title: Function-Based Metagenomic Library Screening and Heterologous Expression Strategy for Genes Encoding Phosphatase Activity.
abstract: The release of phosphate from inorganic and organic phosphorus compounds can be mediated enzymatically. Phosphate-releasing enzymes, comprising acid and alkaline phosphatases, are recognized as useful biocatalysts in applications such as plant and animal nutrition, bioremediation and diagnostic analysis. Metagenomic approaches provide access to novel phosphatase-encoding genes. Here, we describe a function-based screening approach for rapid identification of genes conferring phosphatase activity from small-insert and large-insert metagenomic libraries derived from various environments. This approach bears the potential for discovery of entirely novel phosphatase families or subfamilies and members of known enzyme classes hydrolyzing phosphomonoester bonds such as phytases. In addition, we provide a strategy for efficient heterologous phosphatase gene expression.
authors.affiliation: Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Grisebachstr. 8, 37077, Göttingen, Germany.
authors.forename: Genis A Castillo
authors.initials: GA
authors.lastname: Villamizar
authors.affiliation: Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Grisebachstr. 8, 37077, Göttingen, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: Nacke
authors.affiliation: Göttingen Genomics Laboratory, Department of Genomic and AppliedMicrobiology, Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Grisebachstr. 8, 37077, Göttingen, Germany. rdaniel@gwdg.de.
authors.forename: Rolf
authors.initials: R
authors.lastname: Daniel
mesh_terms: D003001:Cloning, Molecular; D004789:Enzyme Activation; D057075:Enzyme Assays / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D015723:Gene Library; D054892:Metagenome; D056186:Metagenomics / Q000379:methods*; D016254:Mutagenesis, Insertional; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics* / Q000378:metabolism*
chemical_list: D010744:Phosphoric Monoester Hydrolases
keywords: Function-based screening; Metagenomic libraries; Phosphatases; Phosphorus; Phytases