title: Powerful workhorses for antimicrobial peptide expression and characterization.
abstract: Discovery of antimicrobial peptides (AMP) is to a large extent based on screening of fractions of natural samples in bacterial growth inhibition assays. However, the use of bacteria is not limited to screening for antimicrobial substances. In later steps, bioengineered "bugs" can be applied to both production and characterization of AMPs. Here we describe the idea to use genetically modified Escherichia coli strains for both these purposes. This approach allowed us to investigate SpStrongylocins 1 and 2 from the purple sea urchin Strongylocentrotus purpuratus only based on sequence information from a cDNA library and without previous direct isolation or chemical synthesis of these peptides. The recombinant peptides are proved active against all bacterial strains tested. An assay based on a recombinant E. coli sensor strain expressing insect luciferase, revealed that SpStrongylocins are not interfering with membrane integrity and are therefore likely to have intracellular targets.
authors.affiliation: Norwegian College of Fishery Science, Faculty of Bioscience, Fisheries and Economics, University of Tromsø, Breivika, Tromsø, Norway.
authors.forename: Chun
authors.initials: C
authors.lastname: Li
authors.forename: Hans-Matti
authors.initials: HM
authors.lastname: Blencke
authors.forename: Victoria
authors.initials: V
authors.lastname: Paulsen
authors.forename: Tor
authors.initials: T
authors.lastname: Haug
authors.forename: Klara
authors.initials: K
authors.lastname: Stensvåg
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D005818:Genetic Engineering; D047270:Strongylocentrotus purpuratus / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides