Production of dehydroamino acid-containing peptides by Lactococcus lactis.
Nisin is a pentacyclic peptide antibiotic produced by some Lactococcus lactis strains. Nisin contains dehydroresidues and thioether rings that are posttranslationally introduced by a membrane-associated enzyme complex, composed of a serine and threonine dehydratase NisB and the cyclase NisC. In addition, the transporter NisT is necessary for export of the modified peptide. We studied the potential of L. lactis expressing NisB and NisT to produce peptides whose serines and threonines are dehydrated. L. lactis containing the nisBT genes and a plasmid coding for a specific leader peptide fusion construct efficiently produced peptides with a series of non-naturally occurring multiple flanking dehydrobutyrines. We demonstrated NisB-mediated dehydration of serines and threonines in a C-terminal nisin(1-14) extension of nisin, which implies that also residues more distant from the leader peptide than those occurring in prenisin or any other lantibiotic can be modified. Furthermore, the feasibility and efficiency of generating a library of peptides containing dehydroresidues were demonstrated. In view of the particular shape and reactivity of dehydroamino acids, such a library provides a novel source for screening for peptides with desired biological and physicochemical properties.
Rink
Rick
R
BiOMaDe Technology Foundation, Nijenborgh 4, 9747 AG Groningen, The Netherlands. moll@biomade.nl
Wierenga
Jenny
J
Kuipers
Anneke
A
Kluskens
Leon D
LD
Driessen
Arnold J M
AJ
Kuipers
Oscar P
OP
Moll
Gert N
GN
D000613:Aminobutyrates / Q000032:analysis; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism*; D013058:Mass Spectrometry; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000502:physiology*; D009561:Nisin / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012694:Serine / Q000737:chemistry / Q000378:metabolism; D013912:Threonine / Q000737:chemistry / Q000378:metabolism
D000613:Aminobutyrates; D001426:Bacterial Proteins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; C079126:NisB protein, Lactococcus lactis; C094910:NisT protein, Lactococcus lactis; D010455:Peptides; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D009561:Nisin; C088694:dehydrobutyrine; D013912:Threonine; D012694:Serine