Muconic Acid Production Using Engineered <i>Pseudomonas putida</i> KT2440 and a Guaiacol-Rich Fraction Derived from Kraft Lignin

Almqvist, Henrik; Veras, Henrique; Li, Kena; Hidalgo, Javier Garcia; Hulteberg, Christian; Gorwa-Grauslund, Marie-Françoise; Parachin, Nádia Skorupa; Carlquist, Magnus

doi:10.1021/acssuschemeng.1c00933

Cited by 49 publications

(33 citation statements)

References 46 publications

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“…AV and AS are major lignin-derived aromatic monomers produced in alkaline oxidative depolymerization (13)(14)(15)(16)) and base-catalyzed depolymerization of lignin (17)(18)(19)(62)(63)(64)(65)(66)(67)(68). They are also contained in the black liquor produced during the kraft pulping process (20).…”

Section: Discussionmentioning

confidence: 99%

“…In the alkaline oxidation of lignin, aromatic aldehydes (vanillin, syringaldehyde, and p-hydroxybenzaldehyde), aromatic acids (vanillic acid, syringic acid, and phydroxybenzoic acid), and acetophenone-related compounds [acetovanillone (AV), acetosyringone (AS; a syringyl-type acetophenone derivative), and 4'hydroxyacetophenone (a p-hydroxyphenyl-type acetophenone derivative)] are produced as major aromatic monomers (13)(14)(15). Base-catalyzed Indulin AT (a pine kraft lignin) depolymerization resulted in the formation of vanillin, vanillic acid, guaiacol, and AV as major aromatic monomers, similar to the alkaline oxidation of Lignoboost lignin (a softwood kraft technical lignin) (8,(16)(17)(18)(19). A black liquor produced in a softwood kraft pulping process, Lignoforce, contained guaiacol, vanillin, and AV as major aromatic monomers (20).…”

Section: Introductionmentioning

confidence: 99%

“…Among these major aromatic monomers, microbial catabolic systems of vanillin, vanillic acid, and guaiacol have been characterized (21)(22)(23)(24)(25)(26)(27)(28)(29). Additionally, value-added chemical production from vanillin, vanillic acid, and guaiacol has also been explored (12,18,(30)(31)(32)(33)(34)(35)(36). However, due to the lack of information on the microbial AV catabolic system, chemical production from AV has not been reported (16,18,19).…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, value-added chemical production from vanillin, vanillic acid, and guaiacol has also been explored (12,18,(30)(31)(32)(33)(34)(35)(36). However, due to the lack of information on the microbial AV catabolic system, chemical production from AV has not been reported (16,18,19).…”

Section: Introductionmentioning

confidence: 99%

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Bacterial catabolic system of acetovanillone and acetosyringone useful for upgrading aromatic compounds obtained through chemical lignin depolymerization

Higuchi

Kamimura²,

Takenami³

et al. 2022

Preprint

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Acetovanillone is a major aromatic monomer produced in oxidative/base-catalyzed lignin depolymerization. However, the production of chemical products from acetovanillone has not been explored due to the lack of information on the microbial acetovanillone catabolic system. Here acvABCDEF was identified as specifically induced genes during the growth of Sphingobium sp. strain SYK-6 cells with acetovanillone and these genes were essential for SYK-6 growth on acetovanillone and acetosyringone (a syringyl-type acetophenone derivative). AcvAB and AcvF produced in Escherichia coli phosphorylated acetovanillone/acetosyringone and dephosphorylated the phosphorylated acetovanillone/acetosyringone, respectively. AcvCDE produced in Sphingobium japonicum UT26S converted the dephosphorylated phosphorylated acetovanillone/acetosyringone intermediate into vanilloyl acetic acid/3- (4-hydroxy-3,5-dimethoxyphenyl)-3-oxopropanoic acid through carboxylation. To demonstrate the feasibility of producing cis,cis-muconic acid from acetovanillone, a metabolic modification on a mutant of Pseudomonas sp. strain NGC7 that accumulates cis,cis-muconic acid from catechol was performed. The resulting strain expressing vceA and vceB required for converting vanilloyl acetic acid to vanillic acid and aroY encoding protocatechuic acid decarboxylase in addition to acvABCDEF successfully converted 1.2 mM acetovanillone to approximate equimolar cis,cis-muconic acid. Our results are expected to help improve the yield and purity of value-added chemical production from lignin through biological funneling.IMPORTANCEIn the alkaline oxidation of lignin, aromatic aldehydes (vanillin, syringaldehyde, and p-hydroxybenzaldehyde), aromatic acids (vanillic acid, syringic acid, and p- hydroxybenzoic acid), and acetophenone-related compounds (acetovanillone, acetosyringone, and 4’-hydroxyacetophenone) are produced as major aromatic monomers. Also, base-catalyzed depolymerization of guaiacyl lignin resulted in vanillin, vanillic acid, guaiacol, and acetovanillone as primary aromatic monomers. To date, microbial catabolic systems of vanillin, vanillic acid, and guaiacol have been well characterized, and the production of value-added chemicals from them has also been explored. However, due to the lack of information on the microbial acetovanillone and acetosyringone catabolic system, chemical production from acetovanillone and acetosyringone has not been achieved. This is the first study to elucidate the acetovanillone/acetosyringone catabolic system, and to demonstrate the potential of using these genes for value-added chemicals production from these compounds.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Bacterial catabolic system of acetovanillone and acetosyringone useful for upgrading aromatic compounds obtained through chemical lignin depolymerization

Higuchi

Kamimura²,

Takenami³

et al. 2022

Preprint

View full text Add to dashboard Cite

show abstract

“…Samples taken from the acceptor side of the diffusion cells were first neutralized with HCl and then analyzed for vanillin, vanillic acid, and guaiacol in a Waters H-Class UPLC system using a method described in the literature [ 28 ]. A gradient method with water (A) and acetonitrile (B), both with 10 mM formic acid, was used to separate the lignin monomers in an Agilent InfinityLab Poroshell 120 EC-C18 column (100 × 4.6 mm, 4 μm).…”

Section: Methodsmentioning

confidence: 99%

Mass Transport of Lignin in Confined Pores

et al. 2022

Self Cite

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A crucial step in the chemical delignification of wood is the transport of lignin fragments into free liquor; this step is believed to be the rate-limiting step. This study has investigated the diffusion of kraft lignin molecules through model cellulose membranes of various pore sizes (1–200 nm) by diffusion cells, where the lignin molecules diffuse from donor to acceptor cells through a membrane, where diffusion rate increases by pore size. UV–vis spectra of the donor solutions showed greater absorbance at higher wavelengths (~450 nm), which was probably induced by scattering due to presence of large molecules/clusters, while acceptor samples passed through small pore membranes did not. The UV–vis spectra of acceptor solutions show a characteristic peak at around 350 nm, which corresponds to ionized conjugated molecules: indicating that a chemical fractionation has occurred. Size exclusion chromatography (SEC) showed a difference in the molecular weight (Mw) distribution between lignin from the donor and acceptor chambers. The results show that small pore sizes enable the diffusion of small individual molecules and hinder the transport of large lignin molecules or possible lignin clusters. This study provides more detail in understanding the mass transfer events of pulping processes.

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Surface Display and Engineering of Laccase CotA for Increased Growth of Pseudomonas putida on Lignin

Gesing,

Lenz,

Schreiber

et al. 2024

ChemCatChem

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Lignin is an underutilized raw material. Existing processes in which bacteria degrade lignin and produce added value products are not economically feasible yet due to limited lignin degradation. The aim of our study was to enhance microbial lignin degradation by surface‐display of the bacterial laccase CotA from the thermophilic Bacillus coagulans. Enzyme engineering of surface‐displayed CotA was used to increase its activity at 30 °C for the application at moderate temperatures. Libraries of CotA were created using error‐prone PCR and site saturation mutagenesis. Two different substrates with distinct binding properties in CotA were utilized for activity screening to detect variants with enhanced activity regardless of substrate specificity. Combination of favorable mutations resulted in the variant CotA T260S/L385K/F416R which showed 1.95‐fold and 13.4‐fold activity of CotA with 2,2’‐azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) and 2,6‐dimethoxyphenol (2,6‐DMP) as substrate, respectively. All mutations were located within the substrate binding site. The KM‐value of CotA T260S/L385K/F416R was increased with ABTS (2.08 mM versus 0.44 mM of ancestor CotA) and decreased with 2,6‐DMP (0.31 mM versus 3.79 mM of ancestor CotA). Surface display of the identified variant CotA T260S/L385K/F416R on Pseudomonasputida enhanced its growth on lignin.

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Muconic Acid Production Using Engineered Pseudomonas putida KT2440 and a Guaiacol-Rich Fraction Derived from Kraft Lignin

Cited by 49 publications

References 46 publications

Bacterial catabolic system of acetovanillone and acetosyringone useful for upgrading aromatic compounds obtained through chemical lignin depolymerization

Bacterial catabolic system of acetovanillone and acetosyringone useful for upgrading aromatic compounds obtained through chemical lignin depolymerization

Mass Transport of Lignin in Confined Pores

Surface Display and Engineering of Laccase CotA for Increased Growth of Pseudomonas putida on Lignin

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