Screening of neutrophil activating factors from a metagenome library of sponge-associated bacteria

Yoshiko Okamura*, Hirokazu Takahashi, Atsuyuki Shiida, Yuto Hirata, Haruko Takeyama, Katsuhiko Suzuki

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Marine sponge-associated bacteria are known as bio-active compound produce. We have constructed metagenome libraries of the bacteria and developed a metagenomic screening approach. Activity-based screening successfully identified novel genes and novel enzymes; however, the efficiency was only in 1 out of 104 clones. Therefore, in this study, we thought that bioinformatics could help to reduce screening efforts, and combined activity-based screening with database search. Neutrophils play an important role for the immune system to recognize excreted bacterial by-products as chemotactic factors and are recruited to infection sites to kill pathogens via phagocytosis. These excreted by-products are considered critical triggers that engage the immune system to mount a defense against infection, and identifying these factors may guide developments in medicine and diagnostics. We focused on genes encoding amino acid ligase and peptide synthetase and selected from an in-house sponge metagenome database. Cell-free culture medium of each was used in a neutrophil chemiluminescence assay in luminol reaction. The clone showing maximum activity had a genomic sequence expected to produce a molecule like a phospho-N-acetylmuramyl pentapeptide by the metagenome fragment analysis.

Original languageEnglish
Article number427
JournalMarine Drugs
Volume19
Issue number8
DOIs
Publication statusPublished - 2021 Aug

Keywords

  • Activity-based screening
  • Bioactive compound
  • Metagenome analysis
  • Neutrophil activating peptide

ASJC Scopus subject areas

  • Drug Discovery

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