Pathway discovery and engineering for cleavage of a β-1 lignin-derived biaryl compound

Presley, Gerald N.; Werner, Allison Z.; Katahira, Rui; Garcia, David C.; Haugen, Stefan J.; Ramirez, Kelsey J.; Giannone, Richard J.; Beckham, Gregg T.; Michener, Joshua K.

doi:10.1016/j.ymben.2021.02.003

Title: Pathway discovery and engineering for cleavage of a β-1 lignin-derived biaryl compound

Journal Article · Tue Feb 23 00:00:00 EST 2021 · Metabolic Engineering

DOI:https://doi.org/10.1016/j.ymben.2021.02.003· OSTI ID:1771901

Presley, Gerald N. ^[1];

^[2]; Katahira, Rui ^[3];

^[4]; Haugen, Stefan J. ^[3];

^[3];

^[1]; Beckham, Gregg T. ^[2];

^[1]

Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); National Renewable Energy Lab. (NREL), Golden, CO (United States)
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States)

Lignin biosynthesis typically results in a polymer with several inter-monomer bond linkages, and the heterogeneity of linkages presents a challenge for depolymerization processes. While several enzyme classes have been shown to cleave common dimer linkages in lignin, the pathway of bacterial ß-1 spirodienone linkage cleavage has not been elucidated. Here, we identified a pathway for cleavage of 1,2-diguaiacylpropane-1,3-diol (DGPD), a ß-1 linked biaryl representative of a ring-opened spirodienone linkage, in Novosphingobium aromaticivorans DSM12444. In vitro assays using cell lysates demonstrated that RS14230 (LsdE) converts DGPD to a lignostilbene intermediate, which the carotenoid oxygenase, LsdA, then converts to vanillin. A Pseudomonas putida KT2440 strain engineered with lsdEA expression catabolizes erythro-DGPD, but not threo-DGPD. We further engineered P. putida to convert DGPD to a product, cis,cis-muconic acid. Overall, this work demonstrates the potential to identify new enzymatic reactions in N. aromaticivorans and expands the biological funnel of P. putida for microbial lignin valorization.

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Research Organization:: Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL); National Renewable Energy Lab. (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); USDOE Office of Energy Efficiency and Renewable Energy (EERE), Transportation Office. Bioenergy Technologies Office; National Science Foundation (NSF); USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office

Grant/Contract Number:: AC05-00OR22725; AC36-08GO28308

OSTI ID:: 1771901

Alternate ID(s):: OSTI ID: 1772445; OSTI ID: 1815315

Report Number(s):: NREL/JA-2A00-78148

Journal Information:: Metabolic Engineering, Vol. 65; ISSN 1096-7176

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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