Poly-3-Hydroxybutyrate Functionalization with BioF-Tagged Recombinant Proteins

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 KT2440 PhaF phasin (BioF polypeptide) was...

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Bibliographic Details
Published in:Applied and environmental microbiology 2018-02, Vol.84 (4)
Main Authors: Bello-Gil, Daniel, Maestro, Beatriz, Fonseca, Jennifer, Dinjaski, Nina, Prieto, M Auxiliadora, Sanz, Jesús M
Format: Article
Language:English
Subjects:
Bacteria
Bacterial Proteins - metabolism
Binding
Biocompatibility
Biodegradability
Biodegradation
Biomedical materials
Biopolymers
Biotechnology - methods
Choline
Cytoplasm
Enzymatic activity
Enzymology and Protein Engineering
Galactosidase
Hydrogen-Ion Concentration
Hydroxybutyrates - metabolism
Immobilization
Immobilized Proteins
Plant Lectins - chemistry
Polyester resins
Polyesters
Polyesters - metabolism
Polyhydroxyalkanoates
Polyhydroxyalkanoates - metabolism
Polyhydroxybutyrate
Polyhydroxybutyric acid
Polymers
Protective coatings
Proteins
Pseudomonas putida
Pseudomonas putida - metabolism
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - metabolism
Recombinant Proteins - metabolism
β-Galactosidase
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Summary: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 KT2440 PhaF phasin (BioF polypeptide) was investigated as an affinity tag for the 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. 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 wide substrate range of the BioF tag presumably enables protein immobilization of virtually all natural PHAs as well as blends, copolymers, or artificial chemically modified derivatives with novel physicochemical 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.
ISSN:0099-2240
1098-5336
DOI:10.1128/AEM.02595-17