2026-06-27T12:38:26Zhttps://docta.ucm.es/rest/oai/request

oai:docta.ucm.es:20.500.14352/92515.22024-09-03T00:08:39Zcom_20.500.14352_14col_20.500.14352_15

Bello-Gil, Daniel Maestro García-Donas, María Beatriz Fonseca, Jennifer Dinjaski, Nina Prieto, Auxiliadora Sanz, Jesús 2024-01-23T15:01:43Z 2024-01-11T11:50:03Z 2018 Bello-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.02595-17 0099-2240 10.1128/AEM.02595-17 https://hdl.handle.net/20.500.14352/92515.2 1098-5336 https://doi.org/10.1128/AEM.02595-17 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. eng open access Poly-3-Hydroxybutyrate Functionalization with BioF-Tagged Recombinant Proteins journal article