Summary: The genes encoding pyruvate dehydrogenase (PDH) of Thiobacillus ferrooxidans were previously located by cloning and sequence analysis of the region upstream of the genes encoding the citrate synthase and -glutamylcysteine synthetase genes. The pdh genes of T. ferrooxidans were able to complement an Escherichia coli aroP-lpd mutant for growth on minimal medium lacking acetate, indicating that the T. ferrooxidans PDH complex was functional in E. coli. The predicted amino acid sequence of the T. ferrooxidans PDH complex contained three ORFs. The first ORF encoded a 36.7 kDa homologue of the PDH complex E1α subunit, the second ORF a 37.4 kDa E1α subunit and the third ORF an unusual 102 kDa fusion of the E2 and E3 subunits. In spite of T. ferrooxidans being a Gram-negative bacterium, its PDH complex had more features in common with Gram-positive bacteria and eukaryotes.
GuestJ. R.1987; Functional implications and structural homologies between chloramphenicol acetyltransferase and dihydrolipoamide dehydrogenase. FEMS Microbial Lett 44:417–422
GuestJ. R.,
LewisH. M.1985; Genetic reconstruction and functional analysis of the repeating lipoyl domains in the pyruvate dehydrogenase multienzyme complex of Escherichia coli
. J Mol Biol 185:743–754
GuestJ. R.,
RobertsR. E.,
StephensP. E.1983; Hybrid plasmids containing the pyruvate dehydrogenase complex genes and gene-DNA relationships in the 2 to 3 minute region of the Escherichia coli chromosome. J Gen Microbiol 129:671–680
HanemaaijerR.,
JanssenA.,
de KokA.,
VeegerC.1988; The dihydrolipoyltransacetylase component of the pyruvate dehydrogenase complex from Azotobacter vinelandii
. Eur J Biochem 174:593–599
HeinS.,
SteinbüchelA.1994; Biochemical and molecular characterization of the Alcaligenes eutrophus pyruvate dehydrogenase complex and identfication of a new type of dihydrolipoamide dehydrogenase. J Bacteriol 176:4394–4408
HendleJ.,
MatteviA.,
WestphalA. H.,
SpeeJ.,
de KokA.,
TeplyakovA.,
HoIW. G.1995; Crystallographic and enzymatic investigations on the role of Ser558, His610 and Asn614 in the catalytic mechanism of Azotobacter vinelandii dihydrolipoamide acetyltransferase (E2p). Biochemistry 34:4287–4298
KellyD. P.,
HarrisonA. P.1989; Genus Thiobacillus Beijerinck. . In Bergey’s Manual of Systematic Bacteriology , vol. 3 , pp. 1842–1858 . Edited by
StaleyJ. T.,
ByrantM. P.,
PfennigN.,
HoltJ. G.
Baltimore: Williams & Wilkins;
MatteviA.,
ObmolovaG.,
SchulzeE.,
KalkK. H.,
WestphalA. H.,
de KokA.,
HoIW. G. J.1992b; The atomic structure of the cubic core of the pyruvate dehydrogenase complex. Science 255:1544–1550
OliverR. M.,
ReedL. J.1982; Multienzyme complexes. . In Electron Microscopy of Proteins , vol. 2 , pp. 1–48 . Edited by
HarrisJ. R.
London: Academic Press;
PowlesR.,
DeaneS.,
RawlingsD.1996; The gene for γ- glutamylcysteine synthetase from Thiobacillus ferrooxidans has low homology to its Escherichia coli equivalent and is linked to the gene for citrate synthase. Microbiology 142:2543–2548
ThompsonJ. D.,
HigginsD. G.,
GibsonT. J.1994; clustalw: improving the sensitivity of multiple sequence alignment through sequence weighting position specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680