Purification and characterization of the E1 component of the Clostridium magnum acetoin dehydrogenase enzyme system.
In Clostridium magnum strain Wo Bd P1 the formation of the enzyme components of the acetoin dehydrogenase enzyme system E1 (acetoin:2,6-dichlorophenolindophenol oxidoreductase Ao:DCPIP OR), E2 (dihydrolipoamide acetyltransferase DHLTA) and E3 (dihydrolipoamide dehydrogenase DHLDH) were induced during growth on acetoin. Ao:DCPIP OR was purified from acetoin-grown cells in two steps by chromatography on DEAE-Sephacel and on Mono Q HR. Native Ao:DCPIP OR exhibited a M(r) of 138,000; it consisted of two different subunits of M(r) alpha 38,500 and M(r) beta 34,000, and it occurred most probably in a tetrameric alpha 2 beta 2 structure. The N-terminal amino acid sequences of the alpha- and beta-subunits revealed homologies to the N-termini of the corresponding subunits of Ao:DCPIP OR from Pelobacter carbinolicus and from Alcaligenes eutrophus; furthermore, the N-terminus of the beta-subunit exhibited homologies to the N-termini of beta-subunits from different 2-oxo acid dehydrogenases.
Lorenzl
H
H
Institut für Mikrobiologie, Georg-August-Universität zu Göttingen, Germany.
Oppermann
F B
FB
Schmidt
B
B
Steinbüchel
A
A
D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D000093:Acetoin / Q000378:metabolism; D000094:Acetoin Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000217:Acyltransferases / Q000302:isolation & purification; D000421:Alcaligenes / Q000201:enzymology; D000595:Amino Acid Sequence; D002852:Chromatography, Ion Exchange; D003013:Clostridium / Q000201:enzymology*; D003470:Culture Media; D008058:Dihydrolipoamide Dehydrogenase / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005947:Glucose / Q000378:metabolism; D006089:Gram-Negative Anaerobic Bacteria / Q000201:enzymology; D007658:Ketone Oxidoreductases; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000737:chemistry; D010088:Oxidoreductases / Q000737:chemistry; D016415:Sequence Alignment
D003470:Culture Media; D009097:Multienzyme Complexes; D000093:Acetoin; D010088:Oxidoreductases; D000094:Acetoin Dehydrogenase; D007658:Ketone Oxidoreductases; D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); C066825:acetoin-2,6-dichlorophenolindophenol oxidoreductase; D008058:Dihydrolipoamide Dehydrogenase; D000217:Acyltransferases; C033588:dihydrolipoamide acyltransferase; D005947:Glucose