13C-NMR 분광법을 이용하여 SAM 합성효소 반응의 정량 분석법 개발

박정호; 이수진; Jeong Ho Park; Su Jin Lee

Journal of the Korean Industrial and Engineering Chemistry, Vol.13, No.5, 476-481, August, 2002

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13C-NMR 분광법을 이용하여 SAM 합성효소 반응의 정량 분석법 개발

Development of a Simple Quantitative Method for SAM Synthetase Reaction by using 13C-NMR Spectroscopy

박정호^†, 이수진

국립한밭대학교 공업화학과, 대전광역시 유성구 덕명동 305-719

Jeong Ho Park^†, and Su Jin Lee

Department of Chemical Technology, Hanbat National University, Daejeon 305-719, Korea

E-mail:

초록

13C-NMR 분광법을 이용하여 pTsONa, 13CH3-Met과 13CH3-SAM에서 나타나는 특정피크와 내부 기준 피크(100 mM TrisㆍHCl)의 농도에 따른 상대 비를 구하는 검정곡선을 통해 10%의 오차 내에서 이들의 정량 분석을 할 수 있었다. 이러한 새로운 정량 분석방법을 효소 반응 에세이로 이용하여 시간에 따른 SAM 합성효소 반응과정의 정량 분석을 수행하였다.

A simple method of estimating the concentration of pTsONa, 13CH3-Met, and 13CH3-SAM was developed based on the correlation between the concentration of each chemical and the relative intensity of 13C-NMR peak. An internal standard was used and the error range was estimated less than 10%. This new essay method was applied to the quantitative enzyme-assay at the time course experiment of SAM synthetase-enzymatic reaction.

Keywords:13C-NMR spectroscopy;SAM synthetase;enzymatic reaction;quantitative assay

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[1] Markham GD, Parkin DW, Mentch F, Schramm VL, J. Biol. Chem., 262, 5609 (1987)

[2] Markham GD, Parasis JD, Gatmaitan J, E. Coli. J. Biol. Chem., 259, 14505 (1984)

[3] Markam GD, J. Biol. Chem., 256, 1903 (1981)

[4] Zhang C, Markham GD, Brutto RL, Biochemistry, 32, 9866 (1993)

[5] Tabor CW, Tabor H, Adv. Enzyme., 56, 252 (1984)

[6] Tabor CH, Tabor H, Microbiol. Rev., 49, 81 (1985)

[7] Satoh S, Yang SF, Plant Physiol., 91, 1036 (1989)

[8] Slany RK, Bosl M, Crain PF, Kersten H, Biochemistry, 32, 7811 (1993)

[9] Frey M, Royhe M, Wagner AFV, Knappe J, J. Biol. Chem., 269, 12432 (1994)

[10] Moss M, Frey PA, J. Biol. Chem., 262, 14859 (1987)

[11] Kilogore JL, Aberhart DJ, J. Chem. Soc.-Perkin Trans. 1, 79 (1991)

[12] Harber J, Elissson R, Pontis E, Ballinger MD, Reichard P, J. Biol. Chem., 267, 25548 (1992)

[13] Pascale RM, Marras V, Simile MM, Daino L, Pinna G, Bennati S, Carta M, Seddaiu MA, Massarelli G, Feo F, Cancer Res., 52, 4979 (1992)

[14] Kagan BL, Shltzer DL, Rosenlicht N, Gerner H, Am. J. Psychiatry, 147(5), 591 (1990)

[15] Metz J, Nutrition Rev., 51, 12 (1993)

[16] Mitsui KI, Teraoka H, Tsukada K, J. Biol. Chem., 263, 23 (1988)

[17] Chou TC, Lombardini JB, Biochim. Biophy. Acta, 276, 399 (1972)

[18] Lee BH, Choi BY, Ryu GS, Lee ES, Kang KJ, Hwang DY, Hong ND, Korean J. Pharmacogn., 28(4), 167 (1997)

[19] Matos JR, Raushel FM, Wong CH, Biotechnol. Appl. Biochem., 9, 39 (1987)

[20] Morana A, Lernia ID, Carteni M, Rosa RD, Rosa MD, Int. J. Pharm., 194, 61 (2000)

[21] Revelle LK, d'Avignon DA, Reepmeyer JC, Zerfing R, J. AOAC Int., 78, 353 (1995)

[22] Warren MJ, Roessner CA, Ozaki SI, Stolowich NJ, Santander PJ, Scott AI, Biochemistry, 31, 603 (1992)