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

2021; Elsevier BV; Volume: 65; Linguagem: Inglês

10.1016/j.ymben.2021.02.003

1096-7184

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

Enzyme-mediated dye degradation

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 β-1 spirodienone linkage cleavage has not been elucidated. Here, we identified a pathway for cleavage of 1,2-diguaiacylpropane-1,3-diol (DGPD), a β-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.