여지전기영동조에서 유기용매를 이용한 단백질이용

홍현철; 류우권; 박중곤; 송방호

HWAHAK KONGHAK, Vol.29, No.4, 457-462, August, 1991

Export Citation

여지전기영동조에서 유기용매를 이용한 단백질이용

Protein Transport by the Paper Electrophoresis in Organic Solvent Medium

홍현철, 류우권, 박중곤, 송방호

초록

생물고분자의 분석에 이용되어온 전기영동법은 비교적 장치가 간단하고 값이 싸서 주로 임상 분야에 널리 사용되어 왔다. 그러나 완충용액을 이용한 전기영동에서는 증발, 열효과, 완충용액의 농도구배 및 영동조 양단의 pH 변화 등과 같은 제한이 따르게 된다. 본 연구에서는 기존의 전기영동조에서 완충용액대신 유기용매를 매질로 사용하였으며, 유기용매 내에서의 전기분자 추진에 의하여 lysozyme, α-amylase, pepsin을 이동시켰다. 단백질의 이동도는 완충용액을 이용한 전기영동의 경우 보다 약 2배 가량 빠르게 나타났으며 높은 전위 구배에 의한 열발생도 무시할 수 있을 정도로 감소하였다. 미수용매계에서의 단백질의 실활은 α-amylase의 역가 측정 실험으로 검토하였다. 미수용매계내에서 전분을 분해할 때의 α-amylase의 역가 측정 실험결과 1시간 이내에서는 유기용매로 인한 효소의 실활은 없는 것으로 나타났다.

Electrophoresis method has been widely used to analyze proteim molecules in the clinical and biological field because of low cost and simple equipment. However, there are some problems in the application of the conventional electrophoresis such as evaportation of the buffer solution, formation of concentration gradient, pH changes and heat generation during the operation. To reduce the heat generation we used an organic solvent as a medium instead of buffer solution for the electromolecular propulsion, which has been previously used for the separation of dyes. We could increase the rate of movements of lysozyme, α-amylase and pepsin and reduce the heat generation in the electromolecular propulsion. The effect of the organic solvent on the activity of the protein was verified to be negligible by the experimental analysis of the activity of α-amylase.

[References]

Moore DH, "Electrophoresis in Physical Techniques in Biological Research," 2nd ed., Academic Press, New York, 121 (1968)
O'Farrel PH, J. Biol. Chem., 250, 4007 (1975)
Mullon C, Mason N, Spark R, Biotechnol. Bioeng., 30, 123 (1987)
Yamamoto S, Nomura M, Sano Y, AIChE J., 39(9), 1426 (1987)
Ruckenstein E, Lesins V, Biotechnol. Bioeng., 28, 432 (1986)
Weber K, Osborn M, J. Biol. Chem., 244, 4406 (1969)
DeWachter R, Fiers W, Methods Enzymol., 21, 167 (1971)
Longsworth LG, Maclnnes DA, J. Am. Chem. Soc., 62, 705 (1940)
Larson DL, Feinberg R, Science, 120, 426 (1954)
Nielsen LE, Kirkwood JG, J. Am. Chem. Sci., 68, 181 (1946)
Landstein K, Longsworth LG, vanderScheer J, Science, 88, 83 (1938)
Durrum EL, J. Am. Chem. Soc., 72, 2939 (1950)
Durrum EL, J. Am. Chem. Soc., 73, 4875 (1951)
Durrum EL, Paul MH, Smith EB, Science, 116, 428 (1952)
Common RH, Mckinley WP, Maw WA, Science, 118, 86 (1953)
Block RJ, Durrum EL, Zweig G, "A Manual of Paper Chromatography and Paper Electrophoresis," 2nd ed., Academic Press, Inc. (1958)
Haber N, Biochem. J., 79, 272 (1982)
Yoo YJ, Hong J, Hatch RT, Biotechnol. Bioeng., 30, 147 (1987)

[Referenced By]

[1] Moore DH, "Electrophoresis in Physical Techniques in Biological Research," 2nd ed., Academic Press, New York, 121 (1968)

[2] O'Farrel PH, J. Biol. Chem., 250, 4007 (1975)

[3] Mullon C, Mason N, Spark R, Biotechnol. Bioeng., 30, 123 (1987)

[4] Yamamoto S, Nomura M, Sano Y, AIChE J., 39(9), 1426 (1987)

[5] Ruckenstein E, Lesins V, Biotechnol. Bioeng., 28, 432 (1986)

[6] Weber K, Osborn M, J. Biol. Chem., 244, 4406 (1969)

[7] DeWachter R, Fiers W, Methods Enzymol., 21, 167 (1971)

[8] Longsworth LG, Maclnnes DA, J. Am. Chem. Soc., 62, 705 (1940)

[9] Larson DL, Feinberg R, Science, 120, 426 (1954)

[10] Nielsen LE, Kirkwood JG, J. Am. Chem. Sci., 68, 181 (1946)

[11] Landstein K, Longsworth LG, vanderScheer J, Science, 88, 83 (1938)

[12] Durrum EL, J. Am. Chem. Soc., 72, 2939 (1950)

[13] Durrum EL, J. Am. Chem. Soc., 73, 4875 (1951)

[14] Durrum EL, Paul MH, Smith EB, Science, 116, 428 (1952)

[15] Common RH, Mckinley WP, Maw WA, Science, 118, 86 (1953)

[16] Block RJ, Durrum EL, Zweig G, "A Manual of Paper Chromatography and Paper Electrophoresis," 2nd ed., Academic Press, Inc. (1958)

[17] Haber N, Biochem. J., 79, 272 (1982)

[18] Yoo YJ, Hong J, Hatch RT, Biotechnol. Bioeng., 30, 147 (1987)