Identification of a novel 4-hydroxyphenylpyruvate dioxygenase from the soil metagenome.
Biochem Biophys Res Commun
; 370(2): 322-6, 2008 May 30.
Article
em En
| MEDLINE
| ID: mdl-18381065
ABSTRACT
4-Hydroxyphenylpyruvate dioxygenase (HPPD) is a Fe(II)-dependent, non-heme oxygenase that converts 4-hydroxyphenylpyruvate to homogentisate. Essential cofactors, such as plastoquinone and tocopherol, are produced by HPPD-dependent anabolic pathways in plants. To isolate a novel hppd using culture-independent method, a cosmid metagenomic library was constructed from soil in Korea. Screening of Escherichia coli metagenomic libraries led to the identification of a positive clone, YS103B, producing dark brown pigment in Luria-Bertani medium supplemented with l-tyrosine. In vitro transposon mutagenesis of YS103B showed that the 1.3kb insert was sufficient to produce the hemolytic brown pigment. Sequence analysis of YS103B disclosed one open reading frame encoding a 41.4kDa protein with the well-conserved prokaryotic oxygenase motif of the HPPD family of enzymes. The HPPD-specific beta-triketone herbicide, sulcotrione, inhibited YS103B pigmentation. The recombinant protein expressed in E. coli generated homogentisic acid. Thus, we present the successful heterologous expression of a previously uncharacterized hppd gene from an uncultured soil bacterium.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Microbiologia do Solo
/
Proteínas de Bactérias
/
4-Hidroxifenilpiruvato Dioxigenase
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Ano de publicação:
2008
Tipo de documento:
Article