KR20180099259A - Food composition for enhancing function of immunity comprising fermented extract from leek by lactic acid bacteria and preparation method thereof - Google Patents
Food composition for enhancing function of immunity comprising fermented extract from leek by lactic acid bacteria and preparation method thereof Download PDFInfo
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- KR20180099259A KR20180099259A KR1020170026464A KR20170026464A KR20180099259A KR 20180099259 A KR20180099259 A KR 20180099259A KR 1020170026464 A KR1020170026464 A KR 1020170026464A KR 20170026464 A KR20170026464 A KR 20170026464A KR 20180099259 A KR20180099259 A KR 20180099259A
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- KR
- South Korea
- Prior art keywords
- lactic acid
- fermented
- acid bacteria
- broth
- fermentation broth
- Prior art date
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- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
- A23L19/09—Mashed or comminuted products, e.g. pulp, purée, sauce, or products made therefrom, e.g. snacks
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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Abstract
Description
본 발명은 유산균을 이용한 항산화 기능성 및 혈류개선에 효과가 있는 부추 발효액에 관한 것으로, 더욱 상세하게는 상기 부추 발효액에 유산균 사균을 첨가하여 면역활성이 증진된 부추 유산균 발효액을 유효성분으로 함유하는 식품 조성물 및 그 제조방법에 관한 것이다. The present invention relates to a leek fermentation broth having an antioxidative activity and blood flow improvement using lactic acid bacteria. More particularly, the present invention relates to a leek fermentation broth having an active ingredient, And a manufacturing method thereof.
부추는 백합과의 여러해살이풀. 봄에 땅속의 작은 비늘줄기로부터 길이 30cm 정도 되는 선 모양의 두툼한 잎이 무더기로 뭉쳐난다. 8~9월에 긴 꽃줄기가 나와 산형 화서로 흰색의 작은 꽃이 핀다. 열매는 삭과를 맺으며 익으면 저절로 터져서 까만 씨가 나온다. 비늘줄기는 건위ㆍ화상 따위에 쓰고, 잎은 식용한다. 중국과 인도가 원산지로 한국, 일본 등지에 분포한다.It is a perennial herb of lilies. In the spring, a line of thick leaves that are about 30cm long from the small scale stem of the earth are gathered together. A long flower stalk appears from August to September, and a small white flower blooms with a phloem flower. The fruit will bear capsules, and if it is ripe, it will burst itself, and black seed will come out. The stem of the scales should be written on the surface, the image, and the leaves edible. It is distributed in Korea and Japan as origin of China and India.
부추의 효능으로는 비타민 A, B, C, 카로틴, 철 등이 풍부해서 혈액 순환을 원활하게 해주고 소화기관을 튼튼하게 해주고 자율신경을 자극하여 에너지 대사를 활발하게 해준다.The beneficial effect of leek is rich in vitamins A, B, C, carotene, and iron, which smoothes blood circulation, strengthens digestive organs, stimulates autonomic nerves, and stimulates energy metabolism.
본 발명과 관련된 선행기술로는 대한민국 등록특허 10-1018765호에 부추 추출물을 유효성분으로 함유하는 약리적 조성물 및 그 제조방법에 대하여 공지되어 있고 대한민국 공개특허 10-2014-0067242호에 부추의 유산균발효 추출물 및 이를 함유하는 항당뇨 조성물에 관한 것이 공지되어 있다. 그러나 현재까지 부추 유산균 발효액에 사균을 첨가하여 제조한 면역활성 증진 기능성 조성물 및 그 제조방법에 관해서는 공지된 바 없다.As a prior art related to the present invention, Korean Patent No. 10-1018765 discloses a pharmacological composition containing an extract of Leek extract as an active ingredient and a method for producing the same, and Korean Patent Publication No. 10-2014-0067242 discloses a fermented extract of lactic acid bacteria And an antidiabetic composition containing the same. However, to date, there has been no known method for producing an immunostimulatory functional composition prepared by adding dead bacteria to a fermentation broth of Lactobacillus lacunae and a preparation method thereof.
따라서 본 발명의 목적은 항산화 기능성 및 혈류개선 효과가 있는 부추 발효액에 유산균 사균을 첨가하여 면역활성이 증진된 부추 유산균 발효액을 유효성분으로 함유하는 식품 조성물 및 그 제조방법을 제공하는 데 있다.Accordingly, an object of the present invention is to provide a food composition containing as an active ingredient a fermentation broth of fermented Lactobacillus acidus having enhanced immunity activity by adding killed microbes to a fermented broth having antioxidative function and blood flow improving effect, and a method for producing the same.
본 발명의 상기 목적은 (a) 부추 당침액을 pH 6.0으로 보정 후 농도를 바람직하게는 10~20°Brix로 가장 바람직하게는 13°Brix로 조정하는 단계와; 상기 단계에서 얻은 부추 당침액에 영양원으로 yeast extract를 0~5% 첨가하고 pH 6.0으로 보정한 후 121℃에서 15분 동안 멸균하는 단계와; The above object of the present invention is achieved by a method for preparing a fermented beverage comprising the steps of: (a) adjusting the concentration of the leek slurry to pH 6.0, preferably to 10 to 20 Brix, most preferably to 13 Brix; Adding 0 to 5% of yeast extract as a nutrient source to the leek slurry obtained in the above step, correcting it to pH 6.0, and sterilizing at 121 ° C for 15 minutes;
이와 별도로 (b) 유산균 전배양 배지를 121℃에서 15분 동안 멸균하는 단계와; 상기 단계에서 얻은 배지를 냉각 후 유산균 Lactobacillus plantarum , Lactobacillus brevis , Enterococcus faecium 또는 Pediococcus acidilactici를 접종하여 37℃에서 바람직하게는 24~32시간, 가장 바람직하게는 28시간 동안 진탕 배양하여 유산균 전배양액을 제조하는 단계와;Separately, (b) sterilizing the lactic acid pre-culture medium at 121 占 폚 for 15 minutes; After cooling the medium obtained in the above step, the lactic acid bacteria Lactobacillus plantarum , Lactobacillus brevis , Enterococcus faecium or Pediococcus acidilactici at 37 ° C. for 24 to 32 hours, most preferably for 28 hours to produce lactic acid bacteria pre-culture;
(c) 상기 (a)단계에서 얻은 멸균한 부추 당침액에 상기 (b)단계에서 얻은 유산균 배양액을 1~5중량% 접종한 다음 37℃에서 68~76시간 동안 발효시켜 부추 당침액 유산균 발효액을 제조하는 단계와; 상기 단계에서 얻은 발효액에 사균 Enterococcus faecalis를 바람직하게는 100~160mg/g이 가장 바람직하게는 134mg/g을 첨가하여 부추 유산균 발효액 조성물을 수득하는 단계를 통해 이루어지고 상기에서 얻은 부추 유산균 발효액 조성물을 공시재료로 면역활성 증진 효과를 평가함으로써 달성하였다.(c) 1 to 5% by weight of the culture solution of lactic acid bacteria obtained in step (b) is inoculated into the sterilized leek slices obtained in step (a), and then fermented at 37 ° C for 68 to 76 hours, ; ≪ / RTI > The fermentation broth of the fermented broccoli fermented broth obtained by the step of obtaining the fermented broth fermented broth by adding 100 to 160 mg / g of fungus Enterococcus faecalis , most preferably 134 mg / g, And evaluating the effect of enhancing the immune activity with the material.
본 발명은 항산화 기능성 및 혈류개선 효과가 있는 부추 유산균 발효액에 유산균 사균을 첨가하여 발효시킨 조성물은 면역활성 증진에 뛰어난 효과가 있다.The present invention has an excellent effect in enhancing the immune activity of a composition obtained by fermenting lactic acid bacteria with fermented Lactobacillus fermentation broth which has antioxidative function and blood flow improving effect.
도1은 본 발명 부추 당침액 유산균 발효액의 제조공정을 나타낸 모식도이다.
도2는 본 발명 부추 당침액 유산균 발효액의 총 플라보노이드 함량을 나타낸 그래프이다.
도3은 본 발명 부추 당침액 유산균 발효액의 총 폴리페놀함량을 나타낸 그래프이다.
도4는 본 발명 부추 당침액 유산균 발효액의 항산화활성 중 DPPH 활성을 나타낸 그래프이다.
도5는 본 발명 부추 당침액 유산균 발효액의 항산화활성 중 SOD 유사활성을 나타낸 그래프이다.
도6은 본 발명 부추 당침액 유산균 발효액의 HMG-CoA 환원효소억제활성 결과를 나타낸 그래프이다.
도7은 본 발명 부추 당침액 유산균 발효액의 ACE 저해활성 결과를 나타낸 그래프이다.
도8은 본 발명 부추 당침액 유산균 발효액에 사균 E. faecalis 균주 134mg/g을 첨가하여 NO 생성 결과를 나타낸 그래프이다.BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view showing a process for producing a fermentation broth of submerged lactic acid fermentation broth perch of the present invention. FIG.
2 is a graph showing the total flavonoid content of the submerged lactic acid fermentation broth according to the present invention.
FIG. 3 is a graph showing the total polyphenol content of the submerged lactic acid fermentation broth of the present invention.
4 is a graph showing DPPH activity among the antioxidant activities of fermentation broth of submerged lactic acid fermentation broth per day of the present invention.
FIG. 5 is a graph showing SOD-like activity of the fermentation broth of submerged lactic acid fermentation broth of the present invention.
6 is a graph showing the results of HMG-CoA reductase inhibitory activity of the submerged lactic acid fermentation broth of the present invention.
7 is a graph showing the results of ACE inhibition activity of the submerged lactic acid fermentation broth of the present invention.
Fig. 8 is a graph showing the results of the fermentation of E. faecalis And the results of NO production by adding 134 mg / g of the strain.
본 발명은 항산화 기능성 및 혈류개선 효과가 있는 부추 발효액에 유산균 사균을 첨가하여 발효시켜 면역활성 증진에 뛰어난 효과가 있는 부추 유산균 발효액 조성물 및 그 제조방법을 제공한다.The present invention provides a fermented broccoli fermentation broth composition having excellent antioxidative activity and blood flow improving effect by fermenting fermented Lactobacillus japonica with fermentation broth and improving its immunity.
본 발명에 따르면 상기 부추의 발효에 사용할 수 있는 유산균은 바람직하게는 Lactobacillus속, Enterococcus속 또는 Pediococcus속 균주를 가장 바람직하게는 Lactobacillus plantarum , Lactobacillus brevis , Enterococcus faecium 또는 Pediococcus acidilactici 균주이다.According to the present invention, the lactic acid bacteria which can be used for the fermentation of the leek are preferably Lactobacillus , Enterococcus or The Pediococcus sp. Strain is most preferably selected from the group consisting of Lactobacillus plantarum , Lactobacillus brevis , Enterococcus faecium Or Pediococcus acidilactici Lt; / RTI >
본 발명에서 사용되는 유산균 사균은 관능효과 및 면역활성 증대상 Enterococcus faecalis 균주가 가장 바람직하고 첨가량은 100~160mg/g이 가장 바람직하다.The strain of Enterococcus faecalis is most preferably selected from the group consisting of 100-160 mg / g of insecticidal activity and immunological activity.
이하, 본 발명의 구체적인 내용을 실시예, 실험예 및 제조예를 들어 상세하게 설명한다. 하기 실시예, 실험예 및 제조예는 본 발명을 예시하기 위한 것으로 본 발명의 권리범위가 이에 한정되는 것은 아니다.
Hereinafter, the present invention will be described in detail with reference to Examples, Experimental Examples and Preparation Examples. The following Examples, Experimental Examples and Preparation Examples are for illustrating the present invention, and the scope of the present invention is not limited thereto.
실시예1. 유산균 전배양액 제조Example 1. Manufacture of pre-culture of lactic acid bacteria
본 발명 부추 당침액 유산균 발효액 제조에 사용할 수 있는 유산균 전배양액을 제조하기 위하여 유산균 전배양 배지를 121℃에서 15분 동안 멸균 후 냉각시키고 유산균 Lactobacillus plantarum , Lactobacillus brevis , Enterococcus faecium 또는 Pediococcus acidilactici 중에서 선택된 1종의 유산균주를 상기 배지에 접종하여 37℃, 90rpm에서 28시간 동안 진탕배양하여 유산균 전배양액을 수득하였다.
In order to prepare lactic acid bacteria pre-culture solution which can be used for the fermentation of submerged lactic acid fermentation broth per the present invention, lactic acid bacteria pre-culture medium was sterilized at 121 ° C for 15 minutes, and then lactic acid bacteria Lactobacillus plantarum , Lactobacillus brevis , Enterococcus faecium or Pediococcus acidilactici Was inoculated on the medium and cultured with shaking at 37 DEG C and 90 rpm for 28 hours to obtain a lactic acid bacteria pre-culture solution.
실시예2. 본 발명 부추 당침액 유산균 발효액 제조Example 2. The present invention relates to a fermentation broth of fermented lactic acid bacteria
본 발명에 사용되는 부추 당침액 유산균 발효액은 하기와 같이 제조하였다. 먼저, 부추 당침액의 pH를 6.0으로 보정 후 농도를 10~20°Brix로 조정하였다. 상기 단계에서 얻은 부추 당침액에 0~5% yeast extract를 첨가하고 121℃에서 15분 동안 고압멸균하였다. 상기 단계에서 얻은 고압멸균한 부추 당침액을 냉각하고 상기 실시예1에서 얻은 유산균 전배양액을 1~5% 접종하고 37℃, 90rpm에서 68~76시간 진탕배양하여 부추 당침액 유산균 발효액을 수득하였다.
The submerged lactic acid fermentation broth for use in the present invention was prepared as follows. First, the pH of the submerged saliva was adjusted to 6.0, and the concentration was adjusted to 10-20 ° Brix. 0 to 5% yeast extract was added to the leek slurry obtained in the above step and autoclaved at 121 ° C for 15 minutes. The fermented broth fermentation broth perch was obtained by cooling the autoclaved leek slurry obtained in the above step and incubating the fermentation broth obtained in Example 1 at 1 to 5% at 37 ° C and 90 rpm for 68 to 76 hours with shaking.
실시예3. 본 발명 부추 유산균 발효액 조성물 제조Example 3. The present invention provides a fermentation broth composition
본 발명 부추 유산균 발효액 조성물을 제조하기 위해 상기 실시예2에서 얻은 부추 당침액 유산균 발효액 총 중량에 사균 Enterococcus faecalis 균주를 100~160mg/g 범위 내에서 첨가하여 부추 유산균 발효액 조성물을 수득하였다.
To prepare the present invention encourage lactic acid fermentation liquor composition dead cells of lactic acid bacteria in the fermentation broth per immersion leek total weight obtained in Example 2 Enterococcus faecalis The strain was added in a range of 100 to 160 mg / g to obtain a fermented broth fermentation broth.
실험예1. 본 발명 부추 당침액 유산균 발효액의 생리활성 측정Experimental Example 1 Measurement of physiological activity of lactic acid fermentation broth of submerged lactic acid bacteria according to the present invention
본 발명 부추 당침액 유산균 발효액의 생리활성 검정을 위한 Total Flavonoid 함량 측정을 위해 상기 실시예2에서 수득한 본 발명 부추 당침액 유산균 발효액 300uL에 증류수 1200uL를 첨가하고 다시 5% NaNO2 90uL 첨가 후 실온에서 5분 동안 반응시키고 10% AlCl36H2O를 첨가하고 6분 동안 반응시킨 후 1N NaOH 300uL를 첨가하여 반응정지시키고 UV-spectrophotometer를 이용하여 510nm에서 흡광도를 측정하였다. 표준물질로는 rutin을 사용하였다. 대조군으로는 본 발명 부추 당침액 유산균 발효액 대신 생부추를 사용하였다.To measure the total flavonoid content for the fermentation of the fermentation broth of the submerged lactic acid fermentation broth of the present invention, 1200 μL of distilled water was added to 300 μL of the submerged lactic acid fermentation broth perch of the present invention obtained in Example 2, and then 90 μL of 5% NaNO 2 was added, After reacting for 5 minutes, adding 10% AlCl 3 6H 2 O and reacting for 6 minutes, 300 μL of 1N NaOH was added and the absorbance was measured at 510 nm using UV-spectrophotometer. Rutin was used as a reference material. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention.
실험 결과, 도2에서 알 수 있듯이 Lactobacillus plantarum , Lactobacillus brevis, Enterococcus faecium 또는 Pediococcus acidilactici 균주를 접종한 본 발명 부추 당침액 유산균 발효액 모두 생부추에 비해 총 폴라보노이드 함량이 증가하였다.As shown in FIG. 2, Lactobacillus plantarum , Lactobacillus brevis, Enterococcus faecium or Pediococcus acidilactici Inoculated with the strain All of the fermented broth of submerged lactic acid bacteria of the present invention showed a higher content of total polaronolide than that of green leek.
또, Total polyphenol 함량 측정을 위해 상기 실시예2에서 수득한 본 발명 부추 당침액 유산균 발효액 1mL에 1N Folin & Ciocalteu's phenol 1mL를 첨가하고 실온에서 3분 동안 반응시킨 후 10% Na2CO3 1mL 첨가하고 실온에서 1시간 동안 반응시킨 후 700nm에서 흡광도를 측정하였다. 표준물질로는 gallic acid(Sigma co., USA)를 사용하였다. 대조군으로는 본 발명 부추 당침액 유산균 발효액 대신 생부추를 사용하였다.For measurement of the total polyphenol content, 1 mL of 1N Folin and Ciocalteu's phenol was added to 1 mL of the submerged lactic acid fermentation broth per pullulan of the present invention obtained in Example 2, reacted at room temperature for 3 minutes, and then 1 mL of 10% Na 2 CO 3 was added After reacting at room temperature for 1 hour, absorbance was measured at 700 nm. Gallic acid (Sigma co., USA) was used as a reference material. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention.
실험 결과, 도3에서 알 수 있듯이 본 발명 부추 당침액 유산균 발효액 모두 생부추에 비해 총 폴리페놀 함량이 증가하였다.
As shown in FIG. 3, the total polyphenol content of the fermented broth of submerged lactic acid fermentation broth of the present invention was higher than that of green leek.
실험예2. 본 발명 부추 당침액 유산균 발효액의 항산화활성 측정Experimental Example 2 Antioxidant Activity of Fermented Lactic Acid Bacteria from Pepper
본 발명 부추 당침액 유산균 발효액의 항산화능을 측정하기 위해 DPPH(1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity를 측정하였다. 이를 위해 200uM DPPH 1mL와 상기 실시예2에서 수득한 본 발명 부추 유산균 발효액 1mL를 혼합하여 상온, 암소에서 30분 동안 반응시킨 후 517nm에서 흡광도를 측정하였다. 대조군으로는 본 발명 부추 당침액 유산균 발효액 대신 생부추를 사용하였다. DPPH radical 소거능(%)은 하기 수학식1의 방법으로 계산하였다.The scavenging activity of DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity was measured in order to measure the antioxidative activity of the submerged lactic acid fermentation broth. To this end, 1 mL of 200 uM DPPH and 1 mL of the fermentation broth of the present invention obtained in Example 2 were mixed and reacted at room temperature and in a dark place for 30 minutes, and the absorbance was measured at 517 nm. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention. The DPPH radical scavenging activity (%) was calculated by the following equation (1).
실험 결과, 도4에서 알 수 있듯이 본 발명 부추 당침액 유산균 발효액 모두 생부추에 비해 DPPH 라디칼 소거능이 증가하였다.As shown in FIG. 4, the DPPH radical scavenging ability of the fermented broth of submerged lactic acid bacteria of the present invention was higher than that of green leek.
또, 본 발명 부추 당침액 유산균 발효액의 항산화능을 측정하기 위해 SOD 유사활성을 측정하였다. 이를 위해 상기 실시예2에서 수득한 본 발명 부추 당침액 유산균 발효액 200uL에 pH 8.5의 tris-HCl buffer 3mL와 7.2mM pyrogallol 200uL를 첨가하여 25℃에서 10분 동안 반응시킨 후 1N HCl 1mL를 가하여 반응을 정지시키고 흡광도 420nm에서 측정하였다. 대조군으로는 본 발명 부추 당침액 유산균 발효액 대신 생부추를 사용하였다. 발효 부추액의 첨가구와 무첨가구 사이의 흡광도 차이는 하기 수학식2의 방법으로 백분율로 계산하였다. In addition, SOD-like activity was measured to measure the antioxidant activity of the submerged lactic acid fermentation broth of the present invention. For this, 3 μL of tris-HCl buffer (pH 8.5) and 200 μL of 7.2 mM pyrogallol were added to the fermentation broth of fermented lactic acid fermentation broth obtained in Example 2, and the reaction was carried out at 25 ° C. for 10 minutes. And the absorbance was measured at 420 nm. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention. The difference in absorbance between the addition of the fermented vermiculite solution and the no addition was calculated by the following formula (2).
실험 결과, 도5에서 알 수 있듯이 본 발명 부추 당침액 유산균 발효액 모두 생부추에 비해 SOD 유사활성능이 증가하였다.As shown in FIG. 5, the SOD-like activity of the fermented broth of the present invention was higher than that of the fermented broth.
실험예3. 본 발명 부추 당침액 유산균 발효액의 HMG-CoA 환원효소억제활성 측정Experimental Example 3. The inhibitory activity of HMG-CoA reductase in fermentation broth of submerged lactic acid bacteria
본 발명 부추 당침액 유산균 발효액의 혈류개선 효과를 확인하기 위해 HMG=CoA 환원효소억제활성을 HMG-CoA reductase assay kit(Sigma-Aldrich)를 이용하여 측정하였다. 이를 위해 Assay buffer 182uL에 상기 실시예2에서 수득한 본 발명 부추 당침액 유산균 발효액 1uL, HMG-CoA reductase 2uL, NADPH 4uL 및 HMG-CoA 12uL를 첨가한 후 혼합하여 37℃에서 10분 동안 반응시키고 340nm 흡광도에서 측정하였다. 대조군으로는 본 발명 부추 당침액 유산균 발효액 대신 생부추를 사용하였다.The inhibitory activity of HMG = CoA reductase was measured using the HMG-CoA reductase assay kit (Sigma-Aldrich) to confirm the blood flow improvement effect of the submerged lactic acid fermentation broth per pullulan of the present invention. For this, 1 uL of the submerged lactic acid fermentation broth of the present invention obtained in Example 2, 2 uL of HMG-CoA reductase, 4 uL of NADPH and 12 uL of HMG-CoA were added to 182 uL of the assay buffer obtained in Example 2, followed by reaction at 37 ° C for 10 minutes, Absorbance was measured. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention.
실험 결과, 도6에서 알 수 있듯이 본 발명 부추 당침액 유산균 발효액 모두 생부추에 비해 HMG-CoA 환원효소억제활성이 증가하였다.
As shown in FIG. 6, the inhibitory activity of HMG-CoA reductase was increased in the fermented broth of the present invention.
실험예4. 본 발명 부추 당침액 유산균 발효액의 ACE 저해활성 측정Experimental Example 4. ACE inhibitory activity of submerged lactic acid fermentation broth
본 발명 부추 당침액 유산균 발효액의 심혈관질환 및 뇌혈관질환 등 고혈압과 관련된 질환의 치료효과를 측정하기 위해 ACE(Angiotensin converting enzyme) 저해활성을 측정하였다. 이를 위해 상기 실시예2에서 수득한 본 발명 부추 당침액 유산균 발효액 20uL에 substrate buffer 20uL, Enzyme working solution 20uL를 첨가한 후 혼합하여 37℃에서 1시간 동안 반응시킨 후 Indicator working solution 200uL 첨가한 후 10분 동안 반응시켰다. 그 후 450nm에서 흡광도를 측정하였다. 대조군으로는 본 발명 부추 당침액 유산균 발효액 대신 생부추를 사용하였다.Angiotensin converting enzyme (ACE) inhibitory activity was measured in order to measure the therapeutic effect of hypertension related diseases such as cardiovascular diseases and cerebrovascular diseases in fermented broth of submerged lactic acid bacteria according to the present invention. To this, 20 uL of substrate buffer and 20 uL of enzyme buffer solution were added to 20 uL of fermented lactic acid fermentation broth obtained from Example 2, and the mixture was incubated at 37 ° C for 1 hour, and 200 uL of indicator working solution was added thereto. Lt; / RTI > The absorbance was then measured at 450 nm. As a control group, raw leek was used in place of fermented broth of submerged lactic acid fermentation broth of the present invention.
실험 결과, 도7에서 알 수 있듯이 ACE 저해활성이 15.6uL/mL 농도에서 본 발명 부추 당침액 유산균 발효액 모두 생부추에 비해 증가하였다.
As a result, as shown in FIG. 7, the ACE inhibitory activity was increased at a concentration of 15.6 uL / mL compared with that of the live leucocephalic broth of the present invention.
실험예5. 본 발명 부추 유산균 발효액의 면역활성 측정Experimental Example 5 Measuring the Immune Activity of the Fermented Lactic Acid Fermentation Solution of the Invention
본 발명 부추 유산균 발효액의 면역활성을 측정하기 위해 상기 실시예2에서 수득한 L. plantarum 균주를 접종한 부추 당침액 유산균 발효액과 상기 실시예 3에서 수득한 사균 Enterococcus faecalis 균주를 가장 바람직하게는 134mg/g이 첨가된 부추 유산균 발효액 조성물을 이용하여 NO 방출량을 측정하여 면역활성을 확인하였다. 이를 위해 RAW 264.7 세포를 LPS 100ng/mL 농도로 자극을 준 후 생부추, L. plantarum 균주를 접종한 부추 당침액 유산균 발효액, 상기 사균을 첨가한 유산균 발효액을 각각 0.1~1mg/mL 농도로 처리하여 NO 방출량을 측정하였다. 대조군으로는 LPS 무처리군을 이용하였다.To measure the immune activity of the present invention a fermentation broth of lactic acid bacteria L. plantarum leek obtained in Example 2 Inoculated with the strain The fermentation broth of submerged lactic acid bacteria in leek and the Enterococcus fungus obtained in Example 3 faecalis The amount of NO released was determined by using the fermented broth lactic acid fermentation broth composition containing the strain, most preferably 134 mg / g, to confirm immunological activity. RAW 264.7 cells were stimulated with LPS at a concentration of 100 ng / mL and then inoculated with L. plantarum The amount of nitric oxide (NO) released was measured at 0.1 ~ 1mg / mL concentration of lactic acid fermentation broth of leek fermented broth per day and fermented broth of lactic acid fermentation broth supplemented with the above killed bacteria. As a control group, LPS-untreated group was used.
실험 결과, 도8에서 알 수 있듯이 1mg/mL 농도에서 생부추의 NO 방출량이 88.5%인데 비해 부추 당침액 유산균 발효액은 8.17%, 사균을 첨가한 부추 유산균 발효액 조성물은 74.3%로 생부추에 비해 많은 량의 NO를 방출시켜 면역 활성이 증진됨을 확인하였다.
As shown in FIG. 8, at the concentration of 1 mg / mL, the release of NO was 88.5% at the concentration of 1 mg / mL, compared with 8.17% of the fermented broth per leavened broth and 74.3% of leucine fermented broth containing yeast NO was released to enhance the immune activity.
본 발명에 따르면 부추 유산균 발효액을 유효성분으로 함유하는 식품 조성물에 포함할 수 있는 식용가능한 담체, 부형제 또는 희석제로는 락토즈, 덱스트로즈, 슈크로즈, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산칼슘, 알지네이트, 젤라틴, 규산칼슘, 미세결정성 셀룰로즈, 폴리비닐피롤리돈, 셀룰로즈, 폴리비닐피롤리돈, 셀룰로즈, 물, 설탕시럽, 메틸셀룰로즈, 메틸 하이드록시 벤조에이트, 프로필 하이드록시 벤조에이트, 활석, 스테아트산 마그네슘 및 미네랄 오일이 바람직하다.According to the present invention, the edible carrier, excipient or diluent which can be contained in the food composition containing the fermented Lactobacillus fermentation broth as an active ingredient include lactose, dextrose, sucrose, sorbitol, mannitol, starch, , Alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, cellulose, water, sugar syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc , Magnesium stearate and mineral oil are preferable.
또, 상기 성분들 외에 상업적으로 이용가능한 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 중에서 선택하여 그 어느 하나의 성분을 추가로 더 포함할 수 있다.In addition to the above components, the composition may further include any one of a commercially available lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifying agent, a suspending agent, and a preservative.
이하에는 본 발명 방법에 따라 제조된 부추 유산균 발효액을 함유하는 건강식품 조성물 제제의 구체적인 조성을 설명한다.
Hereinafter, a specific composition of a health food composition preparation containing the fermentation broth of L. fermented liquor prepared according to the method of the present invention will be described.
제조예1. 건강 보조 식품의 제조Production Example 1 Manufacture of health supplements
부추 유산균 발효액을 함유하는 건강 보조 식품은 하기 표1의 함량으로 통상의 건강식품 제조방법을 이용하여 제조하였다.The health supplement foods containing the fermentation broth of Lactobacillus acidus lactifera were prepared by the usual health food production method in the contents shown in Table 1 below.
제조예2. 부추 유산균 발효 음료 제조Production Example 2 Manufacture of fermented beverage lactic acid bacteria
본 발명 실시예3에 따라 수득한 부추 유산균 발효액이 함유된 음료를 하기 표2의 함량으로 통상의 음료 제조방법을 이용하여 제조하였다.The beverage containing the fermented liquorice fermented liquor obtained according to Example 3 of the present invention was prepared by the ordinary beverage preparation method in the contents shown in the following Table 2.
이상 설명한 바와 같이 본 발명 유산균을 이용한 항산화 기능성 및 혈류개선에 효과가 있는 부추 발효액에 유산균 사균을 첨가하여 면역활성 증진에 뛰어난 효과가 있으므로 기능성 건강 식품 산업상 매우 유용한 발명이다.
As described above, since the lactic acid bacteria are added to the fermented broth, which has an antioxidative function and blood flow improvement effect using the lactic acid bacteria of the present invention, it is an extremely useful invention in the functional health food industry because it has an excellent effect in promoting the immune activity.
Claims (7)
이와 별도로 (b) 유산균 전배양 배지를 121℃에서 15분 동안 멸균하는 단계와; 상기 단계에서 얻은 배지를 냉각 후 유산균을 접종 후 진탕 배양하여 유산균 전배양액을 제조하는 단계와;
(c) 상기 (a)단계에서 얻은 멸균한 부추 당침액에 상기 (b)단계에서 얻은 유산균 배양액을 접종한 다음 발효시켜 부추 당침액 유산균 발효액을 제조하는 단계와; 상기 단계에서 얻은 발효액에 사균을 첨가하는 단계로 이루어진 것을 특징으로 하는 면역활성 증진 기능성 부추 유산균 발효액의 제조방법.(a) adjusting the submerged saliva to 10-20 [deg.] Brix; Adding yeast extract as a nutrient source to the leek slurry obtained in the above step and sterilizing at 121 ° C for 15 minutes;
Separately, (b) sterilizing the lactic acid pre-culture medium at 121 占 폚 for 15 minutes; Culturing the medium obtained in the above step by shaking after inoculation of the lactic acid bacteria after cooling the lactic acid bacteria;
(c) preparing a fermented broth of fermented lactic acid fermentation broth per spoon by inoculating the fermented broth fermented broth obtained in step (b) into the sterilized fermented broth fermented broth obtained in step (a); And adding dead bacteria to the fermentation broth obtained in the above step.
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