Purification and characterization of acetoin:2,6-dichlorophenolindophenol oxidoreductase, dihydrolipoamide dehydrogenase, and dihydrolipoamide acetyltransferase of the Pelobacter carbinolicus acetoin dehydrogenase enzyme system

Oppermann, Fred Bernd; Schmidt, Bernhard; Steinbüchel, Alexander

doi:10.1128/jb.173.2.757-767.1991

Cited by 75 publications

(81 citation statements)

References 67 publications

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“…In these bacteria, acetoin (3-hydroxy-2-butanone) is formed from pyruvate in two enzymatic steps (191), providing the substrate for AoDH. Reconstituted AoDH containing the E1, E2, and E3 subunits from the bacterium Pelobacter carbinolicus is specific for acetoin and does not use pyruvate or ␣-ketoglutarate as substrate (162). The E1␣ protein contains a region of divergent sequence compared to other ␣-ketoacid dehydrogenases and appears to be responsible for the substrate specificity of AoDH (115).…”

Section: Lipoylated Complexesmentioning

confidence: 99%

Lipoic Acid Metabolism in Microbial Pathogens

Spalding

Prigge

2010

Microbiol Mol Biol Rev

164

204

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SUMMARY Lipoic acid [(R)-5-(1,2-dithiolan-3-yl)pentanoic acid] is an enzyme cofactor required for intermediate metabolism in free-living cells. Lipoic acid was discovered nearly 60 years ago and was shown to be covalently attached to proteins in several multicomponent dehydrogenases. Cells can acquire lipoate (the deprotonated charge form of lipoic acid that dominates at physiological pH) through either scavenging or de novo synthesis. Microbial pathogens implement these basic lipoylation strategies with a surprising variety of adaptations which can affect pathogenesis and virulence. Similarly, lipoylated proteins are responsible for effects beyond their classical roles in catalysis. These include roles in oxidative defense, bacterial sporulation, and gene expression. This review surveys the role of lipoate metabolism in bacterial, fungal, and protozoan pathogens and how these organisms have employed this metabolism to adapt to niche environments.

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Section: Lipoylated Complexesmentioning

confidence: 99%

Lipoic Acid Metabolism in Microbial Pathogens

Spalding

Prigge

2010

Microbiol Mol Biol Rev

164

204

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“…22) In diaphorase activity, the apparent K m values of rBfmBC for DCPIP and NADH were 0.12 mM and 0.25 mM respectively. No diaphorase activity was detected when NADH was replaced by NADPH, as observed in the DLD of halophilic archaebacteria 20) and the hyperthermophilic archaeon Thermococcus kodakaraensis.…”

Section: Kinetic Parametersmentioning

confidence: 99%

Characterization of a Dihydrolipoyl Dehydrogenase Having Diaphorase Activity ofClostridium kluyveri

Chakraborty¹,

Sakka²,

Kimura³

et al. 2008

Bioscience, Biotechnology, and Biochemistry

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“…Therefore, this bacterium lacks the 2,3-butanedi oil cycle which was described by Juni and Heym (36) in Aci netobacter calcoaceticus. In contrast, A. eutrophus degr&Ldes acetoin directly by cleavage into two C2 compounds , like Bacillus subtilis (35,43,44) or Pelobacter car binolicus (48)(49)(50).…”

mentioning

confidence: 99%

Identification and molecular characterization of the Alcaligenes eutrophus H16 aco operon genes involved in acetoin catabolism

et al. 1991

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Acetoin:dichlorophenolindophenol oxidoreductase (Ao:DCPIP OR) and the fast-migrating protein (FMP) were purified to homogeneity from crude ex tracts of acetoin-grown cells of Alcaligenes eutrophus. Ao:DCPIP OR consisted of a and , subunits (Mrs, 35,5 00 and 36,000, respectively), and a tetrameric a2,12 structure was most likely for the native protein. The mole cular weight of FMP subunits was 39,000. The N-terminal amino acid sequences of the three proteins were det ermined, and oligonucleotides were synthesized on the basis of the codon usage of A. eutrophus. With these, the structural genes for the a and ,1 subunits of Ao:DCPIP OR and FMP, which were referred to as acoA, aco)B, and acoC, respectively, were localized on one single EcoRI restriction fragment which has been clonei recently (C. Frund, H. Priefert, A. Steinbuchel, and H. G. Schlegel, J. Bacteriol. 171:6539-6548, 1989). The nucleotide sequences of a 5.3-kbp region of this fragment and one adjacent fragment were determined, an d the structural genes for acoA (1,002 bp), acoB (1,017 bp), and acoC (1,125 bp) were identified. Together woith the gene acoX, whose function is still unknown and which is represented by a 1,080-bp open reading fr ame, these genes are probably organized in one single operon (acoXABC). The transcription start site was identified 27 bp upstream of acoX; this site was preceded by a region which exhibited complete homology to the enterobacterial a54-dependent promoter consensus sequence.The amino acid sequences deduced from aco. 4 and acoB for the a subunit (Mr,35,243) and the 0 subunit (Mr, 35,788) exhibited significant homologies to th e primary structures of the dehydrogenase components of various 2-oxo acid dehydrogenase complexes, whei 'eas those deduced from acoC for FMP (Mr, 38,941) revealed homology to the dihydrolipoamide acetyltrarw,ferase of Escherichia coli. The occurrence of a new enzyme type for the degradation of acetoin is discussed.Alcaligenes eutrophus has been studied with respe ct to the degradation of acetoin. Acetoin-grown cells of A. eb rtrophus are devoid of 2,3-butanediol dehydrogenase and acetoin dehydrogenase (diacetyl forming; EC 1.1.1.5), and mutants which lack 2,3-butanediol dehydrogenase (73) are stilil able to utilize acetoin as the sole carbon source for grovvth (72). Therefore, this bacterium lacks the 2,3-butanedi oil cycle which was described by Juni and Heym (36) (23,59). As insertional inactivation of this gene caused a pleiotropic effect and as these mutants were unable to synthesize proteins which are essential for degradation of acetoin, it was concluded that expression of the corresponding genes is rpoN dependent (23).Physiological studies localized the genes for Ao:DCPIP OR and FMP on restriction fragments A and C, which are closely linked in the genome (23), whereas the gene for acetaldehyde dehydrogenase II was localized on restriction fragment D (52a). The present study was aimed at isolation of FMP and Ao:DCPIP OR and identification and characterization of their structural genes. ...

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Purification and characterization of acetoin:2,6-dichlorophenolindophenol oxidoreductase, dihydrolipoamide dehydrogenase, and dihydrolipoamide acetyltransferase of the Pelobacter carbinolicus acetoin dehydrogenase enzyme system

Cited by 75 publications

References 67 publications

Lipoic Acid Metabolism in Microbial Pathogens

Lipoic Acid Metabolism in Microbial Pathogens

Characterization of a Dihydrolipoyl Dehydrogenase Having Diaphorase Activity ofClostridium kluyveri

Identification and molecular characterization of the Alcaligenes eutrophus H16 aco operon genes involved in acetoin catabolism

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