𝗙𝗿𝗼𝗺 𝗪𝗶𝗹𝗱 𝗧𝘆𝗽𝗲 𝘁𝗼 𝗣𝗛𝗔 𝗙𝗮𝗰𝘁𝗼𝗿𝘆: 𝗘𝗻𝗴𝗶𝗻𝗲𝗲𝗿𝗶𝗻𝗴 𝑪𝒂𝒍𝒅𝒊𝒎𝒐𝒏𝒂𝒔 𝒕𝒉𝒆𝒓𝒎𝒐𝒅𝒆𝒑𝒐𝒍𝒚𝒎𝒆𝒓𝒂𝒏𝒔

𝗙𝗿𝗼𝗺 𝗪𝗶𝗹𝗱 𝗧𝘆𝗽𝗲 𝘁𝗼 𝗣𝗛𝗔 𝗙𝗮𝗰𝘁𝗼𝗿𝘆: 𝗘𝗻𝗴𝗶𝗻𝗲𝗲𝗿𝗶𝗻𝗴 𝑪𝒂𝒍𝒅𝒊𝒎𝒐𝒏𝒂𝒔 𝒕𝒉𝒆𝒓𝒎𝒐𝒅𝒆𝒑𝒐𝒍𝒚𝒎𝒆𝒓𝒂𝒏𝒔

Abstract

Polyhydroxyalkanoates (PHA) are a diverse family of natural, biodegradable polymers that have gained increasing attention as sustainable alternatives to conventional plastics in certain industrial areas. Thermophilic bacteria fitting into the Next Generation Industrial Biotechnology (NGIB) concept represent promising microbial platforms for efficient PHA production. One notable example is the Gram-negative bacterium Caldimonas thermodepolymerans DSM 15344, capable of synthesizing poly(3-hydroxybutyrate) (PHB) on 20 g/L xylose at levels reaching up to 87 wt. % of CDM. To further enhance its industrial applicability, genetic engineering approaches following principles of synthetic biology have been employed. Using homologous recombination, targeted gene deletions were performed to control monomer incorporation and improve the overall PHA extensibility. In this work, we compare the biotechnological characteristics along with stress robustness testing of four deletion mutants derived from the wild-type C. thermodepolymerans DSM 15344, providing insights into strain improvement strategies for sustainable biopolymer manufacturing. The genetically modified strains analyzed in this study include: C. thermodepolymerans ΔphaC (derivative of DSM 15344 with deleted gene of PHA synthase IS481_08630); C. thermodepolymerans AI01 (derivative of DSM 15344, deleted genes of restriction endonucleases: ∆IS481_08585, ∆IS481_14855, ∆IS481_14025); C. thermodepolymerans ΔphaZ (derivative of C. thermodepolymerans AI01, deleted gene of PHA intracellular depolymerase IS481_07130); and C. thermodepolymerans KS01 (derivative of C. thermodepolymerans ΔphaZ with deleted gene of 2-methylcitrate synthase IS481_11870).

Polyhydroxyalkanoates (PHA) are a diverse family of natural, biodegradable polymers that have gained increasing attention as sustainable alternatives to conventional plastics in certain industrial areas. Thermophilic bacteria fitting into the Next Generation Industrial Biotechnology (NGIB) concept represent promising microbial platforms for efficient PHA production. One notable example is the Gram-negative bacterium Caldimonas thermodepolymerans DSM 15344, capable of synthesizing poly(3-hydroxybutyrate) (PHB) on 20 g/L xylose at levels reaching up to 87 wt. % of CDM. To further enhance its industrial applicability, genetic engineering approaches following principles of synthetic biology have been employed. Using homologous recombination, targeted gene deletions were performed to control monomer incorporation and improve the overall PHA extensibility. In this work, we compare the biotechnological characteristics along with stress robustness testing of four deletion mutants derived from the wild-type C. thermodepolymerans DSM 15344, providing insights into strain improvement strategies for sustainable biopolymer manufacturing. The genetically modified strains analyzed in this study include: C. thermodepolymerans ΔphaC (derivative of DSM 15344 with deleted gene of PHA synthase IS481_08630); C. thermodepolymerans AI01 (derivative of DSM 15344, deleted genes of restriction endonucleases: ∆IS481_08585, ∆IS481_14855, ∆IS481_14025); C. thermodepolymerans ΔphaZ (derivative of C. thermodepolymerans AI01, deleted gene of PHA intracellular depolymerase IS481_07130); and C. thermodepolymerans KS01 (derivative of C. thermodepolymerans ΔphaZ with deleted gene of 2-methylcitrate synthase IS481_11870).

polyhydroxyalkanoates, thermophilic bacteria, synthetic biology

polyhydroxyalkanoates, thermophilic bacteria, synthetic biology

ŠLOSÁROVÁ, K.; IEREMENKO, A.; PACASOVÁ, V.; BERGER, M.; DVOŘÁK, P.; OBRUČA, S.

03.10.2025

NOVA School of Science and Technoloígy

Lisbon

88

88

1

https://media.sci-meet.com/www.esbp2025.org/5668be5d-0ba1-4400-9612-ad0388fbe354/BookofAbstractsESBP2025.pdf