Type

Journal Article

Authors

R Paul Ross
Colin Hill
Paul D. Cotter
Barry Collins
Des Field

Subjects

Biochemistry

Topics
plasmids anti bacterial agents promoter regions genetic genetics chemistry amino acid sequence lacticin 481 mutagenesis lactococcus lactis bacteriocins genes bacterial pharmacology molecular sequence data

A system for the random mutagenesis of the two-peptide lantibiotic lacticin 3147: analysis of mutants producing reduced antibacterial activities. (2007)

Abstract Lantibiotics are antimicrobial peptides that contain several unusual amino acids resulting from a series of enzyme-mediated posttranslational modifications. As a consequence of being gene-encoded, the implementation of peptide bioengineering systems has the potential to yield lantibiotic variants with enhanced chemical and physical properties. Here we describe a functional two-plasmid expression system which has been developed to allow random mutagenesis of the two-component lantibiotic, lacticin 3147. One of these plasmids contains a randomly mutated version of the two structural genes, ltnA1 and ltnA2, and the associated promoter, Pbac, while the other encodes the remainder of the proteins required for the biosynthesis of, and immunity to, lacticin 3147. To test this system, a bank of approximately 1,500 mutant strains was generated and screened to identify mutations that have a detrimental impact on the bioactivity of lacticin 3147. This strategy established/confirmed the importance of specific residues in the structural peptides and their associated leaders and revealed that a number of alterations which mapped to the -10 or -35 regions of Pbac abolished promoter activity.
Collections Ireland -> University College Cork -> PubMed

Full list of authors on original publication

R Paul Ross, Colin Hill, Paul D. Cotter, Barry Collins, Des Field

Experts in our system

1
R Paul Ross
Teagasc
Total Publications: 441
 
2
Colin Hill
University College Cork
Total Publications: 351
 
3
Paul D. Cotter
Teagasc
Total Publications: 253
 
4
Barry Collins
University College Cork
Total Publications: 11
 
5
Des Field
Teagasc
Total Publications: 30