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PMID: 8206829 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Acetoin catabolic system of Klebsiella pneumoniae CG43: sequence, expression, and organization of the aco operon.

Journal of bacteriology ·Vol. 176 ·No. 12 ·1994-06-00 ·Pages 3527-35

Deng WL, Chang HY, Peng HL

Abstract

A cosmid clone which was capable of depleting acetoin in vivo was isolated from a library of Klebsiella pneumoniae CG43 cosmids. The smallest functional subclone contained a 3.9-kb DNA fragment of the cosmid clone. Sequencing of the DNA fragment revealed three open reading frames (ORFs A, B, and C) encoding polypeptides of 34, 36, and 52 kDa, respectively. The presence of these proteins was demonstrated by expression of the recombinant DNA clone in Escherichia coli. Considerable similarities between the deduced amino acid sequences of the ORFs and those of the following enzymes were found: acetoin dissimilation enzymes, pyruvate dehydrogenase complex, 2-oxoglutarate dehydrogenase complex, and branched-chain 2-oxo acid dehydrogenase complex of various origins. Activities of these enzymes, including acetoin-dependent dichlorophenolin-dohenol oxidoreductase and dihydrolipoamide acetyltransferase, were detected in the extracts of E. coli harboring the genes encoding products of the three ORFs. Although not required for acetoin depletion in vivo, a possible fourth ORF (ORF D), located 39 nucleotides downstream of ORF C, was also identified. The deduced N-terminal sequence of the ORF D product was highly homologous to the dihydrolipoamide dehydrogenases of several organisms. Primer extension analysis identified the transcriptional start of the operon as an A residue 72 nucleotides upstream of ORF A.

MeSH Terms
3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide) Acetoin/metabolism Amino Acid Sequence Base Sequence Butylene Glycols/metabolism Escherichia coli/genetics Genes, Bacterial/genetics Ketoglutarate Dehydrogenase Complex/biosynthesis,genetics Ketone Oxidoreductases/biosynthesis,genetics Klebsiella pneumoniae/enzymology,genetics Molecular Sequence Data Multienzyme Complexes/biosynthesis,genetics Operon/genetics Pyruvate Dehydrogenase Complex/biosynthesis,genetics Recombinant Proteins/biosynthesis Restriction Mapping Sequence Analysis, DNA Sequence Homology, Amino Acid Transcription, Genetic
Chemicals
Butylene Glycols Multienzyme Complexes Pyruvate Dehydrogenase Complex Recombinant Proteins 2,3-butylene glycol Acetoin Ketone Oxidoreductases Ketoglutarate Dehydrogenase Complex 3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Deng W L
Department of Microbiology and Immunology, Chang Gung College of Medicine and Technology, Tao Yuan, Taiwan.
Chang H Y
Peng H L
References (45)
45 references, click to expand
  1. Diacetyl oxidation by Streptococcus faecalis, a lipoic acid dependent reaction.
    J Bacteriol. 1955 Jan;69(1):51-8 PMID: 13233166
  2. Identification of genes involved in utilization of acetate and acetoin in Bacillus subtilis.
    Mol Microbiol. 1993 Oct;10(2):259-71 PMID: 7934817
  3. Studies on the nature and reactions of protein-bound lipoic acid.
    J Biol Chem. 1958 May;232(1):143-58 PMID: 13549405
  4. A simple method for extraction of RNA from E. coli utilizing diethyl pyrocarbonate.
    Anal Biochem. 1970 Feb;33(2):459-63 PMID: 4910776
  5. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  6. Mutants of Bacillus subtilis blocked in acetoin reductase.
    Eur J Biochem. 1973 Dec 17;40(2):479-83 PMID: 4205556
  7. 2,3-Butanediol biosynthetic system in Aerobacter aerogenes.
    Methods Enzymol. 1975;41:518-32 PMID: 236481
  8. Physiological and biochemical role of the butanediol pathway in Aerobacter (Enterobacter) aerogenes.
    J Bacteriol. 1975 Sep;123(3):1124-30 PMID: 239921
  9. Acetoin degradation in Bacillus subtilis by direct oxidative cleavage.
    Eur J Biochem. 1975 Sep 15;57(2):425-30 PMID: 809287
  10. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  11. Regulatory sequences involved in the promotion and termination of RNA transcription.
    Annu Rev Genet. 1979;13:319-53 PMID: 94251
  12. The pyruvate dehydrogenase complex of Escherichia coli K12. Nucleotide sequence encoding the pyruvate dehydrogenase component.
    Eur J Biochem. 1983 Jun 1;133(1):155-62 PMID: 6343085
  13. The pyruvate dehydrogenase complex of Escherichia coli K12. Nucleotide sequence encoding the dihydrolipoamide acetyltransferase component.
    Eur J Biochem. 1983 Jul 1;133(3):481-9 PMID: 6345153
  14. Nucleotide sequence of the lipoamide dehydrogenase gene of Escherichia coli K12.
    Eur J Biochem. 1983 Oct 3;135(3):519-27 PMID: 6352260
  15. Nucleotide sequence of the sucB gene encoding the dihydrolipoamide succinyltransferase of Escherichia coli K12 and homology with the corresponding acetyltransferase.
    Eur J Biochem. 1984 Jun 1;141(2):361-74 PMID: 6376124
  16. Repeating functional domains in the pyruvate dehydrogenase multienzyme complex of Escherichia coli.
    EMBO J. 1984 Jun;3(6):1315-9 PMID: 6430694
  17. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes.
    J Mol Biol. 1986 May 5;189(1):113-30 PMID: 3537305
  18. Molecular cloning of genes encoding branched-chain keto acid dehydrogenase of Pseudomonas putida.
    J Bacteriol. 1987 Apr;169(4):1619-25 PMID: 3549697
  19. Similarity of the E1 subunits of branched-chain-oxoacid dehydrogenase from Pseudomonas putida to the corresponding subunits of mammalian branched-chain-oxoacid and pyruvate dehydrogenases.
    Eur J Biochem. 1988 Sep 15;176(2):311-7 PMID: 3416875
  20. Sequence analysis of the lpdV gene for lipoamide dehydrogenase of branched-chain-oxoacid dehydrogenase of Pseudomonas putida.
    Eur J Biochem. 1989 Jan 15;179(1):61-9 PMID: 2917566
  21. MalT, the regulatory protein of the Escherichia coli maltose system, is an ATP-dependent transcriptional activator.
    EMBO J. 1989 Mar;8(3):981-7 PMID: 2524384
  22. Molecular cloning and heterologous expression of a Klebsiella pneumoniae gene encoding alginate lyase.
    Gene. 1989 Jan 30;75(1):127-34 PMID: 2542130
  23. A common structural motif in thiamin pyrophosphate-binding enzymes.
    FEBS Lett. 1989 Sep 11;255(1):77-82 PMID: 2792374
  24. Reductive acetylation of tandemly repeated lipoyl domains in the pyruvate dehydrogenase multienzyme complex of Escherichia coli is random order.
    J Mol Biol. 1989 Aug 20;208(4):623-33 PMID: 2509711
  25. Biochemical and genetic analyses of acetoin catabolism in Alcaligenes eutrophus.
    J Bacteriol. 1989 Dec;171(12):6539-48 PMID: 2556366
  26. The 2-oxoglutarate dehydrogenase complex from Azotobacter vinelandii. 2. Molecular cloning and sequence analysis of the gene encoding the succinyltransferase component.
    Eur J Biochem. 1990 Jan 12;187(1):235-9 PMID: 2404760
  27. Structure and regulation of KGD2, the structural gene for yeast dihydrolipoyl transsuccinylase.
    Mol Cell Biol. 1990 Aug;10(8):4221-32 PMID: 2115121
  28. Cloning and sequence analysis of the genes encoding the alpha and beta subunits of the E1 component of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus.
    Eur J Biochem. 1990 Jul 31;191(2):337-46 PMID: 2200674
  29. Secretory S complex of Bacillus subtilis: sequence analysis and identity to pyruvate dehydrogenase.
    J Bacteriol. 1990 Sep;172(9):5052-63 PMID: 1697575
  30. Transcriptional analysis of the promoter region of the Pseudomonas putida branched-chain keto acid dehydrogenase operon.
    J Bacteriol. 1990 Oct;172(10):5655-63 PMID: 2211503
  31. Purification and characterization of acetoin:2,6-dichlorophenolindophenol oxidoreductase, dihydrolipoamide dehydrogenase, and dihydrolipoamide acetyltransferase of the Pelobacter carbinolicus acetoin dehydrogenase enzyme system.
    J Bacteriol. 1991 Jan;173(2):757-67 PMID: 1898934
  32. Identification and molecular characterization of the Alcaligenes eutrophus H16 aco operon genes involved in acetoin catabolism.
    J Bacteriol. 1991 Jul;173(13):4056-71 PMID: 2061286
  33. Domains, motifs, and linkers in 2-oxo acid dehydrogenase multienzyme complexes: a paradigm in the design of a multifunctional protein.
    Biochemistry. 1991 Sep 3;30(35):8501-12 PMID: 1888719
  34. Lipoamide dehydrogenase of Staphylococcus aureus: nucleotide sequence and sequence analysis.
    Biochim Biophys Acta. 1991 Dec 2;1129(1):119-23 PMID: 1756171
  35. Cloning, sequence and transcriptional analysis of the structural gene for LPD-3, the third lipoamide dehydrogenase of Pseudomonas putida.
    Eur J Biochem. 1991 Dec 5;202(2):231-40 PMID: 1722146
  36. Cloning and expression in Escherichia coli of mature E1 beta subunit of bovine mitochondrial branched-chain alpha-keto acid dehydrogenase complex. Mapping of the E1 beta-binding region on E2.
    J Biol Chem. 1992 Jan 25;267(3):1881-7 PMID: 1730724
  37. Identification and molecular characterization of the gene coding for acetaldehyde dehydrogenase II (acoD) of Alcaligenes eutrophus.
    J Bacteriol. 1992 Feb;174(3):899-907 PMID: 1732222
  38. Molecular epidemiology of Klebsiella pneumoniae.
    Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi. 1991 Aug;24(3):264-71 PMID: 1726368
  39. Identification of acoR, a regulatory gene for the expression of genes essential for acetoin catabolism in Alcaligenes eutrophus H16.
    J Bacteriol. 1992 Jul;174(13):4391-400 PMID: 1378052
  40. Cloning, sequencing and heterologous expression of a Klebsiella pneumoniae gene encoding an FAD-independent acetolactate synthase.
    Gene. 1992 Aug 1;117(1):125-30 PMID: 1644303
  41. Expression and assembly of a functional E1 component (alpha 2 beta 2) of mammalian branched-chain alpha-ketoacid dehydrogenase complex in Escherichia coli.
    J Biol Chem. 1992 Aug 15;267(23):16601-6 PMID: 1644840
  42. Characterization of the genes of the 2,3-butanediol operons from Klebsiella terrigena and Enterobacter aerogenes.
    J Bacteriol. 1993 Mar;175(5):1392-404 PMID: 8444801
  43. Thioredoxin-thioredoxin reductase system of Streptomyces clavuligerus: sequences, expression, and organization of the genes.
    J Bacteriol. 1993 Aug;175(16):5159-67 PMID: 8349555
  44. Identification and molecular characterization of the aco genes encoding the Pelobacter carbinolicus acetoin dehydrogenase enzyme system.
    J Bacteriol. 1994 Jan;176(2):469-85 PMID: 8110297
  45. A cyclic pathway for the bacterial dissimilation of 2, 3-butanediol, acetylmethylcarbinol and diacetyl. II. The synthesis of diacetylmethylcarbinol from diacetyl, a new diphosphothiamin catalyzed reaction.
    J Bacteriol. 1956 Dec;72(6):746-53 PMID: 13398360
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.176.12.3527-3535.1994
Published
1994-06-00
Pages
3527-35
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC205540
Subset
IM
Databases
GENBANK
U00985
External Links
PubMed source DOI: 10.1128/jb.176.12.3527-3535.1994 PMC Full Text View journal details
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