Abstract
BackgroundThe efficient depolymerization and utilization of lignin are one of the most important goals for the renewable use of lignocelluloses. The degradation and complete mineralization of lignin by bacteria represent a key step for carbon recycling in land ecosystems as well. However, many aspects of this process remain unclear, for example, the complex network of metabolic pathways involved in the degradation of lignin and the catabolic pathway of intermediate aromatic metabolites. To address these subjects, we characterized the deconstruction and mineralization of lignin with milled wood lignin (MWL, the most representative molecule of lignin in its native state) and alkali lignin (AL), and elucidated metabolic pathways of their intermediate metabolites by a bacterium named Comamonas serinivorans SP-35.ResultsThe degradation rate of MWL reached 30.9%, and its particle size range was decreased from 6 to 30 µm to 2–4 µm—when cultured with C. serinivorans SP35 over 7 days. FTIR analysis showed that the C–C and C–O–C bonds between the phenyl propane structures of lignin were oxidized and cleaved and the side chain structure was modified. More than twenty intermediate aromatic metabolites were identified in the MWL and AL cultures based on GC–MS analysis. Through genome sequencing and annotation, and from GC–MS analysis, 93 genes encoding 33 enzymes and 5 regulatory factors that may be involved in lignin degradation were identified and more than nine metabolic pathways of lignin and its intermediates were predicted. Of particular note is that the metabolic pathway to form the powerful antioxidant 3,4-dihydroxyphenylglycol is described for the first time in bacteria.ConclusionElucidation of the β-aryl ether cleavage pathway in the strain SP-35 indicates that the β-aryl ether catabolic system is not only present in the family of Sphingomonadaceae, but also other species of bacteria kingdom. These newly elucidated catabolic pathways of lignin in strain SP-35 and the enzymes responsible for them provide exciting biotechnological opportunities for lignin valorization in future.
Highlights
The efficient depolymerization and utilization of lignin are one of the most important goals for the renewable use of lignocelluloses
Growth of SP‐35 and removal of chemical oxygen demand (COD) The strain SP-35 was able to slightly grow (OD600 from 0.52 increased to 0.78) with milled wood lignin (MWL) as the sole carbon source and had a faster proliferation when glucose was added as mixed carbon source; cells quickly entered into the decline phase without supplies of carbon sources (Fig. 1)
The degradation rate of lignin by strain SP-35 can be determined by COD removal analysis because MWL contributes almost the entire COD loading in the culture medium when it was the sole carbon source
Summary
The efficient depolymerization and utilization of lignin are one of the most important goals for the renewable use of lignocelluloses. Many aspects of this process remain unclear, for exam‐ ple, the complex network of metabolic pathways involved in the degradation of lignin and the catabolic pathway of intermediate aromatic metabolites. To address these subjects, we characterized the deconstruction and mineraliza‐ tion of lignin with milled wood lignin (MWL, the most representative molecule of lignin in its native state) and alkali lignin (AL), and elucidated metabolic pathways of their intermediate metabolites by a bacterium named Comamonas serinivorans SP-35. It has been reported that Lacc and DyPs, were observed as oxidases in lignin degrading bacteria, but without LiP, MnP and VP [2, 3]. Bacteria do not possess the typical peroxidases that fungi employ for lignin degradation, which suggests that the ligninolytic enzymatic system of bacteria might be different from that of fungus
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