Metagenomic discovery of novel enzymes and biosurfactants in a slaughterhouse biofilm microbial community.
DNA derived from environmental samples is a rich source of novel bioactive molecules. The choice of the habitat to be sampled predefines the properties of the biomolecules to be discovered due to the physiological adaptation of the microbial community to the prevailing environmental conditions. We have constructed a metagenomic library in Escherichia coli DH10b with environmental DNA (eDNA) isolated from the microbial community of a slaughterhouse drain biofilm consisting mainly of species from the family Flavobacteriaceae. By functional screening of this library we have identified several lipases, proteases and two clones (SA343 and SA354) with biosurfactant and hemolytic activities. Sequence analysis of the respective eDNA fragments and subsequent structure homology modelling identified genes encoding putative N-acyl amino acid synthases with a unique two-domain organisation. The produced biosurfactants were identified by NMR spectroscopy as N-acyltyrosines with N-myristoyltyrosine as the predominant species. Critical micelle concentration and reduction of surface tension were similar to those of chemically synthesised N-myristoyltyrosine. Furthermore, we showed that the newly isolated N-acyltyrosines exhibit antibiotic activity against various bacteria. This is the first report describing the successful application of functional high-throughput screening assays for the identification of biosurfactant producing clones within a metagenomic library.
Institut für Molekulare Enzymtechnologie, Heinrich-Heine-Universität Düsseldorf, Jülich, Germany.
Stephan
S
Thies
Institut für Molekulare Enzymtechnologie, Heinrich-Heine-Universität Düsseldorf, Jülich, Germany.
Sonja Christina
SC
Rausch
Institut für Molekulare Enzymtechnologie, Heinrich-Heine-Universität Düsseldorf, Jülich, Germany.
Filip
F
Kovacic
Institut für Bioorganische Chemie, Heinrich-Heine-Universität Düsseldorf, Jülich, Germany.
Alexandra
A
Schmidt-Thaler
Institut für Molekulare Enzymtechnologie, Heinrich-Heine-Universität Düsseldorf, Jülich, Germany.
Susanne
S
Wilhelm
Zentrum für Peptidpharmazeutika, Universität Ulm, Ulm, Germany.
Frank
F
Rosenau
Institut für Mikrobiologie und Genetik, Abteilung für Genomik und Angewandte Mikrobiologie &Göttingen Genomics Laboratory, Georg-August-Universität Göttingen, Germany.
Rolf
R
Daniel
Biocenter Klein Flottbek, Department of Microbiology and Biotechnology, University of Hamburg, Hamburg, Germany.
Wolfgang
W
Streit
Institut für Bioorganische Chemie, Heinrich-Heine-Universität Düsseldorf, Jülich, Germany.|Institut für Bio- und Geowissenschaften IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany.
Jörg
J
Pietruszka
Institut für Molekulare Enzymtechnologie, Heinrich-Heine-Universität Düsseldorf, Jülich, Germany.|Institut für Bio- und Geowissenschaften IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, Jülich, Germany.
Karl-Erich
KE
Jaeger
D000003:Abattoirs; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics; D018441:Biofilms; D053898:Biosynthetic Pathways; D004783:Environmental Microbiology; D004926:Escherichia coli / Q000235:genetics; D005417:Flavobacterium / Q000235:genetics; D005798:Genes, Bacterial; D054892:Metagenome; D059013:Microbial Consortia / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D016366:Open Reading Frames; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D013501:Surface-Active Agents / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D013501:Surface-Active Agents