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Appl Environ Microbiol. 2004 Jun;70(6):3205-12.

In vivo immobilization of fusion proteins on bioplastics by the novel tag BioF.

Author information

1
Department of Molecular Microbiology, Centro de Investigaciones Biológicas, CSIC, 28040 Madrid, Spain.

Abstract

A new protein immobilization and purification system has been developed based on the use of polyhydroxyalkanoates (PHAs, or bioplastics), which are biodegradable polymers accumulated as reserve granules in the cytoplasm of certain bacteria. The N-terminal domain of the PhaF phasin (a PHA-granule-associated protein) from Pseudomonas putida GPo1 was used as a polypeptide tag (BioF) to anchor fusion proteins to PHAs. This tag provides a novel way to immobilize proteins in vivo by using bioplastics as supports. The granules carrying the BioF fusion proteins can be isolated by a simple centrifugation step and used directly for some applications. Moreover, when required, a practically pure preparation of the soluble BioF fusion protein can be obtained by a mild detergent treatment of the granule. The efficiency of this system has been demonstrated by constructing two BioF fusion products, including a functional BioF-beta-galactosidase. This is the first example of an active bioplastic consisting of a biodegradable matrix carrying an active enzyme.

PMID:
15184113
PMCID:
PMC427747
DOI:
10.1128/AEM.70.6.3205-3212.2004
[Indexed for MEDLINE]
Free PMC Article
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