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Export of Thermus thermophilus cytoplasmic beta-glycosidase via the E. coli Tat pathway.
- Source :
-
Journal of molecular microbiology and biotechnology [J Mol Microbiol Biotechnol] 2002 Nov; Vol. 4 (6), pp. 533-8. - Publication Year :
- 2002
-
Abstract
- The Tat pathway is distinct from the Sec machinery given its unusual capacity to export folded proteins, which contain a twin-arginine (RR) signal peptide, across the plasma membrane. The functionality of the Tat pathway has been demonstrated for several Gram-negative and Gram-positive mesophilic bacteria. To assess the specificity of the Tat system, and to analyze the capacity of a mesophilic bacterial Tat system to translocate cytoplasmic proteins from hyperthermophilic bacteria, we fused the Thermus thermophilus beta-glycosidase (Glc) to the twin-arginine signal peptide of the E. coli TorA protein. When expressed in E. coli, the thermophilic RR-Glc chimera was successfully synthesized and efficiently translocated into the periplasm of the wild type strain. In contrast, the beta-glycosidase accumulated within the cytoplasm of all the tat mutants analyzed. The beta-glycosidase synthesized in these strains exhibited thermophilic properties. These results demonstrated, for the first time, the capacity of the E. coli Tat system to export cytoplasmic hyperthermophilic protein, implying an important potential of the Tat system for the production of thermostable enzymes used in bioprocessing applications.
- Subjects :
- Biotechnology methods
Culture Media
Escherichia coli genetics
Escherichia coli Proteins genetics
Glycoside Hydrolases genetics
Hot Temperature
Membrane Transport Proteins genetics
Oxidoreductases, N-Demethylating genetics
Oxidoreductases, N-Demethylating metabolism
Periplasm enzymology
Protein Transport
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins metabolism
Thermus thermophilus genetics
Cytoplasm enzymology
Escherichia coli enzymology
Escherichia coli Proteins metabolism
Glycoside Hydrolases metabolism
Membrane Transport Proteins metabolism
Oxidoreductases, N-Demethylating chemistry
Thermus thermophilus enzymology
Subjects
Details
- Language :
- English
- ISSN :
- 1464-1801
- Volume :
- 4
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Journal of molecular microbiology and biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 12432953