KR20100020582A - Carrot juice residues fermented materials and manufacturing method thereof - Google Patents
Carrot juice residues fermented materials and manufacturing method thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
- A23L19/03—Products from fruits or vegetables; Preparation or treatment thereof consisting of whole pieces or fragments without mashing the original pieces
- A23L19/07—Fruit waste products, e.g. from citrus peel or seeds
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
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- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
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- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
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Abstract
Description
본 발명은 당근 즙 제조 공정 후 나오는 부산물인 당근박의 활용 방안에 관한 것으로, 당근박을 발효시킨 당근박 발효물 및 그 제조방법을 제공함으로서 기능성 점질물을 비롯한 혈전 분해효소 등 다양한 효소들을 생산하며 동시에 프로바이오틱으로서 젖산균 함유 및 뇌 개선물질인 GABA를 포함하는 기능성을 함유한 당근박 발효물 및 그 제조방법에 관한 것이다.The present invention relates to a method of utilizing carrot foil which is a by-product of the carrot juice manufacturing process, and provides various fermented products including thrombolytic enzymes including functional viscous products by providing a fermented carrot foil and a method of preparing the same. It relates to a carrot foil fermented product containing the functionalities including the lactic acid bacteria as a probiotic and GABA which is a brain improving substance, and a manufacturing method thereof.
당근은 풍부한 식이섬유와 인체의 영양원으로 중요한 카로틴을 많이 함유하고 있다. 카로틴은 비타민 A의 전구체로서 항암작용, 성인병 예방 등의 기능을 가지고 있다고 알려져 있다. 카로틴은 당근의 색깔과 영양을 결정하는 주요물질로 체내에 흡수되었을 때 약 30 %가 비타민 A로 전환된다. Carrots are rich in dietary fiber and carotene, which is important for human nutrition. Carotene is a precursor of vitamin A and is known to have functions such as anticancer activity and prevention of adult diseases. Carotene is a major determinant of carrot color and nutrition, and when absorbed into the body, about 30% is converted to vitamin A.
종래 당근은 샐러드, 생식, 라면스프 등으로 많이 이용되었으나, 최근에는 주스 원료로 활용이 크게 증가하고 있는 실정이다. 특히, 식생활의 다양한 변화와 함께 바쁜 생활 속에서 야채 생즙으로 제품화된 것을 섭취하는 경우가 늘고 있으므로 이에 따른 생즙의 재료로 그 수요가 빠르게 증가하고 있다. 특히 당근 생즙의 독특한 색상과 맛 때문에 다른 생즙의 혼합원료로 사용되고 있어서 대량의 당근이 생즙으로 가공되고 있는 실정이다.Conventionally, carrots have been used as salads, raw foods, ramen soups, etc., but the use of juice as a raw material has recently increased. In particular, with the changes in the diet, the consumption of the product of vegetable juice in a busy life is increasing, so the demand for the raw juice is increasing accordingly. In particular, due to the unique color and taste of carrot juice, it is used as a mixed raw material of other juice, so that a large amount of carrots are processed into fresh juice.
그러나 당근 즙을 생산하는 과정에서 착즙방법에 따라 screw를 이용하는 방법이나 원심력에 의한 방법이 가능하여, 상업적으로 당근 즙은 50% 정도의 수율을 보이면서 50%정도가 부산물인 당근박으로 배출되고 있는 실정이다. 특히 중견업체로서 당근박의 매달 10톤 이상 생산되는 시점에서 당근 박을 사료로 사용하는 것은 자원의 낭비이며, 기업의 경쟁력을 높이기 위해서는 당근박의 고부가가치화가 절실히 요구된다. 또한 당근박은 수분함량이 50%이상으로 당근 생즙이 갖는 색소성분과 영양성분을 가지고 있으며, 특히 식이섬유의 주원료로 사용될 수 있다. 이런 영양성분과 수분함량에 기인하여 생산되는 당근박은 쉽게 변질될 수 있어서, 식품소재로 활용이 어려운 실정이다. However, in the process of producing carrot juice, it is possible to use a screw method or centrifugal force depending on the juice method. Commercially, carrot juice has a yield of about 50% and is discharged as a byproduct of carrot foil. to be. In particular, as a medium-sized company, the use of carrot gourd as feed for the production of more than 10 tons of carrot gourd every month is a waste of resources. In addition, carrot foil has more than 50% moisture content and has nutritional components and pigments of carrot juice, especially can be used as the main raw material of dietary fiber. Carrot foil produced due to the nutritional content and moisture content can be easily deteriorated, it is difficult to use as a food material.
지금까지 보고된 당근 박 발효에 관한 연구로는 젖산발효에 의한 혼합과채음료 제조방법, 혼합과채의 품질 등 기능성을 가진 젖산 발효식품을 개발함으로써 용도의 개발이 가능하다는 연구가 있었다. As a study on the fermentation of carrot gourd reported so far, there was a study that the development of lactic acid fermented foods with functionalities such as the method of manufacturing mixed fruit drink by lactic acid fermentation and the quality of mixed fruit was possible.
이처럼 최근까지 당근박을 소재로한 연구는 많이 있으나, 당근박을 발효를 통해서 저장성 향상, 기능성 성분 강화 및 probiotic 소재로의 전환에 대한 연구는 미비한 실정이다.As such, there have been many studies on carrot foils until recently, but studies on improving the shelf life, enhancing functional ingredients, and converting them into probiotic materials have been insufficient.
본 발명은 당근박을 활용하여 점질물을 증진시킴으로써 식품 및 화장품의 소재로 활용할 수 있는 당근박 발효물 및 그 제조방법을 제공하고자 한다.The present invention is to provide a carrot foil fermented product and a method of manufacturing the same that can be utilized as a material of food and cosmetics by promoting a viscous substance using carrot foil.
또한 본 발명은 당근박의 저장성을 향상시키며, 유용 발효 균주로부터 생산된 점질물, 혈전분해효소 및 뇌 개선물질인 GABA 등을 함유하는 기능성 제품으로 활용 가능한 당근박 발효물 및 그 제조방법을 제공하고자 한다.In another aspect, the present invention is to improve the shelf life of carrot foil, and to provide a carrot foil fermented product and a method for producing the same as a functional product containing a viscous substance, a thrombolytic enzyme and a brain improving substance GABA produced from useful fermentation strains .
본 발명은 바람직한 제1구현예로서, 당근박 혼합물을 제조하는 단계; 제조된 당근박 혼합물에 바실러스 스타터 배양액을 0.1~5중량% 접종하여 발효시키는 1차 발효 단계를 포함하는 당근박 발효물의 제조방법을 제공한다.As a first preferred embodiment of the present invention, preparing a carrot foil mixture; It provides a method for producing a carrot foil fermentation product comprising a primary fermentation step of fermentation by inoculating 0.1 ~ 5% by weight of Bacillus starter culture in the prepared carrot foil mixture.
상기 당근박 혼합물은 수분함량이 50~95%인 반고체 상태인 것일 수 있다.The carrot foil mixture may be in the semi-solid state of 50 to 95% moisture content.
상기 1차 발효 단계는 30~50℃에서 10~72시간동안 고체발효시키는 것일 수 있다.The first fermentation step may be a solid fermentation for 10 to 72 hours at 30 ~ 50 ℃.
상기 1차 발효 단계 후 발효된 원료무게에 대하여 발효성 당을 1~10% 첨가하고 젖산균 스타터 배양액을 1~10중량% 접종하여 발효시키는 2차 발효 단계를 더 포함하는 것일 수 있다.After the first fermentation step, the fermented sugar may be further added to the fermented sugar by incorporating 1-10% of the fermented sugar and inoculating 1-10% by weight of the lactic acid bacteria starter culture.
상기 2차 발효 단계는 25~48℃에서 11~72시간 고체발효시키는 것일 수 있다.The secondary fermentation step may be a solid fermentation for 11 to 72 hours at 25 ~ 48 ℃.
상기 당근박 혼합물은 글루타민산염(glutamate)을 당근박 무게에 대하여 1~15중량% 더 포함하는 것일 수 있다.The carrot foil mixture may further comprise 1 to 15% by weight of glutamate (glutamate) with respect to the carrot foil weight.
본 발명은 바람직한 제2구현예로서 상기의 제조방법으로 제조된 당근박 발효 물을 제공한다.The present invention provides a carrot foil fermented product prepared by the above production method as a second preferred embodiment.
본 발명은 바람직한 제3구현예로서 상기의 당근박 발효물을 포함하는 식품을 제공한다.The present invention provides a food comprising the carrot foil fermented product as a third preferred embodiment.
본 발명은 바람직한 제4구현예로서 제 7 항의 당근박 발효물을 포함하는 화장품을 제공한다.The present invention provides a cosmetic comprising the carrot foil fermentation product of claim 7 as a fourth preferred embodiment.
이하, 본 발명을 보다 상세히 설명한다.Hereinafter, the present invention will be described in more detail.
본 발명의 당근박 발효물의 제조방법은 당근박 혼합물을 제조하는 단계; 제조된 당근박 혼합물에 바실러스 스타터 배양액을 0.1~5중량% 접종하여 발효시키는 1차 발효 단계를 포함한다.Carrot foil fermentation method of the present invention comprises the steps of preparing a carrot foil mixture; It comprises a primary fermentation step of fermentation by inoculating 0.1 ~ 5% by weight of Bacillus starter culture in the prepared carrot foil mixture.
상기 당근박 혼합물은 수분함량이 50~95%이 되도록 물로 수분함량을 조절한 반고체 상태인 것일 수 있다.The carrot foil mixture may be in a semi-solid state in which the water content is adjusted to water so that the water content is 50 to 95%.
전통 콩발효 식품의 주된 미생물인 고초균(Bacillus subtilis)은 단백질, 탄수화물 등의 가수분해효소를 생산하면서, 발효 중에 기능성 펩타이드, 점질물 등의 생리활성물질의 생산에 기여하며, 유산균과 함께 프로바이오틱스(probiotics)로서 작용하면서 장 내 균총의 개선과 건강유지 등의 효과를 수행한다. 특히 고초균 발효시에 기질로서 글루타민산염(glutamate)을 이용하여 고분자 점질물인 감마폴리글루타민산(polyglutamic acid)을 생산하는 균주는 액체 및 고체배양에서 효과적으로 고분자 점질물을 생산할 수 있다. 따라서 상기 당근박 혼합물에는 글루탐산나트 륨(monosodium glutamate)을 더 포함할 수 있다. 이 때 당근박 혼합물은 글루타민산염(glutamate)을 당근박 무게에 대하여 1~15중량% 더 포함하는 것일 수 있다. Bacillus subtilis , a major microorganism of traditional soybean fermented foods, produces hydrolase such as proteins and carbohydrates, and contributes to the production of bioactive substances such as functional peptides and viscous substances during fermentation, along with probiotics It acts as a function of improving intestinal flora and maintaining health. In particular, strains that produce polymer viscous gamma polyglutamic acid using glutamate as a substrate during Bacillus subtilis fermentation can effectively produce polymeric viscosities in liquid and solid cultures. Therefore, the carrot foil mixture may further include sodium glutamate (monosodium glutamate). At this time, the carrot foil mixture may be one containing 1 to 15% by weight of glutamate (glutamate) with respect to the carrot foil weight.
상기 1차 발효 단계는 점질물이 적정량 생성되도록 하기 위하여 30~50℃에서 10~72시간동안 고체발효시키는 것일 수 있으며, 이후 2차 발효 단계를 더 포함하는지의 여부에 따라 발효 조건의 조절은 가능하다. The first fermentation step may be a solid fermentation at 30 ~ 50 ℃ for 10-72 hours in order to produce a proper amount of viscous material, after which the fermentation conditions can be adjusted depending on whether or not further comprising a secondary fermentation step .
상기 1차 발효 단계 후 발효된 원료무게에 대하여 발효성 당을 1~10% 첨가하고 젖산균 스타터 배양액을 1~10중량% 접종하여 발효시키는 2차 발효 단계를 더 포함하는 것일 수 있다. After the first fermentation step, the fermented sugar may be further added to the fermented sugar by incorporating 1-10% of the fermented sugar and inoculating 1-10% by weight of the lactic acid bacteria starter culture.
상기 발효성 당으로는 발효를 통하여 젖산을 생성할 수 있는 발효성 당이면 특별히 제한 없이 사용가능한데, 예컨대 포도당, 과당, 유당, 올리고당 등을 사용할 수 있으며, 젖산균으로는 GABA 생성이 가능한 락토바실러스속 또는 스프렙토코커스속 균주를 사용할 수 있다.As the fermentable sugar, any fermentable sugar capable of producing lactic acid through fermentation may be used without particular limitation. For example, glucose, fructose, lactose, oligosaccharide, etc. may be used, and as lactic acid bacteria, Lactobacillus genus or Spreptococcus strains can be used.
젖산균은 글루코오스 등 당류를 대사시켜 젖산을 생성하는 세균으로 유산균이라고도 한다. 젖산발효에 의해 생성되는 젖산에 의해서 병원균과 유해세균의 생육이 저지되는 성질을 유제품ㅇ김치류ㅇ양조식품 등의 식품제조에 이용한다. 또, 포유류의 장내에 서식하여 잡균에 의한 이상발효를 방지하여 정장제(整腸劑)로도 이용되는 중요한 세균이다. Lactic acid bacteria are bacteria that produce lactic acid by metabolizing sugars such as glucose, also known as lactic acid bacteria. The lactic acid produced by lactic acid fermentation prevents the growth of pathogens and harmful bacteria in food production such as dairy products, kimchi and brewed foods. In addition, it is an important bacterium that lives in the intestine of mammals and prevents abnormal fermentation by various bacteria, and is also used as a formal drug.
상기 2차 발효 단계는 GABA 생성율을 고려하여 25~48℃에서 11~72시간 고체발효시키는 것일 수 있다.The second fermentation step may be a solid fermentation for 11-72 hours at 25 ~ 48 ℃ in consideration of the GABA production rate.
이상과 같이 식이섬유가 풍부하며, 당근의 일부 성분, 색소 및 풍미를 함유하고 있는 당근박을 고초균 및 젖산균 2단 혼합발효를 거침으로써, 점질물 PGA 생산 원료, 혈전 분해효소, probiotic을 포함하게 함으로서 식품, 기능성 식품 및 화장품의 소재로 활용이 기대된다. 특히 점질물의 생산을 최적화하기 위해서 사용되는 유용 고초균은 glutamate 의존형 고초균을 이용하여 발효를 수행하였다.As mentioned above, the carrot gourd, which is rich in dietary fiber and contains some components of carrots, pigments and flavors, is subjected to two-stage fermentation of Bacillus subtilis and lactic acid bacteria, so that it contains viscous material PGA production raw materials, thrombolytic enzymes and probiotic. It is expected to be used as a material for functional foods and cosmetics. In particular, useful Bacillus subtilis used to optimize the production of viscous material was fermented using glutamate-dependent Bacillus subtilis.
이와 같이 제조된 당근박 발효물은 도 1과 같이 점질물을 형성하는 것일 수 있다.The carrot foil fermented product thus prepared may be to form a viscous substance as shown in FIG. 1.
이하, 본 발명을 실시예를 통하여 보다 상세히 설명하나, 본 발명의 범위가 하기 실시예로 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples, but the scope of the present invention is not limited to the following Examples.
<제조예 1> 원료처리Preparation Example 1 Raw Material Treatment
당근 즙 추출 제조 공정 후 나오는 부산물인 당근박의 수분함량은 57%로 높은 수분을 함유한 상태이다. 당근 박은 121℃에서 15분 동안 살균처리한 후 발효에 사용하였다. Carrot juice, a by-product of the carrot juice extract manufacturing process, has a high moisture content of 57%. Carrot gourd was sterilized for 15 minutes at 121 ℃ was used for fermentation.
<제조예 2> 바실러스 스타터 제조Preparation Example 2 Bacillus Starter Manufacture
바실러스 서브틸리스 (glutamate 의존형 균주)를 MRS agar plate에서 42℃ 항온기를 이용하여 24시간 활성화시킨 후, 한 콜로니를 따서 121℃에서 15분간 멸균된 5% 콩분말용액에 접종한 후 42℃ 항온기에서 24시간 150rpm에서 진탕 배양시켜 스타터 종균으로 사용하였다. Bacillus subtilis (glutamate dependent strain) was activated on a MRS agar plate for 24 hours using a 42 ° C incubator, and then inoculated with one colony in a sterilized 5% soybean powder solution at 121 ° C for 15 minutes and then in a 42 ° C incubator. Shake incubation at 150 rpm for 24 hours was used as starter spawn.
<제조예 3> 젖산균 스타터 제조Preparation Example 3 Preparation of Lactic Acid Bacteria Starter
젖산균을 MRS agar plate에서 30℃ 항온기를 이용하여 24시간 활성화시킨 후, 한 콜로니를 따서 121℃에서 15분간 멸균된 당근즙 용액에 접종한 후 30℃ 항온기에서 24시간 배양시켜 스타터 종균으로 사용하였다.After lactic acid bacteria were activated for 24 hours using a 30 ℃ thermostat in an MRS agar plate, one colony was inoculated in a sterilized carrot juice solution at 121 ℃ for 15 minutes and then incubated for 24 hours at 30 ℃ constant temperature was used as starter spawn.
<실시예 1~5> 당근박의 고초균에 의한 부재료 함량별 고체발효<Examples 1 to 5> solid fermentation by the amount of subsidiary materials by Bacillus subtilis of carrot foil
제조예 1에서 준비된 당근박 50g을 발효용기에 담은 다음 glutamate를 당근 박 원료무게에 대비하여 0,1,3,5,7% 농도별로 첨가하고 물로 수분함량을 67%로 조절한 반고체상태의 당근박 혼합물에 제조예 2에서 준비된 바실러스 스타터 배양액을 1중량%씩 접종한 후 37~45℃에서 24시간 고체발효하여 당근박 발효물을 제조하였다.50 g of carrot gourd prepared in Preparation Example 1 was placed in a fermentation vessel, and then glutamate was added in 0,1,3,5,7% concentration in preparation for the weight of the carrot gourd, and the water content was adjusted to 67% with water. Carrot foil fermented product was prepared by inoculating 1% by weight of the Bacillus starter culture solution prepared in Preparation Example 2 into the gourd mixture at 37-45 ° C. for 24 hours.
실시예 1~5에서 제조된 당근박 발효물에 대하여 다음과 같이 물성을 측정하였다.The physical properties of the carrot foil fermented product prepared in Examples 1 to 5 were measured as follows.
1. 당근박 발효물의 점질물 함량 측정 1.Measurement of Viscous Content of Carrot Foil Fermentation
당근박 발효물 10g 증류수 20mL을 첨가하여 불용성 고형분을 여과시켜 제거한 후 원심분리하여 상등액을 회수하였다. 회수된 상등액에 2배의 isopropanol을 첨가하여 고형분을 침전시킨후 회수하여 진공 감압 건조기를 이용하여 건조하여 무게를 측정하여 보았다. 당근박을 고초균을 이용하여 발효시 점질물 함량은 표 2에 나타내었다. 점질물 함량은 실시예 5에서 4.24%로 가장 높은 함량을 나타내었다. 10 g of carrot gourd fermented distilled water was added to remove insoluble solids, followed by centrifugation to recover a supernatant. Two times isopropanol was added to the recovered supernatant to precipitate solids, which was recovered, dried using a vacuum decompression dryer, and weighed. The viscous content of fermented carrot gourd using Bacillus subtilis is shown in Table 2. The viscosity content was the highest in Example 5, at 4.24%.
2. 당근박 발효물의 점조도 측정2. Measurement of the consistency of carrot foil fermented products
당근박 발효물 5g에 증류수 10mL 첨가하여 균질화 후에 여과 채(sieve, 0.99 mm)를 이용하여 여과시킨 시료를 제조하였다. 점조도는 Rheometer System(HAAKE RheosStress 1, Germany)에 cone plate device(Plate PP35Ti, 3.5cm diameter)를 장착하여 측정하였다. 시료 1.2mL을 plate에 올려 구간 당 10초 동안의 평균값이 측정되어 얻은 값을 shear rate(1/s)와 shear stress(Pa)로 나타내어 점조도를 측정하였다. 당근박을 고초균을 이용하여 발효시 점조도는 표 3 나타내었다. 점조도는 실시예3에서 18.65±0.06Pa·sn로 가장 높은 함량을 나타내었다. 도 1은 당근박 발효물의 점조성을 나타내는 시료의 예이다.10 mL of distilled water was added to 5 g of carrot foil fermented product, and homogenized and then filtered using a sieve (0.99 mm) to prepare a sample. Viscosity was measured by mounting a cone plate device (Plate PP35Ti, 3.5cm diameter) on a rheometer system (HAAKE Rheos Stress 1, Germany). 1.2mL of the sample was placed on the plate, and the average value for 10 seconds was measured per section. The obtained value was expressed as shear rate (1 / s) and shear stress (Pa), and the consistency was measured. The consistency of the carrot gourd using Bacillus subtilis is shown in Table 3. The consistency was the highest in Example 3, 18.65 ± 0.06 Pa.s n . 1 is an example of a sample showing the consistency of carrot foil fermented product.
3. 혈전용해효소의 활성측정3. Measurement of activity of thrombolytic enzyme
혈전용해효소 활성은 fibrin plate method로 측정하였다. Fibrin plate는 0.5% fibrinogen을 0.067 M sodium phosphate buffer(pH7.4)에 용해시켜서 직경 9cm인 petridish에 10mL을 가하였다. 여기에 0.067M sodium phosphate buffer(pH7.4)에 용해된 thrombin (100NIH/mL) 0.1 mL을 가하고 신속하게 혼합하고 균일한 평판을 제조한 후 실온에서 30분 방치하여 고형화 시켰다. Fibrin plate에 시료 점적 위치를 표시하고 발효당근박 추출액을 20 ㎕씩 각 표시한 위치에 점적하여 37℃에서 2시간 반응시킨 후 용해 면적으로 효소 활성을 구하였다, 당근박을 고초균을 이용하여 발효시 혈전용해효소의 활성은 표 4에서 나타내었다. 혈전용해효소의 활성은 실시예 3에서 13.66unit/g으로 가장 높은 수치를 나타내었다. 도 2는 혈전분해효소의 활성을 나타내는 혈전분해정도를 나타내는 혈전분해환을 나타내고 있다(A: 실시예1, B: 실시예2, C: 실시예3, D: 실시예4, E: 실시예5). Thrombolytic activity was measured by the fibrin plate method. Fibrin plates were dissolved in 0.067 M sodium phosphate buffer (pH7.4) in 0.5% fibrinogen and 10 mL were added to petridish with a diameter of 9 cm. 0.1 mL of thrombin (100NIH / mL) dissolved in 0.067M sodium phosphate buffer (pH7.4) was added thereto, mixed quickly, and a uniform plate was prepared and allowed to stand at room temperature for 30 minutes to solidify. Mark the sample drop position on the fibrin plate, and add 20 µl of the fermented carrot extract to each marked position for 2 hours at 37 ° C, and then determine the enzyme activity by dissolving the area. The activity of thrombolytic enzymes is shown in Table 4. The activity of the thrombolytic enzyme showed the highest value of 13.66 units / g in Example 3. Figure 2 shows the thrombolytic ring showing the degree of thrombolysis indicating the activity of the thrombolytic enzyme (A: Example 1, B: Example 2, C: Example 3, D: Example 4, E: Example 5).
4. L-glutamic acid 잔존량 측정 4. Measurement of L-glutamic acid remaining amount
당근박 발효물에 존재하는 L-glutamic acid의 분석은 상기의 iso-propanol 함유 용액을 10mL로 농축하여 원심분리 한 후 TLC(Thin Layer Chromatography)를 사용하여 잔존량을 확인하였다. 분석조건은 표준물질(1mg/mL, 5mg/mL, 10mg/mL L-glutamic acid)과 여과액을 각각 2 ㎕ 씩 cellulose plate에 점적하여 전개용매(1-butanol : acetate : water = 5 : 4 : 3, ethanol : water = 63 : 37)로 전개하고 건조한 후 발색시약(0.2% ninhydrin in acetone)을 spraying 한 후 105℃에서 30초~1분간 건조한 후 확인하였다. In the analysis of L-glutamic acid in the carrot gourd fermentation, the iso-propanol-containing solution was concentrated to 10 mL and centrifuged to confirm the residual amount using TLC (Thin Layer Chromatography). Analytical conditions were obtained by injecting a standard material (1 mg / mL, 5 mg / mL, 10 mg / mL L-glutamic acid) and 2 μl of the filtrate into the cellulose plate, and then using a developing solvent (1-butanol: acetate: water = 5: 4: 3, ethanol: water = 63: 37) was developed and dried and sprayed with a coloring reagent (0.2% ninhydrin in acetone) and dried at 105 ℃ for 30 seconds to 1 minute and confirmed.
당근 박 발효물에 존재하는 glutamate의 잔존량은 도 3에서 보여주고 있으며, glutamate첨가량 1%, 3% 수준에서는 당근박 발효물에 잔존하는 MSG함량은 매우 미비하여 첨가된 glutamate의 대부분은 점질물 polyglutamate로 전환되었음을 알 수 있었다. 반면에 5% glutamate첨가에서는 전체 첨가중량의 50% 정도의 glutamate가 당근 박의 발효물에 존재하는 것으로 나타났다. The residual amount of glutamate present in the carrot gourd fermentation is shown in FIG. 3. At the level of glutamate addition 1% and 3%, the amount of MSG remaining in the carrot gourd fermentation is very insignificant. It was found that the transition. On the other hand, in the 5% glutamate addition, about 50% of the total weight of glutamate was found in the fermentation of carrot gourd.
<실시예 6~10> MSG 첨가농도에 따른 당근박 혼합발효 <Example 6-10> Carrot foil mixed fermentation according to the concentration of MSG addition
제조예 1에서 제조된 당근박 50g을 발효용기에 담은 다음 mono sodium glutamate를 당근박 원료무게에 대비하여 0,1,3,5,7% 농도별로 첨가하고 수분함량 67%이 되도록 조절한 반고체상태의 당근박 혼합물에 제조예 2에서 제조한 바실러스 스타터 배양액을 1중량%씩 접종하여 37~45℃에서 15시간 1차 발효 후, 1차 발효된 원료무게에 대비하여 포도당을 각각 2%씩 첨가하고 제조예 3에서 제조한 젖산균 스타터 배약액을 4중량%씩 접종한 후 30℃에서 15시간 고체 발효하여 당근박 발효물을 제조하였다. 50 g of carrot foil prepared in Preparation Example 1 was placed in a fermentation container, and then the mono sodium glutamate was added at a concentration of 0,1,3,5,7% relative to the weight of the carrot foil and adjusted to a moisture content of 67%. Inoculated with 1% by weight of the Bacillus starter culture solution prepared in Preparation Example 2 to the carrot foil mixture of 15 hours after primary fermentation at 37 ~ 45 ℃ for 15 hours, 2% of glucose was added to the primary fermented raw material weight Carrot foil fermented product was prepared by inoculating 4 wt% of the lactic acid bacteria starter solution prepared in Preparation Example 3 and then solid fermentation at 30 ° C. for 15 hours.
실시예 6~10에서 제조된 당근박 발효물은 고초균 발효시 glutamate 첨가 농도가 높아질수록 당근 고유의 향은 감소하고 청국장 냄새가 증가하였지만 2단계 젖산발효 후 청국장 냄새는 감소하고 유기산 냄새가 증가하여 향미가 향상되었다. 또한 pH가 감소하면서 점조도 값이 감소하는 경향을 나타내었다. In the carrot foil fermented products prepared in Examples 6-10, the inherent flavor of carrot decreased and the smell of Cheonggukjang increased as the concentration of glutamate was increased during fermentation of Bacillus subtilis. Was improved. In addition, the consistency value decreased with decreasing pH.
상기 실시예 6~10에서 제조된 당근박 발효물에 대하여 다음과 같이 물성을 측정하였다.The physical properties of the carrot foil fermented product prepared in Examples 6 to 10 were measured as follows.
1. 당근박 발효물의 점조도 측정1. Measurement of the consistency of carrot foil fermented products
당근박 발효물의 점도는 Rheometer System(HAAKE RheoStress 1,Germany)에 spindle을 장착하여 cone plate device(Platte PP35 Ti, 3.5㎝ diamete)를 사용하여 측정하였다. 시료 적당량을 plate위에 올려놓고 구간 당 10초 동안의 평균값이 측정되어 얻은 값을 shear rate(1/s)와 shear stress(Pa)로 나타내어 점도가 높아짐에 따라 층밀림 변형력이 높아지는 효과를 측정하였다. 측정온도 20℃에서 전단속도는 1~100 s-1의 범위로 유동특성을 알아보았고, 점조도지수와 유동지수 값은 Power law model로 측정하였다. 표 6에서 2차 발효물의 점조성은 발효물의 pH가 산성으로 변함에 따라 점조성이 감소하는 경향을 보였다. 그러나 2차 발효시에 당의 첨가농도에 따라 산생성은 조절이 가능하여 점조성의 조절도 가능하다고 사료된다. Viscosity of the carrot foil fermentation was measured using a cone plate device (Platte PP35 Ti, 3.5 cm diamete) by mounting a spindle on a Rheometer system (HAAKE RheoStress 1, Germany). The appropriate amount of sample was placed on the plate, and the average value for 10 seconds per section was measured. The value obtained was expressed as shear rate (1 / s) and shear stress (Pa). Shear velocity was measured in the range of 1 ~ 100 s -1 at 20 ℃, and the viscosity index and the flow index were measured by the power law model. In Table 6, the viscosity of the secondary fermentation tended to decrease as the pH of the fermentation product changed to acidic. However, according to the concentration of sugars in the secondary fermentation, the acid production can be controlled, so that the viscosity can be controlled.
2. 당근박 발효물의 점질물 함량2. Viscous content of carrot foil fermentation
당근박 발효물 10g 증류수 20mL을 첨가하여 불용성 고형분을 여과시켜 제거한 후 원심분리하여 상등액을 회수하였다. 회수된 상등액에 2배의 isopropanol을 첨가하여 고형분을 침전시킨 후 회수하여 진공 감압 건조기를 이용하여 건조하여 무게를 측정하여 보았다. 당근박 발효물의 점조성 및 점질물 함량은 하기 표 6에 나타내었다. 당근박의 2차 발효에 의해서도 초기 glutamate함량이 증가함에 따라서 점질물 생성량은 증가하는 경향을 보였으며, 1차 발효에 의해 생산된 양과 유사하였다. 10 g of carrot gourd fermented distilled water was added to remove insoluble solids, followed by centrifugation to recover a supernatant. Two times isopropanol was added to the recovered supernatant to precipitate solids, which was recovered, dried using a vacuum decompression dryer, and weighed. Viscosity and viscous content of the carrot foil fermentation are shown in Table 6 below. The secondary glutamate fermentation increased the initial glutamate content and increased the amount of viscous product, which was similar to the amount produced by the primary fermentation.
3. pH 측정3. pH measurement
pH meter (Model 420A+, Thermo Orion. USA)를 이용하여 측정하였으며, 당근박 10g을 채취하여 20mL의 멸균수에 혼합한 후 vortex로 혼합한 후 시료의 pH를 측정하였다. 당근박 2차 발효시 pH값은 표 7에서 나타내었다. pH값은 1차 발효시 6.5정도를 나타났으며 2차 발효시 당근박에 glutamate 3% 수준으로 첨가된 실시예 8까지는 4.5정도를 보였으며, glutamate 함량이 3% 초과인 실시예 9부터는 비교적 높은 5.5이상을 나타내었다. The pH was measured using a pH meter (Model 420A + , Thermo Orion. USA), and 10 g of carrot foil was collected and mixed in 20 mL of sterile water, mixed with vortex, and then the pH of the sample was measured. The pH value of the carrot foil secondary fermentation is shown in Table 7. The pH value was about 6.5 at the first fermentation, and the level was 4.5 up to Example 8 when glutamate 3% was added to carrot foil during the second fermentation. It showed 5.5 or more.
4. 생균수 측정4. Viable cell count
당근박 발효물 1g을 멸균수 9mL에 혼합하여 얻은 당근박 발효물 추출액을 104, 105, 106배로 단계별 희석하여 MRS agar 배지에 20㎕ 도말한 후, 37℃ 항온배양기에서 24시간 배양한 후 생균수를 측정하였다. 젖산균은 MRS agar 배지(pH4.5)에서 배양 후 30℃ 항온배양기에 위와 동일한 방법으로 측정하였다. 표 8에서 보는 것처럼 젖산균은 1x 109이상을 함유하였다. 1 g of carrot foil fermented product was mixed with 9 mL of sterile water, and the carrot foil fermented extract obtained by diluting stepwise was 10 4 , 10 5 , 10 6 times, 20 μl smeared on MRS agar medium, and then incubated in a 37 ° C. incubator for 24 hours. The viable cell count was then measured. Lactic acid bacteria were measured in the same manner as above in a 30 ℃ incubator after incubation in MRS agar medium (pH4.5). As shown in Table 8, lactic acid bacteria contained more than 1 × 10 9 .
5. GABA(γ-aminobutryric Acid) 생성율5. Production Rate of GABA (γ-aminobutryric Acid)
당근박 발효물에 존재하는 GABA(γ-Aminobutryric Acid) 분석은 상기 isopropanol 함유 용액을 10mL로 농축하여 원심분리 한 후 TLC(Thin Layer Chromatography)를 사용하여 잔존량을 확인하였다. 분석조건은 표준물질(1mg/mL, 5mg/mL, 10mg/mLγ-Aminobutryric Acid)과 여과액을 각각 2㎕씩 silica gel plate에 직적하여 전개용매(Acetic acid : 1-butanol : water = 1 : 4 : 5 )로 전개하고 건조한 후 발색시약(0.2% ninhydrin in acetone)을 spraying 한 후 105℃에서 30초~1분간 건조한 후 확인하였다. 표 8에서 보는 바와 같이 젖산균이 생산하는 GABA함량은 150mg% 정도임을 알 수 있다. GABA (γ-Aminobutryric Acid) analysis present in the carrot gourd fermentation was concentrated by 10 mL of the isopropanol-containing solution was centrifuged to determine the residual amount using TLC (Thin Layer Chromatography). Analytical conditions were developed by applying standard solution (1mg / mL, 5mg / mL, 10mg / mLγ-Aminobutryric Acid) and 2µl each of the filtrate directly onto a silica gel plate (Acetic acid: 1-butanol: water = 1: 4) : 5) After developing and drying, spraying the color development reagent (0.2% ninhydrin in acetone) and dried at 105 ℃ for 30 seconds ~ 1 minute and confirmed. As shown in Table 8, it can be seen that the GABA content produced by lactic acid bacteria is about 150 mg%.
CFU: colony forming unit (생균수)CFU: colony forming unit
도 1은 본 발명의 점질물을 갖는 발효물 시료인 당근박 발효물을 나타낸 사진, 1 is a photograph showing a carrot foil fermented product which is a fermented product sample having a viscosity of the present invention,
도 2는 본 발명의 실시예 1 내지 5에서의 당근박 발효물의 혈전용해효소의 활성을 측정한 결과인 혈전분해환을 나타낸 사진,Figure 2 is a photograph showing a thrombolytic ring which is a result of measuring the activity of the thrombolytic enzyme of carrot foil fermentation in Examples 1 to 5 of the present invention,
도 3은 본 발명의 실시예 1 내지 5에서의 당근박 발효물의 glutamate 잔존함량을 나타내는 TLC 분석 결과를 나타낸 사진이다.Figure 3 is a photograph showing the results of TLC analysis showing the remaining glutamate content of carrot foil fermentation in Examples 1 to 5 of the present invention.
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