Industrial biotechnology of Pseudomonas putida and related species

Poblete‐Castro, Ignacio; Becker, Jörg; Dohnt, Katrin; Santos, Vítor Martins dos; Wittmann, Christoph

doi:10.1007/s00253-012-3928-0

Cited by 306 publications

(211 citation statements)

References 111 publications

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“…An alternative to this approach is to identify alternative production hosts that are naturally tolerant to higher concentrations of the chemicals of interest. P. putida is one such strain that is often utilized for bioremediation and production of more toxic chemicals (Loeschcke & Thies, 2015; Nikel, Martínez‐García, & de Lorenzo, 2014; Poblete‐Castro, Becker, Dohnt, dos Santos, & Wittmann, 2012; Verhoef, Ruijssenaars, de Bont, & Wery, 2007; Verhoef, Wierckx, Westerhof, De Winde, & Ruijssenaars, 2009). …”

Section: Introductionmentioning

confidence: 99%

Genome‐wide identification of tolerance mechanisms toward p‐coumaric acid in Pseudomonas putida

Calero

Jensen

Bojanovič

et al. 2017

Biotech & Bioengineering

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The soil bacterium Pseudomonas putida KT2440 has gained increasing biotechnological interest due to its ability to tolerate different types of stress. Here, the tolerance of P. putida KT2440 toward eleven toxic chemical compounds was investigated. P. putida was found to be significantly more tolerant toward three of the eleven compounds when compared to Escherichia coli. Increased tolerance was for example found toward p‐coumaric acid, an interesting precursor for polymerization with a significant industrial relevance. The tolerance mechanism was therefore investigated using the genome‐wide approach, Tn‐seq. Libraries containing a large number of miniTn5‐Km transposon insertion mutants were grown in the presence and absence of p‐coumaric acid, and the enrichment or depletion of mutants was quantified by high‐throughput sequencing. Several genes, including the ABC transporter Ttg2ABC and the cytochrome c maturation system (ccm), were identified to play an important role in the tolerance toward p‐coumaric acid of this bacterium. Most of the identified genes were involved in membrane stability, suggesting that tolerance toward p‐coumaric acid is related to transport and membrane integrity.

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Section: Introductionmentioning

confidence: 99%

Genome‐wide identification of tolerance mechanisms toward p‐coumaric acid in Pseudomonas putida

Calero

Jensen

Bojanovič

et al. 2017

Biotech & Bioengineering

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“…The saprotrophic soil bacterium Pseudomonas putida is known for its tolerance to a variety of environmental stresses, including but not limited to organic solvents and reactive oxygen species (Poblete‐Castro et al ., 2012; Nikel et al ., 2014; Ramos et al ., 2015). P. putida derives this sturdiness from its EDEMP cycle, a series of metabolic pathways that facilitate NAD(P)H production via carbon recycling processes (Nikel et al ., 2015).…”

Section: Introductionmentioning

confidence: 99%

“…P. putida derives this sturdiness from its EDEMP cycle, a series of metabolic pathways that facilitate NAD(P)H production via carbon recycling processes (Nikel et al ., 2015). An adaptability to a myriad of harsh conditions has made P. putida a popular focus of biosynthetic studies aimed at industrial biotransformations as well as soil and water bioremediation (Garmendia et al ., 2008; Puchałka et al ., 2008; Poblete‐Castro et al ., 2012; Loeschcke and Thies, 2015). Unfortunately, the collection of genetic tools available in Pseudomonads is limited in scope and, when used in conjunction, is subject to bottlenecks that hinder the generation of a biotechnological chassis (Martínez‐García and de Lorenzo, 2011; Martínez‐García et al ., 2011, 2014).…”

Section: Introductionmentioning

confidence: 99%

A standardized workflow for surveying recombinases expands bacterial genome‐editing capabilities

Ricaurte

Martínez‐García

Nyerges

et al. 2017

Microbial Biotechnology

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SummaryBacterial recombineering typically relies on genomic incorporation of synthetic oligonucleotides as mediated by Escherichia coli λ phage recombinase β – an occurrence largely limited to enterobacterial strains. While a handful of similar recombinases have been documented, recombineering efficiencies usually fall short of expectations for practical use. In this work, we aimed to find an efficient Recβ homologue demonstrating activity in model soil bacterium Pseudomonas putida EM42. To this end, a genus‐wide protein survey was conducted to identify putative recombinase candidates for study. Selected novel proteins were assayed in a standardized test to reveal their ability to introduce the K43T substitution into the rpsL gene of P. putida. An ERF superfamily protein, here termed Rec2, exhibited activity eightfold greater than that of the previous leading recombinase. To bolster these results, we demonstrated Rec2 ability to enter a range of mutations into the pyrF gene of P. putida at similar frequencies. Our results not only confirm the utility of Rec2 as a Recβ functional analogue within the P. putida model system, but also set a complete workflow for deploying recombineering in other bacterial strains/species. Implications range from genome editing of P. putida for metabolic engineering to extended applications within other Pseudomonads – and beyond.

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“…An exception is P. oleovorans (ATCC 8062) where its cultivation on 4-hydroxyhexanoic acid resulted in a copolymer predominated by 3HB (92.4 mol%) [56]. Pseudomonads are also well-known for their bioremediation properties including the biodegradation recalcitrant and/or toxic aromatic carbon substrates [98], and have been successfully applied in the treatment of contaminated effluents, exhaust gas and soils [99][100][101]. Recent studies demonstrated that aromatic-degraders P. putida F1 (DSM 6899), P. putida mt-2 (NCIMB 10432), and P. putida CA-3 (NCIMB 41162) could bioconvert toxic pollutants benzene, toluene, ethylbenzene, xylene (BTEX) and styrene to mcl-PHA [86] [76,77,80], which offers the potential benefit to off-set waste treatment cost through PHA recovery.…”

Section: Gram-negative Bacteriamentioning

confidence: 99%