Ivana Novackova

The terpolymer of 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), and 4-hydroxybutyrate (4HB) was produced employing Cupriavidus sp. DSM 19379. Growth in the presence of γ-butyrolactone, ε-caprolactone, 1,4-butanediol, and 1,6-hexanediol resulted in the synthesis of a polymer consisting of 3HB and 4HB monomers. Single and two-stage terpolymer production strategies were utilized to incorporate the 3HV subunit into the polymer structure. At the single-stage cultivation mode, γ-butyrolactone or 1,4-butanediol served as the primary substrate and propionic and valeric acid as the precursor of 3HV. In the two-stage production, glycerol was used in the growth phase, and precursors for the formation of the terpolymer in combination with the nitrogen limitation in the medium were used in the second phase. The aim of this work was to maximize the Polyhydroxyalkanoates (PHA) yields with a high proportion of 3HV and 4HB using different culture strategies. The obtained polymers contained 0–29 mol% of 3HV and 16–32 mol% of 4HB. Selected polymers were subjected to a material properties analysis such as differential scanning calorimetry (DSC), thermogravimetry, and size exclusion chromatography coupled with multi angle light scattering (SEC-MALS) for determination of the molecular weight. The number of polymers in the biomass, as well as the monomer composition of the polymer were determined by gas chromatography.

Polyhydroxyalkanoates (PHA) are biodegradable polyesters, which are produced by various bacteria including numerous halophiles. Employment of halophilic strain for PHA production brings numerous benefits such as robustness of the process against contamination by ubiquitous mesophiles or possibility to isolate polymer from bacterial biomass via hypotonic lysis. In this work, we screened three moderate halophiles – Halomonas halophila, Halomonas organivorans and Halomonas salina for the presence of phaC gene encoding for PHA synthase and, subsequently, we have investigated their PHA production potential on various sugars. Among tested strains, H. organivorans demonstrated the extraordinary capacity of PHA production in particular on galactose and mannose since on these saccharides PHA content in dried bacterial cells reached 83 and 90 wt. % on mannose and galactose, respectively. Therefore, H. organivoras can be considered being promising PHA producing strain in particular suitable for the valorization of lignocellulose materials rich in galactomannans such as spent coffee grounds.