Novosphingobium aromaticivorans uses a Nu-class glutathione S-transferase as a glutathione lyase in breaking the β-aryl ether bond of lignin

Kontur, Wayne S.; Bingman, C.A.; Olmsted, Charles N.; Wassarman, Douglas R.; Ulbrich, Arne; Gall, Daniel L.; Smith, Robert W.; Yusko, Larissa M.; Fox, Brian G.; Noguera, Daniel R.; Coon, Joshua J.; Donohue, Timothy J.

doi:10.1074/jbc.ra117.001268

Cited by 53 publications

(89 citation statements)

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“…15 The present study addresses each of these issues using mutant strains of N. aromaticivorans DSM12444, a microbe naturally capable of degrading S, G, and H type aromatic compounds, as a well as lignin derived aromatic dimers. 25,33 We chose N. aromaticivorans DSM12444 due to its known or predicted ability to grow in the presence of multiple aromatic compounds, its suitability for genetic analysis and modifi-cation, its ability to co-metabolize aromatics in the presence of other organic compounds (such as sugars, which are another plentiful product of plant biomass degradation), and the potential to produce single valuable products using defined mutants.…”

Section: Discussionmentioning

confidence: 99%

“…4 Sphingomonad bacteria, such as N. aromaticivorans DSM12444, are known or predicted to be capable of breaking most of the linkages found between aromatic subunits in natural lignin in defined ways that yield predictable mono-aromatic products that can be further metabolized. 1,36 N. aromaticivorans, specifically, is known to be capable of degrading model aromatic dimers containing β-aryl-ether bonds 25 and its genome contains homologs of genes that code for the degradation of other aromatic dimers in Sphingobium sp. SYK-6.…”

Section: Discussionmentioning

confidence: 99%

“…A variant of N. aromaticivorans DSM12444 (strain 12444Δ1879) that lacks the gene Saro_1879 ( putative sacB; SARO_RS09410 in the recently reannotated genome in NCBI) 25 was used as a parent strain to create the deletion mutant strains 12444ΔligI (lacks gene Saro_2819; SARO_RS14300), 12444ΔdesCD (lacks the genes Saro_2864 and Saro_2865; SARO_RS14525 and SARO_RS14530), and 12444ΔligIΔdesCD (lacks genes Saro_2819, Saro_2864, and Saro_2865). All genetic modifications used a variant of the plasmid pk18mobsacB, 37 which contains sacB and a kanamycin resistance gene.…”

Section: Bacterial Strains Growth Media and Culturing Conditionsmentioning

confidence: 99%

“…26 A complete genome sequence is available for this α-proteobacterium (GenBank NC_007794.1), and the organism is amenable to genetic and genomic techniques needed to test the role of individual genes in aromatic metabolism, and model, engineer, or improve its pathways. 25 Specifically, we show that by using a defined set of mutations, N. aromaticivorans can be engineered to simultaneously produce PDC from all three major types of plant-derived phenolic compounds (S, G, and H). In addition, we find that this organism can metabolize aromatics simultaneously with the use of other organic carbon sources (such as glucose), a feature that allows mutant strains to excrete compounds derived from the incomplete metabolism of the aromatics.…”

Section: Introductionmentioning

confidence: 97%

“…16 Some of these approaches require extensive metabolic re-routing and introduction of foreign pathways, 19,22 while others rely on a small number of mutations that redirect aromatic metabolism to the product of interest. 17,18 Here we report on the impact of gene deletions in the central aromatic catabolic pathways of Novosphingobium aromaticivorans DSM12444, an organism known to degrade aromatic compounds 24 and to break down interlinkages in lignin, 25 that allow it to funnel a large diversity of plant-derived phenolics into PDC, a potential bioplastic and epoxy adhesives precursor. 26 A complete genome sequence is available for this α-proteobacterium (GenBank NC_007794.1), and the organism is amenable to genetic and genomic techniques needed to test the role of individual genes in aromatic metabolism, and model, engineer, or improve its pathways.…”

Section: Introductionmentioning

confidence: 99%

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Funneling aromatic products of chemically depolymerized lignin into 2-pyrone-4-6-dicarboxylic acid withNovosphingobium aromaticivorans

et al. 2019

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Bacterial Strains Growth Media and Culturing Conditionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Funneling aromatic products of chemically depolymerized lignin into 2-pyrone-4-6-dicarboxylic acid withNovosphingobium aromaticivorans

et al. 2019

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Novosphingobium

Kumar¹,

Kumari²,

Kumar³

et al. 2020

Bergey's Manual of Systematics of Archaea and Bacteria

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No.vo.sphin.go'bi.um. L. masc. adj. novus new; N.L. neut. n. Sphingobium a bacterial genus; N.L. neut. n. Novosphingobium a new Sphingobium . Proteobacteria / Alphaproteobacteria / Sphingomonadales / Sphingomonadaceae / Novosphingobium The genus Novosphingobium accommodates Gram‐negative, aerobic, nonsporulating, chemoorganotrophic, and rod‐shaped bacteria which are metabolically versatile. The members of this genus have been isolated from a wide range of ecological habitats including soil, coastal and freshwater sediments, deep‐sea environments, groundwater, wastewater treatment plants, activated sludge, contaminated groundwater, bioremediation reactor, hexachlorocyclohexane‐contaminated soil, oil‐contaminated sites, polychlorinated‐dioxin‐contaminated environments, rice seeds, rhizosphere, and plants. Morphologically, the colonies are mostly yellow in color, convex, circular, and smooth. Most of these isolates can be subcultured on R2A agar and may or may not be catalase‐ and oxidase‐positive. Similarly, they may or may not reduce nitrate into nitrite. C 18:1 ω7 c is the major fatty acid present in the membranes of the genus Novosphingobium . The major quinone is ubiquinone‐10, while the major polyamine is spermidine. The major polar lipids observed in Novosphingobium are diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmethylethanolamine, sphingoglycolipid, and phosphatidylcholine. The DNA G + C content ( mol% ): varies between 57.7 and 69.1, lowest in the case of Novosphingobium marinum and highest in the case of Novosphingobium bradum . DNA G + C content (mol%): 57.7–69.1. Type species: Novosphingobium capsulatum (Leifson 1962) Takeuchi et al. 2001.

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Engineering Pseudomonas putida for improved utilization of syringyl aromatics

Mueller

Willett

Feist

et al. 2022

Biotech & Bioengineering

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Lignin is a largely untapped source for the bioproduction of value-added chemicals.Pseudomonas putida KT2440 has emerged as a strong candidate for bioprocessing of lignin feedstocks due to its resistance to several industrial solvents, broad metabolic capabilities, and genetic amenability. Here we demonstrate the engineering of P. putida for the ability to metabolize syringic acid, one of the major products that comes from the breakdown of the syringyl component of lignin. The rational design was first applied for the construction of strain Sy-1 by overexpressing a native vanillate demethylase. Subsequent adaptive laboratory evolution (ALE) led to the generation of mutations that achieved robust growth on syringic acid as a sole carbon source. The best mutant showed a 30% increase in the growth rate over the original engineered strain. Genomic sequencing revealed multiple mutations repeated in separate evolved replicates. Reverse engineering of mutations identified in agmR, gbdR, fleQ, and the intergenic region of gstB and yadG into the parental strain recaptured the improved growth of the evolved strains to varied extent. These findings thus reveal the ability of P. putida to utilize lignin more fully as a feedstock and make it a more economically viable chassis for chemical production.adaptive lab evolution (ALE), Pseudomonas putida KT2440, syringyl lignin-derived aromatics | INTRODUCTIONLignin is a complex organic polymer that makes up 20%-35% of plant cell walls. It is polymerized from three monolignols: p-coumaryl alcohol, coniferyl alcohol, and sinapyl alcohol, which are categorized as the corresponding hydroxy (H), guaiacyl (G), and syringyl (S) lignin components (Figure 1a;Feofilova & Mysyakina, 2016). Depending on the type of plants, the ratio of H, G, and S subunits in lignin varies, where the grass lignin consists of all subunits, the softwood lignin often has low S component

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Novosphingobium aromaticivorans uses a Nu-class glutathione S-transferase as a glutathione lyase in breaking the β-aryl ether bond of lignin

Cited by 53 publications

References 45 publications

Funneling aromatic products of chemically depolymerized lignin into 2-pyrone-4-6-dicarboxylic acid withNovosphingobium aromaticivorans

Funneling aromatic products of chemically depolymerized lignin into 2-pyrone-4-6-dicarboxylic acid withNovosphingobium aromaticivorans

Novosphingobium

Engineering Pseudomonas putida for improved utilization of syringyl aromatics

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