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PMID: 12957931 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Genetic and biochemical characterization of the phosphoenolpyruvate:glucose/mannose phosphotransferase system of Streptococcus thermophilus.

Applied and environmental microbiology ·Vol. 69 ·No. 9 ·2003-09-00 ·Pages 5423-32

Cochu A, Vadeboncoeur C, Moineau S, Frenette M

Abstract

In most streptococci, glucose is transported by the phosphoenolpyruvate (PEP):glucose/mannose phosphotransferase system (PTS) via HPr and IIAB(Man), two proteins involved in regulatory mechanisms. While most strains of Streptococcus thermophilus do not or poorly metabolize glucose, compelling evidence suggests that S. thermophilus possesses the genes that encode the glucose/mannose general and specific PTS proteins. The purposes of this study were to determine (i) whether these PTS genes are expressed, (ii) whether the PTS proteins encoded by these genes are able to transfer a phosphate group from PEP to glucose/mannose PTS substrates, and (iii) whether these proteins catalyze sugar transport. The pts operon is made up of the genes encoding HPr (ptsH) and enzyme I (EI) (ptsI), which are transcribed into a 0.6-kb ptsH mRNA and a 2.3-kb ptsHI mRNA. The specific glucose/mannose PTS proteins, IIAB(Man), IIC(Man), IID(Man), and the ManO protein, are encoded by manL, manM, manN, and manO, respectively, which make up the man operon. The man operon is transcribed into a single 3.5-kb mRNA. To assess the phosphotransfer competence of these PTS proteins, in vitro PEP-dependent phosphorylation experiments were conducted with purified HPr, EI, and IIAB(Man) as well as membrane fragments containing IIC(Man) and IID(Man). These PTS components efficiently transferred a phosphate group from PEP to glucose, mannose, 2-deoxyglucose, and (to a lesser extent) fructose, which are common streptococcal glucose/mannose PTS substrates. Whole cells were unable to catalyze the uptake of mannose and 2-deoxyglucose, demonstrating the inability of the S. thermophilus PTS proteins to operate as a proficient transport system. This inability to transport mannose and 2-deoxyglucose may be due to a defective IIC domain. We propose that in S. thermophilus, the general and specific glucose/mannose PTS proteins are not involved in glucose transport but might have regulatory functions associated with the phosphotransfer properties of HPr and IIAB(Man).

MeSH Terms
Amino Acid Sequence Base Sequence DNA Primers Genotype Glucose/metabolism Kinetics Mannose/metabolism Molecular Sequence Data Operon Phenotype Phosphoenolpyruvate Sugar Phosphotransferase System/genetics,metabolism RNA, Messenger/genetics Restriction Mapping Streptococcus/enzymology Transcription, Genetic
Chemicals
DNA Primers RNA, Messenger Phosphoenolpyruvate Sugar Phosphotransferase System Glucose Mannose
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Cochu Armelle
Groupe de Recherche en Ecologie Buccale, Faculté de Médecine Dentaire, Université Laval, Québec, Québec G1K 7P4, Canada.
Vadeboncoeur Christian
Moineau Sylvain
Frenette Michel
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/AEM.69.9.5423-5432.2003
Published
2003-09-00
Pages
5423-32
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC194979
Subset
IM
External Links
PubMed source DOI: 10.1128/AEM.69.9.5423-5432.2003 PMC Full Text View journal details
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