KR101284217B1 - A methohd of solubilizing muscle protein and a method of producing protein for food - Google Patents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/04—Animal proteins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/001—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste
- A23J1/002—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from waste materials, e.g. kitchen waste from animal waste materials
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/10—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from hair, feathers, horn, skins, leather, bones, or the like
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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
- A23V2250/00—Food ingredients
- A23V2250/54—Proteins
- A23V2250/542—Animal Protein
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Abstract
본 발명은 심근 단백질의 가용화 방법 및 식품용 단백질의 제조방법에 관한 것으로 좀 더 자세하게는 심근을 pH 8.0 내지 9.5의 나트륨염 또는 인산염에서 현탁시키는 단계; 및 상기 심근 단백질을 초음파 처리하는 단계를 포함하는 심근 단백질을 가용화 시키는 방법이다. 본 발명에 의한 초음파를 이용한 심근 단백질을 가용화 시키는 방법은 저농도 및 저온에서도 충분히 심근 단백질을 가용화 할 수 있으며, 초음파의 처리시간과 진폭(amplitude)을 적당히 조절하여 원하는 분자량의 단백질을 제조할 수 있다.The present invention relates to a method for solubilizing myocardial protein and to a method for preparing food protein, and more particularly, suspending the myocardium in sodium salt or phosphate of pH 8.0 to 9.5; And sonicating the myocardial protein. In the method for solubilizing myocardial protein using ultrasound according to the present invention, the myocardial protein can be solubilized sufficiently even at low concentration and low temperature, and a protein having a desired molecular weight can be prepared by appropriately adjusting the processing time and amplitude of the ultrasonic wave.
Description
본 발명은 심근 단백질의 가용화 방법 및 식품용 단백질의 제조방법에 관한 것이다.The present invention relates to a method for solubilizing myocardial protein and a method for preparing a protein for food.
심장 근육단백질은 근원 섬유를 구성하고 있는 단백질로 미오신, 악틴, 트로포미오신, 트로포닌, α-actinin, M-단백질, C-단백질, 이외에 많은 미량조절성 단백질로 이루어진다. 이들은 의 근육세포(근섬유)의 옆줄무늬 구조형성에 직접 관련되어 배치되어 있고 기질 단백질과 동시에 근육 구조 단백질이라고도 부르며 생화학적으로는 근육의 기능 즉, 수축, 이완의 주역을 행하고 있다.Cardiac muscle proteins are proteins that make up myofibrils and consist of myosin, actin, tropomyosin, troponin, α-actinin, M-protein, C-protein, and many other microregulatory proteins. They are directly related to the formation of the lateral stripe structure of muscle cells (muscle fibers) and are also called muscle structure proteins at the same time as matrix proteins. Biochemically, they play a role in muscle function, namely contraction and relaxation.
한편, 돼지 심근은 근원섬유로 이루어져 있으며 이것을 구성하는 근원섬유 단백질은 대부분 0.5 M 이상의 KCl 또는 NaCl의 고염농도 용액에서 추출되는 염가용성 단백질로 이루어져 있다. 염가용성 단백질은 육을 원료로 하는 햄이나 소시지 제조 시 겔을 형성하는데 필수적인 가장 중요한 역할을 담당하고 있으며, 우리들은 주로 이러한 단백질을 섭취하고 있다. 그러나 심근의 경우는 햄이나 소시지와 같은 가공품에는 적합하지 않은 것으로 알려져 있다. 그 이유는 단백질의 분자레벨에서는 겔 형성능이 우수하나 근육레벨에서는 그렇지 못하기 때문이다. 따라서, 심근의 경우는 우리나라의 전통식품인 순대 등에 일부 쓰여 지고 있을 뿐 대부분 폐기처분 되고 있다.On the other hand, pig myocardium is composed of myofibrils and myofibrillar proteins are composed of salt-soluble proteins extracted from high salt solution of KCl or NaCl of 0.5 M or more. Salt-soluble proteins play the most important role in the formation of gels in the production of meat-based hams and sausages, and we consume these proteins primarily. However, myocardium is not suitable for processed products such as ham and sausage. This is because the gel formation ability at the molecular level of the protein is excellent, but not at the muscle level. Therefore, myocardium is only partially used in Sundae, a traditional food of Korea, and is mostly disposed of.
이에 발명자는 효율적인 식품용 돼지 심근 단백질의 가용화를 위하여 본 발명에 이르렀다.Therefore, the inventors have come to the present invention for efficient solubilization of porcine myocardial protein for food.
본 발명의 목적은, 염가용성 단백질인 심근 단백질을 염류 및 초음파 처리를 하여 가용화 시킴으로써 대부분 폐기되고 있는 심근의 건강 기능성 단백질 식품소재로서의 이용 가능성에 대하여 확인하는 것이다.An object of the present invention is to confirm the applicability of myocardial protein, which is mostly discarded, by solubilizing myocardial protein, a salt-soluble protein, by salting and sonication.
상기 목적을 달성하기 위하여 일 구체예에서 심근 단백질을 pH 8.0 내지 9.5의 나트륨염 또는 인산염에서 현탁시키는 단계; 및 상기 심근 단백질을 초음파 처리하는 단계를 포함하는 심근 단백질을 가용화 시키는 방법을 제공한다. 다른 구체예에서, 상기 나트륨염은 염화나트륨인 심근 단백질을 가용화 시키는 방법을 제공한다. 또 다른 구체예에서, 인산염은 피로린산(Pyrophosphate) 또는 인산(phosphate)인 심근 단백질을 가용화 시키는 방법을 제공한다. 또 다른 구체예에서, 상기 피로린산은 5 내지 20mM 농도인 것을 특징으로 하는 심근 단백질을 가용화 시키는 방법을 제공한다. 또 다른 구체예에서, 상기 인산염이 pH 9.0 내지 9.2인 것을 특징으로 하는 심근 단백질을 가용화 시키는 방법을 제공한다. 또 다른 구체예에서, 상기 초음파 처리 단계는 20 내지 80분간 실행하는 것을 특징으로 하는 심근 단백질을 가용화 시키는 방법을 제공한다. 또 다른 구체예에서, 상기 단백질은 돼지 심근으로부터 추출된 것을 특징으로 하는 심근 단백질을 가용화 시키는 방법을 제공한다. 또 다른 구체예에서, 상기 초음파 처리는 20kHz의 진동수 및 40 내지 80%의 진폭에서 이루어지는 것을 특징으로 하는 심근 단백질을 가용화 시키는 방법을 제공한다.
In one embodiment suspending the myocardial protein in sodium salt or phosphate at pH 8.0 to 9.5; And it provides a method for solubilizing myocardial protein comprising the step of sonicating the myocardial protein. In another embodiment, the sodium salt provides a method of solubilizing myocardial protein that is sodium chloride. In another embodiment, phosphate provides a method of solubilizing myocardial protein that is pyrophosphate or phosphate. In another embodiment, the pyroic acid provides a method for solubilizing myocardial protein, characterized in that the concentration of 5 to 20mM. In another embodiment, there is provided a method for solubilizing myocardial protein, characterized in that the phosphate is pH 9.0 to 9.2. In another embodiment, the sonication step provides a method for solubilizing myocardial protein, which is performed for 20 to 80 minutes. In another embodiment, the protein provides a method for solubilizing myocardial protein, characterized in that it is extracted from porcine myocardium. In another embodiment, the sonication provides a method for solubilizing myocardial protein, characterized in that at a frequency of 20 kHz and an amplitude of 40 to 80%.
일 구체예에서 심근 단백질을 pH 8.0 내지 9.5의 나트륨염 또는 인산염에서 현탁시키는 단계; 및 상기 심근 단백질을 초음파 처리하는 단계를 포함하는 식품용 단백질의 제조 방법을 제공한다. 다른 구체예에서, 상기 나트륨염은 염화나트륨인 식품용 단백질의 제조 방법을 제공한다. 또 다른 구체예에서, 상기 인산염은 피로린산(Pyrophosphate) 또는 인산(phosphate)인 식품용 단백질의 제조 방법을 제공한다. 또 다른 구체예에서, 상기 피로린산은 5 내지 20mM농도인 것을 특징으로 하는 식품용 단백질의 제조 방법을 제공한다. 또 다른 구체예에서, 상기 인산염이 pH 9.0 내지 9.2인 것을 특징으로 식품용 단백질의 제조 방법을 제공한다. 또 다른 구체예에서, 상기 초음파 처리 단계는 20 내지80분간 실행하는 것을 특징으로 하는 식품용 단백질의 제조방법을 제공한다. 또 다른 구체예에서, 상기 심근 단백질은 돼지 심근으로부터 추출된 것을 특징으로 하는 식품용 단백질의 제조 방법을 제공한다. 또 다른 구체예에서, 상기 초음파 처리는 20kHz의 진동수 및 40 내지 80%의 진폭에서 이루어지는 것을 특징으로 하는 식품용 단백질의 제조 방법을 제공한다.
In one embodiment suspending the myocardial protein in sodium salt or phosphate at pH 8.0-9.5; And it provides a method for producing a protein for food comprising the step of sonicating the myocardial protein. In another embodiment, the sodium salt provides a method of preparing a food protein that is sodium chloride. In another embodiment, the phosphate provides a method for producing a food protein that is pyrophosphate or phosphate. In another embodiment, the pyroic acid provides a method for producing a protein for food, characterized in that the concentration of 5 to 20mM. In another embodiment, the phosphate is pH 9.0 to 9.2 provides a method for producing a food protein. In another embodiment, the sonication step provides a method for producing a protein for food, characterized in that performed for 20 to 80 minutes. In another embodiment, the myocardial protein provides a method for producing a food protein, characterized in that extracted from porcine myocardium. In another embodiment, the ultrasonic treatment provides a method for producing a protein for food, characterized in that at a frequency of 20kHz and an amplitude of 40 to 80%.
본 발명에서 “심근 단백질”은 심장의 근원섬유를 구성하고 있는 단백질로 미오신, 악틴, 트로포미오신, 트로포닌, α-actinin, M-단백질, C-단백질, 이외에 많은 미량조절성 단백질로 이루어진 것을 말한다.In the present invention, "myocardial protein" refers to a protein constituting the myofibril of the heart, composed of myosin, actin, tropomyocin, troponin, α -actinin, M-protein, C-protein, and other microregulatory proteins. .
본 발명에 의한 초음파를 이용한 심근 단백질을 가용화 시키는 방법은 저농도 및 저온에서도 충분히 단백질을 가용화 할 수 있으며, 초음파의 처리시간과 진폭(amplitude)를 적당히 조절하여 원하는 분자량의 단백질을 제조할 수 있다.In the method for solubilizing myocardial protein using ultrasound according to the present invention, the protein can be solubilized sufficiently even at low concentration and low temperature, and a protein having a desired molecular weight can be prepared by appropriately adjusting the processing time and amplitude of the ultrasound.
도 1은 초음파 처리시 염류에 따른 심근단백질의 용해도를 나타낸 그래프이다.
도 2는 돼지 심근으로부터 염류 및 초음파에 의해 분리된 단백질의 SDS-PAGE 패턴 결과이다.
도 3은 심근 현탁액에 첨가한 피로린산 양에 따른 pH의 변화를 나타낸 그래프이다.
도 4는 피로린산 양에 따른 심근 단백질의 용해도를 나타낸 그래프이다.1 is a graph showing the solubility of myocardial protein according to salts during sonication.
Figure 2 shows the SDS-PAGE pattern of the protein isolated by salt and ultrasound from porcine myocardium.
Figure 3 is a graph showing the change in pH according to the amount of pyrroline acid added to the myocardial suspension.
Figure 4 is a graph showing the solubility of the myocardial protein according to the amount of pyrroline acid.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .
실시예Example
실시예Example 1. One. 근원섬유단백질의 추출 Extraction of Myofibrillar Proteins 조건 Condition
1-1. 시료의 혈액 제거1-1. Sample Blood Removal
돼지 심근(한일부산물 제공)을 약 1.0㎝의 육면체로 세절한 후, 심근 중량에 대하여 10배의 증류수(4℃)를 가한 후, 균질기(Ultra turrax T20, IKA, Germany)를 이용하여 13,500rpm에서 균질화 하였으며, 아이스배스 상태에서 1분 처리, 1분 정지를 3회 반복하였다. 균질화한 시료는 초음파기로 20kHz, 20% 진폭(amplitude)에서 10분간 처리한 후 6,000 x g에서 10분간 원심분리(High speed centrifuge, Avanti J-E, Beckman. USA)하여 상등액(혈액)을 제거하였다. 이와 같은 조작을 4회 반복하여 혈액을 제거하였다.
Porcine myocardium (provided by Hanil by-product) was cut into cubes of about 1.0 cm, 10 times distilled water (4 ° C) was added to the myocardial weight, and then 13,500 rpm using a homogenizer (Ultra turrax T20, IKA, Germany). Homogenized at 1 minute treatment, 1 minute stop in ice bath was repeated three times. The homogenized sample was treated with an ultrasonic wave at 20 kHz, 20% amplitude for 10 minutes, and then centrifuged at 6,000 xg for 10 minutes (High speed centrifuge, Avanti JE, Beckman. USA) to remove the supernatant (blood). This operation was repeated four times to remove blood.
1-2. 초음파 처리 조건1-2. Ultrasonic treatment conditions
염가용성 단백질인 근원섬유 단백질을 물리적 기법인 초음파 처리를 하여 가용화 시켰다. 초음파 처리조건은 다음과 같았다.Myofibrillar proteins, salt-soluble proteins, were solubilized by sonication, a physical technique. Ultrasonic treatment conditions were as follows.
초음파기는 20kHz의 고정형을 사용하였으며 (VCX 750, 20kHz, Sonic and Material Inc) 표 1에서와 같은 범위 내에서 기기를 조절하여 처리하였다.The sonicator used a fixed type of 20 kHz (
진폭
펄스 on/off
처리시간
온도Frequency
amplitude
Pulse on / off
Processing time
0~100%
20sec/20sec
0~240min
4℃20 kHz
0-100%
20sec / 20sec
0-240 min
4 ℃
심근현탁액의 초음파 처리는 농도를 10mg/ml로 조절한 후 염농도, pH, 진폭 및 온도 등에 변화를 주어 처리하였다. 즉 NaCl 농도를 0~0.2M, pH5.0~8.0으로 하였으며 초음파 처리는 20kHz의 20~80% 진폭의 범위에서 처리하였다. 또한 가열에 의한 영향을 초음파 처리 전 심근의 단백질 농도를 10mg/ml로 조절한 후 20~100℃까지 30분간 열처리하였다. 그런 다음 4℃까지 냉각시킨 후 초음파 처리 하였다. 이 때의 염농도는 0.2M NaCl, pH8.0에서 수행하였으며, 초음파는 20kHz의 20~80% 진폭에서 처리하였다.Ultrasonic treatment of myocardial suspension was treated by adjusting the concentration to 10 mg / ml and then changing the salt concentration, pH, amplitude and temperature. That is, NaCl concentration was 0-0.2M, pH5.0-8.0, and the ultrasonication was performed in the range of 20-80% amplitude of 20kHz. In addition, the effect of heating was adjusted to a protein concentration of myocardium before the sonication to 10mg / ml and heat-treated for 20 minutes to 20 ~ 100 ℃. Then it was cooled to 4 ℃ and sonicated. At this time, the salt concentration was performed at 0.2 M NaCl, pH 8.0, and the ultrasonic wave was treated at 20 to 80% amplitude of 20 kHz.
또한, 각종 염류의 영향에 대해 조사하기 위하여 염화나트륨(NaCl), 피로린산(pyrophosphate) 및 인산(phosphate)을 심근현탁액에 각각 5~20mM을 첨가한 후 상기와 같은 방법으로 초음파 처리를 하였다.
In addition, in order to investigate the effects of various salts, sodium chloride (NaCl), pyrophosphate and phosphoric acid (phosphate) were added to the myocardial suspension, respectively, 5-20 mM, followed by sonication.
1-3. 단백질 용해도 측정1-3. Protein Solubility Measurement
각각의 조건에 따라 초음파 처리한 시료를 20,000×g에서 20분간 원심분리하여 상등액 중의 단백질 농도를 Biurret법(Gornall, A, G.등, 1949)으로 측정하였다. 용해도는 총 단백질 함량에 대한 초음파 처리 후 상등액의 단백질 함량의 비로 계산하였다.
The sonicated samples were centrifuged at 20,000 × g for 20 minutes and the protein concentration in the supernatant was measured by the Biurret method (Gornall, A, G. et al., 1949). Solubility was calculated as the ratio of the protein content of the supernatant after sonication to the total protein content.
1-4. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)1-4. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)
SDS-PAGE는 Lammli법(Lammli, V.K. 등, 1970)에 의하여 10% slab 겔을 이용하여 실험하였다. 초음파에 의해서 분리된 시료에 8M 유리아, 2% 머캅토에탄올, 2% SDS와 20mM Tris-HCl(pH 8.0)을 첨가한 후, 100℃에서 2분간 가열하였다. 고정화 및 염색은 Neuhoff(Neuhoff V. 등 1988)의 방법에 의해 코마시 브릴리언트 블루 R250으로 실시하였다. 탈색은 50% MeOH, 7% 아세트산으로 30분간, 9.2% 아세트산으로 겔의 배경이 투명화 될 때까지 수행하였다.
SDS-PAGE was tested using 10% slab gel by Lammli method (Lammli, VK et al., 1970). To the sample separated by ultrasonic wave, 8M free-form, 2% mercaptoethanol, 2% SDS and 20 mM Tris-HCl (pH 8.0) were added, followed by heating at 100 ° C for 2 minutes. Immobilization and staining were performed with Coomassie Brilliant Blue R250 by the method of Neuhoff (Neuhoff V. et al. 1988). Decolorization was performed for 30 minutes with 50% MeOH, 7% acetic acid and until the background of the gel became clear with 9.2% acetic acid.
실시예Example 2. 2. 초음파 처리에 의한 심근 단백질의 Of myocardial protein by sonication 가용화Solubilization
2-1. 심근 단백질의 가용화에 영향을 주는 요소2-1. Factors Affecting Solubilization of Myocardial Protein
초음파에 의한 심근단백질의 가용화는 염농도(NaCl), pH, 온도, 초음파 및 초음파의 진폭에 영향을 받았다. 이 중, 염농도는 생리적인 염농도에 가까운 0.1~0.2M의 NaCl을 최소한의 양을 필요로 하였으며, 심근 단백질을 식품용으로 사용하기 위하여 식품첨가물로 활용되고 있는 다른 이종의 염류에 대해서도 검토하였다.
The solubilization of myocardial protein by ultrasound was influenced by salt concentration (NaCl), pH, temperature, ultrasound and amplitude of ultrasound. Among them, the salt concentration required a minimum amount of NaCl of 0.1-0.2M, which is close to the physiological salt concentration, and also examined other heterologous salts that are used as food additives for the use of myocardial protein for food.
2-2. 염류의 영향2-2. Salt Effect
심근 현탁액에 최종농도가 10mM이 되도록 피로린산(Pyrophosphate, pH 8.0) 및 인산(Phosphate, pH 8.0)을 첨가하여 20 kHz의 60% 진폭에서 초음파처리를 하였을 때의 용해도를 측정하였다(도 1). 대조구로서 10mM NaCl(pH 8.0)을 첨가하여 초음파 처리를 하였다. 이 결과에 의하면 대조구(NaCl)의 경우 초음파처리 120분 후에 약 15% 정도가 분리된 반면에 인산 처리구는 약 38%, 피로린산은 약 60%의 용해도를 나타내었다. 한편, 피로린산 처리구 만큼의 단백질을 분리시키기 위해서는 NaCl의 경우 0.2M 정도를 필요로 하였다. 이와 같은 첨가량을 중량비로 나타내었을 경우 10mM 피로린산은 0.13%, 0.2M NaCl은 1.16%에 상당하는 것으로 나타나 그 차이는 약 8.9배인 것으로 확인되었다. 그러나 피로린산의 경우 0.05M(0.065%)만 첨가하여도 충분히 단백질을 분리할 수 있는 것으로 확인되었다. 그러면 그 차이는 17.8배에 이른다.Pyrophosphate (Pyrophosphate, pH 8.0) and phosphoric acid (Phosphate, pH 8.0) were added to the myocardial suspension so that the final concentration was 10 mM, and the solubility of the myocardial suspension at the 60% amplitude of 20 kHz was measured (Fig. 1). As a control, 10 mM NaCl (pH 8.0) was added to sonication. According to these results, the control (NaCl) was about 15% after 120 minutes of sonication, while the phosphate treatment showed about 38% solubility and about 60% solubility. On the other hand, in order to separate the protein as much as the pyridonic acid treatment, NaCl required about 0.2M. When the addition amount is represented by the weight ratio, 10mM pyroic acid is equivalent to 0.13%, 0.2M NaCl is 1.16%, and the difference is about 8.9 times. However, it was confirmed that the addition of 0.05M (0.065%) in the case of pyroic acid can fully separate the protein. The difference is then 17.8 times.
이들 3종의 염류에 의해서 분리된 단백질의 성분을 조사하기 위하여 SDS-PAGE로 분석하였다(도 2). NaCl 첨가구의 경우는 초음파 처리 시간이 길어짐에 따라서 각각의 성분들이 늘어나는 것으로 관찰되었지만 그 양은 매우 적었다. 그러나 인산 첨가구의 경우는 초음파 처리 30분만에 150KDa, 43KDa, 34KDa에 상당하는 성분들이 관찰되기 시작하였으며, 이들 성분은 초음파 처리시간이 늘어남에 따라서 증가하는 것으로 나타났다. 한편, 피로린산의 경우는 초음파 처리 초기에 이미 200KDa(미오신중쇄로 추정됨)에 상당하는 성분이 다량 검출되었으며, 그 외에 150KDa, 43KDa 및 34KDa에 상당하는 성분이 관찰되기 시작하였다. 또한 초음파 처리 시간이 길어짐에 따라서 200KDa에 상당하는 성분이 분해되는 양상을 보였으며, 이들의 분해된 성분들은 규칙적인 작용에 의해서 분해가 일어나는 것이 아니라 분자량 200KDa과 43KDa의 사이로 이동되는 불규칙적인 형태로 분해된 성분들이 관찰되었다. 또한 전체적으로 분리된 양을 관찰하였을 때 도 1에서의 결과와 마찬가지로 피로린산(Pyrophosphate) > 인산(Phosphate) > NaCl의 순으로 많이 관찰되었다.
In order to investigate the components of the protein separated by these three salts, it was analyzed by SDS-PAGE (Fig. 2). In the case of NaCl addition, each component was observed to increase as the sonication time increased, but the amount was very small. However, in the case of the phosphate added group, components equivalent to 150KDa, 43KDa, and 34KDa began to be observed after 30 minutes of sonication, and these components increased as the sonication time increased. On the other hand, in the case of pyrochloric acid, a large amount of components corresponding to 200 KDa (presumed as myosin heavy chain) was already detected at the initial stage of sonication, and components corresponding to 150 KDa, 43 KDa, and 34 KDa began to be observed. In addition, as the sonication time increased, the components equivalent to 200 KDa were decomposed, and these decomposed components were decomposed into irregular shapes that were moved between the molecular weights of 200 KDa and 43 KDa, rather than being decomposed by regular action. Components were observed. Also, when the total amount was separated, as in the result of FIG.
2-3. 피로린산(Pyrophosphate)의 농도 의존성2-3. Concentration dependence of pyrophosphate
앞서 제시한 바와 같이 초음파에 의한 심근단백질의 분리능에 있어서 다른 염류보다 피로린산이 가장 효과적인 것으로 나타났다. 그러나 피로린산 첨가 시 pH를 8.0으로 고정하여 첨가하였으므로 pH를 고정하지 않고 그대로 농도별로 첨가하였을 때는 어떠한 현상이 일어나는지 확인하기 위하여 농도별 첨가 시 pH 변화 및 이에 따른 심근단백질의 용해도를 검토하였다. 우선 심근현탁액에 피로린산을 농도별로 첨가하였을 때의 pH를 도 3에 나타내었다. 이 결과에 의하면 5mM의 피로린산을 첨가하였을 때는 pH 8.2 정도를 나타내었으나 10mM의 피로린산을 첨가하였을 때는 pH 9.1 정도인 것으로 나타났다. 그러나 20mM 이상의 피로린산을 첨가하였을 때는 pH 9.2 정도로 큰 변화는 인정되지 않았다.As shown above, pyroic acid was more effective than other salts in the resolution of myocardial protein by ultrasound. However, since the pH was fixed at 8.0 when the pyroic acid was added, the pH change and the solubility of the myocardial protein were examined when the concentration was added in order to check what happens when the concentration was added without changing the pH. First, the pH at the time of adding pyroic acid by concentration to the myocardial suspension is shown in FIG. 3. According to these results, the pH was about 8.2 when 5 mM pyrolic acid was added, but the pH was about 9.1 when 10 mM pyroic acid was added. However, when a pyrolic acid of 20 mM or more was added, a change as large as pH 9.2 was not recognized.
다음은 이와 같이 피로린산의 첨가량 및 이에 따라 pH가 변화하였을 때의 심근의 초음파에 의한 용해도를 측정하여 도 4에 제시하였다. 이 결과에 의하면 5mM의 경우 초음파 처리시간 20분 까지는 급격히 증가하였다가 그 후 서서히 증가였으며, 초음파 처리 80분만에 90% 이상의 용해도를 나타내었다. 한편 10mM 이상의 피로린산 첨가구에서는 전체적인 경향은 같았으나 용해속도가 매우 빨라졌으며, 불과 초음파 처리 40분만에 90% 이상의 용해도를 나타내었다.Next, the solubility of the myocardium at the time when the addition amount of pyroic acid and thus the pH was changed is measured and presented in FIG. 4. According to the results, 5mM of the sonication time increased rapidly up to 20 minutes and then gradually increased, and showed solubility of 90% or more in 80 minutes of sonication. On the other hand, in the addition of 10mM or more pyrolic acid, the overall tendency was the same, but the dissolution rate was very fast, and showed solubility of 90% or more in only 40 minutes of sonication.
Claims (16)
Homogenizing porcine myocardium to prepare homogenized porcine myocardium; Heat-treating the homogenized porcine myocardium for 30 minutes to 20-40 ° C .; Cooling the heat treated pork myocardium to 4 ° C .; First sonicating the cooled porcine myocardium at an amplitude of 60 to 80% for 10 minutes; Suspending the sonicated porcine myocardial protein in pyrroline salt at a concentration of 5-20 mM; And second sonicating the suspended myocardial protein at an amplitude of 60 to 80% for 20 to 60 minutes.
Preparing homogenized porcine myocardium by mixing and homogenizing porcine myocardium; Heat-treating the homogenized porcine myocardium for 30 minutes to 20-40 ° C .; Cooling the heat treated pork myocardium to 4 ° C .; First sonicating the cooled porcine myocardium at an amplitude of 60 to 80% for 10 minutes; Suspending the sonicated porcine myocardial protein in pyrroline acid at a concentration of 5-20 mM; And a second sonication of the suspended myocardial protein at an amplitude of 60 to 80% for 20 to 60 minutes.
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003144097A (en) * | 2001-09-03 | 2003-05-20 | Nippon Meat Packers Inc | Water-soluble processed meat product |
| KR20090043104A (en) * | 2007-10-29 | 2009-05-06 | 한국식품연구원 | Method of making myoprotein water-soluble by using sonication |
| KR20100034421A (en) * | 2008-09-24 | 2010-04-01 | 고려대학교 산학협력단 | Surimi-like materials containing whey protein and manufacturing methods of the same |
Non-Patent Citations (2)
| Title |
|---|
| 한국축산식품학회지 제27권제4호(초음파처리가 노계 가슴육 근원섬유단백질의 수용화에 미치는 영향) * |
| 한국축산식품학회지 제27권제4호(초음파처리가 노계 가슴육 근원섬유단백질의 수용화에 미치는 영향)* |
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| KR20120105239A (en) | 2012-09-25 |
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