks black.

The fermentation microorganism is fermented using lactic acid bacteria, Bacillus subtilis or mixed strains mixed with lactic acid bacteria and Bacillus subtilis, the lactic acid bacteria is Lactobacillus casei (hereinafter referred to as " L. casei "), Lactobacillus plantarium ( Lactobacillus plantarium, hereinafter referred to as "L. plantarium"), Lactobacillus lactis (Lactobacillus lactis , hereinafter referred to as “ L. lactis ”), Lactobacillus acidophilus , hereinafter referred to as “ L. acidophilus ”), Lactobacillus brevis , hereinafter referred to as " L. brevis "), Lactobacillus bulgaricus (hereinafter referred to as " L. bulgaricus "), Lactobacillus rhamnosus ( Lactobacillus) rhamnosus , hereinafter referred to as “ L. rhamnosus ”, Pediococcus pentosaceus pentosaseus , hereinafter, “ Pe. pentosaseus ”, Streptococcus thermophillus , hereinafter, “ St. thermophillus ”, Bifidobacterium ( Bifidobacterium) buchneri , hereinafter, “ Bi . buchneri ”, Bifidobacterium infantis , hereinafter, “ Bi . La infantis "and) rongum Bifidobacterium (Bifidobacterium longum , hereinafter “ Bi . longum ”) and one or two or more mixed strains selected from the group consisting of Lactobacillus casei, Lactobacillus plantarium, Lactobacillus lactis, Lactobacillus asidophilus, Use of one or two or more mixed strains selected from the group consisting of Lactobacillus brevis, Lactobacillus vulgaris, Lactobacillus rhamnosus, Pediococcus pentosaceus, Streptococcus thermophyllus or Bifidobacterium longum Preferably, it is preferable to use one or two or more mixed strains selected from the group consisting of Lactobacillus cassia, Lactobacillus plantarium, Lactobacillus lactis or Lactobacillus asidophilus.

The Bacillus subtilis Bacillus subtillis , hereinafter referred to as " B. subtillis ", Bacillus licheniformis (hereinafter referred to as " B. lichenifomis "), Bacillus natto ( Bacillus) natto , hereinafter referred to as “ B. natto ”), Bacillus cereus, hereinafter referred to as "B. cereus" referred to), Bacillus species (CAMS) (Bacillus species (cham), hereinafter referred to as "chem") and Bacillus RY's (Rika) (Bacillus species (ewha), hereinafter referred to as " ewha ") is preferably used one or two or more mixed strains selected from the group consisting of, preferably ewha, cham, B. subtillis Or B. natto , it is preferable to use one or two or more mixed strains selected from the group consisting of.

The mixed strain of lactic acid bacteria and Bacillus subtilis is a mixed strain of Lactobacillus casei and Bacillus subtilis, a mixed strain of Lactobacillus casei and Bacillus fish (cam), a mixed strain of Lactobacillus casei and Bacillus Spices (Ewha), Lactobacillus One selected from the group consisting of mixed strains of Bacillus plantarium and Bacillus subtilis, mixed strains of Lactobacillus plantarium and Bacillus sp. (Cam) or mixed strains of Lactobacillus plantarium and Bacillus sp. It is preferable to use.

According to the present invention, the inoculation amount of the fermentation microorganisms is preferably inoculated to be 2% (V / V) compared to the mixture, and when the fermentation microorganisms are two kinds of mixed strains, each of the bacterial solutions is mixed at a ratio of 1: 1. It is preferable to inoculate each microbial fluid constituting the mixed strain to be 1% (V / V) of the bokbunja and chlorella mixture, the final inoculation amount of the fermentation microorganisms compared to the bokbunja and chlorella mixture is 2% ( V / V) is preferably inoculated.

According to the present invention, the fermentation period after the appropriate amount of chlorella is mixed with the bokbunja and inoculated with the fermentation microorganism is preferably fermented for 3 to 7 days depending on the characteristics of the inoculated strain.

Hereinafter, the present invention will be described with reference to reference examples and preferred embodiments as follows, but the present invention is not limited to the reference examples and examples disclosed below.

[Test method used in the present invention]

end. Common Ingredients of Bokbunja and Chlorella

General components of bokbunja and chlorella are analyzed for moisture, crude protein, crude fat, crude fiber, crude ash according to AOAC method (36). Moisture was analyzed by 105 ° C atmospheric pressure drying, crude protein was analyzed by semimicro Kjeldahl method, crude fat by ether extraction method, and crude ash by 550 ° C incinerator (37). Crude fiber was analyzed by fritted glass crucible method (38), total dietary fiber (39), total sugar and reducing sugar by DNS (3,5-dinitro-salicylic acid) method (40).

I. Mixing Ratio and Water Content of Bokbunja and Chlorella

After mixing chlorella (5% water) to 5, 10, 20, 30, 35, 40, 50% to Bokbunja (87% water, W / W) and Bokbunja juice (W / V) The content was investigated.

All. Fermentation Characteristics of Bokbunja and Chlorella Mixtures

1) Selection of Microorganisms Suitable for Fermentation of Bokbunja and Chlorella Mixtures

12 kinds of Lactobacillus suitable for fermentation after increasing the mixed group of bokbunja and chlorella casei KCTC 3109 ( L. casei ) , Lactobacillus plantarium KCTC 3108 ( L. plantarium ), Lactobacillus lactis KCCM 11357 ( L. lactis ), Lactobacillus acidophilus KCTC 3111 ( L. acidophilus ), Lactobacillus brevis KCTC 3102 ( L. brevis ), Lactobacillus bulgaricus KCTC 3635 ( L. bulgaricus ), Lactobacillus rhamnosus ( L. rhamnosus ) , Pediococcus pentosaseus KCTC 3507 (Pe. Pentosaseus), Streptococcus thermophillus KCTC 2185 (St. Thermophillus), Bifidobacterium buchneri (Bi. buchneri), Bifidobacterium infantis KCCM 11207 ( Bi . Infantis ), Bifidobacterium longum KCCM 11953 ( Bi . Longum ) and Bacillus sp. wild type A 051221 ( Bacillus species (Cham)), Bacillus subtillis KCTC 3239 ( B. subtillis 3239), Bacillus sp. wild type B 050803 ( Bacillus species (Ewha)), Bacillus licheniformis KCTC 3049 ( B. lichenifomis 3049), Bacillus subtillis KCTC 1028 ( B. subtillis 1028), Bacillus subtillis var. natto 051028 ( B. natto ), Bacillus licheniformis KCTC 1831 ( B. licheniformis 1831) and Bacillus are food hygiene problems Cultivated cereus 051206 ( B. cereus ) and multiplying the bacteria until the absorbance (Abs.) measured at 620 nm reached 1.0, resulting in 2% (v / v) for single culture and 2 for mixed culture, respectively. 1% (v / v) was added to inoculate the cells to add up to 2% (v / v), and the amount of acid produced during the fermentation and the growth rate of the cells were examined to select suitable strains for fermentation of bokbunja and chlorella. It was.

2) optimum temperature

In order to investigate the fermentation temperature of the mixture of bokbunja and chlorella, the fermentation temperature range and the optimum temperature were determined by analyzing the amount of acid produced by 3 days incubation at 20, 25, 30, 35, 40, 45 ℃ conditions.

3) Optimum pH

In order to investigate the pH suitable for the growth and acid production of the strains used for fermentation of bokbunja and chlorella mixtures, the pH range and the optimum pH for fermentation were analyzed as a result of analysis of acid production after 4 days in 4, 5, 6 and 7 conditions. pH was determined.

la. Components and Biological Activities of Fermented Products

1) General Ingredient

General components of mixed bokbunja and chlorella fermentation products are analyzed for moisture, crude protein, crude fat, crude fiber, total dietary fiber, crude ash according to AOAC method (36). Moisture was analyzed by 105 ° C atmospheric pressure drying, crude protein was analyzed by semimicro Kjeldahl method, crude fat by ether extraction method, and crude ash by 550 ° C incinerator (37). Crude fibers were analyzed by fritted glass crucible method (38), and total and reducing sugars were analyzed by DNS (3,5-dinitrosalicylic acid) method (40).

2) pH, acidity, soluble solids and chromaticity

pH and acidity were measured by using a fermented product mixed with bokbunja and chlorella as a sample, and pH was measured by using a pH meter (Mettler-Toledo AG, Switzerland). Acidity was measured by 0.1N NaOH consumption required for pH value of 8.2. The solution is converted into lactic acid and expressed as the total acid content (%). Soluble solids and chromaticity are distilled water diluted with 1: 1 distilled water and filtered. Soluble solids are measured with a digital photometer (PR-32, Atago, Co., Japan), and chromaticity is measured with a colorimeter (JX-777). , Japan) and measure the L, a, b values. In this case, a standard color plate having a value of L: 92.68, a: 0.83, and ｂ: 0.85 was used.

3) free sugar content

For free sugar analysis of dried bokbunja and chlorella fermented products, extract the sample according to AOAC method (36), allow it to cool to room temperature, add ethanol to the initial volume (weight), and filter the extract with 0.45㎛ membrane filter. One filtrate is separated and quantified by HPLC.

4) Amino Acid Content

Constituent amino acid is accurately dried 500mg of dried bokbunja and chlorella fermented product, put into ampule, 15ml of 6N-HCl is added, and it is rapidly sealed by injecting nitrogen gas. After hydrolysis in 110 ℃ oven for 24 hours, it is cooled and dehydrated. 50 mL of deionized water was used, filtered with a 0.2 μm membrane filter, diluted appropriately, separated and quantified by HPLC according to AccQ-Tag's method (41). Amino acid standards (Wako, type H, Japan) are prepared to 0.125 μmol / ml using 0.1 N-HCl as a solvent.

5) organic acid content

The organic acid of the dried bokbunja and chlorella fermented products is pretreated with the sample according to the method of Cristina and Luh (42). That is, 100 ml of 50% ethanol was added to 3 g of the sample, stirred at 150 rpm for about 2 hours, filtered and concentrated under reduced pressure to remove ethanol. This is passed through an amberlite IRA 95 anion exchange resin (Biored, USA) to adsorb organic acids and the resin is washed with about 100 ml of ion exchange water. The adsorbed organic acid was liberated with 6N formic acid, dried under reduced pressure, 10 mL of ion-exchanged water was added thereto, mixed well, filtered through a 0.45 ㎛ membrane filter, separated and quantified by HPLC. Citric acid, lactic acid, formic acid, acetic acid, propionic acid, tartaric acid, malic acid, and succinic acid (Sigma Co., USA) are used as organic acid standards.

6) mineral content

Minerals are analyzed using dried bokbunja and chlorella fermented products as samples. Samples were taken in preliminary crucibles and preliminarily carbonized and incubated in a 560 ° C. incineration furnace until they were white or greyish white. The ashed ash was soaked with a small amount of ion-exchanged water to prevent the ash from scattering, and 5 ml of hydrochloric acid solution (hydrochloric acid: ion-exchanged water = 1: 1) was added and evaporated to dryness on a hot plate, followed by 5 ml of hydrochloric acid solution (hydrochloric acid: Ion-exchanged water = 1: 3) was added thereto, dissolved in heat for 5 minutes, and filtered to make 100 ml. 5 ml of this solution was taken in a 25 ml volumetric flask, and 5 ml of 5% La ₂ O ₃ solution was added to remove the effects of co-existence ions, and then ICP (Inductively Coupled Plasma, Jobin Yvon Co., France) to quantify and analyze.

7) Microbial count

The total bacterial counts of mixed bokbunja and chlorella fermentation products were dispensed by 1 ml of each sample diluted from 10 ⁰ to 10 ⁹ using plate count ager (PCA) medium, followed by standard agar culture (43), and overnight at 37 ° C. After incubation, count. Lactic acid bacteria were counted after incubation at 37 ° C. for 24 to 72 hours using Rogosa SL agar (Difco Co., MI USA) medium adjusted to a final pH of 5.5 by adding 0.133% acetic acid.

8) antimicrobial activity

Escherichia coli 0-157L: H7 , Bacillus cereus , Salmonella Typhimurium is used as the assay strain. The antimicrobial assay was inoculated with 1 ml of platinum in 5 ml of liquid medium and incubated for 1 to 2 days, and then mixed with bokbunja and chlorella fermented products and 0.1 ml of the culture of the assay in a test tube containing 5 ml of fresh medium. After incubating at 37 ° C. for 24 hours, the number of viable cells is measured by standard plate culture. The antimicrobial effect was confirmed by the following formula using the culture solution of the assay bacteria without the added bokbunja sample.

9) Determination of total phenolic compound content

The total phenolic compound content of bokbunja and chlorella mixed fermented product extracts was measured by colorimetric method by Folin-Denis method (44-46). That is, 5 ml of the Folin-Ciocalteu reagent was added to 5 ml of the sample solution diluted 100-fold and mixed. After 3 minutes, 5 ml of 10% Na ₂ CO ₃ was added thereto, shaken and left at room temperature for 1 hour to measure absorbance at 700 nm. . The control was treated in the same way with distilled water instead of the sample solution. At this time, tannic acid is prepared at a concentration of 5 to 50 µg / ml as a standard substance and used to prepare a calibration curve.

10) Measurement of electron donating ability

The electron donating ability (EDA) test of bokbunja and chlorella fermented product extracts was measured by the method (47) using 1,1-diphenyl-2-picrylhydrazyl (DPPH). In other words, dissolve 12 mg of DPPH reagent in 100 ml of absolute ethanol, add 100 ml of distilled water, adjust the absorbance of the DPPH solution to about 1.0 at 517 nm with 50% ethanol solution blank, and then add 5 ml of this solution and 0.5 ml of sample solution. 30 seconds after the addition and mixing, the absorbance at 517 nm is measured. In this case, the electron donating ability was expressed as a percentage (%) of the difference in absorbance between the sample addition and the no addition.

11) Nitrite Scavenging Activity

To investigate the effects of bokbunja and chlorella fermented product extracts on the nitrite, a precursor of carcinogenic nitrosamine formation, to decompose or decompose, color analysis was performed at 520 nm by Do et al. And Gray (48-49). That is, a mixed solution of bokbunja and chlorella fermented product of a predetermined concentration was added to 1 ml of 1 mM NaNO ₂ solution, and 0.1 N HCl (pH 1.2) and 0.2 M citric acid buffer (pH 3.0, 4.2 and 6.0) were used. The pH of the solution was adjusted to 1.2, 3.0, 4.2 and 6.0 to make the volume of the reaction solution 10 ml. Take 1 ml each of the reaction solution obtained by reacting it at 37 ° C for 1 hour, add 5 ml of 2% acetic acid solution, and add 1% sulfanilic acid and 1% α- 0.4 ml of naphtylamine was mixed in a 1: 1 ratio, and mixed immediately before use. The mixture was left to stand at room temperature for 15 minutes, and the absorbance was measured at 520 nm using a spectrophotometer (Shimadzu UV-1601PC, Japan). The nitrite scavenging rate is calculated by the following equation.

12) Sensory test

The sensory evaluation was conducted using a nine-point grading method. The scoring criteria are very strong (very good): 9 points, moderately strong (good): 7 points, moderate (not good or bad): 5 points, moderately low (bad): 3 points, very weak ( Bad): 1 point. The sensory test was performed by Duncan's multiple test using Excel software (Microsoft Corp., Redmond, USA).

[Reference Example 1] Water content and fermentation level according to the mixing ratio of chlorella

Moisture was quantified by the combination of Table 2 in order to investigate the moisture level suitable for fermentation by examining the moisture content of Bokbunja fruit, Bokbunja juice, Chlorella, Bokbunja fruit + Chlorella mixture and Bokbunja juice + Chlorella mixture. 2, as shown in FIG. 1 and FIG.

As shown in Table 2, Figures 1 and 2, the water content tends to decrease as the mixing ratio of chlorella increases, but when inoculated with lactic acid bacteria and Bacillus subtilis, the effect of water content on the growth of the cells is considered to be insignificant.

Reference Example 2 Selection of Lactic Acid Bacteria with Excellent Fermentation Capacity by Addition of Chlorella

3A to 3J show that L. plantarium > L. casei > L. lactis were excellent in lactic acid production after adding chlorella to 0.5 and 1.0% of MRS medium and inoculating 10 lactic acid bacteria, respectively. L. acidophilus > L. brevis > Pe. pentosaseus> L. bulgaricus> St. thermophillus > L. rhamnosus > Bi. I can see that it is in longum order. Among these strains, L. plantarium , L. casei , L. acidophilus and L. lactis showed more than 1% of lactic acid production after 24 hours, and among them, L. plantarium and L. casei were selected It was.

Division such as 33 St. The addition of chlorella extract at a level of 0.25% for yogurt preparation with mixed strains of thermophillus and L. casei was effective in lowering pH and growing bacteria. 0.5% chlorella extract was added to the yogurt preparation . Incubation with only thermophillus showed the lowest pH and optimum titer after 9 hours. L. acidophilus showed a significant decrease in pH at 9 hours for 1% chlorella extract, but 0.5% chlorella extract after 24 hours. the pH and the acid production was good, L. casei, if any phrase is added to the chlorella extract large width is decreased in pH to the culture followed by 6 hours after 24 hours of incubation the culture was added 0.5% chlorella extract sphere acid production, L. bulgaricus also Incubated with 0.5% chlorella extract, the pH was lowered and titratable acidity increased after 9 hours . Although the individual growth characteristics of thermophillus , L. acidophilus , L. casei and L. bulgaricus were slightly different, they all promoted the growth of lactic acid bacteria and promoted the production of acid. This shortening time is shortened by the first, fermented by adding chlorella extract to skim milk powder, lactic acid bacteria can easily use the nutrient source, and secondly, when the fermented by adding chlorella powder directly, lactic acid bacteria are used as a nutrient source chlorella extract It was thought to be a delay.

Reference Example 3 Selection of Lactic Acid Bacteria with Excellent Fermentation Capacity According to Chlorella Addition Level

As shown in Figures 4a to 4f was added to the Chlorella 0.5, 1.0% in MRS medium and inoculated 8 kinds of Bacillus subtilis individually, as a result of the acid-producing bacteria Bacillus species (ewha)> Bacillus species (cham ), B. subtilis > B. licheniformis > B. natto . Among them, B. subtilis , Bacillus species (cham), and Bacillus species (ewha) were selected as fungi.

In 2005, the Korea Food and Drug Administration set the allowable level of B. cereus below 10,000 CFU / g of fermented products, because when mixed with fermented products during the selection process of excellent subtilis, As a result of examining the growth pattern of Bacillus cereus which is a problem, acid production was the highest after 12 hours after inoculation, but gradually decreased afterwards, which can be interpreted as a result of weakening of the activity of bacteria. If so, the hygiene risk was considered low.

[Examples 1 to 11]

L. plantarium and L. casei and Bacillus species (ewha), Bacillus species (cham), B. lactic acid bacteria were selected by adding the chlorella 5, 10, 20% to the bokbunja and selected according to the results of Reference Examples 2 and 3 . subtilis and mixed strains made of these two strain combinations were inoculated to 2% (V / V) compared to the bokbunja and chlorella mixture, and then cultured, and the lactic acid production of the bacteria was examined. The results are shown in FIGS. Lactic acid production was higher in L. planti than in L. casei , followed by B. subtilis < Bacillus species (cham) < Bacillus species (ewha). The yield of lactic acid in the combination of two strains was also higher than that of L. casei + B. subtilis , L. casei + Bacillus species (cham), and L. casei + Bacillus species (ewha). L. plantarium + B. subtilis , L. plantarium + Bacillus higher in species (cham) and L. plantarium + Bacillus species (ewha), in order of L. plantarium + B. subtilis < L. plantarium + Bacillus species (cham) < L. plantarium + Bacillus species (ewha) The combination of relatively good lactic acid production was confirmed to be L. plantarium + Bacillus species (ewha).

On the other hand, the two strain combinations complement each other, namely L. plantarium Since the production of lactic acid is increased and Bacillus species (ewha) has a small amount of lactic acid secreted but secretes fibrin and protease, it is expected that the combination of two strains will complement each other. Inoculation fermentation was expected to increase digestive absorption of bokbunja and chlorella mixture.

[Examples 12 to 17]

After setting the temperature of Bokbunja and Chlorella mixtures (95: 5, 90:10, 80:20) at 20, 25, 30, 35, 40, and 45 ° C, L. plantarium was relatively high in lactic acid production among the selected strains. L. plantarium + Bacillus species (ewha) was inoculated to investigate the amount of lactic acid produced by the fermentation fruit, the results are shown in Figure 6a to 6f.

The mixing ratio of bokbunja and chlorella that had high lactic acid production after inoculation of L. plantarium , that is, the temperature range of (95: 5) and (90:10) was 30-40 ℃ and the optimum temperature was 40 ℃, but (80:20) The temperature range was in the range of 35-40 ℃ and the optimum temperature was 35 ℃. After inoculation of L. plantarium + Bacillus species (ewha), the mixed range of bokbunja and chlorella, that is, the range of lactic acid production at (95: 5) and (90:10) was 30-40 ℃ and the optimum temperature was 40 ℃. The temperature range where lactic acid production was high at (80:20) was 35 to 40 ° C, and the optimum temperature was determined to be 35 ° C. When the amount of chlorella was increased to 20%, the amount of lactic acid produced at the temperature of 35 ° C. was increased, so the amount of chlorella was thought to lower the optimum fermentation temperature.

L. plantarium Bacillus species (ewha) produced slightly less lactic acid than L. plantarium , but Bacillus species (ewha) were thought to be suitable for bokbunja and chlorella mixtures because of their carbohydrate and protease secretion.

[Examples 18 to 21]

L. plantarium and L. plantarium after adjusting the pH of Bokbunja and Chlorella mixtures (95: 5, 90:10, 80:20) to 4, 5, 6, 7 + Bacillus species (ewha) inoculation by the result of the fermentation and the result of the production of lactic acid was investigated as shown in Figure 7a to 7d.

The effect of pH on lactic acid production of bokbunja and chlorella mixtures (90:10) by L. plantarium was in the order of pH 4 <pH 7 <pH 6 <pH 5 until the 2nd day of fermentation. Although the pH of lactic acid production was high in the order of pH 5 <pH 7 <pH 6 pH 4, the pH ranged from 4 to 6, and the optimal pH based on the end of fermentation was determined to be 4.

The effect of pH on lactic acid production of bokbunja and chlorella mixtures (80:20) by L. plantarium was determined until the end of fermentation except for pH 7 <pH 4 <pH 6 <pH 5 on the first day of fermentation. The amount of lactic acid produced in the order of pH 7 <pH 4 <pH 5 <pH 6 was in the range of 4-6, and the optimum pH was 6 based on the end of fermentation.

[Examples 22 to 33]

Bokbunja and chlorella mixtures (95: 5, 90:10, 80:20) were prepared, and the mixtures were inoculated with L. plantarium and L. plantarium. Bacillus inoculated (ewha) inoculated for 4 days at fermentation temperature of 35 ℃, 40 ℃ to prepare a fermented product, and investigated the components and physiological activity of the fermented raw materials and the results are shown in Tables 3 to 14.

1) General Ingredient

General ingredients such as moisture, crude protein, crude fat, crude ash, total sugar, reducing sugar and crude fiber of fermented products are shown in Table 3.

A) water content

The fermented broth, which had the highest water content after the fermentation of Bokbunja and Chlorella mixtures of Table 3, was 66.22 ~ 80.14%, the mixing ratio of Bokbunja and Chlorella was 95: 5, L. plantarium It was fermented No. 4 fermented at 35 ° C by inoculation with Bacillus species (ewha), and the fermented ball with the lowest moisture content was 80:20 mixed with bokbunja and chlorella and fermented at 35 ° C by inoculating L. plantarium. I could see the job.

Bokbunja and chlorella mixtures, the average moisture content of each test group was 72.96% at 40 ° C L. plantarium fermentation at 7, 8, 9 times < 73.06 % at 1, 2, 35 ° C L. plantarium fermentation <35 at 4, 5, 6 times ℃ L. plantarium + Bacillus species (ewha) 75.03% at fermentation L. plantarium at 40 ℃ for 10, 11, 12 + Bacillus species (ewha) The fermentation was in order of 75.43%. These results indicate that L. plantarium In the process of fermentation by equal inoculation of two strains of L. plantarium and Bacillus species (ewha) rather than using only bacteria, the binding water was released in the process of decomposing bokbunja and chlorella by enzymes secreted by the genus Bacillus species (ewha). It was assumed that the growth and enzyme secretion capacity of Bacillus species (ewha) genus was better at 40 ℃.

B) Crude protein content

The fermented broth , which had the highest crude protein content of 2.64 ~ 12.89% after fermentation of bokbunja and chlorella mixtures in Table 3, had a ratio of 80:20 and L. plantarium. It was fermented No. 3 fermented at 35 ℃, and the fermented broth with the lowest content of crude protein had 95: 5 and L. plantarium mixture ratio of bokbunja and chlorella + Bacillus species (ewha) It was found that the fermentation No. 4 fermented to 35 ℃ inoculation.

The crude protein content of fermented test constituents in bokbunja and chlorella mixtures increased more as the chlorella mixture increased. Therefore, the supplementation effect of crude protein was more pronounced than bokbunja alone. There is a difference in the amount of protein according to the fermentation conditions for fermentation by L. plantarium yieoteuna is prone to increase in the case of 40 ℃ than 35 ℃ L. plantarium In the case of fermentation by Bacillus species (ewha), crude protein content tended to decrease at 40 ℃ rather than 35 ℃.

The reduction of crude protein content by fermentation microorganism and temperature used in the present invention is not only predicted synergistic effect in decomposition reactions due to different physiological characteristics of microorganisms and the type and titer of secreted enzymes, It was thought that the degree of activation of the reaction was different.

C) crude fat content

The fermented broth, which had the highest crude fat content of 3.3454 ~ 5.8821% after fermentation of Bokbunja and Chlorella mixtures in Table 3, had a ratio of 80:20, L. plantarium + Bacillus species (ewha) Sample No. 12 was fermented in 40 ℃, a crude fat content of which had the best ever fermentation phrase bokbunja and Chlorella mixing ratio is 95: was found to be 5, L. plantarium + Bacillus species ( ewha) was inoculated to 4 beongu fermentation to 35 ℃.

Crude fat content of fermented broth of bokbunja and chlorella mixtures tended to increase as the amount of chlorella was increased. There is a slight difference in crude fat content according to fermentation conditions, so L. plantarium or L. plantarium + Bacillus species (ewha) at 35 ℃ Crude fat contents tended to increase in L. plantarium and L. plantarium + Bacillus species (ewha) spheres at 40 ° C.

D) ash content

The fermented broth , which had the highest range of ash content of 3.22 ~ 6.37% after fermentation of bokbunja and chlorella mixtures of Table 3, had a mixed ratio of bokbunja and chlorella 80:20, L. plantarium It was found that No. 9 was fermented at 40 ° C, and the fermented ball with the lowest ash content was Bokbunja and Chlorella mixing ratio of 95: 5 and No. 4 fermented at 35 ° C by inoculation with L. plantarium + Bacillus species (ewha).

The crude ash content of fermented broth of bokbunja and chlorella mixtures tended to increase as the amount of chlorella blend increased. L. plantarium at 35 ℃ due to slight difference in contents of crude ash according to fermentation conditions Or L. plantarium + Bacillus species (ewha) L. plantarium and L. plantarium at 40 ℃ + Bacillus species (ewha) tended to increase.

The content of crude ash increased slightly according to the fermentation conditions, but it was not considered to be a problem below the regulation level of general food standards.

E) total sugar content

The fermented broth , which had the highest total sugar content of 5.54-12.37% after fermentation of Bokbunja and Chlorella mixtures in Table 4, had a ratio of 95: 5 and L. plantarium + Bacillus species (ewha) Fermented No. 1 fermented at 35 ℃ and the lowest total sugar content was 80:20, L. plantarium + Bacillus species (ewha) The inoculation was found to be No. 12 fermented at 40 ℃.

The total sugar content of fermented broth of Bokbunbun and Chlorella mixtures decreased as the amount of chlorella blend increased. The total sugar content decreased according to the fermentation conditions of fermented broth by L. plantarium at 35 and 40 ℃, and fermented broth by L. plantarium + Bacillus species (ewha) at 35 and 40 ℃. There was a tendency to decrease, but exceptionally, the fermentation No. 5 and No. 11 fermentation with bokbunja and chlorella mixing ratio were 90:10.

F) reducing sugar content

The fermented broth , which had the highest content of reducing sugar after fermentation of Bokbunja and Chlorella mixtures of Table 3, was 0.51∼2.23 %, and the ratio of Bokbunja and Chlorella was 90:10, L. plantarium. + Bacillus species (ewha) It was found that No. 11 was fermented at 40 ° C., and the fermented ball with the lowest content of reducing sugar was No. 9 fermented at 40 ° C. by inoculating L. plantarium with 80:20 mixed bokbunja and chlorella.

Reducing sugar content of fermented broth of Bokbunja and Chlorella mixtures decreased as the amount of chlorella mixed increased. Reducing sugar content decreased according to the fermentation conditions, fermented by L. plantarium at 35 and 40 ℃, and fermented by L. plantarium + Bacillus species (ewha) at 35 and 40 ℃. With the exception of the tendency to decrease, the bokbunja and chlorella mixing ratios were increased in fermentation No. 5, No. 11 and No. 9 fermentation No. 9 with 80:20.

G) crude fiber content

After fermentation of Bokbunja and Chlorella mixtures , the fermentation broth was 0.38 ～ 2.86%, and the fermentation broth was fermented at 35 ℃ by inoculating L. plantarium at 80:20 and L. plantarium . The fermented broth with the lowest content was found to be 10 fermented broths fermented at 40 ° C by inoculation with L. plantarium + Bacillus species (ewha) at 95: 5.

The crude fiber content of fermented broth was decreased as the amount of chlorella was mixed and as the fermentation temperature was increased. In addition, L. plantarium + Bacillus species (ewha) fermenters tended to decrease more in L. plantarium and L. plantarium + Bacillus species (ewha).

2) pH, acidity and chromaticity

The pH, acidity, Brix and color of the fermentation product are investigated as shown in Table 4 below.

A) pH of fermented product

After fermentation of Bokbunja and Chlorella mixtures in Table 4, the fermentor, which had the highest pH range of 3.31∼3.72 , had a mixture ratio of Bokbunja and Chlorella at 95: 5, L. plantarium. + Bacillus species (ewha) It was found that No. 12 was fermented at 40 ° C., and the lowest pH was fermented at No. 95, 5, L. plantarium , and fermented at 35 ° C. by mixing Bokbunja and Chlorella.

The pH of fermentation broth increased with increasing amount of chlorella, but the fermentation temperature was set to 35, 40 ℃ and L for different ratios of bokbunja and chlorella (95: 5, 90:10, 80:20). . plantarium and L. plantarium It was difficult to recognize the effect of temperature because the result of fermentation by inoculation with Bacillus species (ewha) and the change of pH were not large.

B) Acidity of fermented products

After fermentation of the Bokbunja and Chlorella mixtures, the acidity range was 1.98∼2.72 . The fermented broth was fermented with Bokbunja and Chlorella at 80:20 and L. plantarium at 35 ° C. Chlorella mixing ratio was 95: 5, L. plantarium was inoculated at 1 ℃ fermented at 35 ℃. The acidity of fermented broth by bokbunja and chlorella mixtures tended to increase as the amount of chlorella mixture increased.

Bokbunja and Chlorella specific mixing ratio (95: 5, 90:10, 80:20), the fermentation temperature was 35 ℃ and L. plantarium, and Bacillus species L. plantarium + pH results were respectively inoculated to the fermentation (ewha) is L. plantarium when fermented in a fermentation temperature and showed a minute increase to 40 ℃ and a result of the fermentation by inoculation of L. plantarium, and Bacillus species L. plantarium + (ewha) respectively when fermented with L. plantarium + Bacillus species (ewha) There was a slight increase in acidity. That is, in the 35 ℃ when eoteul to fermentation with L. plantarium, L. plantarium 40 ℃ Fermentation with Bacillus species (ewha) showed a slight increase in acidity.

C) sugar content of fermented products

After fermentation of the mixture of Bokbunja and Chlorella, the sugar content ranged from 4.7 to 11.7 Brix. The highest fermentation broth was No. 6 fermented with Bokbunja and chlorella at 80:20 and L. plantarium 35 ℃. And Chlorella was found to be No. 11 fermented at 40 ℃ by inoculating L. plantarium 90:10, L. plantarium . The sugar content of fermented broth of Bokbunbun and Chlorella mixtures, respectively, tended to increase as the amount of Chlorella blend increased. Mixed ratio of Bokbunja and Chlorella (95: 5, 90:10, 80:20) L. plantarium and L. plantarium + Bacillus species results in 35 ℃ were each inoculated to a fermentation (ewha) the decrease in sugar content, L. plantarium + Bacillus species (ewha) was less, but the sugar content tended to decrease more at 40 ℃. In other words, when fermented with L. plantarium + Bacillus species (ewha) at 35 ℃, it was thought that the sugar consumption was reduced due to the suppressed sugar consumption when fermented with L. plantarium at 40 ℃.

The results of the 35 ℃ condition less the amount of sugar consumed by the L. plantarium more per amount produced by the enzymes secreted by Bacillus species (ewha) a very small amount but has been thought to be accumulated results, in the case of 40 ℃ L. plantarium It was thought that the reduction of sugar produced by suppressing the growth was small and the decrease was small. When inoculated with L. plantarium + Bacillus species (ewha), the growth of L. plantarium is inhibited, but the growth of Bacillus species (ewha) is promoted. As sugar consumption increased, sugar was thought to decrease.

D) color development of fermented products;

The color representation in Table 4 has three independent properties called Value, Chroma, and Hue. In other words, the brightness represents the degree from dark black to bright white, the saturation represents the vividness of the color, and the color becomes dark with gray or colorless as 0, and the color is the five basic colors red, yellow, green, and blue. , Purple and 5 neutral colors.

The color system of undried and lyophilized products after fermentation of bokbunja and chlorella tends to decrease L (brightness), a (+ red,-green) values as chlorella's mixing ratio increases, while b (+ yellow, -Blue) tended to increase. The L, a, b values of Hunter color system of freeze-dried fermented products increased more than undried fermented products, and the color became brighter.

The L-value (brightness) of fermented products ranges from 30.27 to 37.95, with the highest fermentation ratios of 95: 5 and L. plantarium. + Bacillus species (ewha) The fermented broth was fermented at 40 ° C, and the lowest L value was fermented at 40 ° C by inoculating L. plantarium + Bacillus species (ewha) with 90:10 mixed bokbunja and chlorella.

Bokbunja and Chlorella mixed ratio (95: 5, 90:10, 80:20) L. plantarium and L. plantarium + Bacillus species (ewha) were inoculated and fermented at 35 ℃ and 40 ℃, respectively. L value of L. plantarium than L. plantarium + Bacillus species decreased from (ewha), a, b values was a tendency to increase, in 40 ℃ the L and b values than L. plantarium L. plantarium + Values of Bacillus species (ewha) increased, whereas a value tended to decrease.

In addition, as a result of freeze-drying the fermented products by mixing ratio of bokbunja and chlorella, the water content was lowered while the L, a, b values of Hunter color system were increased.

3) the content of free sugar

The results of analyzing the free sugars of the fermented product are as shown in Table 5.

As shown in Table 5, the types of undetected free sugars and the highest and lowest free sugars of fermented samples of bokbunja and chlorella mixtures were determined as L. plantarium . Fermented No. 1 fermented at 35 ℃ was maltose and lactose, and the highest and lowest free sugars were 23.28 mg / kg fructose and 1.62 mg / kg sucrose, and fermented No. 2 was sucrose and maltose. The highest and lowest free sugars were 14.07 mg / kg fructose and 14.04 mg / kg glucose. Three fermenters were maltose and lactose, and the highest and lowest free sugars were 10.66 mg / kg. In case of fructose and 0.85 mg / kg sucrose, but the free sugar analysis of bokbunja and chlorella mixture fermented at 40 ° C. did not detect it, one fermentation No. 7 was glucose, and the highest and lowest free sugar content was 4.25 mg / kg. frucose and 1.38 mg / kg maltose, all 8 fermenters were detected. The highest and lowest free sugars were 27.87 mg / kg glucose and 0.30 mg / kg sucrose, and 9 fermenters were sucrose and maltose. , The highest and lowest free sugar content is 10.01 mg / It was of fructose and 0.93 ㎎ / ㎏ lactose.

L. plantarium The undetected sugars of the bokbunja and chlorella mixtures fermented at 35 ° C with Bacillus species (ewha) were three types of fermentation No. 4, namely sucrose, maltose and lacrose, and the highest and lowest free sugars were 17.09 mg / kg of fructose and The fermented No. 5 was sucrose, maltose and lacrose, and the highest and lowest free sugars were 19.23 mg / kg fructose and 14.70 mg / kg glucose. The highest and lowest free sugars were 10.97 mg / kg fructose and 5.39 mg / kg glucose, but 10 fermenters at 40 ° C were sucrose and lactose and the highest and lowest free sugars were sucrose and maltose. The contents were 18.07 mg / kg fructose and 3.46 mg / kg maltose. The 11 fermentations were two species, sucrose and lactose, and the highest and lowest free sugars were 16.32 mg / kg fructose and 1.72 mg / kg maltose. Fermentation No. 12 is two varieties, maltose and lactose. The sugar content was 12.39 ㎎ / ㎏ fructose and 0.28 ㎎ / ㎏ sucrose.

These results indicate that free sugars according to the fermentation temperature and fermentation strain difference according to the mixing ratio of bokbunja and chlorella tended to decrease as the amount of chlorella was increased, but the contents of free sugar increased in fermentation 5 and 8 fermentation. . In addition, the frequency of undetected sugars in all fermenters increased by 0, 1, 7, 8, 8 in order of fructose <glucose <sucrose <maltose = lactose. If that by varying the mixing ratio at 40 ℃ bokbunja and Chlorella with L. plantarium fermentation was higher than maltose, lactose content of the contrast material, L. plantarium Fermentation with Bacillus species (ewha) resulted in increased maltose.

4) Amino Acid Content

Amino acid content according to fermentation temperature and strain according to the fermentation temperature and strain of bokbunja and chlorella mixture is shown in Table 6.

The undetected amino acid among L. plantarium among 16 kinds of fermented broth obtained by different temperature and species of Bokbunja and Chlorella mixture (95: 5, 90:10, 80:20) The fermentation No. 1 fermented at 35 ° C. was six species, namely, aspartate, histidine, tyrosine, cystine, methionine, and isoleucine. The highest and lowest amino acids were 246.0 mg / kg arginine and 21.95 mg / kg serine Two kinds of bulbs were methionine and isoleucine, and the highest and lowest amino acid contents were 85.06 mg / kg glycine and 9.66 mg / kg cystine, and fermentation No. 3 was one species, isoleucine, and the highest and lowest amino acid content was 104.13. In the case of amino acid analysis of the bokbunja and chlorella mixtures fermented at 40 ° C., the seventh fermenters were cystine, methionine, and isoleucine, and the highest and lowest amino acids were MG / kg glycine and 8.03 mg / kg methionine. The content of was 99.89 mg / kg arginine and 9.74 mg / kg tyrosine, and fermentation No. 8 was five species, histidine, tyrosine, cystine, methionine and isoleucine, and the highest and lowest amino acid contents were 256.54 mg / kg arginine and 11.82 mg / kg Aspartate was fermented No. 9, histidine, tyrosine, cystine, methionine, and isoleucine. The highest and lowest amino acids were 119.31 mg / kg glycine and 10.43 mg / kg methionine.

Also L. plantarium As a result of amino acid analysis of bokbunja and chlorella mixture fermented at 35 ° C with Bacillus species (ewha), six fermented broths were aspartate, histidine, tyrosine, cystine, methionine and isoleucine. The content of was 247.67 mg / kg arginine and 21.73 mg / kg valine, and fermentation No. 5 was three species, cystine, methionine and isoleucine, and the highest and lowest amino acid contents were 116.97 mg / kg glycine and 12.45 mg / kg. fermentation No. 6 was cystine and methionine, and the highest and lowest amino acid contents were 105.33 mg / kg glycine and 9.46 mg / kg isoleucine, but the amino acid analysis of the bokbunja and chlorella mixtures fermented at 40 ° C. In the undetected cases, fermented No. 10 was two species, cystine and methionine, and the highest and lowest amino acid contents were 103.84 mg / kg glycine and 6.64 mg / kg isoleucine, and ferment No. 11 was three species, cystine and methionine. , The highest and lowest amino acids were 188.56 mg / kg arginine and 11.77 mg / kg tyrosine. The fermentation No. 12 was one species, cystine, and the highest and lowest amino acids were 123.39 mg / kg glycine and 8.79. Mg / kg methionine.

  These results showed that the fermentation temperature and amino acid content level according to the fermentation temperature and cultivation strain ratio of bokbunja and chlorella did not tend to be constant, but the frequency ranking of insufficient amino acids in all fermenters was aspartate <histidine <tyrosine <isoleucine <methionine <cystine. Complementing amino acids is thought to be able to maintain the composition of amino acids that are nutritionally superior.

Total amino acid content in the range bokbunja and Chlorella honbap fermented product according to the fermentation temperature and fermentation strain is fermented with the highest phrase bokbunja and Chlorella mixing ratio to 502.3 ~ 727.1 ㎎ / ㎏ 90:10, L. plantarium It was found that No. 8 was fermented at 40 ° C., and the lowest fermentation was No. 2 ball fermented at 35 ° C. by inoculating L. plantarium at 90:10 and Bokbunja and Chlorella.

The total amino acid content of the fermented broth of Bokbunbun and Chlorella mixtures did not show any correlation with the increase of the amount of chlorella mixed.

Bokbunja and chlorella mixed ratio (95: 5, 90:10, 80:20) L. plantarium and L. plantarium + Bacillus species (ewha) were inoculated respectively and fermented at 35 ℃. Total amino acid content was 20% mixed with chlorella with the exception of one case it increased when the inoculated with the bacteria of L. plantarium + Bacillus species (ewha) than L. plantarium.

Bokbunja and Chlorella mixed ratio (95: 5, 90:10, 80:20) L. plantarium and L. plantarium + Bacillus species (ewha) were inoculated respectively and fermented at 40 ℃. Total amino acid content was 5% mixed with chlorella Except in one case, L. plantarium fermenters tended to be higher.

That is, L. plantarium at 35 ℃ When fermented with Bacillus species (ewha), the total amino acid content was high when fermented with L. plantarium at 40 ℃.

5) organic acid content

Citric acid, lactic acid, formic acid, acetic acid, propionic acid, tartaric acid, malic acid, after fermentation at different fermentation temperatures and strains of mixed bokbunja and chlorella (95: 5, 90:10, 80:20) The results of analyzing the content of succinic acid are shown in Table 7.

As seen from table 7 The ferionic fermenters obtained by inoculating L. plantarium with the bokbunja and chlorella mixtures at 35 ° C and 40 ° C were not detected in the propionic acid and succinic acid. 2.96 g / kg malic acid and 0.05 g / kg acetic acid, and the highest and lowest organic acid contents were 2.57 g / kg malic acid and 0.05 g / kg acetic acid, respectively. The lowest organic acid content was 2.97 g / kg lactic acid and 0.07 g / kg acetic acid. However, in fermentation No. 7 fermented at 40 ° C, the highest and lowest organic acid content was 2.75 g / kg malic acid and 0.06 g / kg. acetic acid, and the highest and lowest organic acids were 2.74 g / kg malic acid and 0.22 g / kg formic acid in fermented No. 8, and the highest and lowest organic acids were 3.13 g / kg in lactic acid. And 0.07 g / kg acetic acid.

In the case of undetected amino acid analysis of bokbunja and chlorella mixture fermented at 35 ° C with L. plantarium + Bacillus species (ewha), six fermented broths were aspartate, histidine, tyrosine, cystine, methionine, and isoleucine. The highest and lowest amino acid contents were 247.67 mg / kg arginine and 21.73 mg / kg valine, and fermentation No. 5 was three species, cystine, methionine and isoleucine. The highest and lowest amino acid contents were 116.97 mg / kg glycine and 12.45 mg / kg valine, fermented No. 6 were two species, cystine and methionine, and the highest and lowest amino acid contents were 105.33 mg / kg glycine and 9.46 mg / kg isoleucine, but the mixture of bokbunja and chlorella fermented at 40 ° C. As a result of undetected amino acid analysis, 10 fermented broths were two species, cystine and methionine, and the highest and lowest amino acid contents were 103.84 mg / kg glycine and 6.64 mg / kg isoleucine. , cystine, methionine, and isoleucine, and the highest and lowest amino acid contents were 188.56 mg / kg arginine and 11.77 mg / kg tyrosine. The fermentation No. 12 was one species, cystine, and the highest and lowest amino acid content was 123.39 mg / kg. Glycine and 8.79 mg / kg methionine.

These results showed that the fermentation temperature and amino acid content level according to the fermentation temperature and cultivation strain ratio of bokbunja and chlorella did not tend to be constant, but the frequency ranking of insufficient amino acids in all fermenters was aspartate <histidine <tyrosine <isoleucine <methionine <cystine. Complementing amino acids is thought to be able to maintain the composition of amino acids that are nutritionally superior.

Total amino acid content in the range bokbunja and Chlorella honbap fermented product according to the fermentation temperature and fermentation strain is fermented with the highest phrase bokbunja and Chlorella mixing ratio to 502.3 ~ 727.1 ㎎ / ㎏ 90:10, L. plantarium It was found that No. 8 was fermented at 40 ° C., and the lowest fermentation was No. 2 ball fermented at 35 ° C. by inoculating L. plantarium at 90:10 and Bokbunja and Chlorella.

The total amino acid content of the fermented broth of Bokbunbun and Chlorella mixtures did not show any correlation with the increase of the amount of chlorella mixed.

Bokbunja and chlorella mixed ratio (95: 5, 90:10, 80:20) L. plantarium and L. plantarium + Bacillus species (ewha) were inoculated respectively and fermented at 35 ℃. Total amino acid content was 20% mixed with chlorella with the exception of one case it increased when the inoculated with the bacteria of L. plantarium + Bacillus species (ewha) than L. plantarium.

Mixed ratio of Bokbunja and Chlorella (95: 5, 90:10, 80:20) L. plantarium and L. plantarium Bacillus species (ewha) were inoculated respectively and fermented at 40 ℃, resulting in total amino acid content of L. plantarium except 5% of chlorella Fermented broth tended to be rather high.

That is, L. plantarium at 35 ℃ When fermented with Bacillus species (ewha), the total amino acid content was high when fermented with L. plantarium at 40 ℃.

6) mineral content

The results of analysis of the mineral content after fermentation at different fermentation temperatures and strains of bokbunja and chlorella mixtures (95: 5, 90:10, 80:20) are shown in Table 8.

As seen from table 8 Bokbunja and chlorella mixtures were mixed with L. plantarium and L. plantarium at 35 ° C and 40 ° C. + Fermentation sphere content of Ca, K, Mg and Na Bacillus species obtained by fermentation by the inoculation (ewha) was increased according to the chlorella weak mixing amount, the overall trend is L. plantarium> L. plantarium of 35 ℃ of 40 ℃> of 35 ℃ L. plantarium + Bacillus species ( ewha)> of 40 ℃ L. plantarium + Bacillus species ( ewha) was followed.

The contents of Ca, K and Mg according to the fermentation temperature of L. plantarium were 40 ℃ in L. plantarium. + Bacillus species (ewha) showed a slightly higher tendency of 35 ℃, Na content of L. plantarium 40 ℃, L. plantarium + Bacillus species (ewha) did not differ between 35 ℃ and 40 ℃ .

Mineral levels of fermented products are 0.2421 ± 0.0031 to 0.4714 ± 0.0037 mg / g for Ca, 2.1287 ± 0.0356 to 3.7777 ± 0.0706 mg / g for K, and 0.2705 ± 0.0017 to 2.0811 ± 0.0076 mg / g for Na and In the case of 0.1551 ± 0.0039∼1.0422 ± 0.0105 mg / g.

7) Microbial count

The result of measuring the viable cell number after fermentation at 40 ° C. of bokbunja and chlorella mixtures (95: 5, 90:10, 80:20) is shown in Table 9. (Ferments in Examples 28 to 33)

Table 9 shows that L. plantarium viable cell number was increased by 3.3 × 10 ⁸ CFU / g <8 fermentation No. 7 (95: 5) as chlorella mixture was fermented at 40 ℃. 90:10), 2.88 × 10 ¹¹ CFU / g <9 fermented broths (80:20), 1.08 × 10 ¹² CFU / g in order, and viable cell count of L. plantarium + Bacillus species (ewha) Fermentor (90:10), 4.67 × 10 ¹¹ CFU / g <fermentation No. 10 (95: 5), 4.61 × 10 ¹¹ CFU / g <Fermentation No. 12 (80:20), 9.17 × 10 ¹⁰ CFU / g order. As the number of cells of L. plantarium and L. plantarium + Bacillus species (ewha) did not increase due to the increase in the amount of chlorella mixed, it was weak due to the mixed culture of Bacillus species (ewha) but secreted as each strain was grown. It was estimated that the number of viable cells was small due to the inhibitory effect in the 20% chlorella mixed group due to the substances and nutritional ingredients.

8) antimicrobial activity

The antimicrobial activity of harmful bacteria S. aureus , E. coli and Salmonella by fermentation of bokbunja and chlorella (95: 5, 90:10, 80:20) at 40 ° C is shown in Table 10. 28 to 33 fermentation)

Table 10 shows that the antimicrobial activity decreased as the mixture amount of chlorella increased after fermentation of Bokbunja and Chlorella mixtures at 40 ° C.

As shown in Table 10, the antimicrobial activity against S. aureus was 21.28% for fermentation No. 7 fermented with L. plantarium , 18.52% for fermentation No. 8 and -1.20% for fermentation No. 9, and L. plantarium + Bacillus species (ewha ) it was fermented in the fermentation clause 10 1.90% 11 4.60% fermented sphere, and has a higher antimicrobial activity of L. plantarium except 9 to obtain fermented District 2.90% 12 times fermentation L. plantarium + Bacillus species (ewha Fermentation No. 11 was slightly higher when fermented with mixed strain.

Antimicrobial Activity on E. coli is the mixing amount of the chlorella 6, 10, showed a tendency to decrease The more to 20%, fermentation by fermentation sphere 7 L. plantarium + Bacillus species (ewha) fermented with L. plantarium Only the fermented 11 times had antibacterial activity of 3.64% and 1.94%, respectively. Overall, the antimicrobial activity of bokbunja and chlorella mixtures on E. coli was weak.

The antimicrobial activity of Salmonella did not correlate with the amount of chlorella mixed with bokbunja, and the antimicrobial activity was high when 10% chlorella was mixed. However zymogen mainly L. plantarium, and Bacillus species L. plantarium + (ewha) with the result L. plantarium + Bacillus species (ewha) The antimicrobial activity was low, 7 and 8 only in antimicrobial activity, respectively 7.64% and 13.15 fermentation obtain It was%. Overall, the antimicrobial activity of bokbunja and chlorella mixtures on Salmonella was weak.

9) Determination of Total Phenolic Compound Content

The polyphenol contents of the extracts obtained by inoculating L. plantarium , L. plantarium + Bacillus species (ewha) to the mixtures of Bokbunja and Chlorella were fermented at 35 ° C and 40 ° C.

As shown in Table 11, L. plantarium The polyphenol content of the mixed fermented bokbunja and chlorella fermented at 35 ° C and 40 ° C was 0.59 g / kg for fermentation 1, 0.15 g / kg for fermentation 2, and 0.31 g for fermentation 3 / kg, but 0.50 g / kg for fermentation No. 7 fermented at 40 ℃, 0.59 g / kg for fermentation No. 8, 0.41 g / kg for fermentation No. 9, polyphenol content of fermentation at 40 ℃ than 35 ℃ Increased if yes.

The polyphenol content of bokbunja and chlorella mixtures fermented at 35 ° C and 40 ° C with L. plantarium + Bacillus species (ewha) was 0.72 g / kg for fermentation No. 4 and 0.45 g / for fermentation No. 5. kg and 0.43 g / kg for fermentation No. 6, 0.51 g / kg for fermentation No. 10, 0.72 g / kg for Ferment No. 11, and 0.46 g / kg for Fermentation No. 12.

These results showed that the polyphenol content tended to decrease as the amount of chlorella mixed in bokbunja increased. The polyphenol content of fermented with L. plantarium tended to increase at 40 ℃ rather than 35 ℃, and L. plantarium + Bacillus species (ewha) fermentation at 35 ℃ and 40 ℃ tended to slightly increase the polyphenol content, except the fermenters 4 and 11 fermented chlorella mixtures were higher.

Fermentation temperature Stars the polyphenol content of the L. plantarium than L. plantarium Fermentation with Bacillus species (ewha) showed an increasing tendency, and fermentation at 40 ℃ was higher than 35 ℃.

10) Measurement of electron donating ability

Lipid metabolism in the human body is decomposed through the oxidation process at the cellular level, and free radicals generated at this time not only promote the aging of cells but also lower the cell's activity such as lowering the defense mechanism. It is very important to protect the electron donating ability, by donating electrons to the active free radicals to inhibit the oxidation of cellular components (Park Young-seo, Jang Hak-gil). By selecting fermentation products obtained by fermenting Bokbunja and Chlorella mixtures with fermentation temperature of 35 ℃ and 40 ℃, and selecting electron fermenting ability, we select fermentation pots with excellent inhibitory effect on aging of cells and reduction of protective mechanism by free radicals in vivo. The results of investigating the electron donating ability are shown in Table 12.

As shown in Table 12, with L. plantarium The electron donating ability of fermented mixed bokbunja and chlorella fermented at 35 ℃ and 40 ℃ was 71.24% for fermentation 1, 76.33% for fermentation 2 and 80.25% for fermentation 3, but at 40 ℃ The fermented fermented No. 7 was 75.45%, the fermented No. 8 was 76.06%, and the fermented No. 9 was 80.51%.

L. plantarium The electron donating ability of the bokbunja and chlorella mixtures fermented at 35 ° C and 40 ° C with Bacillus species (ewha) was 72.00% for fermentation No. 4, 72.93% for fermentation No. 5 and 81.59 for fermentation No. 6. %, 70.96% for fermentation 10, 72.96% for fermentation 11 and 81.49% for fermentation 12.

Taken together these results the more the increase in the mixing amount Chlorella bokbunja yieoteuna electron donating ability will tend to increase L. plantarium + Fermented products at 40 ℃ by Bacillus species (ewha) inoculation also showed a tendency to increase. The electron donating ability of fermented with L. plantarium and L. plantarium + Bacillus species (ewha) tended to increase at 40 ℃ rather than 35 ℃, indicating that the electron donating ability is different due to the difference between strain and fermentation temperature. At the same fermentation temperature, the electron donating ability of L. plantarium < L. plantarium + Bacillus species (ewha) was high. Therefore, the fermentation broth with the best electron donating ability was found to be 12 fermentation broth mixed with 20% chlorella.

On the other hand, the fermented products showed 70.96 ~ 81.59% of electron donating ability of fermented products, compared to the known result of gourd and intestine whose electron donating ability was 71.1% before fermentation and 69.0% after fermentation. It was found that the electron donating ability is excellent.

11) Nitrite Scavenging Activity

One of the cancer-causing agents, nitrite is contained in food additives, drinks, and saliva in small amounts. Vegetables and fruits contain large amounts of nitrate, which is reduced to nitrite by nitrate redutase during storage of saliva, gastric juice, and food. Reported. The presence of more than a certain level of nitrite in the blood causes oxidative reactions with hemoglobin to form methemo-globin, which is known to cause poisoning by disrupting metabolism in the body. The reaction of amines in protein foods, pharmaceuticals, and pesticides to produce nitrosoamines, alkylating nucleic acids, proteins, and cellular components has resulted in cancer. These nitrosoamines are of interest because the nitrosoamine reactions can easily occur at pH 2 to 3 of the gastric juice secreted by the reaction between food components or the digestion of food in the stomach. In order to investigate whether the nitrite scavenging effect is reduced in the human body, the nitrite scavenging ability of fermented plants obtained by inoculating L. plantarium , L. plantarium + Bacillus species (ewha) into bokbunja and chlorella mixtures was examined in Table 13 and same.

As shown in Table 13, with L. plantarium Nitrite scavenging ability of mixed fermented bokbunja and chlorella fermented at 35 ℃ and 40 ℃ showed a tendency to decrease the nitrite scavenging ability as the amount of chlorella mixed.

Nitrite scavenging ability at pH 1.2 was 1, 2, and 3 fermented fermented at 35 ° C, the highest fermented nitrite scavenging activity was 86.35 ± 6.507% of fermented No. 1 mixed with 5% Chlorella. Among fermenters Nos. 8 and 9, fermenters with high nitrite scavenging ability were 76.40 ± 2.119% with fermentation No. 7 mixed with 5% Chlorella. Therefore, it was found that the scavenging ability of nitrite by fermentation temperature of bokbunja and chlorella mixture was more effective at 35 ℃ than 40 ℃ for L. plantarium . The nitrite scavenging ability of bokbunja and chlorella mixtures fermented at 35 ° C and 40 ° C with L. plantarium + Bacillus species (ewha) was 10, 11, 12 times fermented at 35 ° C and fermented at 40 ° C. Among the fermented broths, 4 and 10% of chlorella and 11% of 11 were higher. The former was 83.77 ± 4.750% and the latter was 85.20 ± 4.731%. The effect of fermentation temperature on nitrite scavenging activity was 40 ℃. High.

Nitrite scavenging ability at pH 3.0, 4.2, 6.0 was the highest fermentation nitrite scavenging ability of fermented Nos. 1, 2, and 3 fermented at 35 ° C, with fermentation No. 1 mixed with 5% Chlorella, 80.96 ± 4.730, 51.11 ± 31.298, Among the fermented broths 7, 8 and 9 fermented at 40 ° C, the fermented broth with high nitrite scavenging ability was 39.66 ± 22.405% compared with 47.14 ± 7.683, 21.39 ± 8.507, and 24.25 ± 8.361%, respectively. Therefore, it was found that the scavenging ability of nitrite by fermentation temperature of bokbunja and chlorella mixture was more effective at 35 ℃ than 40 ℃ for L. plantarium .

The nitrite scavenging ability at pH 3.0, 4.2 and 6.0 of bokbunja and chlorella mixtures fermented at 35 ° C and 40 ° C with L. plantarium + Bacillus species (ewha) was observed at 40 ° C and 4, 5, and 6 fermenters fermented at 35 ° C. Among the fermented fermenters 10, 11 and 12, pH 3.0 was exceptionally high only 4 times with 5% mixed chlorella, while others 5 and 11 with 10% mixed were higher, and pH 4.2 and 6.0 were all 10% chlorella. In the mixed 5 and 11 times, it was found that the nitrite scavenging ability is high. The effect of fermentation temperature of L. plantarium + Bacillus species (ewha) mixed bacteria on nitrite scavenging activity was higher at 35 ℃.

The nitrite scavenging sequence of bokbunja Chlorella fermenters is 1>11>4>2>5>10>7>8>9>12>3> 6 fermenters, pH>1>4>2> 5 >11>7>8>12>10>3>6> 9 fermenters, pH 4.2 1>5>4>2>7>7>3>8>11>9>10> 12 fermenters, pH In case of 6.0 1>5>2>8>7>4>11>3>10>9>12> 6 Fermented No. 1 fermented with L. plantarium at 30 ° C by mixing 5% Chlorella in bokbunja Was the best, but the other fermenters showed no tendency.

Therefore, the level of nitrite scavenging activity of L. plantarium and L. plantarium + Bacillus species (ewha) was 53.01 ～ 86.35% for the former at 30 ℃, 58.46 ～ 76.46% at 40 ℃ for the former, and 30 ℃ for the latter under the reaction conditions of pH 1.2. Is 46.10 ~ 83.77%, 40 ℃ is 56.53 ~ 77.93%, the former is 35.26 ~ 80.96% for 30 ℃, 29.32 ~ 47.14% for 40 ℃, and 33.62 ~ 77.42% for 30 ℃ in the latter , 40 ℃ ranges from 40.35 to 58.99%, pH is 4.2. In the former, 20.70 ~ 51.11% for 30 ℃, 18.08 ~ 21.39% for 40 ℃, 22.87 ~ 37.70% for 30 ℃ and 16.96 for 40 ℃ The former ranged from 20.02 to 39.66% at 40 ° C, 18.79 to 24.25% at 40 ° C, the latter at 18.30 to 36.29% at 40 ° C and 18.58 to 21.18% at 40 ° C. . Nitrite scavenging ability tends to decrease as the pH approaches the neutral region, suggesting that the nitrite scavenging reaction is active in an environment where nitrite or nitrate formation occurs well. That is, it was found that the nitrite scavenging ability decreased as the pH was increased to 1.2, pH 3.0, pH 4.2, and pH 6.0.

  On the other hand, when fermented Bokbunja concentrate with lactic acid bacteria, gourd and intestine (15) were 38.3% at pH 1.2, 13.10% at pH 3.0, 4.56 at pH 4.2 and 1.34% at pH 6.0. 86.35%, 29.32 ~ 80.96% at pH 3.0, 16.96 ~ 51.11% at pH 4.2, 18.30 ~ 39.66% at pH 6.0. Fermented products have excellent nitrous acid at pH 1.2 and 3.0, and in the process of decomposing food in the stomach It was thought that it could effectively reduce the amount of nitrosoamine that can be produced and cause cancer.

12) Sensory test

The results of sensory evaluation of color, aroma, sweetness and sour taste after fermentation at different fermentation temperatures and strains of mixed bokbunja and chlorella (95: 5, 90:10, 80:20) are shown in Table 14.

According to sensory evaluation on the color of fermented products of Bokbunja and Chlorella mixtures (95: 5, 90:10, 80:20), fermented No. 12 had the highest score of 7.49 ± 0.08, and the score of Fermented No. 2 was 6.87 ± 0.35 Was the lowest among the fermenters (p <0.05)).

As a result of sensory evaluation on the aroma of fermented products of Bokbunja and Chlorella mixtures (95: 5, 90:10, 80:20), fermented 10 had the highest score of 6.52 ± 0.18, and fermented 5 had 5.38 ± 0.05 Was the lowest among the fermenters (p <0.05)).

According to the sensory evaluation on the sweetness of fermented products of Bokbunja and Chlorella mixtures (95: 5, 90:10, 80:20), fermented No. 1 had the highest score of 5.93 ± 0.12, while Fermented No. 3 scored 5.08 ± 0.08. Was the lowest among the fermenters (p <0.05)).

According to sensory evaluation on the sour taste of fermented products of Bokbunja and Chlorella mixtures (95: 5, 90:10, 80:20), fermented No. 12 had the highest score of 8.48 ± 0.06, and Fermented No. 3 scored 7.00 ± 0.02. Was the lowest among the fermenters (p <0.05)).

The results of sensory evaluation by combining color, flavor, sweetness and sourness of fermented products of Bokbunja and Chlorella mixture (95: 5, 90:10, 80:20) showed the highest score of 6.97 ± 0.11. The lowest fermentation score was 6.25 ± 0.10 and significant difference was found among fermentations (p <0.05).

The above sensory evaluation showed that plantarium + Bacillus among fermented bokbunja and chlorella fermented with L. plantarium and L. plantarium + Bacillus species (ewha) at 40 ° C (95: 5, 90:10, 80:20) Ideally, the product would be fermented using species (ewha).

1 is a graph showing the change in moisture content according to the mixing ratio of frozen bokbunja and bokbunja juice and chlorella.

Figure 2 is a photograph showing the culture results of lactic acid bacteria and Bacillus subtilis on the water content change according to the mixing ratio of frozen bokbunja and bokbunja juice and chlorella.

Figure 3a to 3j is a graph showing the acid production of lactic acid bacteria according to the chlorella addition level of lactic acid bacteria used in the present invention.

Figures 4a to 4f is a graph showing the acid production of lactic acid bacteria according to the chlorella addition level of Bacillus subtilis used in the present invention.

5a to 5k are graphs showing the acid production amount change of the selection strain according to the level of chlorella added to bokbunja according to an embodiment of the present invention.

6a to 6f are graphs showing the change in lactic acid production by temperature of the bokbunja and chlorella mixture according to an embodiment of the present invention.

7A to 7D are graphs showing changes in lactic acid production by pH of bokbunja and chlorella mixtures according to an embodiment of the present invention.

Claims (12)

After inoculating fermentation microorganisms, which are lactic acid bacteria, Bacillus subtilis or two mixed strains of Bacillus subtilis and Bacillus subtilis and Lactobacillus in a mixture of 95-50% by weight of bokbunja and 5-50% by weight of chlorella, fermented under conditions of temperature of 30-40 ° C for 3-7 days, A method for producing a functional fermentation raw material using a bokbunja and chlorella mixture, characterized in that it comprises the step of formulating the obtained fermentation broth in liquid or powder form. The method of claim 1, wherein the bokbunja is a natural bokbunja, frozen bokbunja or bokbunja juice characterized in that the functional fermentation raw material using the bokbunja and chlorella mixture. The method of claim 1, wherein the lactic acid bacterium is Lactobacillus casei, Lactobacillus plantarium, Lactobacillus lactis, Lactobacillus acidophyllus, Lactobacillus brevis, Lactobacillus vulgaris, Lactobacillus rhamnosus, Pediococcus pen Bokbunja and chlorella, characterized in that one or two or more mixed strains selected from the group consisting of tosaceus, Streptococcus thermophilus, Bifidobacterium butchineri, Bifidobacterium infantis, Bifidobacterium longum Method for producing a functional fermentation raw material using a mixture. The bacterium according to claim 1, wherein the Bacillus subtilis, Bacillus subtilis, Bacillus licheniformis, Bacillus natto, Bacillus sirius, Bacillus Spices (cam) and Bacillus sushi (Ewha) one or two or more mixed strains selected from the group consisting of Method for producing a functional fermentation raw material using a bokbunja and chlorella mixture, characterized in that. The mixed strain of lactic acid bacterium and Bacillus subtilis is a mixed strain of Lactobacillus casei and Bacillus subtilis, a mixed strain of Lactobacillus casei and Bacillus sp. ), Mixed strain of Lactobacillus plantarium and Bacillus subtilis, mixed strain of Lactobacillus plantarium and Bacillus sp. (Cam) or mixed strain of Lactobacillus plantarium and Bacillus sp. Method for producing a functional fermentation raw material using bokbunja and chlorella mixture, characterized in that one species selected from the group. According to claim 1, wherein the fermentation microorganisms are cultured until the absorbance (Abs.) Measured at 620nm by the species culture, bokbunja characterized in that inoculated into the mixture of bokbunja and chlorella mixture Method for producing a functional fermentation raw material using a chlorella mixture. The method of claim 1, wherein the fermentation microorganism is inoculated with 2% (V / V) of the bokbunja and chlorella mixtures. The method of claim 1, wherein the fermentation microorganisms are two mixed strains, each of which is mixed in a ratio of 1: 1 so that each bacteria solution is 1% (V / V) compared to the bokbunja and chlorella mixture Method for producing a functional fermentation raw material using a mixture of bokbunja and chlorella, characterized in that the inoculation. According to claim 1, wherein the pH range after the fermentation of the bokbunja and chlorella mixture is 3 ~ 4, the acidity range is 1.9 ~ 2.7, the sugar range is 4.7 ~ 11.7 Brix functional fermentation using a mixture of bokbunja and chlorella Method of manufacturing raw materials. The method according to claim 1, wherein the bokbunja and chlorella mixtures are made by extracting bokbunja juice by mixing the bokbunja and mixing chlorella in the bokbunja solution. Functional fermentation raw material using a bokbunja and chlorella mixture prepared by the method of any one of claims 1 to 11. 12. The functional fermentation raw material using the bokbunja and chlorella mixture, characterized in that the form of powder, liquid or mucus.

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