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Poly-3-hydroxybutyrate functionalization with BioF-tagged recombinant proteins

Otros títulosProtein Functionalization of PHB
AutorBello-Gil, Daniel; Maestro, Beatriz; Fonseca, Jennifer; Dinjaski, Nina ; Prieto, María Auxiliadora ; Sanz, Jesús M.
Palabras clavePHB
Affinity tag
Protein immobilization
Polyhydro xyalkanoates
Fecha de publicación1-dic-2017
EditorAmerican Society for Microbiology
CitaciónApplied and Environmental Microbiology (2017)
ResumenPolyhydroxyalkanoates (PHAs) are biodegradable polyesters that accumulate in the cytoplasm of certain bacteria. One promising biotechnological application of these biopolymers contemplates its utilization as supports for protein immobilization. Here, the PHA-binding domain of the Pseudomonas putida KT2440 PhaF phasin (BioF polypeptide) was investigated as affinity tag for 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). Protein-biopolyester interaction was strong and stable in a wide range of pH and temperature, and the bound protein was highly protected from self-degradation, while binding strength could be modulated upon particle coating with amphiphilic compounds. Finally, a mini-bioreactor with immobilized BioF- β-galactosidase displayed a very stable enzymatic activity after several continuous activity+washing cycles. Our results demonstrate the potentialities of PHA and the BioF tag for the construction of novel bioactive materials.
Importance: Our results confirm the biotechnological potential of the BioF affinity tag as a versatile tool for functionalizing PHA supports with recombinant proteins, leading to novel bioactive materials. The BioF tag wide substrate range presumably allows protein immobilization in vitro to virtually all natural PHAs as well as blends, copolymers or artificial, chemically-modified derivatives with novel physical-chemical properties. Moreover, the strength of protein adsorption may be easily modulated by varying the coating of the support, providing new perspectives for the engineering of bioactive materials that require a tight control of protein loading.
Descripción44 p.-6 fig.-2 tab.-5 fig.supl.
Versión del editorhttp://dx.doi.org/10.1128/AEM.02595-17
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