KR20050030819A - Production method for organic acid using of kombucha - Google Patents
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Abstract
Description
본 발명은 홍차버섯 종균을 이용한 유기산의 생산방법에 관한 것으로 보다 상세하게는 호기적 조건하에서 당과 홍차의 혼합물에 홍차버섯 종균을 접종하고 발효시키는 공정을 포함하는 유기산의 생산방법에 관한 것이다.The present invention relates to a method for producing an organic acid using a black tea mushroom spawn, and more particularly, to a method for producing an organic acid including a step of inoculating and fermenting a black tea mushroom spawn in a mixture of sugar and black tea under aerobic conditions.
홍차버섯은 홍차 위에 떠 있는 균체의 모양이 버섯의 갓과 비슷하다 하여 이러한 이름으로 불리워지고 있다. 대부분의 버섯류는 버섯 자체를 식용으로 사용하지만 홍차버섯은 홍차를 발효하여 홍차만 마시고 홍차버섯 자체는 식용으로 사용하지 않는다.Black tea mushrooms are called by this name because the shape of the cells floating on the black tea is similar to that of mushrooms. Most mushrooms use mushrooms for food, but black tea mushrooms ferment black tea and drink only black tea, and black tea mushrooms do not use for food.
발명의 홍차버섯 종균은 종래부터 홍버섯으로 알려진 야생균주인 곰부차(Kombucha)를 산사(설악산 신흥사)에서 입수하여 계대배양을 통하여 유기산 생성능력이 우수한 균주를 순수분리하여 얻은 변이주(Kombucha sp. RHS 2003, KFCC - 11323)로 본 발명의 유기산을 제조하였다.The black tea mushroom spawn of the present invention was obtained from a wild strain known as red mushroom, Kombucha ( Kombucha ) from Sansa (Seoraksan Sinheungsa), and obtained by purely separating strains excellent in organic acid generation ability through subcultures ( Kombucha sp . RHS). 2003, KFCC-11323) to prepare the organic acid of the present invention.
홍차버섯은 류마티스, 천식, 대머리, 백발에 효과가 좋은 것으로 알려진 건강식품으로서 미국, 유럽, 호주 등에서 크게 유행하고 있다. 원산지인 중국에서 구소련 바이칼호수 주변으로 전해졌으며 그후 일본과 한국으로 전파되었다고 전해지고 있다. 바이칼호 주변에는 장수하는 사람이 많다는 것은 세계적으로 유명하며 홍차버섯이 불로장수의 음료로 알려지게 되었다. 홍차버섯은 1974년 일본 키타사토대학 사키모토 마사요시교수에 의하여 그의 정체가 밝혀지게 되었는데 콤부차는 효모세포와 박테리아의 공생체로서 버섯류처럼 포자로 생산되는 것이 아니라 발아로 번식하는 균류(菌類)에 의해 생기는 지의류(地衣類)의 피막이라고 알려져 있다. 그러나 일부 학자들의 견해에 따르면 지의류는 조류(藻類)와 균류의 공생체이고 조류의 전형적인 특징인 광합성을 하기 위하여 에너지원으로 빛을 필요로 하는데 반하여 콤부차는 조류를 포함하지 않기 때문에 어두운 곳에서도 잘 살 수 있으므로 지의류로 분류하지 않는 사람도 있으나 홍차를 비롯한 여러 가지 차를 즐겨 생육하므로 홍차버섯이라고 명명하게 되었다. 한편 호주에서는 홍차버섯을 기적의 버섯(Miracle Mushroom)이라고 부르기도 한다.Black tea mushroom is a health food known to be effective for rheumatism, asthma, baldness, and white hair, and is popular in the United States, Europe, and Australia. It is said to have originated from China, around the former Soviet Union Baikal Lake, and then to Japan and Korea. Many people around the Lake Baikal have a long life, and the black tea mushroom is known as a drink of longevity. The black tea mushroom was identified by his professor Masayoshi Sakimoto, University of Kitasato University in 1974. Kombucha is a symbiosis of yeast cells and bacteria. It is known as the film of lichens. However, some scholars believe that lichens are a symbiosis of algae and fungi and require light as an energy source for photosynthesis, which is typical of algae, whereas Kombucha does not contain algae, so it lives well in the dark. Some people do not classify lichens, but many people like tea and grow it. On the other hand, black tea is sometimes called Miracle Mushroom.
홍차버섯의 배지로는 설탕이나 과당 또는 포도당 6-25(v)% 용액에 홍차추출물 0.5-1.0(v)%을 혼합한 후, 홍차버섯의 종균을 접종하고 생육온도로 14-30℃에서 상대습도 50-80%로 조절하면서 호기적으로 배양하면 48-96시간에 걸쳐 유기산을 생성하게 된다. 본 발명의 균주는 생육과정에서 당과 홍차추출물을 주성분으로 이용하며 발효과정을 거쳐 당을 유기산으로 변화시켜 대사산물은 신맛을 내게 되며 배지에는 잔당이 2-3%로 잔존하게 되며 대사과정에서 탄산가스도 발생시켜 유기산과 잔당이 조화를 이루어 독특한 상쾌한 맛과 향을 제공하게 된다. 유기산은 주로 주석산(Tartaric Acid)과 사과산(Malic Acid)을 2-3%로 많이 생성시키고 호박산(Succinic Acid)과 초산(Acetic Acid)도 0.05-0.25% 정도 생성시킨다. As a medium for black tea mushrooms, 0.5-1.0 (v)% of black tea extract was mixed with 6-25 (v)% of sugar, fructose or glucose, and then inoculated with the seeds of black tea mushrooms and grown at 14-30 ° C. at a growth temperature. Aerobic incubation with 50-80% humidity produces organic acids over 48-96 hours. The strain of the present invention uses the sugar and tea extract as a main component in the growth process, the metabolite gives a sour taste by changing the sugar into an organic acid through the fermentation process, the residual sugar in the medium remains 2-3% and carbonic acid in the metabolic process It also produces gas, which blends organic acids with sugar to provide a unique refreshing taste and aroma. Organic acids mainly produce tartaric acid and malic acid at 2-3%, and succinic acid and acetic acid at 0.05-0.25%.
홍차버섯의 효능은 암을 억제하는 작용, 콜레스테롤을 낮추는 작용, 혈압을 낮추는 작용, 숙취를 예방하는 작용, 류마티스를 개선하는 작용, 변비를 개선하는 작용, 체중감량 작용, 스트레스나 천식을 조절하는 작용 등이 있다고 알려져 있다. 이러한 홍차버섯은 변비, 관절염, 고혈압, 비만, 아토피 피부염, 여드름 치료, 암 치료에 큰 효과가 있다고 하여 상당히 주목을 받고 있으며 러시아, 독일, 오스트리아, 유럽의 여러 나라와 호주, 미국에서도 수백만의 사람들이 복용하고 있다.The benefits of black tea mushrooms include: inhibiting cancer, lowering cholesterol, lowering blood pressure, preventing hangovers, improving rheumatism, improving constipation, weight loss, and controlling stress or asthma It is known that there is. These black teas have received considerable attention for their effectiveness in treating constipation, arthritis, high blood pressure, obesity, atopic dermatitis, acne, and cancer. Millions of people in Russia, Germany, Austria, Europe, Australia and the United States I'm taking it.
한편 산업적으로 널리 사용되고 있는 유기산 제조방법은 화학합성, 효소반응 및 미생물의 발효공정을 이용해서 생산하고 있으며, 이러한 생산방법중 공정의 경제성 때문에 화학합성 방법이 주로 이용되고 있다. On the other hand, the organic acid production method widely used industrially is produced by chemical synthesis, enzymatic reaction and fermentation process of microorganisms, and chemical synthesis method is mainly used because of the economical process.
유기산중에서 현재 초산과 구연산은 발효공정을 이용해서 일부 생산하고 있으나, 그 외의 유기산의 발효생산은 상대적으로 생산성이 낮고, 화학합성에 의한 생산공정과 경쟁관계에 있어 화학합성이 주된 생산방법으로 이용되고 있다(참조: Roehr, M., Products of Primary Metabolism, in: Biotechnology, 2nd Ed., Vol.6 (Rehm, H.-J., Reed, G., Eds.), Weinheim: Verlag Chemie, 1996).Among organic acids, acetic acid and citric acid are partially produced by fermentation process, but fermentation production of other organic acids is relatively low in productivity, and chemical synthesis is used as the main production method in competition with chemical synthesis. (Roehr, M., Products of Primary Metabolism, in: Biotechnology, 2nd Ed., Vol. 6 (Rehm, H.-J., Reed, G., Eds.), Weinheim: Verlag Chemie, 1996) .
그러나 유기산의 생산능력을 가지는 미생물의 계속적인 발견 및 고효율 발효공정의 개발, 그리고 환경문제에 대한 인식의 확산 등에 힘입어 유기산의 생산에 있어서 화학합성 보다는 효소반응이나 미생물을 이용한 발효방법이 점차적으로 증가하고 있는 추세이다.However, fermentation methods using microorganisms or microorganisms are gradually increasing in the production of organic acids due to the continuous discovery of microorganisms capable of producing organic acids, the development of high-efficiency fermentation processes, and the spread of awareness of environmental problems. The trend is.
한편 본 발명과 관련된 종래 기술로서 한국특허공개 1995-0000007(차버섯을 배양하는 방법), 한국특허공개 2003-62562(피부재생 및 피부 노화지연 효과를 가지는 콤부차 발효배양액 및 이를 함유하는 조성물), 한국특허등록 제10-372218호에 유기산 생성 균주인 만헤이미아속 55E(KCTC 0769BP) 및 전기 균주를 혐기적 또는 호기적 조건에서 배양하여 유기산을 생산하는 방법이 있다. 그러나 종래기술은 본원발명에서 유기산을 생산하는 균주가 다르므로 그 기술적 구성을 달리한다. Meanwhile, as the related art related to the present invention, Korean Patent Publication No. 1995-0000007 (Method for culturing tea), Korean Patent Publication No. 2003-62562 (Comb tea fermentation broth and composition containing the same), In Korean Patent Registration No. 10-372218, there is a method of producing an organic acid by cultivating an organic acid producing strain of Manhemia genus 55E (KCTC 0769BP) and an electric strain under anaerobic or aerobic conditions. However, in the prior art, since the strain producing the organic acid in the present invention is different, its technical configuration is different.
본 발명은 홍버섯으로 알려진 야생균주인 곰부차(Kombucha. sp.)로부터 유기산 생성능력이 우수한 균주를 순수분리하여 얻은 변이주(Kombucha sp. RHS2003, KFCC - 11323, 한국미생물보존센터에 기탁)의 발효에 의하여 주석산, 사과산, 호박산의 유기산을 제조하는 데 있다.The present invention fermentation of the mutant strain ( Kombucha sp . RHS2003, KFCC-11323, deposited in the Korea Microorganism Conservation Center) obtained by purely separating the strain excellent in organic acid from the wild strain Gombucha ( Kombucha. Sp. ) Known as Hong mushroom To produce organic acids of tartaric acid, malic acid, and succinic acid.
본 발명의 균주를 이용한 최적배지 및 발효공정을 적용하여 유기산의 생산을 최대로 하는 방법을 제공하는 데 또 다른 목적을 제공한다. Another object of the present invention is to provide a method of maximizing the production of organic acids by applying an optimal medium and a fermentation process using the strain of the present invention.
본 발명의 홍차버섯 종균을 이용한 유기산의 생산방법은 우량균주 선별, 배양조건(차추출액, 당액, 온도 및 상대습도)을 확립하여 발효에 의하여 유기산 생산을 최대화하는 데 있다. Organic acid production method using the black tea mushroom seed of the present invention is to maximize the production of organic acid by fermentation by establishing a good strain selection, culture conditions (tea extract, sugar solution, temperature and relative humidity).
<우량균주의 선별><Selection of excellent strains>
시중에서 입수한 야생균주(wild type)의 곰부차(Kombucha. sp.)를 이용하여 유기산 생성능력이 우수한 균주를 얻기 위하여 수 많은 실험과 계대배양을 통하여 유기산 생성능력이 우수한 균주를 순수분리하여 변이주(Kombucha sp. RHS 2003)를 얻고 이를 균주기탁기관(한국미생물보존센터)에 KFCC-11323으로 기탁하게 되었다.In order to obtain strains with excellent organic acid generation ability using commercially available wild type Kombucha. Sp. , The strains with pure organic acid generation ability were purely separated through numerous experiments and passages. ( Kombucha sp .
표 1. 실험 및 계대배양을 통한 균주의 유기산 생산능력 비교(단위;ppm)Table 1.Comparison of Organic Acid Production Capacity of Strains through Experimental and Passage Culture (Unit: ppm)
배지는 홍차추출액 0.7%, 당도 15%가 함유된 액체배지를 멸균하여 야생균주, 변이주, 기탁균주를 접종하고 상대습도를 60%로 조정한 후, 온도 25℃에서 상대습도 70%로 호기적으로 배양하면서 발효시간에 따른 유기산 생성능을 관찰하였다. 상기의 결과로부터 기탁균주가 주석산과 사과산을 많이 축적시킴을 알 수 있다. The medium was sterilized with a liquid medium containing 0.7% black tea extract and 15% sugar, inoculated with wild strains, mutant strains, and deposited strains, and the relative humidity was adjusted to 60%. While culturing, the ability to produce organic acids according to fermentation time was observed. From the above results, it can be seen that the deposited strain accumulates much tartaric acid and malic acid.
<배양조건><Cultivation conditions>
본 발명에서 홍차나 녹차 또는 설록차는 통상적으로 음용하는 것이라면 어떠한 것이라도 사용할 수 있다. 본 발명에서는 차잎을 100∼120℃ 온도에서 끓인 정제수 1ℓ에 차잎 10∼20g을 넣고 10∼20분간 추출하고 상온으로 냉각한 후, 사용하였다. 배지의 당분은 설탕, 과당, 포도당 중에서 사용하였다.In the present invention, any of black tea, green tea, or sulloc tea may be used as long as it is commonly used for drinking. In the present invention, 10 to 20 g of tea leaves were added to 1 liter of purified water boiled at 100 to 120 ° C., extracted for 10 to 20 minutes, and used after cooling to room temperature. The sugar of the medium was used in sugar, fructose and glucose.
표2. 기탁균주의 차추출액에 따른 유기산 생산능력 비교(단위;ppm)Table 2. Comparison of Organic Acid Production Capacity According to Tea Extract of Deposited Strains (Unit: ppm)
배지는 각각의 차추출액 0.6-0.8%에 설탕 15%가 함유된 액체배지를 멸균하여 기탁균주를 접종하고 상대습도를 60%로 조정한 후, 온도 25℃에서 상대습도 70%로 호기적으로 배양하면서 유기산 생성능을 관찰하였던 바, 상기의 결과로부터 홍차추출물의 유기산 생성능력이 우수함을 알 수 있었다.The medium was inoculated with the deposited strain by sterilizing the liquid medium containing 15% of the sugar in 0.6-0.8% of each tea extract, adjusting the relative humidity to 60%, and incubating aerobicly with the relative humidity of 70% at a temperature of 25 ° C. While observing the organic acid generating ability, it can be seen from the above results that the tea extract is excellent in the organic acid generating ability.
표 3-1. 기탁균주의 당류에 따른 유기산 생산능력 비교(단위;ppm)Table 3-1. Comparison of Organic Acid Production Capacity by Sugars of Deposited Strains (Unit: ppm)
표 3-2. 기탁균주의 당류에 따른 유기산 생산능력 비교(단위;ppm)Table 3-2. Comparison of Organic Acid Production Capacity by Sugars of Deposited Strains (Unit: ppm)
배지는 각각의 홍차추출액 0.7%에 각각의 당류 6.0-25%가 함유된 액체배지를 멸균하여 기탁균주를 접종하고 상대습도를 60%로 조정한 후, 온도 25℃에서 상대습도 70%로 호기적으로 배양하면서 유기산 생성능을 관찰하였던 바, 상기의 결과로부터 설탕용액의 유기산 생성능력이 우수함을 알 수 있었다.The medium was sterilized by inoculating the deposited strain by sterilizing the liquid medium containing 6.0-25% of the sugars in 0.7% of each black tea extract, adjusting the relative humidity to 60%, and then aerobic at 70% relative humidity at the temperature of 25 ° C. When the organic acid generating ability was observed while culturing in the above, it can be seen from the above results that the organic acid generating ability of the sugar solution is excellent.
표 4-1. 온도에 따른 유기산 생산능력 비교(단위;ppm)Table 4-1. Comparison of Organic Acid Production Capacity by Temperature (Unit: ppm)
표 4-2. 상대습도에 따른 유기산 생산능력 비교(단위;ppm)Table 4-2. Comparison of Organic Acid Production Capacity by Relative Humidity (Unit: ppm)
배지는 각각의 홍차추출액 0.7%에 각각의 당류 15%가 함유된 액체배지를 멸균하여 기탁균주를 접종하고 상대습도와 온도에 따라 호기적으로 배양하면서 유기산 생성능을 관찰하였던 바, 상기의 결과로부터 온도 25℃에서 상대습도 60%에서 유기산 생산능력이 우수함을 알 수 있었다.The medium was sterilized with a liquid medium containing 15% of each sugar in 0.7% of each black tea extract, inoculated with the deposited strain, and cultivated aerobicly according to relative humidity and temperature. It was found that the organic acid production capacity was excellent at the relative humidity of 60% at 25 ℃.
이상에서 실험을 통하여 얻은 결과에 의하면 본 발명의 균주는 하기의 표 5 같은 생육조건에서 유기산의 생산에 적합함을 알 수 있었다. According to the results obtained through the above experiments, the strain of the present invention was found to be suitable for the production of organic acids under the growth conditions shown in Table 5 below.
표 5 홍차버섯 종균 생육조건Table 5 Growth Conditions of Black Tea Mushroom Seedling
한편, 본 발명의 방법에 의해 생산된 유기산이 함유된 음료, 건강보조식품, 의약품을 포함한다.On the other hand, it includes beverages, health supplements, pharmaceutical products containing organic acids produced by the method of the present invention.
이하 본 발명을 다음의 실시예, 시험예에 의하여 설명하고자 한다. 그러나 이들은 본 발명의 일실시예로서 이들에 의해 본 발명의 권리범위가 한정되는 것은 아니다. Hereinafter, the present invention will be described by the following examples and test examples. However, these are not limited to the scope of the present invention by these as an embodiment of the present invention.
<실시예 1><Example 1>
(1) 100∼120℃ 온도에서 끓인 정제수 1ℓ에 홍차잎 10∼20g을 넣고 10∼20분간 추출하여 얻은 홍차추출물 0.7(v)%를 냉각하여 상온의 온도로 유지한다.(1) 10 to 20 g of black tea leaves are added to 1 L of purified water boiled at 100 to 120 ° C., followed by cooling for 0.7 (v)% of black tea extract, which is extracted for 10 to 20 minutes, and maintained at room temperature.
(2) 100∼110℃ 온도에서 끓인 정제수 1ℓ에 설탕을 서서히 용해시켜 당도계로 당도가 15(v)% 되도록 당액을 만들고 이를 냉각하여 상온의 온도로 유지한다.(2) Slowly dissolve sugar in 1 L of purified water boiled at 100 ~ 110 ℃, make sugar solution so that the sugar content is 15 (v)% with sugar system and cool it and keep it at room temperature.
(3) 상기 (1)공정에서 얻은 홍차와, (2)공정에서 얻은 당액을 혼합한 후 멸균시켜 냉각한 다음 홍차버섯 종균(Kombucha sp. RHS 2003, KFCC - 11323)을 접종하고 호기적으로 온도 25℃, 상대습도 60%의 조건하에서 72시간 발효시켜 유기산을 제조하였다.(3) The black tea obtained in step (1) and the sugar solution obtained in step (2) are mixed, sterilized and cooled, and then inoculated with black tea mushroom seed ( Kombucha sp . RHS 2003, KFCC-11323), and The organic acid was prepared by fermenting for 72 hours under the condition of 25 ° C. and 60% relative humidity.
<실시예 2><Example 2>
실시예 1과 동일하게 하고 홍차잎 추출물 0.6%, 과당용액 15%를 배지로 사용하고 종균을 접종한 후, 20℃, 상대습도 70%로 하는 것을 제외하고는 실시예 1과 동일하게 발효시켜 유기산을 얻었다. In the same manner as in Example 1, 0.6% of black tea leaf extract and 15% fructose solution were used as a medium and inoculated with the spawn seed, followed by fermentation in the same manner as in Example 1 except that 20 ° C. and relative humidity were 70%. Got.
<실시예 3><Example 3>
실시예 1과 동일하게 하고 홍차잎 추출물 0.8%, 설탕용액 12%를 배지로 사용하고 종균을 접종한 후, 28℃, 상대습도 80%로 하는 것을 제외하고는 실시예 1과 동일하게 발효시켜 유기산을 얻었다. In the same manner as in Example 1, using 0.8% black tea leaf extract, 12% sugar solution as a medium and inoculated spawn, and then fermented in the same manner as in Example 1 except that the temperature is 28 ℃, 80% relative humidity, organic acid Got.
<실시예 4><Example 4>
실시예 1과 동일하게 하고 홍차잎 추출물 0.65%, 설탕용액 20%를 배지로 사용하고 종균을 접종한 후, 30℃, 상대습도 50%로 하는 것을 제외하고는 실시예 1과 동일하게 발효시켜 유기산을 얻었다. In the same manner as in Example 1, 0.65% black tea leaf extract, 20% sugar solution was used as a medium and inoculated with the seed, and then fermented in the same manner as in Example 1 except that the temperature is 30 ℃, relative humidity 50% organic acid Got.
표 6. 실시예에 따른 유기산 농도(단위;ppm)Table 6. Organic Acid Concentration in ppm according to the Examples
<시험예 1><Test Example 1>
상기 실시예 1에서 얻은 발효액 중의 유기산 및 그 밖의 성분을 알아 보기 위하여 검사기관인 과학기술분석센타(대전광역시 대덕구 소재)에 의뢰하여 아래의 표 6과 같은 결과를 얻었다.In order to determine the organic acid and other components in the fermentation broth obtained in Example 1, a request was made to the Science and Technology Analysis Center (Daedeok-gu, Daejeon Metropolitan City) as a testing institution, and the results as shown in Table 6 below were obtained.
표 6. 발효액 성분검사Table 6. Fermentation Broth Composition
<적용예 1><Application example 1>
상기 실시예 1의 방법에 의해 제조한 배지에서 상등액을 분리하여 얻은 유기산 용액을 제균필터로 여과하여 착향, 가당 100ml용기에 넣어 음료용으로 제조하였다.The organic acid solution obtained by separating the supernatant liquid from the medium prepared by the method of Example 1 was filtered through a sterilization filter, and prepared in a flavored, sweetened 100 ml container.
<적용예 2><Application example 2>
상기의 적용예 1과 같은 방법에 의해 제조한 유기산을 농축하여 유당과 함께 캡슐로 제조하여 건강보조식품으로 제조하였다.The organic acid prepared by the same method as in Application Example 1 was concentrated to prepare a capsule with lactose and as a dietary supplement.
본 발명은 홍차버섯 종균을 발효시켜 주석산, 초산, 사과산, 호박산을 제조함에 있어서 특히 주석산과 사과산이 대량으로 축적됨을 알 수 있다. 특히 건강에 좋은 유기산을 동시에 축적시킬 수 있는 균주와 발효방법을 제공함으로써 음료, 건강식품, 의약품으로 용도를 확대해 나갈 수 있으리라 전망된다. The present invention can be seen that the fermentation of black tea mushroom spawns in the production of tartaric acid, acetic acid, malic acid, succinic acid, in particular, large amounts of tartaric acid and malic acid. In particular, it is expected to expand the use of beverages, health foods and medicines by providing strains and fermentation methods that can simultaneously accumulate healthy organic acids.
상술한 바와 같이, 본 발명의 발명의 구성 및 실시예를 통하여 설명하였지만 해당 기술 분야의 숙련된 당업자라면 하기의 특허청구범위에 기재된 본 발명의 사상 및 영역으로부터 벗어나지 않는 범위 내에서 본 발명을 다양하게 수정 및 변경시킬 수 있음을 이해할 수 있을 것이다. As described above, the present invention has been described through the configuration and embodiments of the present invention, but those skilled in the art can variously change the present invention without departing from the spirit and scope of the present invention as set forth in the claims below. It will be appreciated that modifications and variations can be made.
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