# Sustainable bioconversion of excess grape must into polyhydroxyalkanoates by Cupriavidus necator DSM 545 and Hydrogenophaga pseudoflava DSM 1034

## TL;DR

This paper explores using surplus grape must to produce bioplastics (PHAs) with two bacterial strains, showing grape must is a sustainable and effective feedstock.

## Contribution

The study is the first to investigate red and white grape musts as substrates for PHAs production by C. necator and H. pseudoflava.

## Key findings

- C. necator DSM 545 achieved up to 61.5% PHB of cell dry weight in batch cultures using grape must.
- H. pseudoflava DSM 1034 reached 67.9% PHB on grape must but performed poorly in fed-batch conditions.
- Grape musts are promising feedstocks for sustainable PHAs production in circular economy frameworks.

## Abstract

Wine production generates significant quantities of by-products each year, among which surplus grape must is notable for its high content of fermentable sugars, organic acids, and polyphenols. As wine consumption declines and grape must surpluses grow, identifying sustainable valorization strategies becomes increasingly critical. This study investigates for the first time the potential use of red and white grape musts as substrates for polyhydroxyalkanoates (PHAs) production by two well-characterized bacterial strains, Cupriavidus necator DSM 545 and Hydrogenophaga pseudoflava DSM 1034, under both batch and fed-batch fermentation regimes. Both musts supported microbial growth and PHAs accumulation. In batch cultures, C. necator DSM 545 achieved a PHB content of up to 61.5% of cell dry weight (CDW), while H. pseudoflava DSM 1034 reached 67.9% PHB on grape must, with yields and biomass comparable to or exceeding those obtained with synthetic sugar-based media. Under fed-batch conditions with red must, C. necator DSM 545 sustained growth and PHB production across multiple feeding feeding periods, outperforming the control medium. Conversely, H. pseudoflava DSM 1034 displayed initial growth but failed to increase biomass over time, suggesting that this strain may be poorly suited for use in fed-bacth-based applications, likely due to nutrient depletion or the accumulation of inhibitory compounds. Overall, these results proved that grape musts are promising feedstocks for sustainable PHAs production. Their integration into circular economy frameworks offers a valuable opportunity for waste recovery and the development of bioplastics within the agri-food industry, especially in light of the increasing grape must surpluses recently experienced worldwide.

Surplus red and white grape musts were tested as substrates for PHAs production

Batch cultures of C. necator DSM 545 and H. pseudoflava DSM 1034 accumulated PHB

Fed-batch setting confirmed C. necator DSM 545 performance with red must

H. pseudoflava DSM 1034 showed limited fitness to prolonged fed-batch conditions

Grape musts are promising, cost-effective feedstocks for PHAs production