Genome‐wide identification of tolerance mechanisms toward <i>p</i>‐coumaric acid in <i>Pseudomonas putida</i>

Calero, Patricia; Jensen, Sheila Ingemann; Bojanovič, Klara; Lennen, Rebecca M.; Koza, Anna; Nielsen, Alex Toftgaard

doi:10.1002/bit.26495

Cited by 63 publications

(66 citation statements)

References 74 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The adapted strain showed not only robust growth in high ferulate concentration (up to 125 mM) while the growth of wild type was completely inhibited, but also more prominent growth than wild type in low ferulate concentration. In a previous study, the genes involved in the tolerance towards coumaric acid, another lignin-derived phenolic compound, were identified in Pseudomonas putida (Calero et al, 2018). Most of the identified genes were related to membrane stability, transport system, and membrane proteins while the genes involved in the degradation of the compounds only play minor roles in the tolerance (Calero et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

Synthetic Metabolic Pathway for the Production of 1-Alkenes from Lignin-derived Molecules

Luo

Lehtinen

Efimova

et al. 2018

Preprint

View full text Add to dashboard Cite

8Integration of synthetic metabolic pathways to catabolically diverse chassis provides new 9 opportunities for sustainable production. One attractive scenario is the use of abundant 10 waste material to produce readily collectable product, minimizing production costs. 11Towards that end, we established the production of semivolatile medium-chain α-olefins 12 from lignin-derived monomers: we constructed 1-undecene synthesis pathway in 13 Acinetobacter baylyi ADP1 using ferulate as the sole carbon source. In order to overcome 14 the toxicity of ferulate, we first applied adaptive laboratory evolution, resulting in a highly 15 ferulate-tolerant strain. Next, we demonstrated the 1-undecene production from glucose by 16 heterologously expressing a fatty acid decarboxylase UndA and a thioesterase 'TesA in the 17 wild type strain. Finally, we constructed the alkene synthesis pathway in the ferulate-18 tolerant strain. We were able to produce 1-undecene from ferulate and collect the product 19 from the culture headspace without downstream processing. This study demonstrates the 20 potential of bacterial lignin upgradation into value-added products. 21 22 65 chain α-olefins, such as 1-undecene, are attractive molecules for their broad use in 66 detergents, plasticizers and monomers for elastomers (Sarria et al., 2017). In addition, the 67 molecules can accumulate extracellularly and are semivolatile, which allow them to be 68 collected directly from the culture vessel without cell harvesting and/or product extraction, 69 significantly reducing the costs and labour of downstream processing. Recently, a single 70 gene undA originated from Pseudomonas has been discovered to be responsible for 1-71 undecene (C11) biosynthesis (Rui et al., 2014). UndA is an oxygen-activating, nonheme 72 iron (II)-dependent decarboxylase that converts free fatty acids (FFAs) to the corresponding 73 terminal alkenes. The enzyme accepts fatty acids with chain length from 10 to 14 as 74 substrates. The gene has been heterologously expressed e.g. in E. coli for 1-undecene 75 production (Rui et al., 2014). Other possible alkene synthesis pathways include polyketide 76 synthase (PKS) pathway and head-to-head hydrocarbon synthesis pathway, in which 77 multiple enzymes and reactions are involved (Beller et al., 2010; Liu et al., 2015). In 78 comparison, the one-step decarboxylation of FFAs catalyzed by UndA is simpler and has a 79 narrower substrate spectrum (Kang and Nielsen, 2017).80In this study, we employed A. baylyi ADP1 for the production of 1-undecene from a lignin 81 derived model compound, ferulate. We applied ALE to improve the tolerance of A. baylyi 82 ADP1 against ferulate, and established a synthetic pathway for the direct conversion of 83 LDMs to 1-undecene ( Figure 1). We demonstrate the potential of catabolically diverse 84 bacteria for the synthesis of industrially relevant compounds from an abundant and 85 sustainable substrate.86 2 Results 87 2.1 Adaptation of A. baylyi ADP1 to high concentration of ferulate 88 89In order to impro...

show abstract

Section: Discussionmentioning

confidence: 99%

Synthetic Metabolic Pathway for the Production of 1-Alkenes from Lignin-derived Molecules

Luo

Lehtinen

Efimova

et al. 2018

Preprint

View full text Add to dashboard Cite

show abstract

“…The orthologous Mla system in P. aeruginosa is encoded by the PA4452-PA4456 operon (locus tags corresponding to PAO1) and the isolated gene PA2800 (MlaA ortholog, also known as VacJ). Proteins encoded by this gene cluster are highly similar to those encoded by operon ttg2 (toluene tolerance genes) in Pseudomonas putida 32, 33 . Although it is unlikely that organic solvents themselves are substrates of this transporter, this system was initially linked to toluene tolerance in that bacterium 34 .…”

Section: Introductionmentioning

confidence: 95%

Evolution toward maximum transport capacity of the Ttg2 ABC system inPseudomonas aeruginosa

Yero

Costenaro

Conchillo-Solé

et al. 2019

Preprint

View full text Add to dashboard Cite

22In Pseudomonas aeruginosa, Ttg2D is the soluble periplasmic phospholipid-binding 23 component of an ABC transport system thought to be involved in maintaining the 24 asymmetry of the outer membrane. The crystallographic structure of Ttg2D at 2.5Å 25 resolution reveals that this protein can bind two diacyl phospholipids. Native and 26 denaturing mass spectrometry experiments confirm that Ttg2D binds two 27 phospholipid molecules, which may have different head groups. Analysis of the 28 available structures of Ttg2D orthologs allowed us to classify this protein family as a 29 novel substrate-binding protein fold and to venture the evolutionary events that 30 differentiated the orthologs binding one or two phospholipids. In addition, gene 31 knockout experiments in P. aeruginosa PAO1 and multidrug-resistant strains show 32 that disruption of this system leads to outer membrane permeabilization. This 33 demonstrates the role of this system in low-level intrinsic resistance against certain 34 antibiotics that use a lipid-mediated pathway to permeate through membranes. 35 36 Introduction 37Pseudomonas aeruginosa are amongst the most important multidrug-resistant (MDR) 38 human pathogens 1 , showing inherent resistance to an important number of the 39 presently available antibiotics 2 . P. aeruginosa are responsible for chronic lung 40 infections in individuals with chronic obstructive pulmonary disease or cystic fibrosis 41 (CF) 3 and account for over a tenth of all nosocomial infections 4 . A number of effective 42 drugs and formulations can treat P. aeruginosa infections, even in CF patients 5 . 43These include frontline antibiotics such as piperazillin-tazobactam, ceftazidime, 44 aztreonam, imipenem, meropenem, ciprofloxacin, levofloxacin, tobramycin, amikacin, 45 and colistin 6 . Yet, resistance to most of these antimicrobials is being increasingly 46 reported 7 . The basis for the inherently high resistance of these microorganisms is 47primarily their low outer-membrane (OM) permeability 8, 9 , complemented by the 48 production of antibiotic-inactivating enzymes (e.g. β-lactamases), the constitutive 49 expression of efflux pumps 10, 11 and the capacity to form biofilms 1, 12 , among other 50 mechanisms. The susceptibility of P. aeruginosa to antimicrobials can be additionally 51 reduced by the acquisition of inheritable traits, including horizontal gene transfers 52 and mutations that decrease uptake and efflux pump overexpression 13, 14, 15 . 53Although a number of genes and mechanisms of resistance to antibiotics are already 54 known in P. aeruginosa, the complex mechanisms controlling the basal, low-level 55 resistance to these compounds are still poorly understood 16, 17 . 56 57The OM of P. aeruginosa is known to be central to its antibiotic-resistance 58 phenotype. Its intrinsically low permeability is partly determined by inefficient OM 59 porin proteins that provide innate resistance to several antimicrobial compounds, 60 mainly hydrophilic 1, 8, 10 . On the other hand, the loss of specific efflux pump ...

show abstract

“…Remarkably, P. putida has a number of mechanisms for tolerance towards high concentrations of aromatic chemicals, for example toluene, xylenes and styrene (note that these toxic compounds can also be used by some P . putida strains), including a wide repertoire of efflux pumps (Inoue and Horikoshi, ; Ramos et al ., ; Santos et al ., ; Domínguez‐Cuevas et al ., ; Calero et al ., ). Toluene, for instance, is an aromatic and highly toxic solvent that kills most microorganisms at concentrations as low as 0.1% (v/v).…”

Section: Bacterial Species Adopted As a Chassis: From Historical Exammentioning

confidence: 99%

Chasing bacterial chassis for metabolic engineering: a perspective review from classical to non‐traditional microorganisms

Calero

Nikel

2018

Microbial Biotechnology

Self Cite

208

183

View full text Add to dashboard Cite

The last few years have witnessed an unprecedented increase in the number of novel bacterial species that hold potential to be used for metabolic engineering. Historically, however, only a handful of bacteria have attained the acceptance and widespread use that are needed to fulfil the needs of industrial bioproduction - and only for the synthesis of very few, structurally simple compounds. One of the reasons for this unfortunate circumstance has been the dearth of tools for targeted genome engineering of bacterial chassis, and, nowadays, synthetic biology is significantly helping to bridge such knowledge gap. Against this background, in this review, we discuss the state of the art in the rational design and construction of robust bacterial chassis for metabolic engineering, presenting key examples of bacterial species that have secured a place in industrial bioproduction. The emergence of novel bacterial chassis is also considered at the light of the unique properties of their physiology and metabolism, and the practical applications in which they are expected to outperform other microbial platforms. Emerging opportunities, essential strategies to enable successful development of industrial phenotypes, and major challenges in the field of bacterial chassis development are also discussed, outlining the solutions that contemporary synthetic biology-guided metabolic engineering offers to tackle these issues.

show abstract

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

Cited by 63 publications

References 74 publications

Synthetic Metabolic Pathway for the Production of 1-Alkenes from Lignin-derived Molecules

Synthetic Metabolic Pathway for the Production of 1-Alkenes from Lignin-derived Molecules

Evolution toward maximum transport capacity of the Ttg2 ABC system inPseudomonas aeruginosa

Chasing bacterial chassis for metabolic engineering: a perspective review from classical to non‐traditional microorganisms

Contact Info

Product

Resources

About