An Initial Genome Editing Toolset for Caldimonas
thermodepolymerans, the First Model of Thermophilic
Polyhydroxyalkanoates Producer

J 2025

An Initial Genome Editing Toolset for Caldimonas thermodepolymerans, the First Model of Thermophilic Polyhydroxyalkanoates Producer

IEREMENKO, Anastasiia; Kristýna LIPOVSKÁ; Xenie KOUŘILOVÁ; Stanislav OBRUČA; Pavel DVOŘÁK et al.

Základní údaje

Originální název

An Initial Genome Editing Toolset for Caldimonas thermodepolymerans, the First Model of Thermophilic Polyhydroxyalkanoates Producer

Autoři

IEREMENKO, Anastasiia; Kristýna LIPOVSKÁ; Xenie KOUŘILOVÁ; Stanislav OBRUČA a Pavel DVOŘÁK

Vydání

Microbial Biotechnology, Hoboken, Wiley, 2025, 1751-7915

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

20801 Environmental biotechnology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 5.200 v roce 2024

Označené pro přenos do RIV

Ano

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

Caldimonas thermodepolymerans; gene deletion; genetic engineering; polyhydroxyalkanoates; thermophiles

Štítky

14314010, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 31. 3. 2025 09:39, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

The limited number of well-characterised model bacteria cannot address all the challenges in a circular bioeconomy. Therefore, there is a growing demand for new production strains with enhanced resistance to extreme conditions, versatile metabolic capabilities and the ability to utilise cost-effective renewable resources while efficiently generating attractive biobased products. Particular thermophilic microorganisms fulfil these requirements. Non-virulent Gram-negative Caldimonas thermodepolymerans DSM15344 is one such attractive thermophile that efficiently converts a spectrum of plant biomass sugars into high quantities of polyhydroxyalkanoates (PHA)—a fully biodegradable substitutes for synthetic plastics. However, to enhance its biotechnological potential, the bacterium needs to be ‘domesticated’. In this study, we established effective homologous recombination and transposon-based genome editing systems for C. thermodepolymerans. By optimising the electroporation protocol and refining counterselection methods, we achieved significant improvements in genetic manipulation and constructed the AI01 chassis strain with improved transformation efficiency and a ΔphaC mutant that will be used to study the importance of PHA synthesis in Caldimonas. The advances described herein highlight the need for tailored approaches when working with thermophilic bacteria and provide a springboard for further genetic and metabolic engineering of C. thermodepolymerans, which can be considered the first model of thermophilic PHA producer.

Návaznosti

MUNI/J/0003/2021, interní kód MU

Název: A new generation bacterial platform for lignocellulose biotechnology (Akronym: NEWGEN)

Investor: Masarykova univerzita, A new generation bacterial platform for lignocellulose biotechnology, MASH JUNIOR - MUNI Award In Science and Humanities JUNIOR

MUNI/R/1266/2022, interní kód MU

Název: Vývoj nástrojů genetického inženýrství a syntetické biologie pro termofilní bakterie

Investor: Masarykova univerzita, Vývoj nástrojů genetického inženýrství a syntetické biologie pro termofilní bakterie, CAREER RESTART

Citovat

IEREMENKO, Anastasiia; Kristýna LIPOVSKÁ; Xenie KOUŘILOVÁ; Stanislav OBRUČA a Pavel DVOŘÁK. An Initial Genome Editing Toolset for Caldimonas thermodepolymerans, the First Model of Thermophilic Polyhydroxyalkanoates Producer. Microbial Biotechnology. Hoboken: Wiley, 2025, roč. 18, č. 2, s. 1-16. ISSN 1751-7915. Dostupné z: https://doi.org/10.1111/1751-7915.70103.

@article{2481403,
   author = {Ieremenko, Anastasiia and Lipovská, Kristýna and Kouřilová, Xenie and Obruča, Stanislav and Dvořák, Pavel},
   article_location = {Hoboken},
   article_number = {2},
   doi = {https://doi.org/10.1111/1751-7915.70103},
   keywords = {Caldimonas thermodepolymerans; gene deletion; genetic engineering; polyhydroxyalkanoates; thermophiles},
   language = {eng},
   issn = {1751-7915},
   journal = {Microbial Biotechnology},
   title = {An Initial Genome Editing Toolset for Caldimonas thermodepolymerans, the First Model of Thermophilic Polyhydroxyalkanoates Producer},
   url = {https://doi.org/10.1111/1751-7915.70103},
   volume = {18},
   year = {2025}
}

TY  - JOUR
ID  - 2481403
AU  - Ieremenko, Anastasiia - Lipovská, Kristýna - Kouřilová, Xenie - Obruča, Stanislav - Dvořák, Pavel
PY  - 2025
TI  - An Initial Genome Editing Toolset for Caldimonas thermodepolymerans, the First Model of Thermophilic Polyhydroxyalkanoates Producer
JF  - Microbial Biotechnology
VL  - 18
IS  - 2
SP  - 1-16
EP  - 1-16
PB  - Wiley
SN  - 17517915
KW  - Caldimonas thermodepolymerans
KW  - gene deletion
KW  - genetic engineering
KW  - polyhydroxyalkanoates
KW  - thermophiles
UR  - https://doi.org/10.1111/1751-7915.70103
N2  - The limited number of well-characterised model bacteria cannot address all the challenges in a circular bioeconomy. Therefore, there is a growing demand for new production strains with enhanced resistance to extreme conditions, versatile metabolic capabilities and the ability to utilise cost-effective renewable resources while efficiently generating attractive biobased products. Particular thermophilic microorganisms fulfil these requirements. Non-virulent Gram-negative Caldimonas thermodepolymerans DSM15344 is one such attractive thermophile that efficiently converts a spectrum of plant biomass sugars into high quantities of polyhydroxyalkanoates (PHA)—a fully biodegradable substitutes for synthetic plastics. However, to enhance its biotechnological potential, the bacterium needs to be ‘domesticated’. In this study, we established effective homologous recombination and transposon-based genome editing systems for C. thermodepolymerans. By optimising the electroporation protocol and refining counterselection methods, we achieved significant improvements in genetic manipulation and constructed the AI01 chassis strain with improved transformation efficiency and a ΔphaC mutant that will be used to study the importance of PHA synthesis in Caldimonas. The advances described herein highlight the need for tailored approaches when working with thermophilic bacteria and provide a springboard for further genetic and metabolic engineering of C. thermodepolymerans, which can be considered the first model of thermophilic PHA producer.
ER  -

IEREMENKO, Anastasiia; Kristýna LIPOVSKÁ; Xenie KOUŘILOVÁ; Stanislav OBRUČA a Pavel DVOŘÁK. An Initial Genome Editing Toolset for Caldimonas thermodepolymerans, the First Model of Thermophilic Polyhydroxyalkanoates Producer. \textit{Microbial Biotechnology}. Hoboken: Wiley, 2025, roč.~18, č.~2, s.~1-16. ISSN~1751-7915. Dostupné z: https://doi.org/10.1111/1751-7915.70103.

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