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Applied and Environmental Microbiology, June 2004, p. 3205-3212, Vol. 70, No. 6
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.6.3205-3212.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

In Vivo Immobilization of Fusion Proteins on Bioplastics by the Novel Tag BioF

Cristina Moldes, Pedro García, José L. García, and María A. Prieto*

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

Received 8 October 2003/ Accepted 31 January 2004

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-ß-galactosidase. This is the first example of an active bioplastic consisting of a biodegradable matrix carrying an active enzyme.

* Corresponding author. Mailing address: Department of Molecular Microbiology, Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040 Madrid, Spain. Phone: 34-918373112. Fax: 34-915360432. E-mail: auxi{at}cib.csic.es.

Applied and Environmental Microbiology, June 2004, p. 3205-3212, Vol. 70, No. 6
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.6.3205-3212.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Jendrossek, D. (2009). Polyhydroxyalkanoate Granules Are Complex Subcellular Organelles (Carbonosomes). J. Bacteriol. 191: 3195-3202 [Full Text]  
  • Neumann, L., Spinozzi, F., Sinibaldi, R., Rustichelli, F., Potter, M., Steinbuchel, A. (2008). Binding of the Major Phasin, PhaP1, from Ralstonia eutropha H16 to Poly(3-Hydroxybutyrate) Granules. J. Bacteriol. 190: 2911-2919 [Abstract] [Full Text]  
  • de Eugenio, L. I., Garcia, P., Luengo, J. M., Sanz, J. M., Roman, J. S., Garcia, J. L., Prieto, M. A. (2007). Biochemical Evidence That phaZ Gene Encodes a Specific Intracellular Medium Chain Length Polyhydroxyalkanoate Depolymerase in Pseudomonas putida KT2442: CHARACTERIZATION OF A PARADIGMATIC ENZYME. J. Biol. Chem. 282: 4951-4962 [Abstract] [Full Text]  
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