Catabolism of β-5 linked aromatics by Novosphingobium aromaticivorans

Journal Article · Wed Aug 14 00:00:00 EDT 2024 · mBio (Online)

DOI:https://doi.org/10.1128/mbio.01718-24· OSTI ID:2403579

Metz, Fletcher ^[1]; Olsen, Abigail M. ^[2]; Lu, Fachuang ^[2]; Myers, Kevin S. ^[2]; Allemann, Marco N. ^[3]; Michener, Joshua K. ^[3]; Noguera, Daniel R. ^[4]; Donohue, Timothy J. ^[5]; Harwood, ed., Caroline S.

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1. DOE Great Lakes Bioenergy Research Center, University of Wisconsin, Madison, Wisconsin, USA, Wisconsin Energy Institute, University of Wisconsin, Madison, Wisconsin, USA, Laboratory of Genetics, University of Wisconsin, Madison, Wisconsin, USA
2. DOE Great Lakes Bioenergy Research Center, University of Wisconsin, Madison, Wisconsin, USA, Wisconsin Energy Institute, University of Wisconsin, Madison, Wisconsin, USA
3. Biosciences Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee, USA
4. DOE Great Lakes Bioenergy Research Center, University of Wisconsin, Madison, Wisconsin, USA, Wisconsin Energy Institute, University of Wisconsin, Madison, Wisconsin, USA, Department of Civil and Environmental Engineering, University of Wisconsin, Madison, Wisconsin, USA
5. DOE Great Lakes Bioenergy Research Center, University of Wisconsin, Madison, Wisconsin, USA, Wisconsin Energy Institute, University of Wisconsin, Madison, Wisconsin, USA, Department of Bacteriology, University of Wisconsin, Madison, Wisconsin, USA

ABSTRACT <p> Aromatic compounds are an important source of commodity chemicals traditionally produced from fossil fuels. Aromatics derived from plant lignin can potentially be converted into commodity chemicals through depolymerization followed by microbial funneling of monomers and low molecular weight oligomers. This study investigates the catabolism of the β-5 linked aromatic dimer dehydrodiconiferyl alcohol (DC-A) by the bacterium <italic>Novosphingobium aromaticivorans</italic> . We used genome-wide screens to identify candidate genes involved in DC-A catabolism. Subsequent <italic>in vivo</italic> and <italic>in vitro</italic> analyses of these candidate genes elucidated a catabolic pathway composed of four required gene products and several partially redundant dehydrogenases that convert DC-A to aromatic monomers that can be funneled into the central aromatic metabolic pathway of <italic>N. aromaticivorans</italic> . Specifically, a newly identified γ-formaldehyde lyase, PcfL, opens the phenylcoumaran ring to form a stilbene and formaldehyde. A lignostilbene dioxygenase, LsdD, then cleaves the stilbene to generate the aromatic monomers vanillin and 5-formylferulate (5-FF). We also showed that the aldehyde dehydrogenase FerD oxidizes 5-FF before it is decarboxylated by LigW, yielding ferulic acid. We found that some enzymes involved in the β-5 catabolism pathway can act on multiple substrates and that some steps in the pathway can be mediated by multiple enzymes, providing new insights into the robust flexibility of aromatic catabolism in <italic>N. aromaticivorans</italic> . A comparative genomic analysis predicted that the newly discovered β-5 aromatic catabolic pathway is common within the order Sphingomonadales. </p> </sec> <sec> <title>IMPORTANCE

In the transition to a circular bioeconomy, the plant polymer lignin holds promise as a renewable source of industrially important aromatic chemicals. However, since lignin contains aromatic subunits joined by various chemical linkages, producing single chemical products from this polymer can be challenging. One strategy to overcome this challenge is using microbes to funnel a mixture of lignin-derived aromatics into target chemical products. This approach requires strategies to cleave the major inter-unit linkages of lignin to release monomers for funneling into valuable products. In this study, we report newly discovered aspects of a pathway by which the Novosphingobium aromaticivorans DSM12444 catabolizes aromatics joined by the second most common inter-unit linkage in lignin, the β-5 linkage. This work advances our knowledge of aromatic catabolic pathways, laying the groundwork for future metabolic engineering of this and other microbes for optimized conversion of lignin into products.

• Research Organization: Great Lakes Bioenergy Research Center (GLBRC), Madison, WI (United States); Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
• Sponsoring Organization: National Institutes of Health (NIH); USDOE; USDOE Office of Science (SC), Biological and Environmental Research (BER)
• Grant/Contract Number: AC05-00OR22725; SC0018409
• OSTI ID: 2403579
• Alternate ID(s): OSTI ID: 2406535; OSTI ID: 2426056; OSTI ID: 2468795
• Journal Information: mBio (Online), Journal Name: mBio (Online) Journal Issue: 8 Vol. 15; ISSN 2150-7511
• Publisher: American Society for Microbiology
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
Enzymes
aromatic compounds
biotechnology
enzymes
ligninolysis
genomics
biotechnology
aromatic compounds
genetics
genetics
genomics
ligninolysis