f Formation of 4-hydroxybenzoate in Escherichia coli: characterization of the ubiC gene and its encoded enzyme chorismate pyruvate-lyase
- Authors: Marion Siebert, Klaus Severin, Lutz Heide1
- 1Author for correspondence: Lutz Heide: Tel: +49 761 2032804. Fax: +49 791 2032803. e-mail: firstname.lastname@example.org
- Microbiology, April 1994 140: 897-904, doi: 10.1099/00221287-140-4-897
- Subject: Biochemistry
- Published Online:
Chorismate pyruvate-lyase from Escherichia coli converts chorismate to 4-hydroxybenzoate. The enzyme was enriched 3000-fold by overexpression and chromatographic purification. It has an apparent K m value for chorismate of 6·1 μM and an isoelectric point of pH 6·45. The enzyme activity did not require metal cofactors. Promoter sequences in the 5′ flanking sequences of the ubiCA operon were localized by transcription and translation of active chorismate pyruvate-lyase in vitro from different PCR fragments. Sequencing of the ubiC gene of the mutant strain AN244 revealed a G→A transition resulting in a change from glutamic acid to lysine. A feeding experiment with [1,7-13C2]shikimate confirmed the chorismate pyruvate-lyase as the sole enzymic source of 4-hydroxybenzoate in vivo.
© Society for General Microbiology 1994 | Published by the Microbiology Society
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