KR20210100802A - Method for producing fermented spinach and fermented soybean with increased effective component content using lactic acid bacteria - Google Patents
Method for producing fermented spinach and fermented soybean with increased effective component content using lactic acid bacteria Download PDFInfo
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- KR20210100802A KR20210100802A KR1020200014587A KR20200014587A KR20210100802A KR 20210100802 A KR20210100802 A KR 20210100802A KR 1020200014587 A KR1020200014587 A KR 1020200014587A KR 20200014587 A KR20200014587 A KR 20200014587A KR 20210100802 A KR20210100802 A KR 20210100802A
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- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A23V2250/00—Food ingredients
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- A23V2250/00—Food ingredients
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- A—HUMAN NECESSITIES
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Abstract
Description
본 발명은 유산균을 이용한 유용성분 함량이 증가된 시금치 및 콩 발효물의 제조방법에 관한 것이다.The present invention relates to a method for producing spinach and soybean fermented products having an increased content of useful ingredients using lactic acid bacteria.
유산균은 인간이 이용할 수 있는 가장 유익한 미생물 중의 한 종류로서, 비병원성균으로 인간의 장내에 서식하면서 정장 및 정균작용을 할 수 있고 당류를 발효해서 다량의 젖산을 생성하고 낮은 pH 및 혐기적인 조건에서도 잘 생육하며 여러 가지 영양물질을 요구하는 등의 특징을 가지고 있다. 유산균에 의해 채소류가 발효되면 독특한 향과 맛을 내게 되어 관능적 특성이 향상되고, 유기산에 의하여 비타민 C(vitamin C)와 여러 생리활성물질이 잘 보존되며, 초기 재료에는 거의 존재하지 않았던 비타민 B12와 비타민 K가 생합성되어 부패균과 병원성균의 성장과 증식을 저해하여 위생적인 식품이 되는 등의 장점을 가지고 있는 것으로 알려져 있다.Lactobacillus is one of the most beneficial microorganisms available to humans. As a non-pathogenic bacteria, it can perform intestinal and bacteriostatic actions while inhabiting the human intestine. It also ferments sugars to produce a large amount of lactic acid, It has the characteristics of growing and requiring various nutrients. When the vegetable is fermented by lactic acid bacteria it is a unique aroma and taste Me is improved sensory properties, by an organic acid and preserved in vitamin C (vitamin C) and various physiologically active substances, and the initial material, the vitamin B 12 that was almost non-existent It is known that vitamin K is biosynthesized to inhibit the growth and proliferation of putrefactive bacteria and pathogenic bacteria, thereby making it a hygienic food.
시금치(Spinacia oleracea)는 남해지방의 특산물로 명아주과에 속하는 1년생 작물로서, 원산지는 중앙아시아이고 재배지는 북부 온대지방으로 널리 분포되어 있으며 내한성이 강하고 서늘한 기후를 좋아하는 특성이 있다. 시금치는 다른 채소에 비해 비타민 A, B1, B2와 비타민 C, 섬유질, 요오드 외에 특히 DNA 합성에 필수적인 영양성분으로 알려진 엽산이 풍부하여 암의 예방에도 관여하는 것으로 알려져 있다. 최근에는 시금치에 비타민과 섬유소뿐만 아니라 항산화, 항암, 면역증강 활성 등을 나타내는 루테인이 다량 함유되어 있는 것이 보고되어 기능성 식품 소재로서의 활용가치가 매우 높은 농산물로 평가되고 있다. Spinach ( Spinacia oleracea ) is a special product of the Namhae region, and is an annual crop belonging to the family Myopodaceae. Compared to other vegetables, spinach is rich in vitamins A, B 1 , B 2 and vitamin C, fiber, and iodine, as well as folic acid, an essential nutrient for DNA synthesis, and is known to be involved in the prevention of cancer. Recently, it has been reported that spinach contains a large amount of lutein, which exhibits antioxidant, anti-cancer, and immune-enhancing activities as well as vitamins and fiber, and is evaluated as an agricultural product with very high utility value as a functional food material.
콩(Glycine max L.)은 우리나라의 주요 식량작물로서 단백질(약 40%)과 지질(약 20%)을 다량 함유하여 영양면에서 우수한 식품이며, 중요한 식물성 기름의 원료이기도 하다. 콩은 된장, 청국장, 고추장, 간장 등의 발효제품, 콩나물 같은 발아제품과 두유, 두부, 식용유 등 기타 제품의 형태로 가공되어 식품으로서의 이용도가 높고, 또한 의약품, 화장품, 비누 등 공업용 제품의 원료로서도 널리 이용되고 있다. 콩은 주요 영양적 기능 외에도 인체에 유익한 다양한 생리활성 물질이 함유되어 있으므로 이들 영양성분과 생리활성 물질의 분석기법 확립 및 국내ㆍ외 유용자원에 대한 체계적 검토는 향후 국내 식량작물의 경쟁력 강화를 위한 주요 요소로 작용할 것이다.Soybean ( Glycine max L.) is a major food crop in Korea and contains a large amount of protein (about 40%) and lipid (about 20%), so it is an excellent food in terms of nutrition and is an important raw material for vegetable oil. Soybeans are processed into fermented products such as soybean paste, cheonggukjang, gochujang, and soy sauce, germinated products such as bean sprouts, and other products such as soy milk, tofu, and edible oil. It is also widely used as As soybeans contain various physiologically active substances beneficial to the human body in addition to their main nutritional functions, establishment of analysis techniques for these nutritional components and physiologically active substances and systematic review of domestic and foreign useful resources are important for strengthening the competitiveness of domestic food crops in the future. will act as a factor.
한편, 한국등록특허 제1820507호에는 '우엉-시금치 복합 발효추출물을 함유하는 건강기능식품 및 그 제조방법'에 대해 개시하고 있으며, 한국공개특허 제2015-0024121호에는 '원적외선을 이용하여 발아된 콩 종자의 이소플라본 함량과 항산화능이 증가된 콩을 제조하는 방법'에 대해 개시하고 있다. 하지만, 본 발명의 '유산균을 이용한 유용성분 함량이 증가된 시금치 및 콩 발효물의 제조방법'에 대해서는 아직까지 개시된 바가 없다.On the other hand, Korea Patent No. 1820507 discloses 'health functional food containing burdock-spinach complex fermented extract and a manufacturing method thereof', and Korean Patent Publication No. 2015-0024121 discloses 'Germination of soybeans using far-infrared rays'. A method for preparing soybeans with increased isoflavone content and antioxidant activity of seeds is disclosed. However, the 'method for producing fermented spinach and soybean with increased content of useful ingredients using lactic acid bacteria' of the present invention has not been disclosed yet.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명자들은 시금치 및 콩에 락토바실러스 플랜타룸 CBG-C21(Lactobacillus plantarum CBG-C21) 균주를 각각 접종하여 시금치 발효물 및 콩 발효물을 제조하였을 때, 발효 전에 비해 시금치 발효물의 경우 L-아스코르브산(L-ascorbic acid) 함량이 현저하게 증가되고, 콩 발효물의 경우 아글리콘형 이소플라본인 다이드제인(daidzein), 제니스테인(genistein) 및 글리시테인(glycitein) 함량이 현저하게 증가되는 것을 확인함으로써, 본 발명을 완성하였다.The present invention has been derived by the above requirements, and the present inventors inoculated spinach and soybean with Lactobacillus plantarum CBG-C21 (Lactobacillus plantarum CBG-C21) strain respectively to prepare spinach fermented product and soybean fermented product , L-ascorbic acid content is significantly increased in the spinach fermented product compared to before fermentation, and the aglycone-type isoflavones daidzein, genistein and glycitein in the case of soybean fermented product By confirming that the (glycitein) content was significantly increased, the present invention was completed.
상기 과제를 해결하기 위하여, 본 발명은 시금치 또는 콩에 유산균을 접종하여 발효하는 단계를 포함하는 유용성분 함량이 증가된 시금치 발효물 또는 콩 발효물의 제조 방법을 제공한다.In order to solve the above problems, the present invention provides a method for producing a spinach fermented product or a soybean fermented product having an increased content of useful ingredients, comprising the step of inoculating and fermenting spinach or soybean.
또한, 본 발명은 상기 제조 방법으로 제조된 유용성분 함량이 증가된 시금치 발효물 또는 콩 발효물을 제공한다.In addition, the present invention provides a spinach fermented product or soybean fermented product having an increased content of useful ingredients prepared by the above manufacturing method.
또한, 본 발명은 상기 유용성분 함량이 증가된 시금치 발효물 또는 콩 발효물을 포함하는 건강기능식품을 제공한다.In addition, the present invention provides a health functional food comprising a spinach fermented product or soybean fermented product having an increased content of the useful ingredients.
본 발명에 따른 유산균을 이용하여 제조한 시금치 발효물은 L-아스코르브산 함량이 증가되고 콩 발효물은 다이드제인, 제니스테인 및 글리시테인 함량이 증가되어, 여성갱년기 증상 완화, 장 기능 또는 비만 개선 등을 위한 건강기능식품에 더욱 유용하게 이용될 수 있다.The spinach fermented product prepared using the lactic acid bacteria according to the present invention has an increased L-ascorbic acid content and the soybean fermented product has an increased daidzein, genistein and glycitein content, thereby alleviating female menopause symptoms, improving intestinal function or obesity. It can be more usefully used in health functional food for etc.
도 1은 시금치 즙액에 락토바실러스 플랜타룸 CBG-C21(Lactobacillus plantarum CBG-C21) 균주를 접종하여 제조한 시금치 발효물 분말의 L-아스코르브산(L-ascorbic acid) 함량 변화를 확인한 결과이다.1 is a result confirming the change in L-ascorbic acid (L-ascorbic acid) content of spinach fermented powder prepared by inoculating Lactobacillus plantarum CBG-C21 strain into spinach juice.
본 발명의 목적을 달성하기 위하여, 본 발명은 시금치 또는 콩에 유산균을 접종하여 발효하는 단계를 포함하는 유용성분 함량이 증가된 시금치 발효물 또는 콩 발효물의 제조 방법을 제공한다.In order to achieve the object of the present invention, the present invention provides a method for producing a fermented spinach or soybean fermented product with an increased content of useful ingredients, comprising the step of inoculating and fermenting spinach or soybean.
본 발명의 유용성분 함량이 증가된 시금치 발효물 또는 콩 발효물의 제조 방법에서, 상기 시금치 및 콩은 추출물, 즙 또는 분말과 같은 다양한 형태로 준비될 수 있고, 준비된 시금치 또는 콩 시료에 유산균을 접종하여 발효함으로써 시금치 발효물 및 콩 발효물을 제조할 수 있다. In the method for producing a fermented spinach or fermented soybean with an increased content of useful ingredients of the present invention, the spinach and beans may be prepared in various forms such as extracts, juices or powders, and by inoculating the prepared spinach or soybean samples with lactic acid bacteria A spinach fermented product and a soybean fermented product can be manufactured by fermentation.
본 발명의 일 구현예에 따른 제조방법에 있어서, 상기 시금치는 시금치 및 물을 혼합하고 분쇄하여 제조한 시금치 즙액에 유산균을 접종한 것일 수 있고, 상기 콩은 콩 분말 및 물을 혼합하여 콩물로 제조하고 유산균을 접종한 것일 수 있으며, 이들을 발효함으로써 시금치 발효물 및 콩 발효물을 제조한 것일 수 있으나, 이에 제한되지 않는다.In the manufacturing method according to an embodiment of the present invention, the spinach may be inoculated with lactic acid bacteria in a spinach juice prepared by mixing and pulverizing spinach and water, and the beans are prepared as soybeans by mixing soybean powder and water and may be inoculated with lactic acid bacteria, and may be prepared by fermenting spinach fermented products and soybean fermented products, but is not limited thereto.
또한, 상기 유산균은 락토바실러스 플랜타룸(Lactobacillus plantarum) 균주일 수 있으며, 바람직하게는 기탁번호가 KACC92083P인 락토바실러스 플랜타룸 CBG-C21(Lactobacillus plantarum CBG-C21) 균주일 수 있으나, 이에 제한되지 않는다.In addition, the lactic acid bacteria may be a Lactobacillus plantarum strain, preferably a Lactobacillus plantarum CBG-C21 strain having an accession number of KACC92083P, but is not limited thereto.
또한, 상기 시금치 발효물 내에 함량이 증가된 유용성분은 L-아스코르브산(L-ascorbic acid)이고, 상기 콩 발효물 내에 함량이 증가된 유용성분은 아글리콘형 이소플라본일 수 있으며, 바람직하게는 다이드제인(daidzein), 제니스테인(genistein) 및 글리시테인(glycitein)일 수 있으나, 이에 제한되지 않는다.In addition, the useful component having an increased content in the spinach fermented product is L-ascorbic acid, and the useful component having an increased content in the soybean fermented product may be an aglycone-type isoflavone, preferably Daidzein (daidzein), genistein (genistein), and may be glycitein (glycitein), but is not limited thereto.
또한, 상기 발효는 32~42℃에서 2~28시간 동안 수행할 수 있으며, 바람직하게는 시금치 발효의 경우 37℃에서 23시간 동안 수행한 것일 수 있고, 콩 발효의 경우 37℃에서 4~6시간 동안 수행한 것일 수 있으나, 이에 제한되지 않는다.In addition, the fermentation may be performed at 32 to 42 ° C for 2 to 28 hours, preferably spinach fermentation may be performed at 37 ° C for 23 hours, and in the case of soybean fermentation, 4 to 6 hours at 37 ° C. It may be carried out during the period, but is not limited thereto.
본 발명에 따른 제조 방법은 바람직하게는,The manufacturing method according to the present invention preferably comprises:
1) 시금치 즙액 또는 콩물을 멸균하는 단계;1) sterilizing spinach juice or soybean water;
2) 상기 단계 1)에서 멸균한 시금치 즙액 또는 콩물에 유산균을 접종하여 발효하는 단계; 및2) fermenting by inoculating lactic acid bacteria in the spinach juice or soybean water sterilized in step 1); and
3) 상기 단계 2)의 시금치 발효물 또는 콩 발효물을 동결건조하여 분말화하는 단계;를 포함할 수 있으며, 더욱 바람직하게는3) freeze-drying the fermented spinach or fermented soybean in step 2) to powder; may include, and more preferably
1) 시금치 즙액 또는 콩물을 100~120℃에서 10~20분 동안 멸균하는 단계;1) Sterilizing spinach juice or soybean water at 100-120° C. for 10-20 minutes;
2) 상기 단계 1)에서 멸균한 시금치 즙액에 2.2×107~2.2×109 CFU/㎖의 락토바실러스 플랜타룸 CBG-C21 균주를 접종하거나 콩물에 5.9×1010~5.9×1012 CFU/g의 락토바실러스 플랜타룸 CBG-C21 균주를 접종하여 32~42℃에서 2~28시간 동안 발효하는 단계; 및 2) Inoculate 2.2×10 7 ~2.2×10 9 CFU/ml Lactobacillus plantarum CBG-C21 strain into the spinach juice sterilized in step 1) or 5.9×10 10 ~5.9×10 12 CFU/g in soybean water Inoculating the Lactobacillus plantarum CBG-C21 strain of 32-42 ℃ fermenting for 2 to 28 hours; and
3) 상기 단계 2)의 시금치 발효물 또는 콩 발효물을 동결건조하여 분말화하는 단계;를 포함할 수 있으며, 가장 바람직하게는3) lyophilizing the fermented spinach or fermented soybean in step 2) to powder it; may include, and most preferably
1) 시금치 즙액 또는 콩물을 121℃에서 15분 동안 멸균하는 단계;1) Sterilizing spinach juice or soybean water at 121° C. for 15 minutes;
2) 상기 단계 1)에서 멸균한 시금치 즙액에 2.2×108 CFU/㎖의 락토바실러스 플랜타룸 CBG-C21 균주를 접종하여 37℃에서 23시간 동안 발효하거나 콩물에 5.9×1011 CFU/g의 락토바실러스 플랜타룸 CBG-C21 균주 농축액을 콩물 중량 대비 10%를 접종하여 37℃에서 4~6시간 동안 발효하는 단계; 및 2) Inoculate 2.2×10 8 CFU/ml Lactobacillus plantarum CBG-C21 strain into the spinach juice sterilized in step 1) and ferment at 37° C. for 23 hours or 5.9×10 11 CFU/g lactobacilli in soybean water Inoculating the Bacillus plantarum CBG-C21 strain concentrate with 10% of the weight of soybean water and fermenting at 37° C. for 4 to 6 hours; and
3) 상기 단계 2)의 시금치 발효물 또는 콩 발효물을 동결건조하여 분말화하는 단계;를 포함할 수 있으나, 이에 제한되지 않는다.3) freeze-drying the fermented spinach or fermented soybean in step 2) to powder; may include, but is not limited thereto.
본 발명은 또한, 상기 제조 방법으로 제조된 유용성분 함량이 증가된 시금치 발효물 또는 콩 발효물을 제공한다.The present invention also provides a spinach fermented product or soybean fermented product having an increased content of useful ingredients prepared by the above manufacturing method.
본 발명은 또한, 상기 유용성분 함량이 증가된 시금치 발효물 또는 콩 발효물을 포함하는 건강기능식품을 제공한다.The present invention also provides a health functional food comprising a spinach fermented product or soybean fermented product having an increased content of the useful ingredients.
상기 건강기능식품의 종류에는 특별한 제한은 없다. 상기 시금치 발효물 또는 콩 발효물을 함유하는 건강기능식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 떡류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다.There is no particular limitation on the type of the health functional food. Examples of health functional foods containing the spinach fermented product or soybean fermented product include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, rice cakes, dairy products including ice cream, various There are soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes, and includes all health functional foods in the ordinary sense.
본 발명에 따른 시금치 발효물은 발효 전 시금치 즙액에 비해 L-아스코르브산의 함량이 증가되고, 콩 발효물은 발효 전 콩물에 비해 다이드제인, 제니스테인 및 글리시테인의 함량이 증가된 것으로, 상기 유용성분들의 여성갱년기 증상 완화, 장 기능 또는 비만 개선 및 항산화 기능을 이용한 다양한 건강기능식품으로 이용될 수 있다.In the spinach fermented product according to the present invention, the content of L-ascorbic acid is increased compared to the spinach juice before fermentation, and the contents of daidzein, genistein and glycitein are increased in the soybean fermented product compared to the soybean water before fermentation. It can be used as various health functional foods using useful ingredients to relieve symptoms of menopause, improve intestinal function or obesity, and use antioxidant functions.
상기 건강기능식품은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽, 발포정 및 음료 중에서 선택된 어느 하나의 제형으로 제조될 수 있으나, 이에 제한되지 않는다.The health functional food may be prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, effervescent tablet and beverage, but is not limited thereto.
본 발명의 시금치 발효물 또는 콩 발효물을 식품첨가물로 사용하는 경우, 상기 시금치 발효물 또는 콩 발효물을 그대로 첨가하거나 다른 식품 또는 식품성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효성분은 그의 사용 목적에 따라 적절하게 사용될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 발명의 시금치 발효물 또는 콩 발효물은 원료에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양의로 첨가된다. 그러나 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 안전성 면에서 아무런 문제가 없는 범위의 양으로 사용될 수 있다.When using the fermented spinach or fermented soybean of the present invention as a food additive, the fermented spinach or fermented soybean may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. have. The active ingredient may be appropriately used depending on the purpose of its use. In general, in the production of food or beverage, the fermented spinach or fermented soybean of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health control, it can be used in an amount within a range that does not cause any problems in terms of safety.
본 발명의 건강기능식품은 식품 제조 시에 통상적으로 첨가되는 성분을 포함하며, 예를 들어, 단백질, 탄수화물, 지방, 영양소 및 조미제를 포함한다. 예컨대, 드링크제로 제조되는 경우에는 유효성분 이외에 천연 탄수화물 또는 향미제를 추가 성분으로서 포함시킬 수 있다. 상기 천연 탄수화물은 모노사카라이드(예컨대, 글루코오스, 프럭토오스 등), 디사카라이드(예컨대, 말토스, 수크로오스 등), 올리고당, 폴리사카라이드(예컨대, 덱스트린, 시클로덱스트린 등) 또는 당알코올(예컨대, 자일리톨, 소르비톨, 에리쓰리톨 등)인 것이 바람직하다. 상기 향미제는 천연 향미제(예컨대, 타우마틴, 스테비아 추출물 등)와 합성 향미제(예컨대, 사카린, 아스파르탐 등)를 이용할 수 있다. 상기 건강기능식품 조성물 외에 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 더 함유할 수 있다. The health functional food of the present invention includes ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when manufactured as a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrin, cyclodextrin, etc.) or sugar alcohols (eg, , xylitol, sorbitol, erythritol, etc.). As the flavoring agent, natural flavoring agents (eg, taumatine, stevia extract, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) may be used. In addition to the health functional food composition, various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonic acid It may further contain a carbonation agent and the like used in beverages.
상기 첨가되는 성분의 비율은 크게 중요하진 않지만 본 발명의 건강기능식품 100 중량부에 대하여, 0.01 내지 0.1 중량부의 범위에서 선택되는 것이 일반적이다.Although the ratio of the added component is not very important, it is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food of the present invention.
이하, 본 발명의 실시예에 의해 상세히 설명한다. 단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.Hereinafter, an embodiment of the present invention will be described in detail. However, the following examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following examples.
제조예 1. 시금치 발효물 제조Preparation Example 1. Preparation of fermented spinach
90.97g의 시금치를 칭량하여 210㎖의 정제수와 혼합하여 시금치의 함량이 30%(w/v)가 되도록 한 후 믹서기로 10분간 분쇄하여 획득한 시금치 즙액을 pH 7.0으로 보정하였다. 보정한 시금치 즙액에 2%(w/v) 포도당 및 1%(w/v) 카제인 펩톤을 첨가한 후 121℃에서 15분간 가압멸균하였다. 발효 균주인 락토바실러스 플랜타룸(Lactobacillus plantarum) CBG-C21 균주를 24시간 동안 배양하여 획득한 배양액(1.1×1010 CFU/㎖)을 이용하였다. 상기 멸균한 시금치 즙액에 CBG-C21 균주 배양액 2%(2.2×108 CFU/㎖)를 접종하여 37℃에서 23시간 동안 정치배양하여 발효를 수행하였다. 배양이 완료된 시금치 발효물에서 CBG-C21 균주의 생균수(3.9×109 CFU/㎖)를 확인하고 시금치 발효물에 덱스트린을 첨가한 후, 동결건조하여 분말화 하였다.After weighing 90.97 g of spinach, mixing it with 210 ml of purified water so that the content of spinach becomes 30% (w/v), and grinding with a mixer for 10 minutes, the obtained spinach juice was corrected to pH 7.0. 2% (w/v) glucose and 1% (w/v) casein peptone were added to the corrected spinach juice, and then autoclaved at 121° C. for 15 minutes. Fermentation strain Lactobacillus plantarum ( Lactobacillus plantarum ) The culture medium (1.1 × 10 10 CFU / ㎖) obtained by culturing the CBG-C21 strain for 24 hours was used. The sterilized spinach juice was inoculated with 2% of the CBG-C21 strain culture medium (2.2×10 8 CFU/ml), and the fermentation was performed by stationary culture at 37° C. for 23 hours. The number of viable cells (3.9×10 9 CFU/ml) of the CBG-C21 strain was confirmed in the fermented spinach, and dextrin was added to the fermented spinach, followed by freeze-drying and powdering.
제조예 2. 콩 발효물 제조Preparation Example 2. Preparation of soybean fermented product
백태 분말을 칭량하여 정제수를 이용하여 10%(w/v) 콩물을 제조하였다. 제조한 10%(w/v) 콩물을 121℃에서 15분간 가압멸균하였다. 발효 균주인 락토바실러스 플랜타룸(Lactobacillus plantarum) CBG-C21 균주를 24시간 동안 배양한 후 원심분리하여 생균수 5.9×1011 CFU/g 농축액을 이용하였다. 콩 발효를 위해 고형분 함량 기준으로 균체 및 콩을 1:9의 혼합비율로 적용하여 실시하였으며 이를 위해 균주 농축액 657g과 10% 콩물 13,920g을 혼합하여 37℃에서 4~6시간동안 정치배양하여 발효를 수행했다. 배양이 완료된 배양액을 동결건조하여 분말화 하였다.By weighing the white powder, purified water was used to prepare 10% (w/v) soybean water. The prepared 10% (w/v) soybean water was autoclaved at 121° C. for 15 minutes. The fermentation strain, Lactobacillus plantarum ( Lactobacillus plantarum ) CBG-C21 strain was cultured for 24 hours and then centrifuged to use a live cell count of 5.9×10 11 CFU/g concentrate. For soybean fermentation, cells and soybeans were applied at a mixing ratio of 1:9 based on the solid content. carried out The culture solution was lyophilized and powdered.
실시예 1. 시금치 발효물 내의 L-아스코르브산 함량 분석Example 1. Analysis of L-Ascorbic Acid Content in Fermented Spinach
시금치 즙액 분말과 제조예 1에서 제조한 시금치 발효물 분말을 대상으로 L-아스코르브산(L-ascorbic acid) 함량을 측정하였다. L-아스코르브산 함량은 Megazyme assay kit(Megazyme Ltd.)를 이용하여 측정하였다. 구체적으로, 100mg의 시료에 3%(w/v) 메타인산(metaphosphoric acid) 및 10mM EDTA(ethylenediaminetetraacetic acid)를 첨가하여 상온에서 60분간 균질화하여 10분간 원심분리한 후 여과한 상등액을 사용하였다. 10㎕의 여과한 상등액에 152㎕의 3차 증류수 및 버퍼 1을 혼합하고 3분 후 MTT[3-(4,5-dimethylthiazolyl-2) -2,5-diphenyltetrazolium bromide] 버퍼 2를 넣고 3분간 반응시켰으며, 분광광도계를 이용하여 578nm에서 1차 흡광도(A1)를 측정하고, PMS(5-methylphenazinium methyl sulfate) 버퍼를 넣고 8분 동안 반응시킨 후 578nm에서 2차 흡광도(A2)를 측정하였다. 그 결과, 도 1에 개시한 바와 같이 시금치 즙액 분말(0.16 g/100g)에 비해 시금치 발효물 분말(0.22 g/100g)에서 L-아스코르브산 함량이 37.5% 증가한 것을 확인할 수 있었다.L-ascorbic acid (L-ascorbic acid) content was measured for the spinach juice powder and the spinach fermented powder prepared in Preparation Example 1. The L-ascorbic acid content was measured using a Megazyme assay kit (Megazyme Ltd.). Specifically, 3% (w/v) metaphosphoric acid and 10 mM EDTA (ethylenediaminetetraacetic acid) were added to a 100 mg sample, homogenized at room temperature for 60 minutes, centrifuged for 10 minutes, and filtered supernatant was used. 152 μl of tertiary distilled water and buffer 1 were mixed with 10 μl of the filtered supernatant, and after 3 minutes, MTT [3-(4,5-dimethylthiazolyl-2) -2,5-diphenyltetrazolium bromide] buffer 2 was added and reacted for 3 minutes The primary absorbance (A1) was measured at 578 nm using a spectrophotometer, and a PMS (5-methylphenazinium methyl sulfate) buffer was added and reacted for 8 minutes, followed by measuring the secondary absorbance (A2) at 578 nm. As a result, as shown in FIG. 1 , it was confirmed that the L-ascorbic acid content was increased by 37.5% in the spinach fermented product powder (0.22 g/100 g) compared to the spinach juice powder (0.16 g/100 g).
실시예 2. 콩 발효물 내의 이소플라본 함량 분석Example 2. Analysis of isoflavone content in fermented soybeans
백태 콩 분말과 제조예 2에서 제조한 콩 발효물 분말을 대상으로 이소플라본 함량을 측정하였다. 이소플라본 함량은 HPLC 분석법을 이용하여 측정하였다. 각 시료를 0.1g씩 개별 칭량한 후 50%(v/v) 에탄올 1ml을 첨가하고 1시간 동안 초음파 추출을 진행하였다. 획득한 추출액을 원심분리를 실시하여 상층액만을 취하였으며, Agilent 1200 HPLC 장비를 통해 분석하였다. 분석은 Multi-wavelength detector를 사용하고 분석 컬럼으로는 Shiseidotkdml CapcellPAK MGII C18(4.6X200mm ID, 3um)를 사용하였으며 이때 컬럼 온도는 35℃를 유지하였다. 15㎕의 추출액을 기기에 주입하고 용매로는 0.5% 포름산 수용액 및 아세토니트릴을 시간에 따라 농도 구배를 실시했으며 이때 용매의 흐름 속도는 0.5ml/min였다. 그 결과, 하기 표 1에 개시한 바와 같이 백태콩 분말에 비해 콩 발효물 분말에서 이소플라본 아글리콘(비배당체) 형태인 다이드제인(daidzein), 제니스테인(genistein) 및 글리시테인(glycitein)의 함량이 각각 약 16배, 10배 및 5배 증가한 것을 확인할 수 있었다.The content of isoflavones was measured for the white soybean powder and the soybean fermented product powder prepared in Preparation Example 2. The isoflavone content was determined using HPLC analysis. After weighing each sample individually by 0.1 g, 1 ml of 50% (v/v) ethanol was added, and ultrasonic extraction was performed for 1 hour. The obtained extract was centrifuged to take only the supernatant, and was analyzed using an Agilent 1200 HPLC instrument. A multi-wavelength detector was used for analysis, and Shiseidotkdml CapcellPAK MGII C18 (4.6X200mm ID, 3um) was used as an analysis column, and the column temperature was maintained at 35°C. 15 μl of the extract was injected into the instrument, and a concentration gradient of 0.5% aqueous formic acid and acetonitrile as a solvent was performed over time, and the flow rate of the solvent was 0.5 ml/min. As a result, as shown in Table 1 below, daidzein, genistein, and glycitein, which are isoflavone aglycone (non-glycoside) forms, in fermented soybean powder compared to white soybean powder, It was confirmed that the content increased about 16 times, 10 times, and 5 times, respectively.
발효 분말white soybean
fermented powder
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