본문 바로가기
HOME> 논문 > 논문 검색상세

논문 상세정보

Journal of microbiology and biotechnology v.15 no.2, 2005년, pp.296 - 301   피인용횟수: 2

Effects of Methanol on the Catalytic Properties of Porcine Pancreatic Lipase

PARK HYUN    (Korea Polar Research institute, Korea Ocean Research and Development Institute   ); LEE KI SEOG    (Division of Biotechnology and Genetic Engineering, Korea University   ); CHI YOUNG MIN    (Division of Biotechnology and Genetic Engineering, Korea University   ); JEONG SEUNG WEON    (Korea Food Research institute  );
  • 초록

    The effect of aqueous methanol on the catalytic properties of porcine pancreatic lipase has been investigated. The k $_{CAT}$ , values for the hydrolysis of N $^{alpha}$ -benzyloxycarbonyl-L­lysine p-nitrophenyl ester at 0 $^{circ}$ C increased in a linear manner with increasing methanol concentration. However, the K $_{M}$ values were not influenced at methanol concentrations lower than $30\%$ and then began to increase at higher concentrations in an exponential fashion. Based on product analysis, the increase in k $_{CAT}$ , with increasing methanol concentration can be accounted for by nucleophilic competition of methanol for the acyl enzyme intermediate, indicating that the rate-limiting step of the porcine pancreatic lipase-catalyzed reaction is deacylation under current experimental conditions. The exponential increase in K $_{M}$ at methanol concentrations higher than $30\%$ is attributed to the hydrophobic partitioning effect on substrate binding. There was no loss of lipase activity over a 4 h period in $60\%$ methanol concentration at pH $^{circ}$ 5.5 and 0 $^{circ}$ C. By monitoring the intrinsic fluorescence and absorbance, no evidence for structural changes by methanol was observed.


  • 참고문헌 (22)

    1. Burdette, R. A. and D. M. Quinn. 1986. Interfacial reaction dynamics and acylenzyme mechanism for lipoprotein lipasecatalyzed hydrolysis of lipid p-nitrophenyl ester. J. Biol. Chem. 261: 12016- 12021 
    2. Kazlauskas, R. J. 1994. Elucidating structure-mechanism relationships in lipases: Prospects for predicting and engineering catalytic properties. Trends Biotechnol. 12: 464- 472 
    3. Park, H. and Y. M. Chi. 2001. The enhancement of electro stricti on caused by lowering the solvent dielectric constant leads to the decrease of activation energy in trypsin catalysis. Biochim. Biophys. Acta 1568: 53- 59 
    4. Peters, G. H., S. Toxvaerd, O. H. Olsen, and A. Svendsen. 1997. Computational studies of the activation of lipase and the effect of a hydrophobic environment. Protein Eng. 10: 137- 147 
    5. Petersen, M. T. N., P. Fojan, and S. B. Petersen. 2001. How do lipases and esterases work: The electrostatic contribution. J. Biotechnol. 85: 115- 147 
    6. Brady, L., A. M. Brzozowski, Z. S. Derewenda, E. Dodson, G. Dodson, S. Tolley, J. P. Turkenburg, L. Christiansen, B. Huge-Jensen, L. Norskov, L. Thim, and U. Menge. 1990. A serine protease triad forms the catalytic centre of a tryacylglycerollipase. Nature 343: 767- 770 
    7. Cartwright, S. J. and S. G. Waley. 1987. Cryoenzymology of $\beta$ lactamases. Biochemistry 26: 5329- 5337 
    8. Stevanato, R., B. Mondovi, O. Befani, M. Scarpa, and A. Rigo. 1994. Electrostatic control of oxidative deamination catalyzed by bovine serum amine oxidase. Biochem. J. 299: 317- 320 
    9. Compton, P. D., R. J. Coll, and A. L. Fink. 1986. Effect of methanol cryosolvents on the structural and catalytic properties of bovine trypsin. J. Biol. Chem. 261: 1248-1252 
    10. Affleck, R., C. A Haynes, and D. S. Clark. 1992. Solvent dielectric effects on protein dynamics. Proc. Natl. Acad. Sci. USA 89: 5167-5170 
    11. Fink, A L. 1974. The trypsin-catalyzed hydrolysis of $N^{\alpha}-benzyloxycarbonyl-L-lysine-p-nitrophenyl$ ester in dimethylsulfoxide at subzero temperature. J. Biol. Chem. 249: 5027- 5032 
    12. Lemke, K., M. Lemke, and F. Theil. 1997. A threedimensional predictive active site model for lipase from Pseudomonas cepacia. J. Org. Chem. 63: 6268- 6273 
    13. Zandonella, G., P. Stadler, L. Haalck, F. Spener, F. Paltauf, and A. Hermetter. 1999. Interactions of fluorescent triacylglycerol analogs covalently bound to the active site of a lipase from Rhizopus oryzae. Eur. J. Biochem. 262: 63- 69 
    14. Svendsen, A. 2000. Lipase protein engineering. Biochim. Biophys. Acta 1543: 223- 238 
    15. Villeneuve, P., J. M. Muderhwa, J. Graille, and M. J. Haas. 2000. Customizing lipases for biocatalysis: A survey of chemical, physical and molecular biological approaches. J. Mol. Catal. B: Enzymatic 9: 113- 148 
    16. Hilton, S. and J. T. Buckley. 1991. Studies on the reaction mechanism of a Microbiol Iipase/acyltransferase using chemical modification and site-directed mutagenesis. J. Biol. Chem. 266: 997- 1000 
    17. Paiva, A. L., V. M. Balcao, and F. X. Malcata, 2000. Kinetics and mechanisms of reactions catalyzed by immobilized lipases, Enzyme Microb. Technol. 27: 187- 204 
    18. Stryer, L. 1995. Biochemistry, 4th Ed. pp. 207-210. W. H. Freeman & Company, NY, U.S.A 
    19. Moreau, H., A. Moulin, Y. Gargouri, J. Noel. and R. Verger. 1991. Inactivation of gastric and pancreatic lipases by diethyl p-nitrophenyl phosphate. Biochemistry 30: 1037-1041 
    20. Peters, G. H., D. M. F. van Aalten, O. Edholm, S. Toxvaerd, and R. Bywater. 1996. Dynamics of proteins in different solvent systems: Analysis of essential motion in lipases. Biophys. J. 71: 2245- 2255 
    21. Reyes, H. R. and C. G. Hill. 1993. Kinetic modeling of interesterification reactions catalyzed by immobilized lipase. Biotechnol. Bioengin. 43: 171- 18 
    22. Warshel, A., G. Naray-Szabo, F. Sussman, and J. K. Hwang. 1989. How do serine proteases work? Biochemistry 28: 3629- 3637 
  • 이 논문을 인용한 문헌 (2)

    1. 2007. "" Journal of microbiology and biotechnology, 17(4): 650~654     
    2. 2007. "" Journal of microbiology and biotechnology, 17(6): 1054~1057     

 저자의 다른 논문

  • 이기석 (1)

    1. 2002 "Effect of Pressure on Catalytic Properties of Glutamate Racemase from Aquifex pyrophilus, an Extremophilic Bacteria" Journal of microbiology and biotechnology 12 (1): 149~152    
  • Chi, Young-Min (7)

  • Jeong, Seung-Weon (1)

 활용도 분석

  • 상세보기

    amChart 영역
  • 원문보기

    amChart 영역

원문보기

무료다운로드
유료다운로드

유료 다운로드의 경우 해당 사이트의 정책에 따라 신규 회원가입, 로그인, 유료 구매 등이 필요할 수 있습니다. 해당 사이트에서 발생하는 귀하의 모든 정보활동은 NDSL의 서비스 정책과 무관합니다.

원문복사신청을 하시면, 일부 해외 인쇄학술지의 경우 외국학술지지원센터(FRIC)에서
무료 원문복사 서비스를 제공합니다.

NDSL에서는 해당 원문을 복사서비스하고 있습니다. 위의 원문복사신청 또는 장바구니 담기를 통하여 원문복사서비스 이용이 가능합니다.

이 논문과 함께 출판된 논문 + 더보기