KR100390529B1 - Method of extracting nucleic acid complex material from tuna testes and nucleic acid complex material obtained therefrom - Google Patents
Method of extracting nucleic acid complex material from tuna testes and nucleic acid complex material obtained therefrom Download PDFInfo
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Abstract
본 발명은 참치정소로부터 핵산복합물질을 추출하는 방법 및 이 방법에 의해 얻어진 핵산복합물질에 관한 것으로서, 참치정소시료의 세포로부터 핵산복합물질을 추출하는 전처리 수행 후 원심분리 및 여과를 통해 얻은 수용액을 분획분자량(MWCO)이 1,000∼100,000인 한외여과막이 설치된 모듈에 연속적으로 통과시켜 통과되지 않는 수용액을 냉동건조나 분무건조 또는 열풍건조를 통해 분말상 제조하여 상기 참치정소로부터 고순도의 핵산복합물질을 얻음으로써, 부산물로 값싸게 폐기되는 참치 정소로부터 한외여과법을 이용하여 고순도의 핵산복합물질을 보다 신속하고 저렴하게 추출할 수 있는 효과를 갖는다.The present invention relates to a method for extracting a nucleic acid complex material from a tuna testament and to a nucleic acid complex material obtained by the method, wherein the aqueous solution obtained through centrifugation and filtration after pretreatment of extracting the nucleic acid complex material from the cells of the tuna testament is performed. An aqueous solution that is not passed by continuously passing through an ultrafiltration membrane with a fraction molecular weight (MWCO) of 1,000 to 100,000 is prepared in powder form by freeze drying, spray drying or hot air drying to obtain a high purity nucleic acid composite material from the tuna refinery. In addition, it is possible to extract high-purity nucleic acid complexes more quickly and cheaply using ultrafiltration from tuna testament which is cheaply discarded as a by-product.
Description
본 발명은 참치정소로부터 핵산복합물질을 추출하는 방법 및 이 방법에 의해 얻어진 핵산복합물질에 관한 것으로, 특히 어획된 참치중 부산물로서 값싸게 폐기되는 정소를 채취하여 균질화한 후 한외여과막법(Ultra Filtration)을 이용한 가공공정을 통해 핵산복합물질을 제조하여 각종 제품의 기능성 원료로 이용함으로써, 부산물의 부가가치를 높이고 새로운 생리활성 소재로서 신물질을 개발하여 산업적으로 이용 가능하도록 하는 핵산복합물질의 제조방법에 관한 것이다.The present invention relates to a method for extracting a nucleic acid complex from a tuna testament and to a nucleic acid complex obtained by the method. In particular, an ultrafiltration membrane method (Ultra Filtration) is obtained after homogenizing by collecting a waste that is cheaply discarded as a by-product of the tuna caught. A method for producing a nucleic acid composite material by manufacturing a nucleic acid composite material through a processing process using) and using it as a functional raw material for various products, thereby increasing the added value of by-products and developing new materials as new bioactive materials. will be.
상기 핵산복합물질은 RNA와 DNA 등의 핵산과 단백질이 혼합된 물질로서 이에대한 생리활성이 활발하게 연구되고 있는데, 이는 체내에 섭취되어 세포를 부활시켜 노화 및 과산화지질 형성을 억제하며, 면역능을 증가시키고 장내 균총의 개선작용을 가짐은 물론 화장품의 원료로서 피부보습효과 및 자외선 흡수효과 등을 가지는 것으로 알려져 있다.The nucleic acid complex is a mixture of nucleic acids and proteins such as RNA and DNA, and its physiological activity is being actively studied. It is ingested in the body to rejuvenate cells, inhibit aging and formation of lipid peroxide, and increase immunity. It is known to have the effect of improving the intestinal flora and of course the skin moisturizing effect and ultraviolet absorption effect as a raw material of cosmetics.
현재 일본에서는 화장품 및 건강보조식품의 소재로서 연어 정소 및 효모로부터 추출한 핵산복합물질을 이용하고 있으며, 또한 중국에서는 경구약과 주사약으로 사용되고 있고, 구미에서는 암 치료 보조용으로 이를 이용하는 연구가 활발하게 진행되고 있는데, 이렇듯 미국, 일본뿐만 아니라 유럽의 많은 국가들은 연어, 청어의 정소나 효모 등에서 핵산복합물질을 분리추출하여 각종 용도로 이용하고 있으나 아직 참치 정소로부터의 추출물에 대한 발표는 이루어지지 않고 있다.Currently, in Japan, nucleic acid complexes extracted from salmon testis and yeast are used as materials for cosmetics and dietary supplements, and in China, they are used as oral and injectable medicines. As described above, many countries in Europe as well as the United States and Japan are extracting and extracting nucleic acid complexes from salmon, herring, yeast, etc. for various purposes, but there are no announcements about extracts from tuna testes.
종래에 이러한 생리활성을 가지는 핵산복합물질을 분리하는 데에는 국외의 경우 연어와 청어를 재료로 사용하여 핵산복합물질을 분리추출하는 방법이 다양하게 시도되어 왔는데 참치로부터 분리추출한 경우는 아직까지 발견할 수 없고, 국내의 경우도 아직 연구된 적이 없으며 참치정소와 관련된 연구로는 유일하게 오뚜기식품 주식회사에서 발표한 참치정소로부터 프로타민 황산염의 분리방법(특허공고 93-8087, 1993. 8. 25자 공고)에 관한 것이 있는데 이는 프로타민이라는 단백질에 관한 특허이며 그 제법 또한 염산, 암모니아수, 메타인산 나트륨과 2M 황산암모늄을 이용하여 추출하는 것으로 그 과정이 복잡하여 산업적으로 이용하기 어렵고 본 발명과는 무관한 것이라고 할 수 있다.Conventionally, in order to separate nucleic acid complexes having such physiological activity, various methods of separating and extracting nucleic acid complexes using salmon and herring as materials have been attempted in foreign countries. No studies have been conducted in Korea, and the only research related to tuna refinement is the separation of protamine sulfate from tuna refinery published by Ottogi Food Co., Ltd. (Patent Publication 93-8087, August 25, 1993). There is a patent on a protein called protamine, and the manufacturing method is also extracted using hydrochloric acid, aqueous ammonia, sodium metaphosphate and 2M ammonium sulfate, and the process is complicated and industrially difficult to use and irrelevant to the present invention. have.
상기 국외의 경우 연어와 청어 등의 어류 정소로부터 핵산복합물질을 분리추출하는 방법으로는 식염수에 의한 추출(J. Biol. Chem., 203, 167(1953)) 또는 유기용제나 SDS와 같은 계면활성제 처리에 의한 방법(J. Chem. Soc., 1129(1947), J. Biol. Chem., 190, 165(1951)) 등의 발표가 있으며, 이렇게 하여 얻어진 수용액을 산으로 침전시키거나 에탄올 등 알코올로 침전시켜 회수하는 방법이 알려져 있지만 이러한 방법에 의하면 원하는 핵산복합물질을 얻는데 대한 수율이 떨어지고 생산하는데 대량의 가수가 필요하거나 에탄올 등 유기용매를 사용함으로써 산업적으로 이용하기 어려운 단점이 있다.In the case of the above, the method of separating and extracting a nucleic acid complex from fish testes such as salmon and herring is performed by using saline (J. Biol. Chem., 203, 167 (1953)) or an organic solvent or a surfactant such as SDS. The method by the treatment (J. Chem. Soc., 1129 (1947), J. Biol. Chem., 190, 165 (1951)), etc. have been published, and the aqueous solution thus obtained is precipitated with acid or alcohol such as ethanol It is known to recover by precipitating with a known method, but this method has a disadvantage in that the yield of obtaining a desired nucleic acid complex material is inferior, and a large amount of water is required to produce or it is difficult to use industrially by using an organic solvent such as ethanol.
또한 핵산관련물질과 단백질 등을 분리추출하는 방법으로 이온교환방법을 이용한 보고가 있지만(일본특개소 54-15488, 일본특개소 57-57197) 이러한 이온교환체를 이용하여 핵산복합물질을 분리하는 데에는 그 선택성이나 결합할 수 있는 용량이 충분하지 않은 경우가 있고, 상기 이온교환체에 흡착된 물질을 고효율로 탈리시켜 회수할 수 없는 경우가 많아 분리추출하는데 높은 설비비가 요구되지만 높은 설비비에 비해 상대적으로 낮은 수율을 나타내어 산업적으로 이용시 어려움이 있다.In addition, there have been reports using ion exchange as a method of separating and extracting nucleic acid-related substances and proteins (Japanese Patent Application Laid-Open No. 54-15488, Japanese Patent Application Laid-Open No. 57-57197). The selectivity and binding capacity are not sufficient, and the material adsorbed on the ion exchanger can not be removed by high efficiency and can not be recovered. Therefore, a high equipment cost is required for separation and extraction. Difficulty in industrial use due to low yield.
한편 일본 특개공 11-253158에 의하면 생체시료로부터 핵산을 용출하는 전처리를 수행한 후 불용성 유기용매 및 고농도 무기염과 완충제를 포함하는 수용액을 가해 핵산을 함유하는 물층과 함유하지 않는 유기용매층으로 나눈 뒤 원심분리를 실시하고 핵산을 함유하는 물층을 분취하여 핵산을 추출하는 방법을 이용하였으나, 이는 유기용매를 이용함으로써 분리공정에 따른 폐액이 다량 발생하여 환경오염을 일으킬 수 있어 산업적으로 이용하기 어려운 단점이 있다.On the other hand, Japanese Patent Application Laid-Open No. 11-253158 shows that after pretreatment of eluting nucleic acid from a biological sample, an insoluble organic solvent and an aqueous solution containing a high concentration of inorganic salts and a buffer are added to the aqueous layer containing the nucleic acid and the organic solvent layer containing no nucleic acid. After centrifugation and the extraction of the nucleic acid by extracting the water layer containing the nucleic acid, this is difficult to use industrially because it can cause environmental pollution by generating a large amount of waste liquid by the separation process by using an organic solvent There is this.
또한 상기 한외여과는 압력차를 추진력으로 하는 막분리법으로 막세공막과 용질간의 크기차에 의해 특정물질을 분리하는 조작으로서, 한외여과막은 막에 의해 90% 이상의 배제도를 나타내는 용질의 최소분자량으로 정의되는 분획분자량(molecular weight cutoff, 이하 MWCO라 칭함)으로서 그 분리성능을 나타내고, 분자량 300 ∼ 300,000 달톤(dalton) 정도의 중분자 및 고분자량을 갖는 당류, 단백질 등의 생체물질, 고분자물질 등이 분리에 주로 이용되는데 1970년대 중반 이후 우수한 막재료 및 막모듈이 개발된 후 현재 그 응용분야가 급속히 확대되어 가고 있는 방법이다.In addition, the ultrafiltration is a membrane separation method using the pressure difference as a driving force to separate a specific substance by the size difference between the membrane pore membrane and the solute, and the ultrafiltration membrane is defined as the minimum molecular weight of the solute having a degree of exclusion of 90% or more by the membrane. Molecular weight cutoff (hereinafter referred to as MWCO), which shows its separation performance, and biomolecules such as saccharides, proteins, and macromolecules having high molecular weight and molecular weight of about 300 to 300,000 daltons, and high molecular materials, and the like. It is mainly used for the development of excellent membrane materials and membrane modules since the mid-1970s, and its applications are rapidly expanding.
국내의 경우, 공개특허 98-76086에서는 상기 한외여과막을 이용한 수처리 장치를 발표하였고, 공개특허 98-27376에서는 도축혈액의 혈장단백질 가수분해물로부터 안지오텐신전환효소 저해제의 제조방법에 한외여과법을 이용하였으며, 공고특허 96-9051에서는 사람의 뇨로부터 유용 단백질을 생산하는 공정에 한외여과법을 이용하였으며, 공고특허 89-3695에서는 초여과막 및 역 삼투막을 이용한 고농축 사과쥬스의 제조에 한외여과법을 이용하였는 바, 이러한 한외여과법을 이용한 특허가 다수 발표되어 있지만 아직까지 어류정소 특히 참치정소로부터 핵산복합물질을 분리하는데 이용된 자료는 알려져 있지 않다.In Korea, Patent Publication No. 98-76086 discloses a water treatment apparatus using the ultrafiltration membrane, and Patent Publication No. 98-27376 uses an ultrafiltration method for preparing an angiotensin converting enzyme inhibitor from plasma protein hydrolysates of slaughtered blood. In patent 96-9051, ultrafiltration was used for the production of useful proteins from human urine. In patent 89-3695, ultrafiltration was used for the preparation of highly concentrated apple juice using ultrafiltration membranes and reverse osmosis membranes. Although a number of patents using filtration have been published, the data used to isolate nucleic acid complexes from fish stocks, especially from tuna, is unknown.
일반적으로 핵산 분리시 전처리를 수행한 후 얻어진 수용액으로부터 핵산복합물질을 분리하는데는 일본 특개공 9-31093(1997. 2. 4 공개)에 나타난 바와 같이 알칼리를 첨가하여 가열처리하고 원심분리하여 상등액을 얻고 다시 산을 첨가하여 수소이온농도(pH)를 저하시킨 후 핵산복합물질을 침전시키고 나서 흐르는 물로 충분히 수세하여 다시 가열하고 알칼리로 중화하여 pH 7의 수용액을 얻는 여러 단계의 분리방법을 행하거나, 일본특개공 11-253158(1999. 9. 21 공개)에 발표된 것과 같이 불용성 유기용매 및 고농도 무기염과 완충제를 포함하는 수용액을 가해 핵산복합물질을 분리하고 알코올을 첨가하여 추출하는 공정을 수행하여 분리하게 되나 지나치게 공정이 많아 핵산복합물질을 분리추출하는데 상당한 시간이 소요되거나 유기용매를 이용함으로써 분리공정에 따른 폐액이 다량 발생하여 환경오염을 일으킬 수 있는 단점이 있다.Generally, nucleic acid complexes are separated from an aqueous solution obtained after performing pretreatment during nucleic acid separation. As shown in Japanese Patent Application Laid-Open No. 9-31093 (published on Feb. 4, 1997), the supernatant is subjected to heat treatment and centrifugation by adding an alkali. After the addition of acid to lower the hydrogen ion concentration (pH), the nucleic acid complex is precipitated, and then washed with running water, heated again and neutralized with alkali to obtain a pH 7 aqueous solution, or As disclosed in Japanese Patent Application Laid-Open No. 11-253158 (published on September 21, 1999), an insoluble organic solvent and an aqueous solution containing a high concentration of inorganic salts and a buffer are added to separate nucleic acid complexes and extract by adding alcohol. However, due to the excessive process, it takes considerable time to separate and extract the nucleic acid complex material or by using an organic solvent. It has the disadvantage that the large amount of the waste liquid generated according to the process may cause environmental pollution.
상기 단점을 해결하기 위해 본 발명은, 핵산복합물질 분리시 한외여과법을 이용하여 보다 빠르고 효율이 높은 고순도의 핵산복합물질을 분리추출하기 위해 참치의 정소를 원료로 한 핵산복합물질을 분리추출하는 방법을 제공하는 것을 목적으로 한다.In order to solve the above disadvantages, the present invention, a method for separating and extracting a nucleic acid composite material based on the testis of tuna in order to separate and extract a faster and more efficient high purity nucleic acid composite material by using ultrafiltration when separating the nucleic acid composite material. The purpose is to provide.
상기 목적을 달성하기 위해 본 발명은, 전처리하여 얻어진 수용성물질을 한외여과막을 통과시켜 분리시킨 분획물을 냉동건조, 열풍건조 및 분무건조를 실시하여 핵산복합물질을 분리추출하는 것을 특징으로 한다.In order to achieve the above object, the present invention is characterized by separating and extracting the nucleic acid composite material by performing freeze drying, hot air drying and spray drying the fractions separated by passing the water-soluble material obtained by pretreatment through an ultrafiltration membrane.
또한 참치정소시료의 세포로부터 핵산복합물질을 추출하는 전처리 수행 후 원심분리 및 여과를 통해 수용액을 얻는 제 1 단계, 상기 수용액을 분획분자량(MWCO)이 1,000∼100,000인 한외여과막이 설치된 모듈에 연속적으로 통과시켜 통과되지 않는 수용액을 얻는 제 2 단계 및 상기 수용액을 냉동건조나 분무건조 또는 열풍건조를 통해 분말상 제조하여 상기 참치정소로부터 고순도의 핵산복합물질을 얻는 제 3 단계를 포함하는 것을 특징으로 한다.In addition, a first step of obtaining an aqueous solution through centrifugation and filtration after performing a pretreatment to extract the nucleic acid complex material from the cells of the tuna testicles, the aqueous solution is continuously installed in a module installed with an ultrafiltration membrane having a molecular weight of 1,000 to 100,000 (MWCO) And a third step of obtaining an aqueous solution that does not pass through, and a third step of obtaining the high-purity nucleic acid composite material from the tuna purifier by preparing the powder form through freeze drying, spray drying, or hot air drying.
본 발명은 전처리하여 얻어진 수용물질을 MWCO 1,000∼100,000인 한외여과막이 설치된 기기에 통과시켜 분획물을 분리하고 분리된 분획물을 냉동건조, 열풍건조 혹은 분무건조 등을 실시하여 핵산복합물질을 분리추출함으로써 공정을 최대한 단순화시키고 핵산복합물질의 분리추출 시간을 줄이며 유기용매 등의 이용을 배제함으로써 환경오염을 발생시키지 않고 핵산복합물질을 얻을 수 있으며, 또한 종래의 일반적인 방법으로 핵산을 분리하는데 이용되는 유기용매를 이용하지 않고 고순도의 핵산복합물질을 얻을 수 있을 뿐만 아니라 이를 통해 보다 값 싼 핵산복합물질을 얻을 수 있다. 즉 상기 한외여과막은 분획분자량(MWCO)이 1,000 미만인 경우 목표로 하는 핵산복합물질 이외의 다른 불순물의 함유 및 크기(size)가 작아 공정상 높은 압력과 긴 시간을 요구하고, 분획분자량이 100,000 초과인 경우에는 목표로 하는 핵산복합물질을 손실시키는 특징이 있다.The present invention is a process by separating the fractions by passing the water-receiving material obtained by pretreatment through an ultrafiltration membrane with a MWCO 1,000 ~ 100,000 and the separated fractions by freeze drying, hot air drying or spray drying to separate and extract the nucleic acid composite material Simplifies and reduces the extraction time of nucleic acid complexes and eliminates the use of organic solvents, thereby obtaining nucleic acid complexes without generating environmental pollution. High purity nucleic acid complexes can be obtained without using them, as well as cheaper nucleic acid complexes. That is, when the ultrafiltration membrane has a fraction molecular weight (MWCO) of less than 1,000, the content and size of impurities other than the target nucleic acid complex are small, requiring high pressure and a long time in the process, and the fraction molecular weight is greater than 100,000. In this case, the target nucleic acid complex is lost.
도 1 은 본 발명이 적용되는 덱스트런(Dextran) 표준품의 표준곡선 및 시료용액을 흡광도로 측정한 결과 파형도.BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a waveform diagram of results obtained by measuring absorbance of a standard curve and a sample solution of a Dextran standard product to which the present invention is applied.
이하 첨부된 도면을 참조하여 본 발명을 상세히 설명하면 다음과 같다.Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.
도 1 은 본 발명이 적용되는 덱스트런(Dextran) 표준품의 표준곡선 및 시료용액을 흡광도로 측정한 결과 파형도이다.1 is a waveform diagram of the results obtained by measuring absorbance of a standard curve and a sample solution of a Dextran standard to which the present invention is applied.
상기 도 1의 측정결과 파형은 냉동된 참치 정소 1㎏을 상온으로 해동시킨 후 쵸퍼(Chopper)로 잘게 균질화하고 나서 교반기가 부착된 플라스크에 옮겨 3,000㎖의 가수를 한 뒤 염화나트륨(NaCl) 300g을 가해 교반하면서 3시간 가열한 후 5,000G, 10분간 원심분리를 실시하여 상등액을 얻으며, 상기 상등액을 MWCO 10,000인 한외여과막이 부착된 기기(Sartocon Slicecrossflow system, Sartorius)에 투입하여 연속적으로 통과시켜 통과되지 않은 농축 수용액을 탱크로부터 분리하고 냉동건조를 실시하여 핵산복합물질 나트륨염 분말 41.3g을 얻는데, 이때 Diphenyl amine 시약을 이용하여 분석한 결과 핵산함량은 69.6% 이었으며, Lowry 방법으로 분석한 결과 단백질 함량은 10.7% 이었다.The measurement result waveform of FIG. 1 is thawed 1 kg of frozen tuna testicles to room temperature, and then homogenized finely with a chopper, and then transferred to a flask equipped with an agitator, followed by 3,000 ml of water, followed by addition of 300 g of sodium chloride (NaCl). After heating for 3 hours with stirring, the supernatant was obtained by centrifugation at 5,000 G for 10 minutes, and the supernatant was introduced into an ultrafiltration membrane (Sartocon Slicecrossflow system, Sartorius) of MWCO 10,000 and continuously passed through it. The concentrated aqueous solution was separated from the tank and lyophilized to obtain 41.3 g of the sodium salt powder of the nucleic acid complex, wherein the content of nucleic acid was 69.6% using Diphenyl amine reagent, and the content of protein was 10.7. Was%.
그 비교 예로 냉동 건조된 참치 정소를 상기 도 1과 같이 처리하여 얻은 상등액을 MWCO 1,000인 한외여과막에 통과시키면 핵산복합물질 나트륨염 48.2g을 얻을 수 있고, 상기 Diphenyl amine 시약을 이용하여 분석한 결과 핵산함량은 60.8%이었으며, Lowry 방법으로 분석한 결과 단백질 함량은 9.3%이었다.As a comparative example, when the supernatant obtained by treating the freeze-dried tuna testis as shown in FIG. 1 was passed through an ultrafiltration membrane of MWCO 1,000, 48.2 g of the nucleic acid sodium salt was obtained, and the result was analyzed using the Diphenyl amine reagent. The content was 60.8% and the protein content was 9.3% as analyzed by Lowry method.
또한 상기 냉동된 참치 정소를 상기 도 1과 같이 처리하여 얻은 상등액을 MWCO 100,000인 한외여과막에 통과시켜 핵산복합물질 나트륨염 9.7g을 얻었으며, 상기 Diphenyl amine 시약을 이용하여 분석한 결과 핵산함량은 9.4%, 상기 Lowry 방법으로 분석한 결과 단백질 함량은 2.3%이었다.In addition, the supernatant obtained by treating the frozen tuna testis as shown in FIG. 1 was passed through an ultrafiltration membrane of MWCO 100,000 to obtain 9.7 g of the nucleic acid complex sodium salt. The nucleic acid content was analyzed using the Diphenyl amine reagent. %, The protein content was 2.3% as analyzed by the Lowry method.
다음 실시예로서 냉동된 참치정소 1㎏을 상온에서 해동시킨 후 균질화하고 나서 교반기가 부착된 플라스크에 옮겨 0.5N NaOH 용액을 3,000㎖ 첨가하여 교반하면서 3시간 동안 가열추출한 후 5,000G 10분간 원심분리를 실시하여 상등액을 얻은 다음 6N 염산(HCl)을 이용하여 pH 7로 중화하고 나서 이후 공정을 상기 도 1에서와 같이 실시하여 핵산복합물질 나트륨염 분말 47.8g을 얻었으며, 상기 Diphenylamine 시약을 이용하여 분석한 결과 핵산함량은 52.2%, 상기 Diphenyl amine 시약을 이용하여 분석한 결과 단백질 함량은 8.9%이었다.In the following example, 1 kg of frozen tuna testine was thawed at room temperature, homogenized, and then transferred to a flask equipped with a stirrer. Then, 3,000 ml of 0.5N NaOH solution was added thereto, followed by heat extraction for 3 hours with stirring, followed by centrifugation at 5,000 G for 10 minutes. A supernatant was obtained, and then neutralized to pH 7 using 6N hydrochloric acid (HCl), and then the process was carried out as in FIG. 1 to obtain 47.8 g of the nucleic acid sodium salt powder, and analyzed using the Diphenylamine reagent. As a result, the nucleic acid content was 52.2%, and the protein content was 8.9% when analyzed using the Diphenyl amine reagent.
상기 실시예에서 얻은 핵산복합물질 나트륨염의 분자량을 Gel Permeation Chromatogratph법으로 측정한 추정 분자량은, Dextran 표준품을 이용해서 Sepharose CL-4B resin을 통과시켜 페놀황산법을 이용하여 흡광도를 측정하고, 분자량과 Tube No.를 plot하여 표준곡선을 작성하였고, 상기 도 1의 시료용액을 동일하게 용출시켜 260㎚의 흡광도를 측정하여 표준곡선과 비교한 뒤 추정분자량을 계산한 결과 대체적으로 약 100,000내외의 분자량을 나타내었다.Molecular weight of the sodium salt of the nucleic acid complex obtained in the above example was measured by Gel Permeation Chromatogratph method, and the absorbance was measured by phenol sulfate method by passing Sepharose CL-4B resin using a Dextran standard. The standard curve was prepared by plotting. The sample solution of FIG. 1 was eluted in the same manner, and the absorbance at 260 nm was measured, compared with the standard curve, and the estimated molecular weight was calculated. The molecular weight was approximately 100,000. .
상술한 바와 같이 본 발명은, 부산물로 값싸게 폐기되는 참치 정소로부터 한외여과법을 이용하여 고순도의 핵산복합물질을 보다 신속하고 저렴하게 추출할 수 있는 효과를 갖는다.As described above, the present invention has an effect that the nucleic acid complex of high purity can be extracted more quickly and cheaply using ultrafiltration from tuna testis which is cheaply discarded as a by-product.
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| KR20160003496A (en) | 2014-07-01 | 2016-01-11 | (주)재원산업기계 | Apparatus for removing provisional seam on welded steel pipe |
| KR20180008059A (en) | 2016-07-15 | 2018-01-24 | 주식회사 글랜젠 | Polydeoxyribonucleotide containing skin filling material having improved durability and antioxidative activity and the maufacturing method thereof |
| KR20180060693A (en) | 2016-11-29 | 2018-06-07 | 주식회사 글랜젠 | Cosmetic composition having improved skin wrinkle elimination and skin elasticity |
| KR20190065676A (en) | 2017-12-04 | 2019-06-12 | 주식회사 비알팜 | Manufacturing method of high purity DNA fragments mixtures |
| KR102053221B1 (en) | 2018-10-23 | 2019-12-06 | 김석순 | Manufacturing method of high purity DNA fragments mixtures |
| KR102190612B1 (en) | 2019-08-27 | 2020-12-14 | (주)모아캠 | Method of Producing Low Molecular Weight DNA derived from Plant Materials, Microbial Materials or Marine Materials |
| KR20230061788A (en) | 2021-10-29 | 2023-05-09 | 주식회사 현대메디텍 | Methods of preparing polynucleotide mixture and suture containing the same |
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