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Controlled Feeding Facilitates the Development of a Robust Production Process for Terpolymeric Polyhydroxyalkanoates.

Birk Achenbach^1,2, Pravesh Tamang², Steven Leonhardt^1,2

¹Institute of Interfacial Process Engineering and Plasma Technology IGVP, University of Stuttgart, Stuttgart, Germany.

Biotechnology Journal

|June 2, 2026

Summary

This study demonstrates scalable production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxyvalerate) (PHBVV) terpolymers using Cupriavidus necator. Controlled feeding strategies enabled high yields of PHBVV with significant 3-hydroxyvalerate content.

Keywords:

C. necator PHA PHBVV controlled fed‐batch levulinic acid pH‐pO2‐stat scale‐up

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High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles

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Published on: July 6, 2012

Area of Science:

Biotechnology and Biopolymer Production
Microbial Fermentation and Bioprocess Engineering
Sustainable Materials Science

Background:

Polyhydroxyalkanoates (PHAs) are biodegradable biopolymers with potential for sustainable applications.
Limited availability and material properties restrict current PHA usage.
Novel PHA variants like terpolymers offer opportunities for enhanced material characteristics.

Purpose of the Study:

To report the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate-co-4-hydroxyvalerate) (PHBVV) terpolymer with high 3-hydroxyvalerate (3HV) content.
To implement and validate a controlled feeding strategy for levulinic acid in bioreactors.
To demonstrate the scalability of PHBVV production in stirred tank bioreactors.

Main Methods:

Utilized Cupriavidus necator for terpolymer production.
Implemented a fully controlled feeding strategy based on pH and pO2 signals for levulinic acid delivery.
Validated the process in 7.5-L and 42-L bioreactors.

Main Results:

Achieved a maximum biomass of 40.2 g L⁻¹ with 75% PHA content (dry cell weight).
Obtained a PHA yield of 0.32 gPHA gLA⁻¹.
Successfully produced PHBVV terpolymers with 38%-46% 3HV and 1%-2% 4HV content at both scales.

Conclusions:

Demonstrated a scalable and automated method for PHBVV terpolymer production.
Provided a comprehensive dataset for optimizing PHA bioprocesses.
Established a framework for the industrial production of novel PHA variants with tailored properties.