Please use this identifier to cite or link to this item: https://hdl.handle.net/11000/4586

Poly-3-Hydroxybutyrate Functionalization with BioF-Tagged Recombinant Proteins


Thumbnail

View/Open:
 1-aem02595-17_Proof.pdf
2,18 MB
Adobe PDF
Share:
Title:
Poly-3-Hydroxybutyrate Functionalization with BioF-Tagged Recombinant Proteins
Authors:
Bello Gil, Daniel
Maestro García-Donas, Beatriz
Fonseca, Jennifer
Dinjaski, Nina
Prieto, M. Auxiliadora
Sanz, Jesús M.
Department:
Departamentos de la UMH::Bioquímica y Biología Molecular
Issue Date:
2017-11-01
Abstract:
Polyhydroxyalkanoates (PHAs) are biodegradable polyesters that accumulate in the cytoplasm of certain bacteria. One promising biotechnological application utilizes these biopolymers as supports for protein immobilization. Here, the PHA-binding domain of the Pseudomonas putida KT2440 PhaF phasin (BioF polypeptide) was investigated as an affinity tag for the in vitro functionalization of poly-3- hydroxybutyrate (PHB) particles with recombinant proteins, namely, full-length PhaF and two fusion proteins tagged to BioF (BioF–C-LytA and BioF–_-galactosidase, containing the choline-binding module C-LytA and the _-galactosidase enzyme, respectively). The protein-biopolyester interaction was strong and stable in a wide range of pHs and temperatures, and the bound protein was highly protected from selfdegradation, while the binding strength could be modulated by coating with amphiphilic compounds. Finally, BioF–_-galactosidase displayed very stable enzymatic activity after several continuous activity-plus-washing cycles when immobilized in a minibioreactor. Our results demonstrate the potentialities of PHA and the BioF tag for the construction of novel bioactive materials.
Keywords/Subjects:
PHB
phasins
affinity tag
protein immobilization
polyhydroxyalkanoates
Type of document:
application/pdf
Access rights:
info:eu-repo/semantics/openAccess
Appears in Collections:
Instituto de Biología Molecular y Celular



Creative Commons ???jsp.display-item.text9???