title: The non-haem chloroperoxidase from Pseudomonas fluorescens and its relationship to pyrrolnitrin biosynthesis.
abstract: The non-haem chloroperoxidase gene (cpoF) from the pyrrolnitrin producer Pseudomonas fluorescens BL914 was cloned using an oligonucleotide derived from part of the N-terminal amino acid sequence of chloroperoxidase (CPO-P) from Pseudomonas pyrrocina as a probe. Based on the overexpression of cpoF in Escherichia coli and the stability of CPO-F against higher temperatures and proteases, the enzyme was purified to homogeneity. Partial characterization of the enzyme showed that it belongs to the class of bacterial non-haem CPOs. To investigate the role of CPO-F in pyrrolnitrin biosynthesis, the cpoF gene was inactivated by insertion of a kanamycin cassette. Exchange of the chromosomal cpoF gene against the disrupted copy had no influence on pyrrolnitrin production demonstrating that CPO-F was not involved in pyrrolnitrin biosynthesis.
authors: Institut für Mikrobiologie, Universität Hohenheim, Stuttgart, Germany.
authors: S
authors: S
authors: Kirner
authors: S
authors: S
authors: Krauss
authors: G
authors: G
authors: Sury
authors: S T
authors: ST
authors: Lam
authors: J M
authors: JM
authors: Ligon
authors: K H
authors: KH
authors: van Pée
mesh_terms: D002711:Chloride Peroxidase / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D004926:Escherichia coli; D007700:Kinetics; D016296:Mutagenesis; D011551:Pseudomonas fluorescens / Q000201:enzymology*; D011764:Pyrrolnitrin / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D017384:Sequence Deletion
chemical_list: D011994:Recombinant Proteins; D002711:Chloride Peroxidase; D011764:Pyrrolnitrin