Inspection of an acyl-enzyme intermediate in a lipase reaction by gas chromatography-mass spectrometry and modelling of the reaction mechanism

Masaya Kawase, Kenji Sonomoto, Atsuo Tanaka

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

An acyl-enzyme intermediate proposed in the reaction mechanism of lipase was inspected by the exchange of oxygen between substrate (oleic acid) and solvent (18O-labelled water). Gas chromatography-mass spectrometry analysis supported the formation of an acyl-enzyme intermediate in the reaction mechanism through the observed incorporation of 18O into oleic acid. The incorporation did not occur in the absence of the lipase. When Ser residues were modified with diisopropylfluorophosphate, the activity of lipase OF 360 was markedly decreased. Photooxidation of His residues also resulted in a decrease in the activity of the lipase. Chemical modification studies suggested the existence of a charge relay system (Ser-His-Asp/Glu) in the active site. Based on these results, a model of the active site and reaction mechanism of the lipase are presented.

Original languageEnglish
Pages (from-to)43-50
Number of pages8
JournalBiocatalysis and Biotransformation
Volume6
Issue number1
DOIs
Publication statusPublished - Jan 1 1992
Externally publishedYes

Fingerprint

Lipases
Lipase
Gas chromatography
Gas Chromatography-Mass Spectrometry
Mass spectrometry
Enzymes
Inspection
Oleic acid
Oleic Acid
Catalytic Domain
Isoflurophate
Photooxidation
Chemical modification
Oxygen
Water
Substrates

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Catalysis
  • Biochemistry

Cite this

Inspection of an acyl-enzyme intermediate in a lipase reaction by gas chromatography-mass spectrometry and modelling of the reaction mechanism. / Kawase, Masaya; Sonomoto, Kenji; Tanaka, Atsuo.

In: Biocatalysis and Biotransformation, Vol. 6, No. 1, 01.01.1992, p. 43-50.

Research output: Contribution to journalArticle

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