Pseudomonas putida as a synthetic biology chassis and a metabolic engineering platform

Martínez-García, Esteban; de Lorenzo, Víctor

doi:10.1016/j.copbio.2023.103025

Cited by 18 publications

(4 citation statements)

References 69 publications

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“…The versatile metabolism of P. putida has increased the interest of the scientific and biotechnology communities in utilizing this microbe in bioproduction and biodegradation applications. To engineer a robust microbial cell factory from this laboratory work-horse, the genome of P. putida has been significantly edited and reduced (Martínez-García et al ., 2014; Aparicio, De Lorenzo and Martínez-García, 2019; Martínez-García and De Lorenzo, 2024). Notably, while P. putida is considered to have high potential for bio-industry, it is curious that there is no knowledge regarding how this bacterium protects itself from its natural enemies, bacteriophages.…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of a phage collection againstPseudomonas putida

Brauer,

Rosendahl,

Kängsep

et al. 2024

Preprint

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The environmental bacterium Pseudomonas putida has a wide range of metabolic pathways and thus entails great promise both as cell factories in biotechnological production and for bioremediation approaches to degrade various aromatic pollutants. To thrive in the environment, P. putida has to be able to endure the constant threat posed by bacteriophages. Surprisingly, up until now, only a few phages have been isolated for the common laboratory strain P. putida KT2440, and no phage defence mechanisms have been characterized yet. Here, we introduce a novel Collection of Environmental Pseudomonas putida Phages from Estonia (CEPEST), consisting of 67 dsDNA phages that belong to 22 phage species and nine phage genera. We show that most of the phages in the CEPEST collection are more infectious at lower temperatures, have a narrow host range, and require intact LPS for P. putida infection. Additionally, we demonstrate that cryptic prophages in the P. putida chromosome strongly protect against the infection of many phages, whereas the chromosomal toxin-antitoxin systems play no role in phage defence of P. putida.

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Section: Introductionmentioning

confidence: 99%

Isolation and characterization of a phage collection againstPseudomonas putida

Brauer,

Rosendahl,

Kängsep

et al. 2024

Preprint

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show abstract

“…The versatile metabolism of P. putida has increased the interest of the scientific and biotechnology communities in utilizing this microbe in bioproduction and biodegradation applications. To engineer a robust microbial cell factory from this laboratory workhorse, the genome of P. putida has been significantly edited and reduced (Aparicio et al, 2019;Martínez-García et al, 2014;Martínez-García & De Lorenzo, 2024). Notably, while P. putida is considered to have high potential for bio-industry, it is curious that there is no knowledge regarding how this bacterium protects itself from bacteriophages.…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of a phage collection against Pseudomonas putida

Brauer,

Rosendahl,

Kängsep

et al. 2024

Environmental Microbiology

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The environmental bacterium, Pseudomonas putida, possesses a broad spectrum of metabolic pathways. This makes it highly promising for use in biotechnological production as a cell factory, as well as in bioremediation strategies to degrade various aromatic pollutants. For P. putida to flourish in its environment, it must withstand the continuous threats posed by bacteriophages. Interestingly, until now, only a handful of phages have been isolated for the commonly used laboratory strain, P. putida KT2440, and no phage defence mechanisms have been characterized. In this study, we present a new Collection of Environmental P. putida Phages from Estonia, or CEPEST. This collection comprises 67 double‐stranded DNA phages, which belong to 22 phage species and 9 phage genera. Our findings reveal that most phages in the CEPEST collection are more infectious at lower temperatures, have a narrow host range, and require an intact lipopolysaccharide for P. putida infection. Furthermore, we show that cryptic prophages present in the P. putida chromosome provide strong protection against the infection of many phages. However, the chromosomal toxin–antitoxin systems do not play a role in the phage defence of P. putida. This research provides valuable insights into the interactions between P. putida and bacteriophages, which could have significant implications for biotechnological and environmental applications.

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“…Pseudomonas putida KT2440, an ideal chassis for industrial biotechnology and bioremediation, has attracted increasing interest from researchers [ 39 , 40 , 41 , 42 ]. Recently, researchers make great efforts to reveal and elucidate the functional gene for tolerance under various stress conditions of P. putida KT2440 and to expand its biotechnological applications, such as a major facilitator superfamily transporter involved in propionic acid tolerance [ 43 ], an ABC transporters involved in furfural tolerance [ 44 ], and the efflux pump protein TtgB involved in p -coumaric and ferulic acid tolerance [ 45 ].…”

Section: Introductionmentioning

confidence: 99%

Improvement in Salt Tolerance Ability of Pseudomonas putida KT2440

Fan,

Tan,

Wang

et al. 2024

Biology

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Pseudomonas putida KT2440 is a popular platform for bioremediation due to its robust tolerance to environmental stress and strong biodegradation capacity. Limited research on the salt tolerance of P. putida KT2440 has hindered its application. In this study, the strain KT2440 was tested to tolerate a maximum of 4% w/v NaCl cultured with minimal salts medium. Transcriptomic data in a high-salinity environment showed significant expression changes in genes in membrane components, redox processes, chemotaxis, and cellular catabolic processes. betB-encoding betaine-aldehyde dehydrogenase was identified from the transcriptome data to overexpress and enhance growth profile of the strain KT2440 in minimal salts medium containing 4% w/v NaCl. Meanwhile, screening for exogenous salt-tolerant genes revealed that the Na+/H+ antiporter EcnhaA from Escherichia coli significantly increased the growth of the strain KT2440 in 4% w/v NaCl. Then, co-expression of EcnhaA and betB (KT2440-EcnhaA-betB) increased the maximum salt tolerance of strain KT2440 to 5% w/v NaCl. Further addition of betaine and proline improved the salt tolerance of the engineered strain to 6% w/v NaCl. Finally, the engineered strain KT2440-EcnhaA-betB was able to degrade 56.70% of benzoic acid and 95.64% of protocatechuic acid in minimal salt medium containing 4% w/v NaCl in 48 h, while no biodegradation was observed in the normal strain KT2440 in the same conditions. However, the strain KT2440-EcnhaA-betB failed to degrade catechol in minimal salt medium containing 3% w/v NaCl. This study illustrated the improvement in the salt tolerance performance of Pseudomonas putida KT2440 and the feasibility of engineered strain KT2440 as a potential salt-tolerant bioremediation platform.

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Pseudomonas putida as a synthetic biology chassis and a metabolic engineering platform

Cited by 18 publications

References 69 publications

Isolation and characterization of a phage collection againstPseudomonas putida

Isolation and characterization of a phage collection againstPseudomonas putida

Isolation and characterization of a phage collection against Pseudomonas putida

Improvement in Salt Tolerance Ability of Pseudomonas putida KT2440

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