The complete nucleotide sequence of two genes from Clostridium thermosulfurogenes EM1 homologous to E. coli genes encoding transport proteins was determined by the dideoxy procedure. The genes were cloned from plasmid pCT4, which contains the alpha-amylase gene from C. thermosulfurogenes EM1 as a 2.9-kbp XbaI fragment, inserted into the XbaI site of pUC18, to yield plasmid pCT401. The proteins encoded by the two identified complete ORFs are very hydrophobic and thus are probably integral membrane proteins. They show over 50% similarity to the maltose transport proteins MalF and MalG and to the glycerol-3-phosphate uptake proteins UgpA and UgpE of Escherichia coli. Since these genes are located immediately upstream of the alpha-amylase gene (amyA) of C. thermosulfurogenes EM1, the encoded proteins might be involved in transport of starch degradation products. The genes were tentatively designated amyC and amyD.
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http://dx.doi.org/10.1016/0378-1097(91)90476-q | DOI Listing |
J Mol Microbiol Biotechnol
July 2004
Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Göttingen, Germany.
Bacteria may possess various kinds of cytoskeleton. In general, bacterial cytoskeletons may play a role in the control and preservation of the cell shape. Such functions become especially evident when the bacteria do not possess a true wall and are nevertheless elongated (e.
View Article and Find Full Text PDFAppl Environ Microbiol
September 1999
Department of Chemical Engineering, Molecular Biology, and Cell Biology, Northwestern University, Evanston, Illinois 60208, USA.
A gene expression reporter system (pHT3) for Clostridium acetobutylicum ATCC 824 was developed by using the lacZ gene from Thermoanaerobacterium thermosulfurogenes EM1 as the reporter gene. In order to test the reporter system, promoters of three key metabolic pathway genes, ptb (coding for phosphotransbutyrylase), thl (coding for thiolase), and adc (coding for acetoacetate decarboxylase), were cloned upstream of the reporter gene in pHT3 in order to construct vectors pHT4, pHT5, and pHTA, respectively. Detection of beta-galactosidase activity in time course studies performed with strains ATCC 824(pHT4), ATCC 824(pHT5), and ATCC 824(pHTA) demonstrated that the reporter gene produced a functional beta-galactosidase in C.
View Article and Find Full Text PDFAppl Environ Microbiol
April 1995
Agrotechnological Research Institute, Wageningen, The Netherlands.
Extensive characterization of the thermostable alpha-amylase of Clostridium thermosulfurogenes EM1, recently reclassified as Thermoanaerobacterium thermosulfurigenes, clearly demonstrated that the enzyme is a cyclodextrin glycosyltransferase (CGTase). Product analysis after incubation of the enzyme with starch revealed formation of alpha-, beta-, and gamma-cyclodextrins, as well as linear sugars. The specific activity for cyclization of this CGTase was similar to those of other CGTases, whereas the specific activity for hydrolysis was relatively high in comparison with other CGTases.
View Article and Find Full Text PDFCurr Microbiol
July 1994
Institut für Mikrobiologic, Georg-August-Universität Göttingen, Germany.
The response to heat stress was examined in Thermoanaerobacterium thermosulfurigenes EM1. Upon a temperature shift-up from 50 degrees to 62 degrees C, four heat shock proteins (hsps) were synthesized at an elevated level. Two proteins were found to be immunologically related to the Escherichia coli GroEL protein and the Mycobacterium tuberculosis hsp71 (DnaK similar protein), and the corresponding groE and dnaK homologous sequences were detected in the chromosome of T.
View Article and Find Full Text PDFJ Bacteriol
June 1994
Institut für Mikrobiologie, Georg-August-Universität Göttingen, Federal Republic of Germany.
The complete pullulanase gene (amyB) from Thermoanaerobacterium thermosulfurigenes EM1 was cloned in Escherichia coli, and the nucleotide sequence was determined. The reading frame of amyB consisted of 5,586 bp encoding an exceptionally large enzyme of 205,991 Da. Sequence analysis revealed a composite structure of the pullulanase consisting of catalytic and noncatalytic domains.
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