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PMID: 8083158 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Trehalose-6-phosphate hydrolase of Escherichia coli.

Journal of bacteriology ·Vol. 176 ·No. 18 ·1994-09-00 ·Pages 5654-64

Rimmele M, Boos W

Abstract

The disaccharide trehalose acts as an osmoprotectant as well as a carbon source in Escherichia coli. At high osmolarity of the growth medium, the cells synthesize large amounts of trehalose internally as an osmoprotectant. However, they can also degrade trehalose as the sole source of carbon under both high- and low-osmolarity growth conditions. The modes of trehalose utilization are different under the two conditions and have to be well regulated (W. Boos, U. Ehmann, H. Forkl, W. Klein, M. Rimmele, and P. Postma, J. Bacteriol. 172:3450-3461, 1990). At low osmolarity, trehalose is transported via a trehalose-specific enzyme II of the phosphotransferase system, encoded by treB. The trehalose-6-phosphate formed internally is hydrolyzed to glucose and glucose 6-phosphate by the key enzyme of the system, trehalose-6-phosphate hydrolase, encoded by treC. We have cloned treC, contained in an operon with treB as the promoter-proximal gene. We have overproduced and purified the treC gene product and identified it as a protein consisting of a single polypeptide with an apparent molecular weight of 62,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme hydrolyzes trehalose-6-phosphate with a Km of 6 mM and a Vmax of at least 5.5 mumol of trehalose-6-phosphate hydrolyzed per min per mg of protein. The enzyme also very effectively hydrolyzes p-nitrophenyl-alpha-D-glucopyranoside, but it does not recognize trehalose, sucrose, maltose, isomaltose, or maltodextrins. treC was sequenced and found to encode a polypeptide with a calculated molecular weight of 63,781. The amino acid sequence deduced from the DNA sequence shows homology (50% identity) with those of oligo-1,6-glucosidases (sucrase-isomaltases) of Bacillus spp. but not with those of other disaccharide phosphate hydrolases. This report corrects our previous view on the function of the treC gene product as an amylotrehalase, which was based on the analysis of the metabolic products of trehalose metabolism in whole cells.

Related Genes
treC
MeSH Terms
Amino Acid Sequence Base Sequence Cloning, Molecular Disaccharidases/chemistry,genetics,isolation & purification,metabolism Escherichia coli/enzymology,genetics Gene Expression Genes, Bacterial/genetics Glucosides/metabolism Hexokinase/antagonists & inhibitors Kinetics Models, Biological Molecular Sequence Data Molecular Weight Sequence Analysis, DNA Sequence Homology, Amino Acid Substrate Specificity Sugar Phosphates/metabolism Trehalose/analogs & derivatives,metabolism
Chemicals
Glucosides Sugar Phosphates 4-nitrophenyl alpha-glucoside trehalose-6-phosphate Trehalose Hexokinase Disaccharidases alpha, alpha-phosphotrehalase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Rimmele M
Department of Biology, University of Konstanz, Germany.
Boos W
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.176.18.5654-5664.1994
Published
1994-09-00
Pages
5654-64
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC196768
Subset
IM
Databases
GENBANK
U06195
SWISSPROT
P28904
External Links
PubMed source DOI: 10.1128/jb.176.18.5654-5664.1994 PMC Full Text View journal details
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