[논문]Effects of Methanol on the Catalytic Properties of Porcine Pancreatic Lipase

PARK HYUN; LEE KI SEOG; CHI YOUNG MIN; JEONG SEUNG WEON

Effects of Methanol on the Catalytic Properties of Porcine Pancreatic Lipase 원문보기

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

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)

Abstract ▼ AI-Helper

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.

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