Affiliation:
1. Alimentary
Pharmabiotic Centre
2. Department of
Microbiology
3. Department of Food and
Nutritional Sciences, National University of
Ireland, Cork, Cork, Ireland
Abstract
ABSTRACT
The
molecular interactions between the bifidobacterial cell and its natural
environment, namely, the gastrointestinal tract of its host, are
particularly important in understanding the presumed positive effects
of
Bifidobacterium
on the health status of the host. In this
study an export-specific reporter system, designed for use in
gram-positive organisms and based on the use of the staphylococcal
nuclease (Nuc) as a reporter, was employed to identify exported
proteins in
Bifidobacterium breve
UCC2003. A
B. breve
genomic library of translational fusions to the Nuc-encoding gene
devoid of its own export signal was established in the shuttle vector
pFUN (I. Poquet, S. D. Ehrlich, and A. Gruss, J. Bacteriol.
180:
1904-1912, 1998) and screened for bifidobacterial
export signals. Sequence analysis of the fusion proteins obtained that
displayed a nuclease-producing phenotype in both
Lactococcus
lactis
and
B. breve
predicted the presence of a classical
signal peptide and/or single or multiple transmembrane domains, thus
indicating that some of the export signals in
B. breve
are
comparable to those used in
L. lactis
. Cell fractionation
studies, zymograms, nuclease assays, and Western blotting were employed
to confirm the function of the predicted signals and to determine the
location and activity of the exported fusion proteins in
B.
breve
and/or
L.
lactis
.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
39 articles.
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