Decoding how a soil bacterium extracts building blocks and metabolic energy from ligninolysis provides road map for lignin valorization

• Published in: Proceedings of the National Academy of Sciences - PNAS 2016-10, Vol.113 (40), p.E5802-E5811
• Main Authors: Varman, Arul M., He, Lian, Follenfant, Rhiannon, Wu, Weihua, Wemmer, Sarah, Wrobel, Steven A., Tang, Yinjie J., Singh, Seema
• Format: Article
• Language: English
• Subjects: 09 BIOMASS FUELS; 13C-MFA; 60 APPLIED LIFE SCIENCES; Amino Acids - metabolism; Bacteria; Bacteria - genetics; Bacteria - growth & development; Bacteria - metabolism; Benzaldehydes - chemistry; Benzaldehydes - metabolism; Biological Sciences; Biosynthesis; Carbon - metabolism; Carbon Isotopes; Energy Metabolism; Escherichia coli; fingerprinting; Gene Expression Profiling; Gene Expression Regulation, Bacterial; gluconeogenesis; Hydrogen-Ion Concentration; INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Lignin; Lignin - metabolism; Metabolic Flux Analysis; Metabolism; NADP - metabolism; NADPH; PNAS Plus; Ribonucleic acid; RNA; Sequence Analysis, RNA; Soil; Soil Microbiology; Sphingobium; TCA; Vanillic Acid - chemistry; Vanillic Acid - metabolism
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1606043113

Sphingobium sp. SYK-6 is a soil bacterium boasting a well-studied ligninolytic pathway and the potential for development into a microbial chassis for lignin valorization. An improved understanding of its metabolism will help researchers in the engineering of SYK-6 for the production of value-added chemicals through lignin valorization. We used 13C-fingerprinting, 13C metabolic flux analysis (13C-MFA), and RNA-sequencing differential expression analysis to uncover the following metabolic traits: (i) SYK-6 prefers alkaline conditions, making it an efficient host for the consolidated bioprocessing of lignin, and it also lacks the ability to metabolize sugars or organic acids; (ii) the CO₂ release (i.e., carbon loss) from the ligninolysis-based metabolism of SYK-6 is significantly greater than the CO₂ release from the sugar-basedmetabolism of Escherichia coli; (iii) the vanillin catabolic pathway (which is the converging point of majority of the lignin catabolic pathways) is coupled with the tetrahydrofolate-dependent C1 pathway that is essential for the biosynthesis of serine, histidine, and methionine; (iv) catabolic end products of lignin (pyruvate and oxaloacetate) must enter the tricarboxylic acid (TCA) cycle first and then use phosphoenolpyruvate carboxykinase to initiate gluconeogenesis; and (v) 13C-MFA together with RNA-sequencing differential expression analysis establishes the vanillin catabolic pathway as the major contributor of NAD(P)H synthesis. Therefore, the vanillin catabolic pathway is essential for SYK-6 to obtain sufficient reducing equivalents for its healthy growth; cosubstrate experiments support this finding. This unique energy feature of SYK-6 is particularly interesting because most heterotrophs rely on the transhydrogenase, the TCA cycle, and the oxidative pentose phosphate pathway to obtain NADPH.