2026-06-01T02:23:31Zhttps://docta.ucm.es/rest/oai/requestoai:docta.ucm.es:20.500.14352/92515.22024-09-03T00:08:39Zcom_20.500.14352_14col_20.500.14352_15
Poly-3-Hydroxybutyrate Functionalization with BioF-Tagged Recombinant Proteins
Bello-Gil, Daniel
Maestro García-Donas, María Beatriz
Fonseca, Jennifer
Dinjaski, Nina
Prieto, Auxiliadora
Sanz, Jesús
577.112
579.8
PHB,affinitytag,proteinimmobilization,polyhydroxyalkanoates
Phasins
Affinity tag
Protein immobilization
Polyhydroxyalkanoates
Bioquímica (Química)
Microbiología (Biología)
2403 Bioquímica
2414 Microbiología
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.
Ministerio de Economía y Competitividad (España)
Comunidad de Madrid
Depto. de Bioquímica y Biología Molecular
Fac. de Ciencias Biológicas
TRUE
pub
2024-01-23T15:01:43Z
2024-01-11T11:50:03Z
2024-01-23T15:01:43Z
2018
journal article
VoR
https://hdl.handle.net/20.500.14352/92515.2
0099-2240
10.1128/AEM.02595-17
1098-5336
eng
(BFU2010-17824, BIO2013-44878-R, BIO2013-47684-R, and BIO2016-79323-R)
(NanoBIOSOMAS2014733/MlT-2807)
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
open access
application/pdf