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Microbial synthesis of polyhydroxyalkanoate blends with engineered Pseudomonas putida.

Minglong Li¹, Khalid Doudin², David B Robins¹

¹School of Chemical, Materials & Biological Engineering, University of Sheffield, Sir Robert Hadfield Building, Mappin Street, Sheffield S1 3JD, United Kingdom.

New Biotechnology

|June 4, 2025

Summary

Engineered microbes can now produce custom polyhydroxyalkanoate (PHA) blends in a single step. This microbial synthesis of PHA blends offers a sustainable alternative to traditional plastic production methods.

Keywords:

Polyhydroxyalkanoate Polymer blends Promoters Pseudomonas putida Synthetic biology

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Published on: October 26, 2016

Area of Science:

Biotechnology
Polymer Science
Microbial Engineering

Background:

Polyhydroxyalkanoates (PHAs) are biodegradable biopolymers serving as sustainable alternatives to petroleum-based plastics.
Current PHA production often involves post-synthesis blending, requiring multiple fermentation and extraction steps.
Direct synthesis of PHA blends offers a more efficient and streamlined production pathway.

Purpose of the Study:

To engineer Pseudomonas putida for the direct microbial synthesis of poly-3-hydroxybutyrate [P3(HB)] and medium-chain-length PHA (mcl-PHA) blends.
To control the composition of PHA blends by utilizing different promoter strengths.
To evaluate the feasibility of one-step biomanufacturing of customizable PHA blends.

Main Methods:

Genetic engineering of Pseudomonas putida to co-produce P3(HB) and mcl-PHA.
Utilizing various promoters to modulate the monomer composition of the PHA blends.
Optimizing fermentation conditions for maximal PHA yield and content.
Characterization of PHA blends using gel permeation chromatography, nuclear magnetic resonance, and diffusion ordered spectroscopy.

Main Results:

Successfully produced PHA blends with 3HB monomer content ranging from 17.9 to 99.6 mol%.
Achieved a maximum PHA production of 1.48 g/L with 52.2 wt% PHA content.
Confirmed higher molecular weight for P(3HB) compared to mcl-PHA in all blends.
Synthesized PHA blends exhibited thermal properties comparable to those produced via melt compounding.

Conclusions:

Demonstrated the microbial synthesis of PHA blends in Pseudomonas putida.
Established promoter selection as a method for controlling PHA blend composition during fermentation.
Paved the way for the one-step biomanufacturing of tailored PHA blends, enhancing sustainability in bioplastics.