Pseudomonas putida—a versatile host for the production of natural products

Loeschcke, Anita; Thies, Stephan

doi:10.1007/s00253-015-6745-4

Cited by 245 publications

(164 citation statements)

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“…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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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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“…205 Thus, P. putida has been employed to produce a broad portfolio of natural products including amino acid derived compounds, non-ribosomal peptides, polyketides, rhamnolipids and terpenoids. 206 …”

Section: Heterologous Production Of Bacterial Natural Productsmentioning

confidence: 99%

Engineering microbial hosts for production of bacterial natural products

et al. 2016

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Microbial fermentation provides as an attractive alternative to chemical synthesis for the production of structurally complex natural products. In most cases, however, production titers are low and need to be improved for compound characterization and/or commercial production. Owing to advances in functional genomics and genetic engineering technologies, microbial hosts can be engineered to overproduce a desired natural product, greatly accelerating the traditionally time-consuming strain improvement process. This review covers recent developments and challenges in the engineering of native and heterologous microbial hosts for production of bacterial natural products, focusing on the genetic tools and strategies for strain improvement. Special emphasis is placed on bioactive secondary metabolites from actinomycetes. The considerations for the choice of host systems will also be discussed in this review.

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“…In recent years, there has been an increasing interest in engineering Pseudomonas putida for the production of various chemicals and natural products, either by de novo synthesis or by bioconversion of precursor molecules (Poblete-Castro et al, 2012;Loeschcke and Thies, 2015;Mi et al, 2014;Cornelissen et al, 2013). Among the reasons are the fast growth of P. putida, its ease of genetic manipulation, its simple nutrient requirements and especially its pronounced ability to cope with toxic and hydrophobic substances.…”

Section: Introductionmentioning

confidence: 99%