Open AccessMonitoring Key Reactions in Degradation of Chloroaromatics by In Situ 1 H Nuclear Magnetic Resonance: Solution Structures of Metabolites Formed from cis -Dienelactone
Author(s) -
Dietmar H. Pieper,
Katrin Pollmann,
Patricia Nikodem,
Bernardo González,
Victor Wray
Publication year - 2002
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.184.5.1466-1470.2002
Subject(s) - decarboxylation , enzyme , nuclear magnetic resonance spectroscopy , in situ , biology , stereochemistry , nuclear magnetic resonance , biochemistry , chemistry , organic chemistry , catalysis , physics
A1 H nuclear magnetic resonance (1 H NMR) assay was used to study the enzymatic transformation ofcis -dienelactone, a central intermediate in the degradation of chloroaromatics. It was shown that the product of thecis -dienelactone hydrolase reaction is maleylacetate, in which there is no evidence for the formation of 3-hydroxymuconate. Under acidic conditions, the product structure was 4-carboxymethyl-4-hydroxybut-2-en-4-olide. Maleylacetate was transformed by maleylacetate reductase into 3-oxoadipate, a reaction competing with spontaneous decarboxylation intocis -acetylacrylate. One-dimensional1 H NMR in1 H2 O could thus be shown to be an excellent noninvasive tool for monitoring enzyme activities and assessing the solution structure of substrates and products.