Bello-Gil, DanielMaestro García-Donas, María BeatrizFonseca, JenniferDinjaski, NinaPrieto, AuxiliadoraSanz, Jesús2024-01-232024-01-112024-01-232018Bello-Gil DMaestro BFonseca J, Dinjaski NPrieto MA, Sanz JM 2018. Poly-3-Hydroxybutyrate Functionalization with BioF-Tagged Recombinant Proteins. Appl Environ Microbiol 84:e02595-17. https://doi.org/10.1128/AEM.02595-170099-224010.1128/AEM.02595-17https://hdl.handle.net/20.500.14352/92515.2Polyhydroxyalkanoates (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.engjournal article1098-5336https://doi.org/10.1128/AEM.02595-17open access577.112579.8PHB,affinitytag,proteinimmobilization,polyhydroxyalkanoatesPhasinsAffinity tagProtein immobilizationPolyhydroxyalkanoatesBioquímica (Química)Microbiología (Biología)2403 Bioquímica2414 Microbiología