2026-06-08T08:27:22Zhttps://docta.ucm.es/rest/oai/requestoai:docta.ucm.es:20.500.14352/63232024-10-02T14:05:23Zcom_20.500.14352_14col_20.500.14352_15
Docta Complutense
author
Mato, Aranzazu
author
Blanco, Francisco G.
author
Maestro García-Donas, María Beatriz
author
Sanz, Jesús M.
author
Pérez-Gil, Jesús
author
Prieto, M. Auxiliadora
2023-06-16T15:19:03Z
2023-06-16T15:19:03Z
2020-06-02
0099-2240, ESSN: 1098-5336
10.1128/AEM.00570-20
https://hdl.handle.net/20.500.14352/6323
https://aem.asm.org/content/86/12/e00570-20
Phasin PhaF from Pseudomonas putida consists of a modular protein whose N-terminal domain (BioF) has been demonstrated to be responsible for binding to the polyhydroxyalkanoate (PHA) granule. BioF has been exploited for biotechnological purposes as an affinity tag in the functionalization of PHA beads with fusion proteins both in vivo and in vitro. The structural model of this domain suggests an amphipathic -helical conformation with the hydrophobic residues facing the PHA granule. In this work, we analyzed the mean hydrophobicity and the hydrophobic moment of the native BioF tag to rationally design shorter versions that maintain affinity for the granule. Hybrid proteins containing the green fluorescent protein (GFP) fused to the BioF derivatives were studied for in vivo localization on PHA, stability on the surface of the PHA granule against pH, temperature, and ionic strength, and their possible influence on PHA synthesis. Based on the results obtained, a minimized BioF tag for PHA functionalization has been proposed (MinP) that retains similar binding properties but possesses an attractive biotechnological potential derived from its reduced size. The MinP tag was further validated by analyzing the functionality and stability of the fusion proteins MinP–-galactosidase and MinP-CueO from Escherichia coli.
eng
Atribución 3.0 España
Dissecting the Polyhydroxyalkanoate-binding domain of the PhaF Phasin: rational design of a minimized affinity tag
journal article
URL
https://docta.ucm.es/bitstreams/6719cc7b-600e-49ee-8e2d-88bf13179948/download
File
MD5
d4ad95dd2acbf07c31af009bff0687b2
2399416
application/pdf
Mato, Aranzazu et al. 2020. Dissecting the polyhydroxyalkanoate-Binnding.....pdf