Multiplexed Fitness Profiling by RB-TnSeq Elucidates Pathways for Lignin-Related Aromatic Catabolism in Sphingobium sp. SYK-6

Alissa Bleem, Ryo Kato, Zoe Kellermyer, Rui Katahira, Masahiro Miyamoto, Koh Niinuma, Naofumi Kamimura, Eiji Masai, Gregg Beckham

Research output: Contribution to journalArticlepeer-review

3 Scopus Citations

Abstract

Bioconversion of lignin-related aromatic compounds relies on robust catabolic pathways in microbes. Sphingobium sp. SYK-6 (SYK-6) is a well-characterized aromatic catabolic organism that has served as a model for microbial lignin conversion, and its utility as a biocatalyst could potentially be further improved by genome-wide metabolic analyses. To this end, we generate a randomly barcoded transposon insertion mutant (RB-TnSeq) library to study gene function in SYK-6. The library is enriched under dozens of enrichment conditions to quantify gene fitness. Several known aromatic catabolic pathways are confirmed, and RB-TnSeq affords additional detail on the genome-wide effects of each enrichment condition. Selected genes are further examined in SYK-6 or Pseudomonas putida KT2440, leading to the identification of new gene functions. The findings from this study further elucidate the metabolism of SYK-6, while also providing targets for future metabolic engineering in this organism or other hosts for the biological valorization of lignin.

Original languageAmerican English
Article number112847
Number of pages20
JournalCell Reports
Volume42
Issue number8
DOIs
StatePublished - 2023

Bibliographical note

Publisher Copyright:
© 2023 The Authors

NREL Publication Number

  • NREL/JA-2A00-84889

Keywords

  • acid tolerance
  • aromatic catabolism
  • bacterial genetics
  • CP: Microbiology
  • lignin
  • Pseudomonas putida
  • Sphingobium sp. SYK-6
  • Transposon insertion sequencing

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