Effects of methanol on the catalytic properties of porcine pancreatic lipase

Hyun Park, Ki Seog Lee, Young Min Chi, Seung Weon Jeong

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27 Citations (Scopus)

Abstract

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 α-benzyloxycarbonyl-L-lysine p-nitrophenyl ester at 0°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* 5.5 and 0°C. By monitoring the intrinsic fluorescence and absorbance, no evidence for structural changes by methanol was observed.

Original languageEnglish
Pages (from-to)296-301
Number of pages6
JournalJournal of Microbiology and Biotechnology
Volume15
Issue number2
Publication statusPublished - 2005 Apr 1

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Keywords

  • Hydrolysis
  • Lipase
  • Methanol solvent
  • Methanolysis
  • Rate-limiting step

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Microbiology
  • Bioengineering

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