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Updated: Jan 20, 2026

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Exploring phasin-polyhydroxyalkanoate interactions through in vivo and in vitro binding assays.

Maria-Tsampika Manoli1, Paula Llamas1, Francisco G Blanco1

  • 1Polymer Biotechnology Group, Department of Biotechnology, Margarita Salas Center for Biological Research (CIB-CSIC), Madrid, Spain.

International Journal of Biological Macromolecules
|January 18, 2026
PubMed
Summary

Two bacterial proteins, PhaP1 and PhaI, influence polyhydroxyalkanoates (PHA) granule formation and surface properties. Their distinct binding affinities for different PHA types offer potential for targeted bio-based applications.

Keywords:
PhasinsPolyhydroxyalkanoatesPseudomonas putida

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Area of Science:

  • Biotechnology and Synthetic Biology
  • Polymer Science and Engineering
  • Microbial Physiology

Background:

  • Polyhydroxyalkanoates (PHAs) are biodegradable polyesters produced by bacteria.
  • Granule-associated proteins (GAPs), particularly phasins, are crucial for PHA granule formation and metabolism.
  • Phasins are being explored for functionalizing PHA surfaces in various biotechnological applications.

Purpose of the Study:

  • To investigate the binding affinity and influence of two specific phasins, PhaP1 and PhaI, on PHA accumulation and granule morphology.
  • To assess these effects in engineered Pseudomonas putida KT2440 strains producing distinct PHA compositions (short-chain-length [scl]-PHA, medium-chain-length [mcl]-PHA, and hybrid scl/mcl-PHA).
  • To explore the potential of phasins as bio-affinity tags for selective PHA functionalization.

Main Methods:

  • Expression of phasin-msfGFP fusion proteins in engineered Pseudomonas putida strains.
  • In vivo fluorescence binding assays to monitor phasin surface binding.
  • Microscopy and Gas Chromatography-Mass Spectrometry (GC-MS) for PHA accumulation analysis.
  • Transmission Electron Microscopy (TEM) for granule morphology assessment.
  • Flow cytometry for differential surface affinity analysis.

Main Results:

  • Phasin expression did not significantly alter overall PHA yield (%PHA/CDW) but influenced granule characteristics.
  • PhaI expression in mcl-PHA-producing strain PP00_01 increased granule number and homogeneity.
  • Flow cytometry revealed differential phasin surface affinities based on PHA type, with PhaP1 preferring scl-PHA and PhaI preferring mcl-PHA.
  • Physicochemical properties of PHA and phasin type collectively impact binding strength.

Conclusions:

  • Phasins play a dual role in modulating PHA production and enabling selective functionalization of PHA granules.
  • The distinct binding preferences of PhaP1 and PhaI for different PHA types highlight their potential as specific bio-affinity tags.
  • This study supports the use of phasins for targeted surface modification of PHA granules in biotechnological applications like enzyme immobilization and drug delivery.