Metasecretome-selective phage display approach for mining the functional potential of a rumen microbial community

Milica Ciric, Christina D. Moon, Sinead C Leahy, Christopher J Creevey, Eric Altermann, Graeme T. Attwood, Jasna Rakonjac, Dragana Gagic

Research output: Contribution to journalArticlepeer-review

14 Citations (Scopus)
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In silico, secretome proteins can be predicted from completely sequenced genomes using various available algorithms that identify membrane-targeting sequences. For metasecretome (collection of surface, secreted and transmembrane proteins from environmental microbial communities) this approach is impractical, considering that the metasecretome open reading frames (ORFs) comprise only 10% to 30% of total metagenome, and are poorly represented in the dataset due to overall low coverage of metagenomic gene pool, even in large-scale projects.

By combining secretome-selective phage display and next-generation sequencing, we focused the sequence analysis of complex rumen microbial community on the metasecretome component of the metagenome. This approach achieved high enrichment (29 fold) of secreted fibrolytic enzymes from the plant-adherent microbial community of the bovine rumen. In particular, we identified hundreds of heretofore rare modules belonging to cellulosomes, cell- surface complexes specialised for recognition and degradation of the plant fibre.

As a method, metasecretome phage display combined with next-generation sequencing has a power to sample the diversity of low-abundance surface and secreted proteins that would otherwise require exceptionally large metagenomic sequencing projects. As a resource, metasecretome display library backed by the dataset obtained by next-generation sequencing is ready for i) affinity selection by standard phage display methodology and ii) easy purification of displayed proteins as part of the virion for individual functional analysis.
Original languageEnglish
Article number356
JournalBMC Genomics
Issue number1
Publication statusPublished - 12 May 2014


  • phage display
  • Nxt generation sequencing
  • metagenomics
  • rumen
  • cellulosome
  • surface and secreted proteins
  • Metagenomics
  • Cellulosome
  • Phage display
  • Surface and secreted proteins
  • Rumen
  • Rumen/microbiology
  • Open Reading Frames
  • Bacteriophages/metabolism
  • Cellulosomes/metabolism
  • Sequence Analysis, DNA
  • Metagenomics/methods
  • Animals
  • Cattle
  • Cell Surface Display Techniques
  • Databases, Protein
  • Metagenome/genetics
  • High-Throughput Nucleotide Sequencing


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