%0 Journal Article
%A Bello-Gil, Daniel
%A Maestro García-Donas, María Beatriz
%A Fonseca, Jennifer
%A Dinjaski, Nina
%A Prieto, Auxiliadora
%A Sanz, Jesús 
%T Poly-3-Hydroxybutyrate Functionalization with BioF-Tagged Recombinant Proteins
%D 2018
%@ 0099-2240
%U https://hdl.handle.net/20.500.14352/92515.2
%X 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–β-galactosidase, containing the choline-binding module C-LytA and the β-galactosidase enzyme, respectively). The protein-biopolyester interaction was strong and stable at a wide range of pHs and temperatures, and the bound protein was highly protected from self-degradation, while the binding strength could be modulated by coating with amphiphilic compounds. Finally, BioF–β-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.
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