The identification of a low molecular mass bacteriocin, rhamnosin A, produced by Lactobacillus rhamnosus strain 68.
OBJECTIVE
This study focuses on the isolation and characterization of a peptide with bacteriocin-like properties isolated from Lactobacillus rhamnosus strain 68, previously identified by 16S rRNA gene sequencing and originating from human gastrointestinal flora.


RESULTS
The peptide was isolated from a supernatant of bacteria maintained under restrictive conditions by a combination of ethanol precipitation and reversed-phase chromatography. The molecular mass of the peptide as assessed by mass spectrometry was 6433.8 Da. An isoelectric point of 9.8 was determined by 2D-PAGE. The peptide designated rhamnosin A inhibited Micrococcus lysodeikticus ATCC 4698 but did not inhibit Lactobacillus plantarum 8014 or Lact. plantarum 39268. Inhibitory activity against M. lysodeikticus at concentrations used in this study was shown to be bacteriostatic rather than bacteriolytic or bactericidal. Rhamnosin A retained biological activity after heat treatment (95 degrees C, 30 min) but was sensitive to proteolytic activity of pepsin and trypsin.


CONCLUSIONS
The N-terminal sequence of rhamnosin A, as determined by Edman degradation and in more detail by blast analysis, did not show identity with any currently available Lact. rhamnosus HN001-translated protein sequences, nor any significant similarity with other sequences in the nonredundant protein sequence database. Being a small, heat-stable, nonlanthionine-containing peptide, rhamnosin A should be categorized as a class II bacteriocin.


CONCLUSIONS
This study describes a partial bacteriocin sequence isolated from Lact. rhamnosus 68 and broadens our understanding of bacteriocins.
Dimitrijević
R
R
Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Belgrade, Serbia. dimitrijevicrajna@gmail.com
Stojanović
M
M
Zivković
I
I
Petersen
A
A
Jankov
R M
RM
Dimitrijević
L
L
Gavrović-Jankulović
M
M
D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000737:chemistry* / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D007526:Isoelectric Point; D048191:Lactiplantibacillus plantarum / Q000235:genetics / Q000378:metabolism; D052201:Lacticaseibacillus rhamnosus / Q000235:genetics / Q000378:metabolism*; D008837:Micrococcus / Q000187:drug effects; D008970:Molecular Weight
D001430:Bacteriocins; C550864:rhamnosin A, Lactobacillus rhamnosus