KR102547999B1 - Method for manufacturing fermented product enhanced free amino acid content comprising multi-stage fermentation of soybean by-products - Google Patents
Method for manufacturing fermented product enhanced free amino acid content comprising multi-stage fermentation of soybean by-products Download PDFInfo
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- KR102547999B1 KR102547999B1 KR1020220148551A KR20220148551A KR102547999B1 KR 102547999 B1 KR102547999 B1 KR 102547999B1 KR 1020220148551 A KR1020220148551 A KR 1020220148551A KR 20220148551 A KR20220148551 A KR 20220148551A KR 102547999 B1 KR102547999 B1 KR 102547999B1
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- bacillus
- fermentation
- soybean
- fermented product
- products
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Abstract
Description
본 발명은 대두부산물과 참깨부산물의 다단발효를 포함하는 발효물의 제조방법에 관한 것으로, 보다 구체적으로 대두 가공 시 부산물로 발생하는 대두피(배아 포함)와 참기름 제조과정에서 발생하는 참깨박을 이용하여 고초균(B.spp)으로 1차 발효하고, 유산균(L.spp)으로 2차 발효하여 얻어지는 발효물의 제조방법에 관한 것이다. The present invention relates to a method for producing a fermented product including multi-stage fermentation of soybean by-products and sesame by-products, and more specifically, by using soybean hulls (including embryos) generated as by-products during soybean processing and sesame meal generated during the sesame oil manufacturing process. It relates to a method for producing a fermented product obtained by primary fermentation with Bacillus subtilis ( B.spp ) and secondary fermentation with lactic acid bacteria (L.spp ).
대두는 단백질의 함량이 높을 뿐만 아니라 포화지방산 함량이 낮고 콜레스테롤이 거의 함유되어 있지 않으며 비타민과 무기질 및 식이섬유소까지 풍부하게 함유되어 있다. Soybeans are high in protein, low in saturated fatty acids, contain almost no cholesterol, and are rich in vitamins, minerals, and dietary fiber.
대두 부산물은 상기 대두를 가공하는 과정에서 발생하는 것으로, 배아, 대두피, 순물, 침지수 등 대두단백질을 제외한 나머지 모두를 포함할 수 있다. Soybean by-products are generated in the process of processing the soybeans, and may include everything except for soybean proteins, such as embryos, soybean hulls, pure water, and steep water.
그 중에서도 대두피 (또는 대두외피, soybean hull)는 대두유(soybean oil), 두유 및 두부 제조를 위한 공정 중 다량발생하고 있는 부산물로 그 성분 중 조섬유질의 함량이 40 ％ 이상이며, 특히 대두피의 조섬유질은 높은 소화흡수율을 보이는 것으로 알려져 있다. Among them, soybean hull (or soybean hull) is a by-product that is generated in large quantities during the manufacturing process of soybean oil, soybean milk, and tofu, and contains more than 40% of crude fiber among its components. Fiber is known to have a high digestibility.
대두피를 탈피하는 과정에서 대두배아가 함께 떨어져 나가는 경우가 많은데, 이때 유실되는 대두배아에는 대두단백질의 13 ~ 15%의 단백질이 함유되어 있는 것으로 알려져 있다. In the process of shedding soybean skin, soybean embryos often fall together, and it is known that the soybean embryos that are lost at this time contain 13 to 15% of soybean protein.
특히 대두피와 탈피시 유실되는 대두배아에서 얻어지는 생리 활성 성분으로는 조섬유질 외에 암을 예방하고 호르몬대사를 원활하게 하는 이소플라본, 지질대사를 돕고 항산화 효과가 있는 사포닌, 콜레스테롤 대사를 조절하는 파이틴, 섬유질과 같이 소화기능을 돕는 올리고당, 생체방어기전에 관여하는 레시틴 등이 있다.In particular, physiologically active ingredients obtained from soybean hulls and soybean embryos that are lost during shedding include, in addition to crude fiber, isoflavones that prevent cancer and facilitate hormone metabolism, saponin that helps lipid metabolism and has an antioxidant effect, and phytin that regulates cholesterol metabolism. , oligosaccharides that help digestion, such as fiber, and lecithin, which is involved in biological defense mechanisms.
대두는 대두유, 대두단백 등의 다양한 형태로 가공되어 사용되고 있으나, 대두의 껍질인 대두피(soybean hull)는 이의 가공방법이 개발되지 않아, 현재까지는 식품으로 활용되지 않고 버려지거나 또는 동물사료의 원료 등으로만 주로 활용되고 있는 실정이다. 또한, 대두피에는 대두배아가 포함되어 있는 경우가 많아 단백질 함량이 높으므로 본 발명의 대두피를 사용하는 경우에 다른 대두부산물에 비해 발효물의 유리아미노산 함량이 우수한 발효물을 얻을 수 있다. Soybeans are processed and used in various forms such as soybean oil and soybean protein, but soybean hull, the soybean husk, has not been developed, so it is not used as food and is discarded or used as a raw material for animal feed. It is mainly used only as a In addition, since soybean hulls often contain soybean embryos and have high protein content, when using the soybean husk of the present invention, a fermented product having a higher free amino acid content than other soybean by-products can be obtained.
참깨는 참깨과에 속하는 1년생 초본식물로써, 원산지는 고온 건조한 에티오피아와 인도로 추정되며, 아프리카에서 동쪽으로, 인도에서 서쪽으로 전파된 식물이다. 참깨의 주요 생산 국가는 인도, 미얀마, 수단, 중국이며, 전세계 재배면적의 70%, 총생산량의 60%를 차지하고 있다. 참깨는 우리나라에서 유지작물로 재배되어 왔으며 깨소금, 참기름 등으로 많이 이용되고 있다. 일반적으로 참깨 종자에는 지방 50%, 단백질 20%, 탄수화물 15%, 리그난 0.5% 정도 함유되어 있으며, 참기름의 지방산 중 주성분은 불포화지방산인 오레인산, 리놀레산으로 이들은 혈액 중의 혈청콜레스테롤의 농도를 낮추어 동맥경화를 예방하고, 혈압 상승을 억제하는 작용을 한다. 특히 참깨 종자에 함유된 리그난 성분은 강력한 항산화 물질로서 참기름의 산패를 억제하여 저장안정성을 높일 뿐만 아니라 인체의 각종 혈관 질환을 예방하고 면역 기능을 강화하는 역할을 한다.Sesame is an annual herbaceous plant belonging to the Sesame family. Its origin is estimated to be hot and dry Ethiopia and India, and it is a plant that has spread from Africa to the east and from India to the west. The main producing countries of sesame are India, Myanmar, Sudan, and China, which account for 70% of the world's cultivation area and 60% of the total production. Sesame has been cultivated as an oil crop in Korea and is widely used as sesame salt and sesame oil. In general, sesame seeds contain 50% fat, 20% protein, 15% carbohydrates, and 0.5% lignans. Among the fatty acids in sesame oil, the main components are oleic acid and linoleic acid, which are unsaturated fatty acids. It prevents and suppresses blood pressure rise. In particular, the lignan component contained in sesame seeds is a powerful antioxidant that inhibits rancidity of sesame oil to increase storage stability, prevents various vascular diseases in the human body, and strengthens the immune function.
이러한 참깨는 아직까지 그 쓰임새가 다양하지 못하다. 현재는 참깨를 고온에서 볶거나 생들깨를 압착 하여 참기름 또는 들기름으로 제조해 식용유로써 가장 많이 사용하고 있다. 그러나 기름을 착유하는 과정에서 부산물이 많이 생성된다 These sesame seeds have not yet been used in a variety of ways. Currently, sesame seeds are roasted at high temperature or raw perilla seeds are pressed to make sesame oil or perilla oil, which is most often used as cooking oil. However, many by-products are produced in the process of milking oil.
상기 참깨 부산물은 참깨박일 수 있다. 참기름 착유 과정에서 특히 깻묵 또는 유박이라고도 불리는 참깨박이 많이 생성된다. 참깨박에는 섬유질을 비롯한 양질의 단백질 및 잔여 지방 성분들이 남아 있는데도 불구하고 그 동안은 사료나 비료 등으로 재활용을 하거나 폐기 처분해왔으나, 최근 들어 각종 식품이나 기능성 조성물의 용도로 활용할 수 있는 방안이 다양하게 연구되고 있다.The sesame by-product may be sesame meal. In the process of milking sesame oil, a lot of sesame meal, also called seed cake or oil cake, is produced. Although sesame meal contains fiber, high-quality protein and residual fat, it has been recycled or discarded as feed or fertilizer, but recently, there are various ways to use it for various foods or functional compositions. are being researched.
발효는 미생물이 가지고 있는 효소를 이용하여 유기물을 분해시켜 유용한 물질을 생성하거나 인체 세포에 흡수가 용이한 성분으로 전환한다. 상기 발효 미생물은 고초균(Bacillus subtilis), 유산균(Lactic acid bacteria), 초산균(Acetic acid bacteria), 낙산균(butyric acid bacteria), 화락균(hiochi bacteria), 곰팡이균(fungus) 및 효모 등이 있다.Fermentation uses enzymes possessed by microorganisms to decompose organic substances to produce useful substances or convert them into components that are easily absorbed by human cells. The fermenting microorganisms include Bacillus subtilis, lactic acid bacteria, acetic acid bacteria, butyric acid bacteria, hiochi bacteria, fungus and yeast.
이들 발효 미생물은 발효원에 따라 생산되는 물질이 상이하고, 경우에 따라서는 인체에 유해한 물질을 생산하므로 발효원 및 발효목적에 따라 유용한 발효 미생물을 선택하는 것이 중요하다.Since these fermenting microorganisms produce different substances depending on the fermentation source and, in some cases, produce substances harmful to the human body, it is important to select useful fermentation microorganisms according to the fermentation source and fermentation purpose.
대한민국 공개특허 제 10-2022-0118682호는 깻묵 메주와 쌈장의 제조방법에 관한 것으로, 대두와 깻묵(참깨박)을 이용하여 혼합 발효시키는 방법을 개시하고 있다. Republic of Korea Patent Publication No. 10-2022-0118682 relates to a method for manufacturing seed cake meju and ssamjang, and discloses a method of mixing and fermenting soybeans and seed cake (sesame meal).
대한민국 등록특허 제 10-1883626호는 발효 사료 조성물을 반추동물에 급여하여 육류에서 불포화지방의 올레산 함량을 증가시키는 반추동물용 단미사료 및 그 제조방법에 관한 것으로, 대두피 및 참깨박을 이용하여 고초균, 유산균, 효모 등 생균제로 발효시킨 발효물 제조 방법을 개시하고 있다. Korean Registered Patent No. 10-1883626 relates to a simple feed for ruminants that increases the oleic acid content of unsaturated fat in meat by feeding a fermented feed composition to ruminants and a method for manufacturing the same, using soybean husk and sesame meal. , Disclosed is a method for producing a fermented product fermented with probiotics such as lactic acid bacteria and yeast.
대한민국 등록특허 제 10-1248572호는 참깨박과 대두의 혼합 및 발효를 이용한 기능성 떡갈비 및 그 제조방법에 관한 것으로, 참깨박과 대두를 고초균으로 발효시켜 발효 과정에서 고초균이 생산하는 효소에 의해 발생하는 불쾌취를 참깨박을 통해 후각적 및 미각적으로 개선시킬 수 있음을 개시하고 있다. Republic of Korea Patent Registration No. 10-1248572 relates to functional tteokgalbi using mixing and fermentation of sesame meal and soybeans and its manufacturing method. It is disclosed that unpleasant odor can be improved olfactory and taste through sesame meal.
이처럼 종래 발효물의 제조에 있어서 대두 부산물이나 참깨 부산물을 이용한 예가 있으나, 이들은 재료를 생균제와 함께 모두 혼합하여 발효하는 것으로, 1, 2차 발효로 나누어 발효하는 방법에 대해서는 개시하고 있지 않다. As such, there are examples of using soybean by-products or sesame by-products in the production of conventional fermented products, but these are fermented by mixing all the materials together with probiotics, and the method of fermentation by dividing into primary and secondary fermentation is not disclosed.
대두 부산물 또는 참깨 부산물의 발효를 통해 유리아미노산을 증가시키는 것은 종래 알려져 있으나, 본 발명의 2단 발효에 의하는 경우에 종래 기술에 비해 유리아미노산 함량이 현저하게 증가되는 효과에 대해서는 알려져 있지 않다. 이에, 본 발명자들은 대두 분말 가공 시 발생하는 대두 부산물과 참기름 제조과정에서 발생하는 참깨 부산물을 이용하여 천연 발효물을 개발하고자 예의 노력한 결과, 2단 발효한 경우에 1단 발효에 비해 아미노산 함량이 5~7배 증가함을 확인하여 본 발명을 완성하기에 이르렀다. It is conventionally known to increase free amino acids through fermentation of soybean by-products or sesame by-products, but the effect of significantly increasing the free amino acid content compared to the prior art in the case of the two-stage fermentation of the present invention is not known. Accordingly, the present inventors have made diligent efforts to develop a natural fermented product using soybean by-products generated during soybean powder processing and sesame by-products generated during the sesame oil manufacturing process. It was confirmed that the increase was ~7 times, and the present invention was completed.
본 발명의 목적은 가공과정에서 발생하는 대두 부산물 및 참깨 부산물을 이용하여 다단발효를 통해 감칠맛이 상승한 발효물을 제조하는 방법 및 그러한 방법에 의해 제조된 발효물을 제공하는 것이다. An object of the present invention is to provide a method for producing a fermented product with increased umami through multi-stage fermentation using soybean by-products and sesame by-products generated in the processing process, and a fermented product produced by the method.
본 발명은the present invention
발효물의 제조방법으로서, As a method for producing a fermented product,
1) 대두 부산물을 발효하여 대두 부산물의 1단 발효물을 제조하는 단계;1) fermenting soybean by-products to prepare a first-stage fermentation product of soybean by-products;
2) 참깨 부산물을 발효하여 참깨 부산물의 1단 발효물을 제조하는 단계;2) fermenting sesame by-products to prepare a first-stage fermentation product of sesame by-products;
3) 상기 대두 부산물의 1단 발효물과 상기 참깨 부산물의 1단 발효물을 혼합하는 단계;3) mixing the first-stage fermentation product of the soybean by-product and the first-stage fermentation product of the sesame by-product;
4) 상기 혼합물에 유산균을 접종하여 2단 발효물을 제조하는 단계; 및4) preparing a two-stage fermented product by inoculating the mixture with lactic acid bacteria; and
5) 상기 2단 발효물을 여과하여 얻은 상층액을 농축하여 발효 농축액을 얻는 단계;를 포함하는 것을 특징으로 하는 유리 아미노산이 증가된 발효물의 제조방법을 제공한다. 5) obtaining a fermentation concentrate by concentrating the supernatant obtained by filtering the two-stage fermentation product; it provides a method for producing a fermentation product with increased free amino acids, characterized in that it comprises a.
또한, 본 발명은 상기 발효물의 제조방법에 의해 제조된 발효물을 제공한다. In addition, the present invention provides a fermented product prepared by the method for producing a fermented product.
또한, 본 발명은 대두부산물과 참깨부산물을 다단발효시키는 단계를 포함하는 것을 특징으로 하는 유리 아미노산 함량을 증가시키는 방법을 제공한다. In addition, the present invention provides a method for increasing the free amino acid content, comprising the step of multi-stage fermentation of soybean by-products and sesame by-products.
본 발명은 대두 및 참깨의 가공과정에서 발생하는 대두 부산물 및 참깨 부산물을 이용하여 발효물을 제조하는 방법을 제공한다. 본 발명의 제조 방법에 의하면 대두 및 참깨의 부산물을 다단발효함으로써 유리아미노산 함량이 증가하며, 특히 감칠맛을 내는 아미노산인 글루타민과 감칠맛 상승 효과가 있는 알라닌의 함량이 현저히 증가함으로써 감칠맛이 우수한 천연발효물을 제조할 수 있다. 본 발명에 의하면 가치가 낮아 폐기의 대상이 되는 부산물을 이용하여 발효를 통해 아미노산 함량이 증가된 천연발효물을 제조할 수 있다. The present invention provides a method for preparing a fermented product using soybean by-products and sesame by-products generated in the processing of soybeans and sesame seeds. According to the manufacturing method of the present invention, the content of free amino acids is increased by multi-stage fermentation of by-products of soybeans and sesame seeds, and in particular, the content of glutamine, an amino acid that gives a umami taste, and alanine, which have a umami-enhancing effect, are remarkably increased, thereby producing a naturally fermented product with excellent umami taste. can be manufactured According to the present invention, a naturally fermented product having an increased amino acid content can be produced through fermentation using by-products that are discarded due to their low value.
도 1은 실험예 1의 최종 발효물에서 필수아미노산별 함량을 비교한 결과를 나타낸다. A는 원료 증자 후 비발효 처리군, B는 유산균 1단 발효 처리군, C는 고초균으로 1단 발효 후 유산균으로 2단 발효한 발효 처리군이며, 각 처리군별로 필수아미노산인 Lysine, Threonine, Leucine, Isoleusine, Valine, Methionine, Phenylalanine, Tryptophan 및 총 필수아미노산의 함량을 나타낸다.
도 2는 실험예 2의 각 처리군별로 감칠맛을 내는 아미노산인 Glutamic acid 및 감칠맛을 상승시키는 Alanine 함량을 측정한 결과를 나타낸다. A는 원료 증자 후 비발효 처리군, B는 유산균 1단 발효 처리군, C는 고초균으로 1단 발효 후 유산균으로 2단 발효한 발효 처리군이다. Figure 1 shows the results of comparing the content of each essential amino acid in the final fermented product of Experimental Example 1. A is a non-fermentation treatment group after raw material increase, B is a lactobacillus first fermentation treatment group, C is a fermentation treatment group obtained by Bacillus subtilis first fermentation followed by lactic acid bacteria second fermentation treatment, and essential amino acids for each treatment group are Lysine, Threonine, and Leucine. , Isoleusine, Valine, Methionine, Phenylalanine, Tryptophan and total essential amino acid content.
Figure 2 shows the results of measuring the content of Glutamic acid, which is an amino acid that gives umami, and Alanine, which increases umami, for each treatment group in Experimental Example 2. A is a non-fermentation treatment group after raw material increase, B is a lactic acid bacteria first fermentation treatment group, C is a fermentation treatment group after first fermentation with Bacillus subtilis and second fermentation with lactic acid bacteria.
다른 식으로 정의되지 않는 한, 본 명세서에서 사용된 기술적 및 과학적 용어들은 그 기술분야에서 통상의 지식을 가진 전문가에게 통상적으로 이해되는 것과 동일한 의미를 갖는다. 또한, 본 명세서에서 명시된 실험 과정은 특별히 설명되지 않는 이상 그 기술분야에서 통상적으로 수행되는 실험 과정과 동일하다. Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, the experimental procedures specified in this specification are the same as those commonly performed in the art unless otherwise specified.
이하, 본 발명에 대하여 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 대두 부산물 및 참깨 부산물을 이용하여 다단발효함으로써 유리아미노산 함량이 증가하여 감칠맛이 우수한 발효물의 제조방법을 제공한다.The present invention provides a method for producing a fermented product having excellent umami by increasing free amino acid content by multi-stage fermentation using soybean by-products and sesame by-products.
본 발명의 일 실시예에 따른 대두 부산물 및 참깨 부산물의 다단발효를 이용한 발효물의 제조방법은Method for producing a fermented product using multi-stage fermentation of soybean by-products and sesame by-products according to an embodiment of the present invention
1) 대두 부산물을 발효하여 대두 부산물의 1단 발효물을 제조하는 단계;1) fermenting soybean by-products to prepare a first-stage fermentation product of soybean by-products;
2) 참깨 부산물을 발효하여 참깨 부산물의 1단 발효물을 제조하는 단계;2) fermenting sesame by-products to prepare a first-stage fermentation product of sesame by-products;
3) 상기 대두 부산물의 1단 발효물과 상기 참깨 부산물의 1단 발효물을 혼합하는 단계;3) mixing the first-stage fermentation product of the soybean by-product and the first-stage fermentation product of the sesame by-product;
4) 상기 혼합물에 유산균을 접종하여 2단 발효물을 제조하는 단계; 및4) preparing a two-stage fermented product by inoculating the mixture with lactic acid bacteria; and
5) 상기 2단 발효물을 여과하여 얻은 상층액을 농축하여 발효 농축액을 얻는 단계;를 포함할 수 있다. 5) obtaining a fermentation concentrate by concentrating the supernatant obtained by filtering the two-stage fermentation product; may include.
본 발명에서 대두 부산물은 배아, 대두피, 순물, 침지수 등 대두단백질을 제외한 나머지 모두를 사용할 수 있으며, 바람직하게는 대두피를 사용할 수 있다. In the present invention, soybean by-products may be used, except for soybean protein, such as embryos, soybean hulls, pure water, and steeping water, and preferably, soybean hulls may be used.
대두피에는 대두피의 탈피 과정에서 함께 떨어져 나가는 대두배아를 포함하고 있는 경우가 많으며, 상기 대두배아에는 대두단백질의 13 ~ 15%의 단백질을 함유하고 있어 대두피를 사용하는 경우에 단백질 발효에 의한 유리아미노산 함량 증가 효과가 우수하다. Soybean hulls often contain soybean embryos that fall together during the shedding process of soybean hulls, and the soybean embryos contain 13 to 15% of soybean protein. The effect of increasing the amino acid content is excellent.
본 발명의 대두 및 참깨 부산물의 1단 발효물 제조 과정은 배지에서 고초균을 접종하여 발효하는 과정을 포함하는 것을 특징으로 한다. The process for producing a one-stage fermented product of soybean and sesame by-products of the present invention is characterized in that it includes a process of inoculating and fermenting Bacillus subtilis in a medium.
상기 배지는 사용하는 미생물의 종류와 성질에 따라 사용 가능한 식물성 또는 동물성의 모든 배지를 사용할 수 있으며, 바람직하게는 식물성 원료인 대두펩톤 1.5% (w/v), 효모추출물 2.5% (w/v), 글루코스 4.0% (w/v), 황산마그네슘 0.05% (w/v), 초산나트륨 1.5% (w/v)을 포함하는 배지를 사용할 수 있다. The medium may be any vegetable or animal medium that can be used depending on the type and nature of the microorganism used, and preferably 1.5% (w / v) of soybean peptone, 2.5% (w / v) of yeast extract, which is a vegetable raw material. , a medium containing 4.0% (w/v) glucose, 0.05% (w/v) magnesium sulfate, and 1.5% (w/v) sodium acetate may be used.
본 발명의 1단 발효는 고초균(B.spp)을 접종하는 것을 특징으로 하며, 상기 고초균의 접종량은 0.1 ~ 5% 바람직하게는 0.5 ~ 3% 더욱 바람직하게는 1% 이상 접종하는 것일 수 있다. The first stage fermentation of the present invention is characterized by inoculating Bacillus subtilis ( B.spp ), and the inoculum of B. subtilis may be 0.1 to 5%, preferably 0.5 to 3%, more preferably 1% or more.
고초균은 발효 과정을 거치면서 단백질 및 탄수화물 가수분해효소, 혈전 분해효소, 기능성 펩타이드 및 고분자 점질물 등을 포함하는 생리활성 물질을 생산하는 것으로 알려져 있다. 따라서, 본 발명에서 단백질을 많이 함유하고 있는 대두부산물 및 참깨부산물을 고초균으로 발효함으로써 대두부산물 및 참깨부산물의 단백질이 아미노산으로 분해된다. Bacillus subtilis is known to produce physiologically active substances including protein and carbohydrate hydrolases, thrombolytic enzymes, functional peptides, and polymeric mucilages through a fermentation process. Therefore, in the present invention, by fermenting soybean by-products and sesame by-products containing a lot of protein with Bacillus subtilis, the proteins of soybean by-products and sesame by-products are decomposed into amino acids.
상기 고초균은 발효에 관여하는 바실러스속 미생물(Bacillus sp.)이면 특별한 제한 없이 모두 사용할 수 있으며, 구체적으로 바실러스 섭틸리스 (B. subtilis), 바실러스 리크네포르미스 (B. lichneformis), 바실러스 메가테리움 (B. megaterium), 바실러스 아밀로리퀘파시엔스 (B. amyloliquefaciens), 바실러스 낫토 (B. natto), 바실러스 안스라시스 (B.antharcis), 바실러스 렌투스 (B.lentus), 바실러스 퍼미러스 (B.pumilus), 바실러스 더링지엔시스 (B.thuringiensis), 바실러스 알베이 (B.alvei), 바실러스 아조토픽산스 (B.azotofixans), 바실러스 매세란스 (B.macerans), 바실러스 포리믹사 (B.polymyxa), 바실러스 파필리에 (B.popilliae), 바실러스 코아글란스 (B.coagulans), 바실러스 스테아로더모필러스 (B.stearothermophilus), 바실러스 파스퇴리 (B.pasteurii), 바실러스 패리커스 (B.sphaericus), 바실러스 패스티디오서스 (B.fastidiosus) 등이 사용될 수 있다. The Bacillus subtilis may be used without particular limitation as long as it is a microorganism of the genus Bacillus involved in fermentation ( Bacillus sp. ), specifically Bacillus subtilis ( B. subtilis ), Bacillus lichneformis ( B. lichneformis ), Bacillus megate Leeum ( B. megaterium ), Bacillus amyloliquefaciens ( B. amyloliquefaciens ), Bacillus natto ( B. natto ), Bacillus anthracis ( B.antharcis ), Bacillus lentus ( B.lentus ), Bacillus fermirus ( B. pumilus ), Bacillus the ring gyensis ( B.thuringiensis ), Bacillus albei (B.alvei ), Bacillus azotopic sanseu ( B.azotofixans ), Bacillus Macerans ( B.macerans ), Bacillus forimixa ( B. polymyxa ), Bacillus papilliae ( B.popilliae ), Bacillus coagulans ( B.coagulans ), Bacillus stearothermophilus ( B.stearothermophilus ), Bacillus pasteurii ( B.pasteurii ), Bacillus paricus ( B.sphaericus ) , Bacillus fastidiosus ( B. fastidiosus ) and the like can be used.
본 발명의 1단 발효는 온도 15 ~ 50℃ 바람직하게는 35 ~ 45℃ 더욱 바람직하게는 45℃일 수 있다. 발효 온도가 위 범위보다 낮을 경우 발효 속도가 느려질 수 있고 또한 원하지 않는 발효 산물이 생겨날 수 있으며, 발효 온도가 위 범위보다 높을 경우도 발효 미생물의 사멸에 따라 마찬가지로 발효 속도가 느려지거나 원하지 않는 발효 산물이 생겨날 수 있다.The first stage fermentation of the present invention may be at a temperature of 15 to 50 ° C, preferably 35 to 45 ° C, more preferably 45 ° C. If the fermentation temperature is lower than the above range, the fermentation rate may slow down and unwanted fermentation products may be produced, and if the fermentation temperature is higher than the above range, the fermentation rate may also slow down or unwanted fermentation products may be produced due to the death of fermenting microorganisms. can arise
본 발명의 1단 발효는 12 ~ 36시간, 바람직하게는 18 ~ 30시간, 더욱 바람직하게는 24시간 동안 일 수 있다. 발효시간이 짧으면 발효가 잘 되지 않아 발효대사물이 적게 만들어지며 감칠맛이 덜하고, 발효시간이 길어지면 대사산물의 함량은 높아질 수 있으나 가스 생성 등으로 감칠맛 보다 신맛이 생길 우려가 있다. 온도가 높거나 발효시간이 길어지면 부패할 우려도 있다. 따라서 적당한 발효조건(온도, 시간)이 중요하다.The first stage fermentation of the present invention may be performed for 12 to 36 hours, preferably 18 to 30 hours, and more preferably 24 hours. If the fermentation time is short, fermentation does not work well, resulting in less fermentation metabolites and less savory taste. If the fermentation time is longer, the content of metabolites may increase, but there is a risk that a sour taste rather than a savory taste may occur due to gas generation. If the temperature is high or the fermentation time is long, there is a risk of spoilage. Therefore, proper fermentation conditions (temperature, time) are important.
또한 본 발명의 고초균에 의한 발효 후에 숙성 단계가 추가로 포함될 수 있는데, 이러한 숙성 단계는 접종된 미생물의 생장이 정지된 후에도 미생물에 의한 발효 산물을 안정화하기 위한 것으로, 상기 숙성 단계는 1일 내지 12개월 정도 수행될 수 있다. In addition, after fermentation by the Bacillus subtilis of the present invention, a maturation step may be additionally included. This maturation step is to stabilize the fermentation product by the microorganism even after the growth of the inoculated microorganism is stopped. It can take about a month.
또한 고초균에 의한 발효는 혐기적 조건과 호기적 조건 모두에서 배양이 가능하기 때문에 배양 조건이 혐기적이거나 호기적인 조건의 어느 것으로도 할 수 있다. In addition, since fermentation by Bacillus subtilis can be cultured under both anaerobic and aerobic conditions, the culture conditions can be either anaerobic or aerobic.
본 발명의 2단 발효는 상기 1단 발효물을 유산균 (L.spp)으로 발효시키는 것을 특징으로 한다. The two-stage fermentation of the present invention is characterized in that the first-stage fermentation product is fermented with lactic acid bacteria ( L.spp ).
상기 2단 발효는 온도 15 ~ 45℃ 바람직하게는 25 ~ 35℃ 더욱 바람직하게는 35℃일 수 있으며, 발효 시간은 6 ~ 24 시간 바람직하게는 9시간 ~ 18시간, 가장 바람직하게는 12시간 동안일 수 있다. 발효 온도가 위 범위보다 낮을 경우 발효 속도가 느려질 수 있고 또한 원하지 않는 발효 산물이 생겨날 수 있으며, 발효 온도가 위 범위보다 높을 경우도 발효 미생물의 사멸에 따라 마찬가지로 발효 속도가 느려지거나 원하지 않는 발효 산물이 생겨날 수 있다. 또한, 발효시간이 짧으면 발효가 잘 되지 않아 발효대사물이 적게 만들어지며 감칠맛이 덜하고, 발효시간이 길어지면 대사산물의 함량은 높아질 수 있으나 가스 생성 등으로 감칠맛 보다 신맛이 생길 우려가 있다. 온도가 높거나 발효시간이 길어지면 부패할 우려도 있다. 따라서 적당한 발효조건(온도, 시간)이 중요하다.The two-stage fermentation may be at a temperature of 15 to 45 ° C, preferably 25 to 35 ° C, more preferably 35 ° C, and the fermentation time is 6 to 24 hours, preferably 9 to 18 hours, and most preferably 12 hours. can be If the fermentation temperature is lower than the above range, the fermentation rate may slow down and unwanted fermentation products may be produced, and if the fermentation temperature is higher than the above range, the fermentation rate may also slow down or unwanted fermentation products may be produced due to the death of fermenting microorganisms. can arise In addition, if the fermentation time is short, fermentation does not proceed well, resulting in less fermentation metabolites and less savory taste. If the fermentation time is longer, the content of metabolites may increase, but there is a risk that a sour taste rather than a savory taste may occur due to gas generation. If the temperature is high or the fermentation time is long, there is a risk of spoilage. Therefore, proper fermentation conditions (temperature, time) are important.
상기 2단 발효에서 배지는 사용하는 미생물의 종류와 성질에 따라 사용 가능한 식물성 또는 동물성의 모든 배지를 사용할 수 있으며, 바람직하게는 식물성 원료인 대두펩톤 1.5% (w/v), 효모추출물 2.5% (w/v), 글루코스 4.0% (w/v), 황산마그네슘 0.05% (w/v), 초산나트륨 1.5% (w/v)을 포함하는 배지를 사용할 수 있다.In the two-stage fermentation, any medium of usable vegetable or animal origin can be used depending on the type and nature of the microorganism used, and preferably 1.5% (w / v) of soybean peptone and 2.5% of yeast extract (w / v), which are vegetable raw materials. w/v), glucose 4.0% (w/v), magnesium sulfate 0.05% (w/v), sodium acetate 1.5% (w/v) may be used.
본 발명의 2단 발효는 전분을 첨가하는 과정을 더 포함할 수 있으며, 상기 전분은 파쇄미를 사용할 수 있다. 상기 파쇄미는 백미, 흑미 등 모든 품종을 사용할 수 있으며, 호화시켜 사용할 수 있다. 상기 흑미는 백미에 비해 라이신 함량이 25 ~ 75% 많은 것으로 알려져 있다. The two-stage fermentation of the present invention may further include a step of adding starch, and crushed rice may be used as the starch. The crushed rice can be used in all varieties, such as white rice and black rice, and can be used luxuriously. The black rice is known to have 25 to 75% more lysine than white rice.
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. 따라서, 유산균을 이용하는 2단 발효 단계에서 전분을 더 첨가하는 것이 바람직하다. Lactobacillus is one of the most beneficial microorganisms available to humans. It is a non-pathogenic bacterium that can live in the intestines of humans and perform bacteriostatic and bacteriostatic action. It has characteristics such as growing and requiring various nutrients. Therefore, it is preferable to further add starch in the second fermentation step using lactic acid bacteria.
상기 유산균은 발효에 관여하는 락토바실러스속 또는 비피도박테리움속 이면 특별한 제한 없이 모두 사용할 수 있으며, 구체적으로 락토바실러스 애시도필러스(Lactobacillus acidophilus), 락토바실러스 람노서스(Lactobacillus rhamnosus), 비피도박테리움 락티스(Bifidobacterium lactis) 및 비피도박테리움 비피덤(Bifidobacterium bifidum)으로 구성된 유산균군에서 선택된 어느 1종 이상일 수 있다. The lactic acid bacteria may be used without particular limitation as long as they are of the genus Lactobacillus or Bifidobacterium involved in fermentation, and specifically , Lactobacillus acidophilus , Lactobacillus rhamnosus , and Bifidobacter It may be at least one selected from the group of lactic acid bacteria consisting of Bifidobacterium lactis and Bifidobacterium bifidum .
본 발명의 발효물의 제조방법은 상기 단계1)의 전단계로 고초균과 유산균을 활성화하는 단계를 더 포함할 수 있다. 균주를 최적 온도와 시간으로 최대 활성화시켜 균량을 늘리기 위함이다. The method for producing a fermented product of the present invention may further include activating Bacillus subtilis and lactic acid bacteria as a step before step 1). This is to increase the amount of strain by maximally activating the strain at the optimum temperature and time.
상기 활성화에 사용되는 배지는 사용하는 미생물의 종류와 성질에 따라 사용 가능한 식물성 또는 동물성의 모든 배지를 사용할 수 있으며, 바람직하게는 식물성 원료인 대두펩톤 1.5% (w/v), 효모추출물 2.5% (w/v), 글루코스 4.0% (w/v), 황산마그네슘 0.05% (w/v), 초산나트륨 1.5% (w/v)을 포함하는 배지를 사용할 수 있다. The medium used for the activation can be any vegetable or animal medium that can be used depending on the type and nature of the microorganism used, and preferably contains soybean peptone 1.5% (w / v) and yeast extract 2.5% (w / v), which are vegetable raw materials. w/v), glucose 4.0% (w/v), magnesium sulfate 0.05% (w/v), sodium acetate 1.5% (w/v) may be used.
상기 활성화 단계에서 온도는 30 ~ 45℃, 바람직하게는 35 ~ 40℃, 가장 바람직하게는 37℃ 일 수 있다. In the activation step, the temperature may be 30 to 45°C, preferably 35 to 40°C, and most preferably 37°C.
상기 활성화 단계에서 고초균은 12 ~ 48시간, 바람직하게는 24 ~ 36시간, 가장 바람직하게는 24시간 동안 배양할 수 있다. In the activation step, Bacillus subtilis can be cultured for 12 to 48 hours, preferably 24 to 36 hours, and most preferably 24 hours.
상기 활성화 단계에서 유산균은 5 ~ 25시간, 바람직하게는 10 ~ 20시간, 가장 바람직하게는 15시간 동안 배양할 수 있다. In the activation step, the lactic acid bacteria may be cultured for 5 to 25 hours, preferably 10 to 20 hours, and most preferably 15 hours.
본 발명의 발효물 제조시, 고초균을 접종·발효시키기 전에 탄소원을 첨가할 수 있다. 이러한 탄소원은 당업계에 알려진 임의의 것을 사용 할 수 있으며, 통상은 올리고당, 유당, 포도당, 과당, 설탕, 이들의 혼합물 등이 사용될 수 있다. 이러한 탄소원은 의도한 발효 시간, 발효 정도 등을 고려하여 임의의 범위로 첨가될 수 있다. 통상은 재료 100 중량부 기준 1 중량부 내지 50 중량부의 범위로 첨가할 수 있다. When preparing the fermented product of the present invention, a carbon source may be added before inoculation and fermentation of Bacillus subtilis. Any carbon source known in the art may be used, and oligosaccharides, lactose, glucose, fructose, sucrose, mixtures thereof, and the like may be used in general. Such a carbon source may be added in an arbitrary range considering the intended fermentation time, fermentation degree, and the like. Usually, it may be added in the range of 1 part by weight to 50 parts by weight based on 100 parts by weight of the material.
본 발명의 발효물은 감압 저온 건조 또는 동결 건조 등의 방식으로 발효물 자체를 농축시켜 얻어지는 액상, 또는 분말상의 형태일 수 있고, 발효물을 여과할 경우 여액과 그 여과 잔사로 분리된 형태일 수 있으며, 그 여액과 여과 잔사를 농축·건조하여 얻은 액상 또는 분말상의 형태일 수도 있다. 또한 그 발효물을 적절한 추출 용매(예컨대 물, 에탄올 또는 이들의 혼합 용매)로 추출하여 사용할 수도 있다.The fermented product of the present invention may be in the form of a liquid or powder obtained by concentrating the fermented product itself in a method such as drying at low temperature under reduced pressure or freeze drying, and may be in the form of separating the filtrate and the filtrate residue when the fermented product is filtered. It may be in liquid or powder form obtained by concentrating and drying the filtrate and filtration residue. In addition, the fermented product may be extracted with an appropriate extraction solvent (for example, water, ethanol or a mixed solvent thereof) and used.
본 발명의 발효물 제조방법에 의할 경우, 유리 아미노산 함량이 증가하는 효과가 있다. 상기 유리아미노산은 기능성 아미노산으로서 황함유 아미노산, 감칠맛을 내는 아미노산, 감칠맛을 증가시키는 아미노산일 수 있다. In the case of the fermented product manufacturing method of the present invention, there is an effect of increasing the free amino acid content. The free amino acid is a functional amino acid and may be a sulfur-containing amino acid, an amino acid that gives umami, or an amino acid that increases umami.
상기 황함유 아미노산은 메티오닌, 시스틴, 시스테인일 수 있으며, 상기 감칠맛을 내는 아미노산은 아스파라긴산, 글루타민일 수 있으며, 상기 감칠맛을 증가시키는 아미노산은 알라닌일 수 있다. The sulfur-containing amino acid may be methionine, cystine, or cysteine, the amino acid that gives the umami taste may be aspartic acid or glutamine, and the amino acid that increases the umami taste may be alanine.
상기 메티오닌은 황을 함유하는 필수 아미노산의 하나로, 단백질의 구성 성분이며 생체 안에서 메틸기 공여체(供與體)로서 중요하다. 카세인(casein), 달걀의 알부민, 효모 등에 함유되어 있으며 영양제나 간장 질환, 중독증 등의 치료에 사용된다. The methionine is one of the essential amino acids containing sulfur, is a component of protein, and is important as a methyl group donor in vivo. It is contained in casein, egg albumin, yeast, etc., and is used as a nutritional supplement or to treat liver disease and poisoning.
상기 시스틴은 황을 함유하는 α-아미노산의 하나로 최초로 발견된 아미노산이다. 천연으로는 모두 L형으로 존재하며, 케라틴에 10% 이상 함유되어 있고 쉽게 환원되어 시스테인이 된다.Cystine is one of the sulfur-containing α-amino acids and is the first amino acid discovered. In nature, all of them exist in the L form, and they are contained in keratin by 10% or more and are easily reduced to cysteine.
상기 시스테인은 황을 함유하는 아미노산으로 필수 아미노산은 아니지만 특히 산화/환원 과정을 담당하며 중금속을 배출하는 역할을 한다.The cysteine is a sulfur-containing amino acid and is not an essential amino acid, but is particularly responsible for oxidation/reduction processes and serves to release heavy metals.
상기 아스파라긴산은 아스파라긴산은 아스파르트산이라고도 하며 산성 α-아미노산의 하나이다. 아미노숙신산에 해당하고 백합과 식물인 아스파라거스 (Asparagus)의 액즙에서 최초로 분리되었다. 식물계에 널리 존재하며 특히 싹이 튼 콩류(콩나물 등)에 많이 함유되어 있다. 아스파라긴산은 숙취해소 효능이 탁월한 물질이며 감미료로도 사용된다.The aspartic acid is also called aspartic acid and is one of acidic α-amino acids. It corresponds to aminosuccinic acid and was first isolated from the sap of Asparagus, a plant of the lily family. It is widely present in the plant kingdom and is especially abundant in sprouted legumes (bean sprouts, etc.). Aspartic acid is an excellent hangover cure and is also used as a sweetener.
상기 글루타민은 식물의 발아 종자(호박·해바라기 등)내에 축적되며, 포유류 혈액 속의 아미노산의 주성분으로, 동식물의 단백질 속에 존재한다. 생체 내에서는 암모니아의 저장 역할을 하며, 핵산의 퓨린핵 생성에 관여하고 페닐아세트산과 결합하여 해독한다. Glutamine is accumulated in germinated seeds of plants (pumpkins, sunflowers, etc.), is a main component of amino acids in mammalian blood, and is present in proteins of animals and plants. In vivo, it serves as a storage of ammonia, is involved in the formation of purine nuclei of nucleic acids, and binds to phenylacetic acid to detoxify it.
상기 알라닌은 일반적으로 피루브산의 환원적 아미노화반응에 의해 생성된다. 그리고 트렌스아미네이션에 의해 쉽게 만들어진다. 알라닌은 주로 의약품이나 식품공업에서 이용되고 있다. 또, D-알라닌은 주로 의약품으로, DL-알라닌은 주로 식품공업 등에 이용되고 있다. 식품첨가물로서의 알라닌은 짠맛, 신맛의 완화효과가 있으며, 다른 감칠맛 물질과의 상승효과에 의해 감칠맛을 증강하는 작용을 한다. 또, 여러 가지 맛을 조화하여 감칠맛을 끌어낸다. 알라닌은 다른 아미노산에 비해 아미노산 특유의 갈변 반응을 낮은 상태로 억제할 수 있다. 이러한 작용으로, 절임류, 진미, 수산가공품, 반찬첨가제재, 양조용첨가제, 청량음료수의 산미교정제 등으로 사용되고 있다. 알라닌은 지방산의 생합성에 관계하여 지방산분해효소를 활성화한다고 알려져있다. 또 알라닌의 생리작용으로는 알코올의 대사촉진, 간 기능 보호, 인슐린의 분비촉진 등을 들 수 있다. 의약품의 원료로는 주로 전립선비대증 약 등의 의약품 성분으로서 이용된다. 또, 다양한 생리작용을 갖고 있기 때문에 보충제로도 사용된다. 알라닌의 생리작용에는 알코올의 대사촉진, 간 기능 보호, 혈당치를 낮추는 인슐린의 분비촉진 등이 있다. 특히, 간장에서 포도당의 생산을 촉진하여 근육으로의 에너지를 주는 작용에 착안한 스포츠용 제품도 있다. 화장품에서는 콜라겐을 재생하는데 필요한 아미노산으로 피부 컨디셔닝제, 킬레이트제, 세정제의 원료로 사용된다. The alanine is generally produced by reductive amination of pyruvic acid. And it is easily made by transamination. Alanine is mainly used in the pharmaceutical and food industries. In addition, D-alanine is mainly used as a medicine, and DL-alanine is mainly used in the food industry and the like. Alanine as a food additive has the effect of alleviating salty and sour tastes, and acts to enhance the umami taste by synergistic effect with other umami substances. In addition, various flavors are harmonized to bring out the umami. Compared to other amino acids, alanine can suppress the browning reaction specific to amino acids to a lower state. Due to this action, it is used as pickles, delicacies, processed fish products, additives for side dishes, additives for brewing, and acidity correctors for soft drinks. It is known that alanine activates lipolytic enzymes related to the biosynthesis of fatty acids. In addition, the physiological actions of alanine include promotion of alcohol metabolism, protection of liver function, and promotion of insulin secretion. As a raw material for pharmaceuticals, it is mainly used as a pharmaceutical ingredient such as a prostatic hyperplasia medicine. It is also used as a supplement because it has various physiological effects. Alanine's physiological actions include promoting alcohol metabolism, protecting liver function, and promoting secretion of insulin that lowers blood sugar levels. In particular, there are products for sports that focus on the action of promoting glucose production in the liver to give energy to muscles. In cosmetics, it is an amino acid necessary for regenerating collagen and is used as a raw material for skin conditioning agents, chelating agents, and detergents.
본 발명의 일 실시 양태는 사용 균주 및 균주의 활성화를 위해 식물성 배지에서 37℃, 150 rpm 조건에서 고초균(Bacillus amyloliquefaciens HJ5-2)은 24시간, 유산균(Lactobacillus plantarum JSA22)은 15시간 배양하여 균주를 활성화 하였다. In one embodiment of the present invention , Bacillus amyloliquefaciens HJ5-2 is cultured for 24 hours and lactic acid bacteria ( Lactobacillus plantarum JSA22) is cultivated for 15 hours at 37 ° C. and 150 rpm in a vegetable medium for activation of strains and strains. Activated.
본 발명의 일 실시 양태는 연구용 배지를 산업용 배지로 대체하기 위해 식물성 배지에서 균주 활성의 최적 조건을 설정하는 실험을 통해 고초균은 24시간 배양 (8.47 log CFU/g), 유산균은 15시간 배양 (9.45 log CFU/g)로 각각 균주를 활성화하였다. In one embodiment of the present invention, Bacillus subtilis was cultured for 24 hours (8.47 log CFU / g) and lactic acid bacteria was cultured for 15 hours (9.45 Each strain was activated as log CFU/g).
본 발명의 일 실시 양태는 1단 발효의 대두피 발효 조건으로서 대두피, 글루코스, 정제수, 고초균을 45℃에서 발효하는 것으로 발효 조건을 결정하였다. In one embodiment of the present invention, the fermentation conditions were determined by fermenting soybean hull, glucose, purified water, and Bacillus subtilis at 45 ° C.
본 발명의 일 실시 양태는 대두피에서 접종 균주 및 발효방법에 따른 유리아미노산의 함량을 측정하여, 유산균 단독으로 발효한 발효물의 아미노산 함량이 고초균 단독 처리군 및 복합균주 처리군에 비해 높은 것을 확인하였다. One embodiment of the present invention measured the content of free amino acids in soybean hulls according to the inoculated strain and fermentation method, and it was confirmed that the amino acid content of the fermented product fermented with lactic acid bacteria alone was higher than that of Bacillus subtilis alone treatment group and complex strain treatment group. .
본 발명의 일 실시 양태는 고초균과 유산균으로 2단 발효한 발효물(C)을 제조하고, 상기 2단 발효물의 유리아미노산 함량을 유산균 1단 발효물(B)과 비교하였다. 그 결과, 본 발명의 고초균을 이용한 1단 발효물에 파쇄미를 혼합하고 유산균으로 2단 발효한 경우(C)가 유산균으로 1단 발효한 경우(B)에 비해 총 유리아미노산, 메티오닌, 알라닌, 글루타민 등의 아미노산 함량이 5~7배 증가함을 확인하였다. In one embodiment of the present invention, a fermented product (C) obtained by two-stage fermentation with Bacillus subtilis and lactic acid bacteria was prepared, and the free amino acid content of the two-stage fermentation was compared with that of the first-stage fermentation product (B). As a result, when crushed rice was mixed with the first-stage fermented product using the Bacillus subtilis of the present invention and two-stage fermentation was performed with lactic acid bacteria (C), total free amino acids, methionine, alanine, It was confirmed that the content of amino acids such as glutamine increased 5 to 7 times.
또한, 본 발명의 일 실시 양태는 본 발명의 최종 발효물의 알라닌 및 글루타민 함량을 측정하여 감칠맛을 내는 글루타민 및 감칠맛 상승 효과가 있는 알라닌의 함량이 증가한 것을 확인 하였으며, 기호도법에 의해 본 발명의 2단 발효에 의한 발효물(C)이 감칠맛 및 전체적인 기호도에 있어서 비발효(A), 유산균 1단 발효(B)에 비해 가장 높은 점수를 나타냄을 확인하였다. In addition, in one embodiment of the present invention, by measuring the alanine and glutamine contents of the final fermented product of the present invention, it was confirmed that the contents of glutamine, which gives a umami taste, and alanine, which has a umami synergistic effect, increased. It was confirmed that the fermented product (C) by fermentation showed the highest score compared to non-fermented (A) and lactic acid bacteria single-stage fermentation (B) in terms of umami and overall acceptability.
이하, 본 발명은 하기 실시예 및 실험예에 의해 상세히 설명한다. Hereinafter, the present invention will be described in detail by the following examples and experimental examples.
단, 하기 실시예 및 실험예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예 및 실험예에 한정되는 것은 아니다. However, the following Examples and Experimental Examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following Examples and Experimental Examples.
<실시예 1> 사용 균주의 활성화<Example 1> Activation of strains used
1-1. 연구용 배지의 균주 활성화1-1. Activation of strains in research media
본 발명의 하기 실험예에 사용한 균주는 농촌진흥청 국립식량과학원에서 개발하여 기탁한 고초균 (Bacillus amyloliquefaciens HJ5-2, 수탁번호 : KACC81027)과 유산균 (Lactobacillus plantarum JSA22, 수탁번호 : KACC91973P)을 사용하였다. The strains used in the following experimental examples of the present invention were Bacillus subtilis ( Bacillus amyloliquefaciens HJ5-2, accession number: KACC81027) and lactic acid bacteria ( Lactobacillus plantarum JSA22, accession number: KACC91973P) developed and deposited at the National Institute of Food Science, Rural Development Administration.
균주 활성화에 사용된 배지는 식물성 원료인 대두펩톤 1.5% (w/v), 효모추출물 2.5% (w/v), 글루코스 4.0% (w/v), 황산마그네슘 0.05% (w/v), 초산나트륨 1.5% (w/v)을 이용하여 37℃, 150 rpm 조건에서 고초균은 24시간, 유산균은 15시간 배양하여 균주를 활성화시켰다. The medium used for strain activation is vegetable raw material soybean peptone 1.5% (w/v), yeast extract 2.5% (w/v), glucose 4.0% (w/v), magnesium sulfate 0.05% (w/v), acetic acid The strains were activated by culturing Bacillus subtilis for 24 hours and Lactobacillus for 15 hours at 37 ° C. and 150 rpm using sodium 1.5% (w / v).
1-2. 산업용배지의 균주 활성화1-2. Activation of strains in industrial media
연구용 배지를 대량생산이 가능한 산업용배지로 대체하기 위하여 식물성 원료로 스크린된 대두펩톤 1.5% (w/v), 효모추출물 2.5% (w/v), 글루코스 4.0% (w/v), 황산마그네슘 0.05% (w/v), 초산나트륨 1.5% (w/v)으로 최종 결정하여 식물성 산업용배지 원료로 사용하였다. 원료계량 및 혼합 교반하여 고압멸균기로 121℃에서 15분 멸균한 다음 스타터를 각각 접종하여 37℃, 150 rpm 조건으로 배양하였다. 하기의 표 1은 배지의 균주 활성화 결과를 나타낸다. Soybean peptone 1.5% (w/v), yeast extract 2.5% (w/v), glucose 4.0% (w/v), magnesium sulfate 0.05 % (w/v) and sodium acetate 1.5% (w/v) were finally determined and used as a raw material for vegetable industrial media. Raw materials were weighed, mixed, and sterilized at 121° C. for 15 minutes with a high-pressure sterilizer, and then each starter was inoculated and cultured at 37° C. and 150 rpm. Table 1 below shows the strain activation results of the medium.
균주 활성 최적 조건은 실험을 통하여 각각 고초균은 24시간 배양 (8.47 log CFU/g), 유산균은 15시간 배양 (9.45 log CFU/g)로 각각 균주를 활성화시켰다. The optimal conditions for strain activity were 24-hour culture (8.47 log CFU/g) for Bacillus subtilis and 15-hour culture (9.45 log CFU/g) for lactic acid bacteria, respectively.
<실시예 2> 1단 발효의 대두피 발효조건 설정<Example 2> Setting conditions for fermentation of soybean hulls for one-stage fermentation
대두피 및 참깨박의 발효 조건을 결정하고자 Sample 1), Sample 2)를 제조하여 미생물수를 측정하였다. Sample 1)은 대두피, 정제수, 고초균이며, Sample 2)는 대두피, 글루코스, 정제수, 고초균이다. 하기의 표 2는 미생물수를 측정한 결과를 나타낸다. To determine the fermentation conditions of soybean husk and sesame meal, Sample 1) and Sample 2) were prepared and the number of microorganisms was measured. Sample 1) is soybean husk, purified water, and Bacillus subtilis, and Sample 2) is soybean husk, glucose, purified water, and Bacillus subtilis. Table 2 below shows the results of measuring the number of microorganisms.
그 결과, 대두피와 글루코스(포도당)를 함께 발효한 발효액이 미생물균수가 높게 나타났다. 따라서, Sample 2)의 대두피, 글루코스, 정제수, 고초균을 45℃에서 발효하는 것으로 발효 조건을 결정하였다.As a result, the fermentation broth fermented with soybean hull and glucose (glucose) showed a high number of microorganisms. Therefore, the fermentation conditions were determined by fermenting the soybean hull, glucose, purified water, and Bacillus subtilis of Sample 2) at 45 ° C.
<실시예 3> 대두피에서 접종균주(고초균/유산균) 및 발효방법(단일/복합)에 따른 유리아미노산 함량 측정 <Example 3> Measurement of free amino acid content in soybean hulls according to inoculated strains (Bacillus subtilis/lactic acid bacteria) and fermentation method (single/complex)
균주 및 발효방법을 결정하기 위하여 대두피 발효시 단일균주로 고초균과 유산균를 각각 접종하여 발효물을 제조하였으며, 복합균주로 고초균과 유산균를 1:1비율로 접종하여 복합발효 발효물을 제조하여 유리아미노산 함량을 측정하였다. 하기의 표 3은 유리아미노산 함량을 측정한 결과를 나타낸다. In order to determine the strain and fermentation method, a fermented product was prepared by inoculating Bacillus subtilis and lactic acid bacteria as a single strain during soybean hull fermentation, and inoculating Bacillus subtilis and lactic acid bacteria as a complex strain at a 1:1 ratio to prepare a fermented product for free amino acid content. was measured. Table 3 below shows the results of measuring the free amino acid content.
단위:mg/kgUnit:mg/kg
발효전은 대두피분말, 쌀가루, 효모추출물, 포도당, 정제수를 혼합한 것으로 대조군에 해당한다. 발효후는 각각 대조군에 고초균을 접종한 단일균주 처리군, 유산균을 접종한 단일균주 처리군, 고초균과 유산균을 1:1 비율로 접종한 복합균주 처리군에 해당한다. Before fermentation, soybean husk powder, rice flour, yeast extract, glucose, and purified water were mixed and corresponded to the control group. After fermentation, the control group corresponds to a single strain treatment group inoculated with Bacillus subtilis, a single strain treatment group inoculated with lactic acid bacteria, and a complex strain treatment group inoculated with Bacillus subtilis and lactic acid bacteria at a 1: 1 ratio.
발효하는 동안, 단백질 분해는 유리아미노산 (FAA)을 생성물로 유리시키는 주요 활성으로 발생한다. 일반적으로, 유리 아미노산은 미각에 직접 기여하고 풍미 향상에 중요한 역할을 한다. During fermentation, proteolysis occurs as the main activity liberating free amino acids (FAAs) into products. In general, free amino acids directly contribute to taste and play an important role in flavor enhancement.
측정 결과, 유산균 단독으로 발효한 발효물의 아미노산 함량이 고초균 단독 처리군 및 복합균주 처리군에 비해 높았다. As a result of the measurement, the amino acid content of the fermented product fermented with lactic acid bacteria alone was higher than that of the Bacillus subtilis alone treatment group and the complex strain treatment group.
<실시예 4> 고초균과 유산균으로 2단 발효한 식물성 발효조미농축액의 제조<Example 4> Preparation of vegetable fermented seasoning concentrate fermented in two stages with Bacillus subtilis and lactic acid bacteria
4-1. 1단 발효4-1. 1 stage fermentation
4-1-1. 대두피 1단 발효물의 제조4-1-1. Preparation of soybean hull fermented product
본 발명의 대두피 1단 발효물은 국산 대두를 구입하여 선별 및 정선한 다음 세척 후 물?兮? 하여 65℃에서 수분함량이 5%가 되도록 열풍건조하였다. 건조된 대두를 거피기를 이용하여 거피한 다음 거피 대두와 대두 배아가 포함된 대두피(껍질)을 얻었다. 대두피는 대두 분말 및 단백질 분획 시 발생하는 부산물이며, 이를 분쇄하여 대두피 분말을 획득하였다. The soybean husk first-stage fermented product of the present invention is purchased from domestic soybeans, sorted and selected, washed, and then washed with water? and dried with hot air at 65° C. to a moisture content of 5%. The dried soybeans were peeled using a peeling machine, and then soybean skins (shells) containing the peeled soybeans and soybean embryos were obtained. Soybean hull is a by-product generated during soybean powder and protein fractionation, and soybean hull powder was obtained by pulverizing it.
대두피 분말 10%와 글루코스 5%, 정제수 85%를 계량하여 혼합한 다음 고압멸균기에서 121℃에서 15분 멸균하였다. 40℃가 되도록 냉각한 후 산업용 식물성 배지로 제조한 고초균(B.spp)을 접종 농도 1% 이상이 되도록 접종한 후 45℃에서 24시간 동안 1단 발효하여 대두피 발효물을 제조하였다. 10% of soybean hull powder, 5% of glucose, and 85% of purified water were weighed and mixed, and then sterilized in a high-pressure sterilizer at 121°C for 15 minutes. After cooling to 40 ° C., Bacillus subtilis ( B.spp ) prepared as an industrial vegetable medium was inoculated to an inoculum concentration of 1% or more, and then fermented in one stage at 45 ° C. for 24 hours to prepare a fermented soybean husk.
4-1-2. 참깨박 1단 발효물의 제조4-1-2. Preparation of one-stage fermented sesame meal
본 발명의 참개박 1단 발효물은 참기름 제조과정에서 발생하는 부산물인 참깨박 (단백질 50%, 지방 10%)을 분쇄하여 분말화하였다. 참깨박 분말 10%와 글루코스 5%, 정제수 85%를 계량하여 혼합한 다음 고압멸균기에서 121℃에서 15분 멸균하였다. 40℃가 되도록 냉각한 후 산업용 식물성 배지로 제조한 고초균(B.spp)을 접종 농도 1% 이상이 되도록 접종한 후 45℃에서 24시간 동안 1단 발효하여 참깨박 발효물을 제조하였다. Sesame meal (protein 50%, fat 10%), a by-product generated in the sesame oil manufacturing process, was pulverized and powdered for the first-stage fermented product of the present invention. 10% of sesame meal powder, 5% of glucose, and 85% of purified water were weighed and mixed, and then sterilized in a high-pressure sterilizer at 121°C for 15 minutes. After cooling to 40 ° C., Bacillus subtilis ( B.spp ) prepared as an industrial vegetable medium was inoculated to an inoculation concentration of 1% or more, and then fermented in one stage at 45 ° C. for 24 hours to prepare fermented sesame meal.
4-2. 2단 발효4-2. 2 stage fermentation
4-2-1. 파쇄미 분말의 제조4-2-1. Manufacture of crushed rice powder
본 발명의 파쇄미 분말은 파쇄미를 세척 및 물빼기한 후 열풍건조한 다음 분쇄하여 파쇄미분말을 제조하였다. The crushed rice powder of the present invention was prepared by washing and draining the crushed rice, drying it with hot air, and then pulverizing it.
4-2-2. 2단 발효에 의한 식물성 발효물의 제조4-2-2. Production of vegetable fermented product by two-stage fermentation
상기 1단 발효물(대두피발효물과 참깨박발효물)에 미리 준비된 파쇄미 분말, 효모추출물, 글루코스, 정제수를 계량하여 100℃에서 60분 동안 증자한 다음 37℃로 냉각한 혼합액을 첨가하여 무균적으로 혼합하였다. 혼합물에 유산균을 g 당 106 CFU 되게 접종하여 37℃에서 12시간 2단 발효한 다음 15℃로 냉각한 후 8시간 동안 숙성시켜 2단 발효한 발효물을 얻었다. 상기 발효물을 여과(원심분리)하여 상층액을 감압농축기 온도 80℃에서 pH 4.3~4.5, 60~65 Brix가 되도록 농축하여 식물성 발효조미농축액을 제조하였다. To the first-stage fermented product (fermented soybean skin and fermented sesame meal), crushed rice powder, yeast extract, glucose, and purified water prepared in advance were weighed, steamed at 100 ° C for 60 minutes, and then cooled to 37 ° C. Mix aseptically. The mixture was inoculated with lactic acid bacteria at 10 6 CFU per g, fermented in two stages at 37° C. for 12 hours, cooled to 15° C., and then aged for 8 hours to obtain a two-stage fermentation product. The fermented product was filtered (centrifuged), and the supernatant was concentrated to pH 4.3-4.5 and 60-65 Brix at a vacuum concentrator temperature of 80 ° C to prepare a vegetable fermented seasoning concentrate.
<실시예 5> 발효물의 원심분리 후 유리아미노산 함량 비교<Example 5> Comparison of free amino acid content after centrifugation of fermented product
제조공정에서 발효 후 대사산물인 아미노산 함량을 증진 시키기 위해 발효물을 원심분리 (10,000rpm, 10min)한 다음 상등액 (supernatant)과 잔사 (pellet)에 대한 유리아미노산 함량을 비교하였다. 상기 실시예 3의 유산균 접종 처리군을 대상으로 원심분리 하였다. 하기의 표 4는 발효물의 원심분리 후 상층액과 잔사의 유리아미노산 함량을 비교한 결과를 나타낸 것이다. In order to increase the amino acid content, which is a metabolite after fermentation in the manufacturing process, the fermented product was centrifuged (10,000 rpm, 10 min), and then the free amino acid content of the supernatant and the pellet was compared. The lactic acid bacteria inoculation treatment group of Example 3 was subjected to centrifugation. Table 4 below shows the results of comparing the free amino acid content of the supernatant and the residue after centrifugation of the fermented product.
단위:mg/kg Unit: mg/kg
그 결과, 원심분리하여 얻은 상층액이 잔사에 비해 유리아미노산 함량이 높아 원심분리 후 상층액을 소재로 사용하기로 결정하였다. As a result, the supernatant obtained by centrifugation had a higher free amino acid content than the residue, so it was decided to use the supernatant after centrifugation as a material.
<실험예 1> 1단 발효 및 2단 발효에 의한 발효물의 유리아미노산 함량 비교<Experimental Example 1> Comparison of free amino acid content of fermented products by first-stage fermentation and second-stage fermentation
본 발명의 2단 발효에 의할 경우, 1단 발효에 비해 유리아미노산 함량이 증가함을 확인하기 위해 비발효, 1단 발효 및 2단 발효물의 유리아미노산 함량을 측정하였다. In the case of the two-stage fermentation of the present invention, in order to confirm that the free amino acid content increased compared to the first-stage fermentation, the free amino acid contents of non-fermented, first-stage fermentation and two-stage fermentation were measured.
최종 발효물에서 필수아미노산별 함량을 비교한 결과를 도 1에 나타내었다. The results of comparing the contents of each essential amino acid in the final fermented product are shown in FIG.
A는 원료 증자 후 비발효 처리군이며, B는 유산균 1단 발효 처리군, C는 고초균으로 1단 발효 후 유산균으로 2단 발효한 2단 발효 처리군에 해당한다. 각 처리군별로 필수아미노산인 Lysine, Threonine, Leucine, Isoleusine, Valine, Methionine, Phenylalanine, Tryptophan 및 총 필수아미노산의 함량을 나타낸다. A is a non-fermentation treatment group after raw material increase, B is a lactic acid bacteria first-stage fermentation treatment group, C corresponds to a two-stage fermentation treatment group in which lactic acid bacteria are fermented in two stages after first fermentation with Bacillus subtilis. The content of essential amino acids Lysine, Threonine, Leucine, Isoleusine, Valine, Methionine, Phenylalanine, Tryptophan and total essential amino acids for each treatment group is shown.
하기의 표 5는 최종 발효 전후 발효물의 유리아미노산 함량을 비교한 결과를 나타낸다. Table 5 below shows the results of comparing the free amino acid content of fermented products before and after final fermentation.
단위:mg/kg Unit: mg/kg
A는 대두피분말, 참깨박분말, 파쇄비(쌀가루), 흑미가루, 효모추출물, 포도당, 정제수를 혼합한 것으로 발효하지 아니한 것이다. A is a mixture of soybean husk powder, sesame meal powder, crushed rice flour, black rice flour, yeast extract, glucose, and purified water, and is not fermented.
B는 대두피분말, 참깨박분말, 파쇄미(쌀가루), 흑미가루, 효모추출물, 포도당, 정제수를 혼합하고 유산균을 접종하여 1단 발효한 처리군에 해당한다. B corresponds to a treatment group in which soybean husk powder, sesame meal powder, crushed rice (rice flour), black rice powder, yeast extract, glucose, and purified water were mixed and lactic acid bacteria were inoculated and fermented in one stage.
C는 고초균으로 1단 발효한 대두피발효액 및 참깨발효액, 파쇄미(쌀가루), 흑미가루, 효모추출물, 포도당, 정제수를 혼합하고 유산균을 접종하여 2단 발효한 처리군에 해당한다. C corresponds to a treatment group in which soybean skin fermented liquid and sesame fermented liquid fermented in one stage with Bacillus subtilis, crushed rice (rice flour), black rice flour, yeast extract, glucose, and purified water were mixed and inoculated with lactic acid bacteria and fermented in two stages.
구체적인 방법은 상기 실시예4의 방법에 따랐다. The specific method followed the method of Example 4 above.
그 결과, 대두피 및 참깨박을 2단 발효(C)함으로써 총유리아미노산 함량이 비발효(A)에 비해 약 5.2배 증가하였으며, 1단 발효물(B)에 비해 약 3배 증가한 것으로 나타났다. As a result, by two-stage fermentation (C) of soybean husk and sesame meal, the total free amino acid content increased by about 5.2 times compared to non-fermented (A), and increased by about 3 times compared to one-stage fermentation (B).
또한, 대두피 및 참깨박을 2단 발효(C)함으로써 아미노산 중 기능성 아미노산으로 황함아미노산인 메티오닌 함량이 비발효(A)에 비해 약 2.1배 증가하였으며, 1단 발효물(B)에 비해 약 1.3배 증가한 것으로 나타났다. 황함아미노산은 고기를 조리할 때 화학적 반응인 마이야르 반응(maillard reaction)에 의해 고기의 풍미 및 감칠맛을 상승시키는 아미노산으로 알려져 있다. In addition, by two-stage fermentation (C) of soybean husk and sesame meal, the content of methionine, a sulfur-containing amino acid among amino acids, increased by about 2.1 times compared to non-fermented (A), and about 1.3 times higher than that of first-stage fermentation (B). appeared to increase twice. Sulfur amino acids are known as amino acids that increase the flavor and umami of meat by a Maillard reaction, which is a chemical reaction when meat is cooked.
또한, 대두피 및 참깨박을 2단 발효(C)함으로써 감칠맛을 내는 아미노산 (Aspartic acid, 글루타민) 함량이 비발효(A)에 비해 약 2배 증가하였으며, 1단 발효물(B)에 비해 약 1.3배 증가한 것으로 나타났다. In addition, by two-stage fermentation of soybean hull and sesame meal (C), the content of amino acids (aspartic acid, glutamine) that gives savory taste increased by about 2 times compared to non-fermentation (A), and about 1-stage fermentation (B) showed a 1.3-fold increase.
또한, 대두피 및 참깨박을 2단 발효(C)함으로써 감칠맛을 상승시키는 아미노산(알라닌) 함량이 비발효(A)에 비해 약 3.9배 증가하였으며, 1단 발효물(B)에 비해 약 1.3배 증가한 것으로 나타났다. In addition, by two-stage fermentation of soybean husk and sesame meal (C), the amino acid (alanine) content that enhances umami increased by about 3.9 times compared to non-fermentation (A), and about 1.3 times compared to one-stage fermentation (B) appeared to increase.
따라서, 본 발명의 고초균을 이용한 1단 발효물을 혼합하고 유산균으로 2단 발효한 경우에 유산균으로 1단 발효한 경우에 비해 총 유리아미노산, 메티오닌, 글루타민, 알라닌 등의 아미노산 함량이 5~7배 증가한 것으로 나타났으며, 이러한 결과를 통해 본 발명의 2단 발효에 의하는 경우에 감칠맛이 현저하게 우수한 발효물을 제조할 수 있음을 확인하였다. Therefore, when the first-stage fermented product using the Bacillus subtilis of the present invention is mixed and the second-stage fermentation is performed with lactic acid bacteria, the content of amino acids such as total free amino acids, methionine, glutamine, and alanine is 5 to 7 times higher than that of single-stage fermentation with lactic acid bacteria. It was found to increase, and through these results, it was confirmed that in the case of the two-stage fermentation of the present invention, a fermented product having a remarkably excellent umami taste could be prepared.
<실험예 2> 1단 발효 및 2단 발효에 의한 발효물의 글루타민 및 알라닌 함량 비교<Experimental Example 2> Comparison of glutamine and alanine contents of fermented products by first-stage fermentation and second-stage fermentation
최종 발효물의 감칠맛을 내는 아미노산인 글루타민 및 감칠맛을 상승시키는 알라닌의 함량을 측정하였다. The contents of glutamine, an amino acid that gives umami, and alanine, which enhances umami, were measured in the final fermented product.
A는 원료 증자 후 비발효 처리군이며, B는 유산균 1단 발효 처리군, C는 고초균으로 1단 발효 후 유산균으로 2단 발효한 2단 발효 처리군에 해당한다.A is a non-fermentation treatment group after raw material increase, B is a lactic acid bacteria first-stage fermentation treatment group, C corresponds to a two-stage fermentation treatment group in which lactic acid bacteria are fermented in two stages after first fermentation with Bacillus subtilis.
하기 표 6은 각 처리군별로 감칠맛을 내는 아미노산인 Glutamic acid 및 감칠맛을 상승시키는 Alanine 함량을 측정한 결과를 나타낸다. 도 2에 그 결과를 그래프로 도시하였다. Table 6 below shows the results of measuring the content of glutamic acid, an amino acid that enhances umami, and alanine, which enhances umami, for each treatment group. 2 shows the results graphically.
그 결과, 2단 발효 처리군(C)에서 비발효(A) 및 유산균 1단 발효 처리군(B)에 비해 글루타민 및 알라닌 모두 증가한 것으로 나타났다.As a result, it was found that both glutamine and alanine increased in the two-stage fermentation group (C) compared to the non-fermentation group (A) and the lactic acid bacteria one-stage fermentation group (B).
한편 알라닌은 식품첨가물로서 사용되는 경우에 짠맛, 신맛의 완화 효과가 있으며, 감칠맛 물질과의 상승효과에 의해 감칠맛을 증가시키는 작용을 하며, 여러가지 맛을 조화하여 감칠맛을 더욱 상승시키는 것으로 알려져 있다. On the other hand, when used as a food additive, alanine has the effect of alleviating salty and sour tastes, acts to increase umami by synergistic effect with umami substances, and is known to further increase umami by harmonizing various tastes.
따라서, 감칠맛을 내는 글루타민 및 감칠맛을 상승시키는 알라닌이 모두 증가한 결과에 의할 때, 본 발명의 2단 발효물이 유산균 1단 발효에 비해 감칠맛이 우수한 효과가 있음을 확인하였다. Therefore, it was confirmed that the two-stage fermented product of the present invention has an excellent umami effect compared to the lactic acid bacteria single-stage fermentation, as a result of the increase in both glutamine, which enhances umami, and alanine, which enhances umami.
<실험예 3> 최종 발효물의 관능평가 <Experimental Example 3> Sensory evaluation of the final fermented product
최종 발효물의 관능평가를 위해 9점 기호도법을 실시하였다. 하기 표 7은 그 결과를 나타낸다. For the sensory evaluation of the final fermented product, a 9-point symbology was performed. Table 7 below shows the results.
A는 대두피분말 1.5%, 참깨박분말 0.5%, 파쇄미분말 3.0%, 효모추출물 1.5%, 포도당 1.5%, 정제수를 혼합하여 발효하지 아니한 것이며,A is a mixture of soybean skin powder 1.5%, sesame meal powder 0.5%, crushed rice powder 3.0%, yeast extract 1.5%, glucose 1.5%, and purified water and is not fermented.
B는 대두피분말 1.5%, 참깨박분말 0.5%, 파쇄미분말 3.0%, 효모추출물 1.5%, 포도당 1.5%, 정제수를 혼합하여 유산균을 접종하여 1단 발효한 발효물이며, B is a fermented product obtained by mixing 1.5% soybean husk powder, 0.5% sesame meal powder, 3.0% crushed rice powder, 1.5% yeast extract, 1.5% glucose, and purified water, inoculating lactic acid bacteria, and fermenting in one stage,
C는 대두피분말 1.5%, 포도당 0.5%, 정제수를 혼합하여 고초균으로 1단 발효한 대두피 1단 발효물과 참깨박분말 1.5%, 포도당 0.5%, 정제수를 혼합하여 고초균으로 1단 발효한 참깨박 1단 발효물을 제조한 후, 상기 1단 발효물 3%에 파쇄미분말 3.0%, 효모추출물 1.5%, 포도당 1.5%, 정제수를 혼합하여 유산균으로 2단 발효한 2단 발효물에 해당한다. C is a mixture of soybean husk powder, 0.5% glucose, and purified water and fermented in one step with Bacillus subtilis, soybean hull fermented in one step, and sesame meal powder, 1.5% glucose, 0.5%, and purified water, and sesame fermented in one step with Bacillus subtilis. After preparing the first-stage fermented product, 3% of the first-stage fermentation product was mixed with 3.0% of crushed fine powder, 1.5% yeast extract, 1.5% glucose, and purified water, and it was fermented in two stages with lactic acid bacteria.
관능평가 결과에 의하면, 감칠맛 및 전체적인 기호도에서 2단 발효물(C)가 가장 높은 점수를 나타냈다. According to the sensory evaluation results, the two-stage fermented product (C) showed the highest score in umami and overall acceptability.
따라서, 본 발명의 2단 발효에 의하는 경우 유산군 1단 발효에 비해 감칠맛을 내는 글루타민 및 글루타민과의 상승효과에 의해 감칠맛을 더욱 증가시키는 알라닌 함량의 증가로 인해 감칠맛이 현저히 우수한 발효물을 제공할 수 있을 것으로 기대된다. Therefore, in the case of the two-stage fermentation of the present invention, compared to the one-stage fermentation of the lactic acid group, glutamine and alanine content, which further increase the umami taste due to the synergistic effect with glutamine, provide a fermented product with remarkably excellent umami taste. Expect to be able to do it.
Claims (23)
1) 대두 부산물을 고초균(B.spp.)으로 발효하여 대두 부산물의 1단 발효물을 제조하는 단계;
2) 참깨 부산물을 고초균(B.spp.)으로 발효하여 참깨 부산물의 1단 발효물을 제조하는 단계;
3) 상기 대두 부산물의 1단 발효물과 상기 참깨 부산물의 1단 발효물을 혼합하는 단계;
4) 상기 혼합물에 유산균을 접종하여 2단 발효물을 제조하는 단계; 및
5) 상기 2단 발효물을 여과하여 얻은 상층액을 농축하여 발효 농축액을 얻는 단계;를 포함하는 것을 특징으로 하는 유리 아미노산이 증가된 발효물의 제조방법.
As a method for producing a fermented product,
1) fermenting soybean by-products with Bacillus subtilis (B.spp.) to prepare a first-stage fermentation product of soybean by-products;
2) fermenting sesame by-products with Bacillus subtilis (B.spp.) to prepare a first-stage fermented product of sesame by-products;
3) mixing the first-stage fermentation product of the soybean by-product and the first-stage fermentation product of the sesame by-product;
4) preparing a two-stage fermented product by inoculating the mixture with lactic acid bacteria; and
5) Concentrating the supernatant obtained by filtering the two-stage fermentation product to obtain a fermentation concentrate; A method for producing a fermentation product with increased free amino acids.
According to claim 1, wherein the Bacillus subtilis is Bacillus subtilis ( B. subtilis ), Bacillus lichneformis ( B. lichneformis ), Bacillus megaterium ( B. megaterium ), Bacillus amyloliquefaciens ( B. amyloliquefaciens ), Bacillus natto ( B. natto ), Bacillus anthracis ( B.antharcis ), Bacillus lentus ( B.lentus ), Bacillus pumilus ( B. pumilus ), Bacillus theringjeensis ( B.thuringiensis ), Bacillus Alvey (B.alvei ), Bacillus azotopic Sans ( B.azotofixans ), Bacillus macerans ( B.macerans ), Bacillus polymyxa ( B.polymyxa ), Bacillus papilliae ( B.popilliae ), Bacillus coagulans ( B.coagulans ), Bacillus stearothermophilus ( B.stearothermophilus ), Bacillus pasteurii ( B.pasteurii ), Bacillus paricus ( B.sphaericus ), Bacillus pastidiosus ( B.fastidiosus ) selected from the group consisting of Method for producing a fermented product, characterized in that one or more.
The method for producing a fermented product according to claim 1, wherein the Bacillus subtilis is inoculated with an inoculum of 0.1 to 5%.
The method of claim 1, wherein the soybean by-product is soybean husk.
The method of claim 5, wherein the soybean husk contains soybean embryos.
The method for producing a fermented product according to claim 1, wherein the first stage fermentation is performed at a temperature of 15 to 50 ° C.
The method of claim 1, wherein the first stage fermentation is performed for 12 to 36 hours.
The method for producing a fermented product according to claim 1, wherein the two-stage fermentation of step 4) is performed at a temperature of 15 to 45 ° C.
The method for producing a fermented product according to claim 1, wherein the two-stage fermentation of step 4) is performed for 6 to 24 hours.
The method of claim 1, wherein the lactic acid bacteria in step 4) are of the genus Lactobacillus or Bifidobacterium.
The method of claim 12, wherein the lactic acid bacteria are Lactobacillus acidophilus , Lactobacillus rhamnosus , Lactobacillus plantarum , Bifidobacterium lactis and Bifidobacterium Method for producing a fermented product, characterized in that at least one selected from the group consisting of Gadomterium bifidum ( Bifidobacterium bifidum ).
The method of claim 1, wherein the two-stage fermentation of step 3) further comprises adding starch.
The method of claim 16, wherein the starch is crushed rice.
The method of claim 17, wherein the crushed rice is black rice or white rice.
The method for producing a fermented product according to claim 18, wherein the crushed rice is gelatinized.
Claims 1, 3 to 8, 10 to 13, and 16 to 19, a fermented product produced by the method of any one of claims 19.
[Claim 21] The fermented product according to claim 20, wherein the fermented product has an increased content of free amino acids and thus has excellent umami.
The fermented product according to claim 21, wherein the free amino acid is at least one selected from the group consisting of methionine, glutamine and alanine.
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| KR20030038132A (en) * | 2001-11-08 | 2003-05-16 | 류충호 | A novel fermented soybeans preparated by three steps fermentation using bacteria and method of preparation thereof |
| KR100891608B1 (en) * | 2008-08-29 | 2009-04-02 | (주)한국파비스 알엔디 | Method of producing fermentation of esculent plants, the fermentation produced thereby, and food comprising the fermentation |
| KR20100052666A (en) * | 2008-11-11 | 2010-05-20 | 경상대학교산학협력단 | Fermented soybean product prepared by rapid multi-step fermentation and method of preparation therefor |
| KR20220118682A (en) * | 2021-02-19 | 2022-08-26 | 농업회사법인 이웅식품 유한회사 | Meju and ssamjang with oil cake and manufacturing method thereof |
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| KR20030038132A (en) * | 2001-11-08 | 2003-05-16 | 류충호 | A novel fermented soybeans preparated by three steps fermentation using bacteria and method of preparation thereof |
| KR100891608B1 (en) * | 2008-08-29 | 2009-04-02 | (주)한국파비스 알엔디 | Method of producing fermentation of esculent plants, the fermentation produced thereby, and food comprising the fermentation |
| KR20100052666A (en) * | 2008-11-11 | 2010-05-20 | 경상대학교산학협력단 | Fermented soybean product prepared by rapid multi-step fermentation and method of preparation therefor |
| KR20220118682A (en) * | 2021-02-19 | 2022-08-26 | 농업회사법인 이웅식품 유한회사 | Meju and ssamjang with oil cake and manufacturing method thereof |
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