KR20160063659A - The producing method of lactic acid fermented cereal grains that increase the availability and preservation of lactic acid using a enzyme pretreatment cereal grains - Google Patents
The producing method of lactic acid fermented cereal grains that increase the availability and preservation of lactic acid using a enzyme pretreatment cereal grains Download PDFInfo
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- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
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- A23L7/00—Cereal-derived products; Malt products; Preparation or treatment thereof
- A23L7/10—Cereal-derived products
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Abstract
Description
본 발명은 유산균 발효곡물 및 그 제조방법에 관한 것으로, 정제화된 고농축 효소의 전(前)처리과정을 통하여 유산균(Lactic acid bacteria)의 곡물 이용성을 높여줌과 동시에 유산균 생균을 고농도로 함유하며 생리활성 성분의 증가와 항균물질의 생성 및 세포 외 다당류(EPS; exopolysaccharide)를 함유하는 유산균 발효곡물 및 그 제조방법에 관한 것이다.
The present invention relates to a lactic acid fermented grain and a method for producing the lactic acid bacterium. The present invention relates to a process for preparing lactic acid bacteria, And the production of antimicrobial substances and EPS (exopolysaccharide), and a process for producing the same.
유산균은 발효식품의 주된 미생물로서 오랫동안 발효유제품의 스타터로 이용되어 왔다. 특히, 최근 유산균이 정장작용, 면역 조절, 항암 및 항돌연변이 효과, 콜레스테롤 저하, 항알레르기 효과, 유당불내증 완화 등의 프로바이오틱으로서 그 기능성이 알려지면서 식품분야는 물론 의약, 화장품, 사료까지 그 범위가 확대되고 있는 실정이다.
Lactic acid bacteria have been used as starters of fermented milk products for a long time as the main microorganisms of fermented foods. In particular, recently, as the function of lactic acid bacteria as probiotics such as a suicide action, immunomodulation, anticancer and antimutagenic effect, cholesterol lowering, antiallergic effect, and lactose intolerance has been known, not only food, medicine, cosmetics, Is expanding.
한국의 전통 발효식품인 김치, 장류, 젓갈 등은 이러한 유산균 발효식품의 대표적인 사례이다. 특히 김치에서 분리되는 유산균에 대한 연구가 다양해지면서 김치의 유익한 효과인 식욕 촉진, 비만 예방, 변비 및 대장암 예방, 콜레스테롤 감소, 항산화 효과, 면역증강 효과, 고지혈증 억제 등이 연구발표 되면서 현재는 세계적으로 유산균 발효식품인 김치의 우수성이 널리 알려지고 있다.
Korean traditional fermented foods such as kimchi, soy sauce, and salted fish are representative examples of these lactic acid fermented foods. In particular, research on lactic acid bacteria isolated from kimchi has been diversified, and studies on the beneficial effects of kimchi such as appetite promotion, prevention of obesity, prevention of constipation and colorectal cancer, reduction of cholesterol, antioxidant effect, immune enhancement effect, The superiority of kimchi, a fermented food of lactic acid bacteria, is widely known.
이러한 유산균 발효식품의 특징은 유산균이 세포 외로 분비하는 유기산, 비단백성 세포 외 다당류(EPS), 항균성 펩타이드 외 여러 대사물질 등에 의한 것이고 특히, EPS에 대한 연구가 활발히 이루어지고 있는 실정이다. 미생물이 생산하는 EPS는 구성당 및 연결구조의 다양성으로 인해 다양한 물성을 가지고 있고, 자연계에서 쉽게 분해될 뿐만 아니라, 배양액 자체로도 쉽게 회수가 되기 때문에 적은 비용으로도 정제가 가능하다. 이런 장점으로 인해 식품산업에서의 유화제, 안정제, 응집제 등의 용도 외에도 다양한 분야에서 많은 주목을 받고 있다.
The characteristics of such lactic acid fermented foods are due to the fact that lactic acid bacteria are secreted by extracellularly, such as organic acids, non-necrotic extracellular polysaccharides (EPS), antimicrobial peptides, and other metabolites. Especially, EPS has been actively studied. The EPS produced by the microorganism has various physical properties due to the diversity of the constituent sugar and the linkage structure, and it is not only easily decomposed in the natural world, but also easily recovered by the culture medium itself, so that it can be purified even at a small cost. These advantages have attracted much attention in various fields besides the use of emulsifiers, stabilizers and coagulants in the food industry.
이러한 배경 하에, 정제화된 고농축 효소의 전처리과정을 거치면 유산균의 곡물 이용성이 증대되고 이로 인해 유산균 생균을 고농도로 함유할 수 있으며 유산균이 분비한 EPS가 안정제 역할을 하여 상온 보관 시 안정적으로 장기보관이 가능하게 되었고 각종 생리활성 성분과 항균물질을 생성함을 확인하여 본 발명을 완성하게 되었다.
Under these circumstances, the pre-treatment of purified high-concentration enzymes increases the availability of lactic acid bacteria to cereals, which can contain high concentrations of lactic acid bacteria, and the EPS secreted by lactic acid bacteria acts as a stabilizer. And it was confirmed that various physiologically active ingredients and antimicrobial substances were produced, and thus the present invention was completed.
따라서, 본 발명의 주된 목적은 유산균 발효곡물 및 그 제조방법에 관한 것으로, 곡물의 정제화된 고농축효소의 전처리과정을 통하여 유산균의 곡물 이용성을 높여줌과 동시에 발효곡물이 유산균 생균을 고농도로 함유, 생리활성 성분의 증가, 항균물질의 생성 및 EPS를 함유하는 유산균 발효곡물 및 그 제조방법을 제공하기 위함이다.
Accordingly, the main object of the present invention is to provide a fermented cereal grains having a high concentration of lactic acid bacteria and a physiological activity An antimicrobial substance, an EPS-containing lactobacillus fermented grain, and a process for producing the same.
본 발명은 상기의 목적을 달성하기 위한 실시방법으로 본 발명자는 원료곡물의 유산균 발효를 위해 정제된 고농축효소를 이용해 전처리과정을 거친 후 유산균 발효를 진행하는 방법으로 유산균 발효곡물을 제조하였다.
The present invention is a method for achieving the above object. The present inventors prepared a fermented lactic acid bacterium by subjecting a raw grain to a pretreatment using a highly concentrated enzyme purified for lactic acid bacteria fermentation, followed by lactic acid fermentation.
본 발명에 있어서, 전처리 단계의 정제화된 고농축효소는 아밀라아제(amylase), 프로테아제(protease), 자일라나제(xylanase), 셀룰라아제(cellulase), 헤미셀룰라아제(hemicellulase), 리파아제(lipase), 펙티나제(pectinase), 글루코아밀라아제(glucoamylase), 인베르타아제(invertase), 파파인(papain), 브로멜라인(bromelain), 디아스타제(diastase), 판크레아틴(pancreatin)으로 구성되는 어느 하나 또는 하나 이상의 것으로 종래의 미생물에 의해 해당기질의 유기물분해 효소를 분비하도록 유도하는 것이 아닌 순수하게 정제된 유기물분해 효소를 전처리 단계에 활용함으로서 단시간 내에 원료곡물의 분해를 촉진하고 유산균 성장에 있어 이용성을 높여줌으로서 유산균 생균은 물론 그 대사물질을 함유하도록 하는 단계를 포함하는 유산균 발효곡물 및 그 제조방법을 제공한다.
In the present invention, the purified highly concentrated enzyme in the pretreatment step may be an amylase, a protease, an xylanase, a cellulase, a hemicellulase, a lipase, a pectinase pectinase, glucoamylase, invertase, papain, bromelain, diastase, pancreatin, and one or more of them, The use of purely purified organolytic enzymes in the pretreatment stage, which does not induce the secretion of the organic degrading enzyme of the corresponding substrate by the microorganisms of the present invention, promotes the decomposition of the raw grains in a short time and improves the availability of the lactic acid bacteria. Comprising a step of allowing the lactic acid bacteria to contain the metabolite, and a method of producing the fermented lactic acid bacterium.
전술한 본 발명의 발효방법은, 국내에서 생산, 유통되는 곡물을 원료로 하는 단계; 준비된 곡물을 중량 대비 2배 부피의 효소액으로 5시간 내지 10시간 동안 침지하는 단계; 침지가 완료된 곡물을 흐르는 물로 세척하는 단계; 스팀을 이용해 100℃ 내지 105℃에서 1시간 내지 5시간 동안 살균하는 단계; 살균이 완료된 곡물을 40℃ 내지 50℃로 냉각하는 단계; 냉각된 곡물에 미리 준비된 유산균 스타터를 접종하여 20시간 내지 50시간 동안 고체배양 하는 단계; 및 고체배양 후 40℃ 내지 50℃에서 열풍으로 건조하는 단계; 를 포함하는 유산균 발효곡물 및 그 제조방법을 제공함으로서 달성 될 수 있다.
The fermentation method of the present invention comprises: a step of using cereals produced and distributed in the country as raw materials; Immersing the prepared cereal grains in an enzyme solution twice the volume of the grains for 5 to 10 hours; Washing the immersed grain with running water; Sterilizing with steam at 100 ° C to 105 ° C for 1 hour to 5 hours; Cooling the sterilized cereal to 40 占 폚 to 50 占 폚; Inoculating the cooled grains with a lactic acid bacteria starter prepared in advance and solid-culturing for 20 to 50 hours; And drying with hot air at 40 ° C to 50 ° C after solid culture; And a method for producing the fermented cereal grains.
또한, 곡물이라 함은 현미, 보리, 율무, 대두, 서리태, 흑미, 밀, 찹쌀, 수수, 옥수수로 이루어진 것으로 선택된 어느 하나 또는 하나 이상인 것을 포함하는 것을 특징으로 한다.
The term " grain " includes any one or more selected from the group consisting of brown rice, barley, yulmu, soybean, black rice, wheat, glutinous rice, sorghum and corn.
또한, 효소액이라 함은 아밀라아제(amylase), 프로테아제(protease), 자일라나제(xylanase), 셀룰라아제(cellulase), 헤미셀룰라아제(hemicellulase), 리파아제(lipase), 펙티나제(pectinase), 글루코아밀라아제(glucoamylase), 인베르타아제(invertase), 파파인(papain), 브로멜라인(bromelain), 디아스타제(diastase), 판크레아틴(pancreatin)으로 이루어진 군에서 선택된 어느 하나 또는 하나 이상인 것을 포함하는 것을 특징으로 한다.
Also, the enzyme solution refers to an amylase, protease, xylanase, cellulase, hemicellulase, lipase, pectinase, glucoamylase, And at least one selected from the group consisting of invertase, papain, bromelain, diastase, pancreatin, and the like. .
또한, 효소액의 제조는 상기에서 제시한 효소 중 선택되어진 어느 하나 또는 하나 이상인 것을 포함하되, 최종 부피의 0.1% 내지 1.0% 로 구성되어 지는 것을 특징으로 한다.
In addition, the preparation of the enzyme solution may include any one selected from the above-mentioned enzymes or one or more selected from the group consisting of 0.1% to 1.0% of the final volume.
또한, 유산균 스타터라 함은 비피도박테리움 비피덤(Bifidobacterium bifidum), 비피도박테리움 브레브(Bifidobacterium breve), 비피도박테리움 락티스(Bifidobacterium lactis), 비피도박테리움 롱검(Bifidobacterium longum), 락토바실러스 애시도필러스(Lactobacillus acidophilus), 락토바실러스 브레비스(Lactobacillus brevis), 락토바실러스 불가리쿠스(Lactobacillus bulgaricus), 락토바실러스 카세이(Lactobacillus casei), 락토바실러스 델브루에키(Lactobacillus delbrueckii), 락토바실러스 퍼멘텀(Lactobacillus fermentum), 락토바실러스 가세리(Lactobacillus gasseri), 락토바실러스 락티스(Lactobacillus lactis), 락토바실러스 파라카세이(Lactobacillus paracasei), 락토바실러스 파라플랜타룸(Lactobacillus paraplantarum), 락토바실루스 펜토서스(Lactobacillus pentosus), 락토바실러스 플란타룸(Lactobacillus plantarum), 락토바실러스 루테리(Lactobacillus reuteri), 락토바실러스 람노서스(Lactobacillus rhamnosus), 락토바실루스 사케이(Lactobacillus sakei), 류코노스톡 시트륨(Leuconostoc citreum), 류코노스톡 메센테로이데스(Leuconostoc mesenteroides), 페디오코커스 매시디락티스(Pediococcus acidilactici), 페디오코쿠스 펜토사세우스(Pediococcus pentosaceus), 바이젤라 콘푸사(Weissella confusa), 바이젤라 코리엔시스(Weissella koreensis)로 이루어진 군에서 선택된 어느 하나 또는 하나 이상인 것을 포함하는 것을 특징으로 한다.
In addition, the lactic acid bacteria starter is Bifidobacterium ( Bifidobacterium < RTI ID = 0.0 > bifidum), Bifidobacterium breather bracket (Bifidobacterium breve), Bifidobacterium lactis (Bifidobacterium lactis ), Bifidobacterium longum ), Lactobacillus acidophilus ( Lactobacillus acidophilus , Lactobacillus < RTI ID = 0.0 > brevis , Lactobacillus bulgaricus , Lactobacillus < RTI ID = 0.0 > casei ), Lactobacillus delbruecki delbrueckii , Lactobacillus < RTI ID = 0.0 > fermentum), Lactobacillus biasing Li (Lactobacillus gasseri , Lactobacillus < RTI ID = 0.0 > lactis ), Lactobacillus paracasei paracasei , Lactobacillus < RTI ID = 0.0 > paraplantarum , Lactobacillus < RTI ID = 0.0 > pentosus , Lactobacillus < RTI ID = 0.0 > plantarum , Lactobacillus reuteri , Lactobacillus < RTI ID = 0.0 > rhamnosus , Lactobacillus < RTI ID = 0.0 > sakei), flow sheets Stock Pocono volume (Leuconostoc citreum), current Kono Stock mesen teroyi Death (Leuconostoc mesenteroides , Pediococcus < RTI ID = 0.0 > acidilactici), Phedi OKO kusu pen soil three mouse (Pediococcus pentosaceus , Weissella confusa , and Weissella koreensis . The present invention is characterized in that it comprises at least one selected from the group consisting of Confusa and Weissella koreensis .
또한, 유산균 스타터의 제조는 상기에서 제시한 유산균 중 선택되어진 어느 하나 또는 하나 이상인 것을 포함하되, 최종 접종액 부피의 1.0% 내지 5.0%, 1.0×107 cfu/mL 내지 5.0×107 cfu/mL 인 것을 특징으로 함으로서 달성 될 수 있다.
The lactic acid bacterial starter may be prepared from any one or more selected lactic acid bacteria selected from the group consisting of 1.0% to 5.0%, 1.0 × 10 7 cfu / mL to 5.0 × 10 7 cfu / mL of the final inoculum volume As shown in FIG.
본 발명에 의하면 정제화된 고농축효소의 전처리과정을 통하여 유산균의 곡물 이용성을 높여줌과 동시에 유산균 생균을 고농도로 함유, 생리활성 성분의 증가, 항균물질의 생성 및 세포 외 다당류(EPS)를 함유하는 유산균 발효곡물을 제조할 수 있는 효과가 있다.
According to the present invention, it is possible to improve the utilization of lactic acid bacteria in cereals by pretreatment of purified highly concentrated enzyme, and also to provide a high concentration of lactic acid bacteria, increase of physiologically active ingredients, production of antibacterial substances and lactic acid bacteria fermentation There is an effect that the grain can be manufactured.
도면 1 : 본 발명의 제조과정을 도시화하여 나타낸 흐름도.
도면 2 : 본 발명으로 제조된 발효곡물의 장기간 저장동안 유산균 생균의 변화 그래프.
도면 3 : 본 발명으로 제조된 발효곡물의 배양시간 변화에 따른 희석액의 바실러스 세레우스 (Bacillus cereus, KCTC 13153)에 대한 항균력 사진.
도면 4 : 본 발명으로 제조된 발효곡물의 배양시간 변화에 따른 희석액의 살모넬라 타이피뮤리움 (Salmonella typhimurium, KCCM 40253)에 대한 항균력 사진.
도면 5 : 본 발명으로 제조된 발효곡물의 배양시간 변화에 따른 희석액의 대장균 (Escherichia coli, KCCM 40405)에 대한 항균력 사진.1 is a flow chart illustrating the manufacturing process of the present invention.
Figure 2: Graph of change in lactobacillus viability during long-term storage of fermented grains prepared according to the present invention.
Figure 3: Bacillus cereus (Bacillus of the diluent according to the culture time variation of the fermented grains prepared in the present invention cereus , KCTC 13153).
Figure 4: Salmonella ties in the diluent according to the culture time variation of the fermented grains prepared in the present invention pimyu Solarium (Salmonella typhimurium , KCCM 40253).
FIG. 5: Photograph of the antimicrobial activity against Escherichia coli (KCCM 40405) of the diluted solution according to the change of culture time of the fermented grain prepared according to the present invention.
본 발명의 이해를 돕고자 바람직한 실시예를 제시한다. 그러나 하기의 실시예는 본 발명을 보다 쉽게 이해하기 위함이며 본 발명은 하기의 실시예에 한정되는 것은 아니다.
Preferred embodiments are provided to facilitate understanding of the present invention. However, the following examples are intended to further illustrate the present invention, and the present invention is not limited to the following examples.
대두 1kg에 2리터의 증류수를 넣고 5시간동안 상온에서 침지 시킨 후 흐르는 물로 세척하고, 100℃에서 2시간 동안 살균하였다. 살균 후 40℃로 냉각하고 중량대비 sucrose 2%(w/w)를 첨가 후 혼합해 주고 0.85%(w/v) 멸균 생리식염수 300mL를 접종하고 40시간 동안 45℃에서 방치 후 건조하여 완성하였다(대조군).
Two liters of distilled water was added to 1 kg of soybeans, dipped at room temperature for 5 hours, washed with flowing water, and sterilized at 100 ° C for 2 hours. After sterilization, the mixture was cooled to 40 ° C., and 2% (w / w) of sucrose was added thereto. The mixture was inoculated with 300 mL of 0.85% (w / v) sterilized physiological saline and allowed to stand at 45 ° C. for 40 hours and dried Control group).
대두 1kg에 2리터의 증류수를 넣고 5시간동안 상온에서 침지 시킨 후 흐르는 물로 세척하고, 100℃에서 2시간 동안 살균하였다. 살균 후 40℃로 냉각하고 중량대비 sucrose 2%(w/w)를 첨가 후 혼합해 주고 미리 전배양한 페디오코쿠스 펜토사세우스(Pediococcus pentosaceus)를 1.0×107 cfu/mL 수준으로 300mL를 접종하고 40시간 동안 고체배양 후 45℃로 건조하여 완성하였다.
Two liters of distilled water was added to 1 kg of soybeans, dipped at room temperature for 5 hours, washed with flowing water, and sterilized at 100 ° C for 2 hours. After sterilization, the mixture was cooled to 40 ° C, and 2% (w / w) sucrose was added to the mixture. Pediococcus pentosaceus , which had been pre-preincubated, was added at a concentration of 1.0 × 10 7 cfu / Inoculated and solidified for 40 hours and then dried at 45 < 0 > C to complete.
대두 1kg에 브로멜라인(1,200 GDU) 0.2%, 아밀라아제(500 Unit) 0.2%, 자일라나제(500 Uint) 0.2%, 셀룰라아제(500 Unit) 0.2%를 포함한 2리터의 증류수를 넣고 5시간동안 상온에서 침지 시킨 후 흐르는 물로 세척하고, 100℃에서 2시간 동안 살균하였다. 살균 후 40℃로 냉각하고 중량대비 sucrose 2%(w/w)를 첨가 후 혼합해 주고 미리 전배양한 페디오코쿠스 펜토사세우스(Pediococcus pentosaceus)를 1.0×107 cfu/mL 수준으로 300mL를 접종하고 40시간 동안 고체배양 후 45℃로 건조하여 완성하였다.
Two liters of distilled water containing 0.2% of bromelain (1,200 GDU), 0.2% of amylase (500 units), 0.2% of xylanase (500 Uint) and 0.2% of cellulase (500 units) was added to 1 kg of soybeans, , Washed with flowing water, and sterilized at 100 DEG C for 2 hours. After sterilization, the mixture was cooled to 40 ° C, and 2% (w / w) sucrose was added to the mixture. Pediococcus pentosaceus , which had been pre-preincubated, was added at a concentration of 1.0 × 10 7 cfu / Inoculated and solidified for 40 hours and then dried at 45 < 0 > C to complete.
상기 실시예 3의 방법으로 제조하되, 고체배양 중 10시간, 20시간, 30시간, 40시간 때 수거 후 건조하여 완성하였다.
The cells were prepared by the method of Example 3, and were collected and dried after 10, 20, 30, and 40 hours in solid culture.
[분석 1] 생균수 측정[Analysis 1] Measurement of viable cell count
실시예 2와 실시예 3으로 제조된 시료를 각 2g씩 0.85% 생리식염수 18mL에 현탁 후 순차적으로 1mL씩 9mL의 생리식염수로 10배씩 희석하여 최종 희석액을 제조하였다. 본 희석액을 0.1mL씩 MRS agar에 도말한 후 37℃, 48시간 동안 배양 후 생균수를 확인하였다(표 1).
The samples prepared in Example 2 and Example 3 were suspended in 18 mL of 0.85% physiological saline every 2 g each, and then diluted 10 mL with 9 mL of physiological saline sequentially by 1 mL each to prepare a final diluted solution. 0.1 ml of this diluted solution was applied to MRS agar and cultured at 37 ° C for 48 hours. The number of viable cells was confirmed (Table 1).
상기와 별도로, 실시예 3으로 제조된 시료를 상온에서 보관하면서 상기에서 언급한 생균수 측정법에 따라 8개월 동안 생균수의 변화를 주기적(매월 마지막 주)으로 측정하였다(표 2).
Apart from the above, the sample prepared in Example 3 was stored at room temperature, and the change in the viable cell count was measured periodically (last week of each month) for 8 months according to the above-mentioned method for measuring viable cell counts (Table 2).
상기 결과에서 효소액 전처리한 발효곡물(실시예 3)에서 효소액 전처리하지 않은 발효곡물(실시예 2)보다 높은 수의 생균수를 나타내었으며, 특히 동결건조가 아닌 열풍건조를 통해서도 높은 수의 생균을 유지할 수 있음을 확인하였다. 또한 표2의 결과에서 보았듯이 상온에서 장기간 보관 하더라도 생균수의 변화가 거의 없는 것으로 확인되었는데 이는 제품 보관이 용이하다는 것과 유통기한 내에는 같은 품질의 유산균을 섭취할 수 있다는 것을 보여준다.
In the above results, the number of viable cells was higher than that of the fermented grains that had not been pretreated with the enzyme solution (Example 3) in the fermented grain pretreated with the enzyme solution (Example 2). In particular, Respectively. In addition, as shown in Table 2, it was confirmed that even when stored at room temperature for a long period of time, the viable cell count was almost unchanged. This indicates that the product is easy to store and that the same quality lactobacillus can be consumed within the shelf life.
[분석 2] 총 이소플라본(isoflavone) 함량[Analysis 2] Total isoflavone content
대조예 1과 실시예 2와 실시예 3으로 제조된 시료 0.5g에 1M HCl 2mL를 첨가한 후 2시간 동안 항온수조에서 95℃ 온도로 가열한 후 냉각하고 8mL의 아세토니트릴을 첨가하여 교반한 다음 2시간 동안 정치하여 최종 이소플라본을 추출하였다. 본 추출액은 다시 감압농축하고 고형물을 80%(v/v) 메탄올에 용해 후 0.2μm 실린지 필터로 여과하여 HPLC에 주입 후 아세토니트릴 : 0.05M KH2PO4용액을 25 : 75로 혼합한 용매를 이동상으로 UV 파장 260nm에서 측정하여 정량하였다.
2 ml of 1M HCl was added to 0.5 g of the sample prepared in Control Example 1, Example 2 and Example 3, and the mixture was heated to 95 ° C in a constant temperature water bath for 2 hours, cooled, added with 8 ml of acetonitrile, The mixture was allowed to stand for 2 hours to extract the final isoflavone. The extract was concentrated under reduced pressure, and the solids were dissolved in 80% (v / v) methanol, filtered through a 0.2 μm syringe filter, injected into HPLC, and then dissolved in a solvent mixture of acetonitrile: 0.05M KH 2 PO 4 Was measured as a mobile phase at a UV wavelength of 260 nm and quantified.
상기 결과에서 효소액 처리하지 않은 발효곡물(실시예 2) 보다 효소액 전처리를 한 발효곡물(실시예 3)에서 생리활성 성분인 총 이소플라본이 높은 함량으로 증가되어 있음을 알 수 있다.
From the above results, it can be seen that the total amount of isoflavone, which is a physiologically active ingredient, is increased in the fermented cereals (Example 3) that were pretreated with the enzyme solution than the fermented cereals without the enzyme solution (Example 2).
[분석 3] EPS(Exopolysaccharide)의 정량[Analysis 3] Quantification of EPS (Exopolysaccharide)
대조예 1을 대조구로 하여 실시예 2와 실시예 3으로 제조된 시료에 10배(v/w) 분량의 멸균 증류수로 현탁 후 10분간 볼텍싱 해준 후 12,000 xg에서 5분간 원심분리하고, 상징액을 회수 후 동일 부피만큼(1:1) 95% 에탄올을 첨가 후 4℃에서 24시간동안 정치하고 다시 1,000 xg에서 5분간 원심분리하여 침전물을 회수한 다음 60℃에서 12시간 동안 건조하여 실시예 1의 건조중량(w/v)을 대조구로 하여 실시예 2와 실시예 3의 건조중량을 비교하였다.
Samples prepared in Example 2 and Example 3 were suspended in sterilized distilled water 10 times (v / w), and vortexed for 10 minutes, then centrifuged at 12,000 xg for 5 minutes, and the supernatant After the recovery, 95% ethanol was added by the same volume (1: 1), and the mixture was allowed to stand at 4 ° C for 24 hours. The mixture was centrifuged at 1,000 xg for 5 minutes to recover the precipitate, followed by drying at 60 ° C for 12 hours. The dry weights of Example 2 and Example 3 were compared using dry weight (w / v) as a control.
(실시예 2 건조중량
- 대조예 1 건조중량)Example 2
(Example 2 Dry weight
- Control 1 Dry weight)
(실시예 3 건조중량
- 대조예 1 건조중량)Example 3
(Example 3 Dry weight
- Control 1 Dry weight)
상기 결과에서 효소액을 처리하지 않은 발효곡물(실시예 2)보다 효소액의 전처리를 한 발효곡물(실시예 3)에서 안정제 역할을 하여 상온 보관 시 안정적으로 장기보관이 가능토록 균체의 보호역할을 하는 EPS의 함유량이 증가되었음을 알 수 있다.
From the above results, it can be seen that as a stabilizer in the fermented cereal (Example 3) which has been pretreated with the enzyme solution than the fermented grain not treated with the enzyme solution (Example 2) Of the total amount of water.
[분석 4][Analysis 4]
실시예 4에서 얻어진 각 시료를 3배(v/w) 분량의 멸균 증류수로 현탁 후 10분간 볼텍싱 해주고 12,000 xg에서 15분간 원심분리한 검액을 4℃에서 보관한 후 미리 전배양한 바실러스 세리우스(Bacillus cereus, KCTC 13153), 살모넬라 타이피뮤리움(Salmonella typhimurium, KCCM 40253), 대장균(Escherichia coli, KCCM 40405)을 1.0×107 cfu/mL로 조정하고, Nutrient agar(Difco사)를 중층배지로 하여 각 1mL씩 분주 후 5회 정도 섞어준 후 굳힌다. 준비된 검액은 paper disc(지름 5mm)에 100μL씩 점적하고 잘 말려준 후 병원성 미생물을 중층한 고체배지에 배양시간 변화로 구분하여 올려주고 완료된 플레이트를 37℃에서 24시간 배양 후 clear zone의 지름(mm)으로 비교하였다.
Each sample obtained in Example 4 was suspended in sterilized distilled water three times (v / w), vortexed for 10 minutes, centrifuged at 12,000 xg for 15 minutes, stored at 4 ° C and pre- ( Bacillus cereus, KCTC 13153), S. typhimurium (Salmonella typhimurium , KCCM 40253), Escherichia coli , KCCM 40405) is adjusted to 1.0 × 10 7 cfu / mL, and Nutrient agar (Difco) is used as an intermediate layer medium. Dissolve each 1 mL each, mix five times, and then harden. 100 μL of the prepared sample solution was added dropwise to the paper disc (diameter 5 mm) and dried. Then, the pathogenic microorganisms were placed on a solid medium in which the culture medium was grown, and the plate was incubated at 37 ° C. for 24 hours. ).
(KCTC 13153) Bacillus cereus
(KCTC 13153)
(KCCM 40253) Salmonella typhimurium
(KCCM 40253)
(KCCM 40405) Escherichia coli
(KCCM 40405)
상기 결과에서 배양시간이 경과하면서 장내의 독소생성세균(바실러스 세리우스, 살모넬라 타이피뮤리움, 대장균)에 대한 항균력을 가진 유산균의 증식이 잘 이루어졌음을 보여준다.
The above results show that the growth of the lactic acid bacteria having the antimicrobial activity against the toxin-producing bacteria (Bacillus cerius, Salmonella typhimurium, Escherichia coli) in the intestines was well accomplished during the incubation time.
이상 본 발명에 따르면, 효소 전처리 하지 않은 발효곡물(실시예 2)과 효소 전처리한 발효곡물(실시예 3)을 비교하여 본 결과 정제 효소 전처리를 통해 곡물 발효를 진행할 경우 상기의 결과들처럼 발효곡물이 높은 수의 유산생균을 함유할 수 있었고 유익한 생리활성물질 및 항균물질을 포함하며 이러한 대사물질로 인해 장기간 저장 시에도 안정한 균수를 유지할 수 있는 것으로 확인되었다.
According to the present invention, when the fermentation of the fermented cereal (Example 2) without enzyme pretreatment and the fermented cereal with enzyme pretreatment (Example 3) are compared with each other, It was confirmed that this high number of lactic acid bacteria could be contained, and beneficial biologically active substances and antimicrobial substances could be contained.
따라서, 본 발명을 통해 유산균 생균은 물론 대사물질과 생리활성 물질을 모두 포함하는 곡물발효 식품을 제공할 수 있음을 보여 주었다.Therefore, it has been shown through the present invention that a fermented food product containing both a metabolite and a physiologically active substance as well as lactic acid bacteria can be provided.
Claims (7)
Grain produced and distributed domestically is used as raw material; Immersing the prepared cereal grains in an enzyme solution twice the volume of the grains for 5 to 10 hours; Washing the immersed grain with running water; Sterilizing with steam at 100 ° C to 105 ° C for 1 hour to 5 hours; Cooling the sterilized cereal to 40 占 폚 to 50 占 폚; Inoculating the cooled grains with a lactic acid bacteria starter prepared in advance and solid-culturing for 20 to 50 hours; And drying with hot air at 40 ° C to 50 ° C after solid culture; Of fermented lactic acid bacteria and method for producing the same
The method according to claim 1, wherein the grain is one or more selected from the group consisting of brown rice, barley, yulmu, soybean, seabream, black rice, wheat, glutinous rice, sorghum and corn.
The enzyme solution according to claim 1, wherein the enzyme solution is selected from the group consisting of amylase, protease, xylanase, cellulase, hemicellulase, lipase, pectinase, But are not limited to, any one or more selected from the group consisting of glucoamylase, invertase, papain, bromelain, diastase, pancreatin, ≪ / RTI >
The method according to claim 1, wherein the preparation of the enzyme solution comprises any one or more than one selected from the group of claim 3, and is comprised of 0.1% to 1.0% of the final volume.
The lactic acid bacteria starter according to claim 1, wherein the lactic acid bacteria starter is Bifidobacterium bifidum (Bifidobacterium bifidum), Bifidobacterium breve (Bifidobacterium breve), Bifidobacterium lactis (Bifidobacterium lactis), Bifidobacterium longum (Bifidobacterium longum), Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus brevis (Lactobacillus brevis), Lactobacillus bulgaricus (Lactobacillus bulgaricus), Lactobacillus casei (Lactobacillus casei), Lactobacillus delbruecki (Lactobacillus delbrueckii), Lactobacillus fermentum (Lactobacillus fermentum), Lactobacillus casei (Lactobacillus gasseri), Lactobacillus lactis (Lactobacillus lactis), Lactobacillus paracasei (Lactobacillus paracasei), Lactobacillus paraplanaceum (Lactobacillus paraplantarum), Lactobacillus pentosus (Lactobacillus pentosus), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus lutea (Lactobacillus reuteri), Lactobacillus lambatus (Lactobacillus rhamnosus), Lactobacillus casei (Lactobacillus sakei), Leuconosistium (Leuconostoc citreum), Leuconostomesenteroids (Leuconostoc mesenteroides), Pediococcus macydiractis (Pediococcus acidilactici), Pediococcus pentosaceus (Pediococcus pentosaceus), Vizela Confusa (Weissella confusa), Vizela coliensis (Weissella koreensis). ≪ / RTI >
6. The process according to claim 5, wherein the preparation of the lactic acid bacteria starter comprises one or more than one selected from the group of claims 5, wherein 1.0% to 5.0%, 1.0x10 7 cfu / mL to 5.0x10 7 cfu / mL. < / RTI >
A fermented lactic acid bacterium produced by any one of claims 1 to 6.
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| KR20190082179A (en) * | 2019-06-28 | 2019-07-09 | 푸른솔푸드 주식회사 | Enzyme food composite manufacture method using Isolated Soy Protein and brown rice |
| KR20200002156A (en) * | 2018-06-29 | 2020-01-08 | 제너럴네이처주식회사 | Brown rice fermented soybean with increased isoflavone aglycone content and producing method thereof |
| KR20210026889A (en) * | 2019-09-02 | 2021-03-10 | 주식회사 비피도 | Weissella confusa VP30 to highly secrete EPS and food composition for improvement of constipation therefrom |
| KR20220064000A (en) * | 2020-11-11 | 2022-05-18 | 박노흥 | Production method of fermented Soybean using Pineapple |
| KR20220108348A (en) * | 2021-01-27 | 2022-08-03 | 단국대학교 천안캠퍼스 산학협력단 | Functional fermented brown rice having enhanced nutritional quality and manufacturing method thereof |
| KR102508665B1 (en) * | 2022-11-07 | 2023-03-15 | 주식회사 보부상바이오팜 | Preparation method for protein preparations with improved digestion |
-
2014
- 2014-11-27 KR KR1020140167173A patent/KR20160063659A/en not_active Application Discontinuation
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20190079836A (en) * | 2017-12-28 | 2019-07-08 | (주)프레쉬벨 | Method for producing fermented barley sprout with enhanced saponarin, isovitexin and luteolin using novel Lactobacillus fermentum strain |
| KR20200002156A (en) * | 2018-06-29 | 2020-01-08 | 제너럴네이처주식회사 | Brown rice fermented soybean with increased isoflavone aglycone content and producing method thereof |
| KR20190082179A (en) * | 2019-06-28 | 2019-07-09 | 푸른솔푸드 주식회사 | Enzyme food composite manufacture method using Isolated Soy Protein and brown rice |
| KR20210026889A (en) * | 2019-09-02 | 2021-03-10 | 주식회사 비피도 | Weissella confusa VP30 to highly secrete EPS and food composition for improvement of constipation therefrom |
| KR20220064000A (en) * | 2020-11-11 | 2022-05-18 | 박노흥 | Production method of fermented Soybean using Pineapple |
| KR20220108348A (en) * | 2021-01-27 | 2022-08-03 | 단국대학교 천안캠퍼스 산학협력단 | Functional fermented brown rice having enhanced nutritional quality and manufacturing method thereof |
| KR102508665B1 (en) * | 2022-11-07 | 2023-03-15 | 주식회사 보부상바이오팜 | Preparation method for protein preparations with improved digestion |
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