1887

Abstract

Acetate kinase (ATP:phosphotransferase, EC 2.7.2.1) has been purified 294-fold from acid-producing cells of DSM 1731 to a specific activity of 1087 U mg (ADP-forming direction). The dimeric enzyme consisted of subunits with a molecular mass of 43 kDa. The molecular mass of the native acetate kinase was in the range 87-94 kDa as judged by gel filtration and native gel electrophoresis. The enzyme showed high specificity for the substrates acetate and ATP, and maximal activity was obtained with Mn as divalent cation. The presence of mercury compounds such as HgCl and -hydroxymercuribenzoate resulted in an essential loss of activity. The apparent values for acetate, Mg-ATP, acetyl phosphate, and Mg-ADP were 73, 0.37, 0.58 and 0.71 mM. An activity-staining procedure for detection of acetate kinase in crude cell extracts after separation on native polyacrylamide gels was developed. A DNA fragment encoding 246 bp of the acetate kinase gene of DSM 792 was cloned by a PCR-based approach. Northern blot analysis revealed transcription of the gene under acid- and solvent-producing conditions with no significant differences at the level of transcription.

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1997-10-01
2021-05-08
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References

  1. Aceti D.J., Ferry J.G. 1988; Purification and characterization of acetate kinase from acetate-grown Methanosarcina thermophila . J Biol Chem 263:15444–15448
    [Google Scholar]
  2. Andersch W., Bahl H., Gottschalk G. 1983; Level of enzymes involved in acetate, butyrate, acetone and butanol formation by Clostridium acetobutylicum . Eur J Appl Microbiol Biotechnol 18:327–332
    [Google Scholar]
  3. Anthony R.S., Spector L.B. 1971; Exchange reactions catalyzed by acetate kinase. J Biol Chem 246:6129–6135
    [Google Scholar]
  4. Anthony R.S., Spector L.B. 1972; Phosphorylated acetate kinase. J Biol Chem 247:2120–2125
    [Google Scholar]
  5. Bahl H., Ersch W., Gottschalk G. 1982; Continuous production of acetone and butanol by clostridium acetobutylicum in a two-stage phosphate limited chemostat. Eur J Appl Microbiol Biotechnol 15:201–205 (erratum 17, p. 73, 1983)
    [Google Scholar]
  6. Bertram J., DUrre P. 1989; Conjugal transfer and expression of streptococcal transposons in clostridium acetobutylicum . Arch Microbiol 151:551–557
    [Google Scholar]
  7. Blum H., Beier H., Gross H.J. 1987; Improved silver staining of plant proteins, rna and dna in polyacrylamide gels. Electrophoresis 8:93–99
    [Google Scholar]
  8. Boynton Z.L., Bennett G.N., Rudolph F.B. 1996; Cloning, sequencing, and expression of genes encoding phosphotransacetylase and acetate kinase from clostridium acetobutylicum atcc 824. Appl Environ Microbiol 62:2758–2766
    [Google Scholar]
  9. Bradford M.M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
    [Google Scholar]
  10. Brown M.S., Akagi J.M. 1966; Purification of acetokinase from Desulfovibrio desulfuricans . J Bacteriol 92:1273–1274
    [Google Scholar]
  11. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the vax. Nucleic Acids Res 12:387–395
    [Google Scholar]
  12. DUrre P., Bahl H. 1996; Microbial Production of Acetone/Butanol/Isopropanol. In Biotechnology, 2nd edn. vol. 6 pp. 229–268 Edited by Röhr M. Weinheim: VCH;
    [Google Scholar]
  13. Fox D.K., Roseman S. 1986; Isolation and characterization of homogeneous acetate kinase from salmonella typhimurium and escherichia coli . J Biol Chem 261:13487–13497
    [Google Scholar]
  14. Gerischer U., Dürre P. 1992; Mrna analysis of the adc gene region of clostridium acetobutylicum during the shift to solventogenesis. J Bacteriol 174:426–433
    [Google Scholar]
  15. Griffith M.J., Nishimura J.S. 1979; Acetate kinase from veillonella alcalescens . J Biol Chem 254:442–446
    [Google Scholar]
  16. Grundy F.J., Waters D.A., Allen S.H.G., Henkin T.M. 1993; Regulation of the bacillus subtilis acetate kinase gene by ccpa. J Bacteriol 175:7348–7355
    [Google Scholar]
  17. Hartmanis M.G.N. 1987; Butyrate kinase from clostridium acetobutylicum . J Biol Chem 262:617–621
    [Google Scholar]
  18. Hartmanis M.G.N., Gatenbeck S. 1984; Intermediary metabolism in clostridium acetobutylicum: Levels of enzymes involved in the formation of acetate and butyrate. Appl Environ Microbiol 47:1277–1283
    [Google Scholar]
  19. Hartmanis M.G.N., Klason T., 8T Gatenbeck S. 1984; Uptake and activation of acetate and butyrate in clostridium acetobutylicum . Appl Microbiol Biotechnol 20:66–71
    [Google Scholar]
  20. Hüsemann M.H.W., Papoutsakis E.T. 1989; Comparison between in vivo and in vitro enzyme activities in continuous and batch fermentations of clostridium acetobutylicum . Appl Microbiol Biotechnol 30:585–595
    [Google Scholar]
  21. Jones D.T., Woods D.R. 1986; Acetone-butanol fermentation revisited. Microbiol Rev 50:484–524
    [Google Scholar]
  22. Kahane I., Muhlrad A. 1979; Purification and properties of acetate kinase from. J Bacteriol 137:764–772
    [Google Scholar]
  23. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage t4. Nature 227:680–685
    [Google Scholar]
  24. Latimer M.T., Ferry J.G. 1993; Cloning, sequence analysis, and hyperexpression of the genes encoding phosphotransacetylase and acetate kinase from methanosarcina thermophila . J Bacteriol 175:6822–6829
    [Google Scholar]
  25. Marck C. 1988; Dna strider: A ‘c’ program for the fast analysis of dna and protein sequences on the apple macintosh family of computers. Nucleic Acids Res 16:1829–1836
    [Google Scholar]
  26. Marmur J. 1960; A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3:208–218
    [Google Scholar]
  27. Matsuyama A., Yamamoto H., Nakano E. 1989; Cloning, expression, and nucleotide sequence of the escherichia coli k-12 acka gene. J Bacteriol 171:577–580
    [Google Scholar]
  28. Matsuyama A., Yamamoto-Otake H., Hewitt J., MacGillivray R.T.A., Nakano E. 1994; Nucleotide sequence of the phosphotransacetylase gene of escherichia coli strain k12. Biochim Biophys Acta 1219:559–562
    [Google Scholar]
  29. Morrison J.F. 1979; Approaches to kinetic studies on metalactivated enzymes. Methods Enzymol 63:257–294
    [Google Scholar]
  30. Nakajima H., Suzuki K., Imahori K. 1978; Purification and properties of acetate kinase from bacillus stearothermophilus . J Biochem 84:193–203
    [Google Scholar]
  31. O'Brien R.W., Morris J.G. 1971; Oxygen and the growth and metabolism of. J Gen Microbiol 68:307–318
    [Google Scholar]
  32. Oelmdller U., Krüger N., SteinbUchel A., Friedrich C. 1990; Isolation of prokaryotic rna and detection of specific mrna with biotinylated probes. J Microbiol Methods 11:73–81
    [Google Scholar]
  33. Oultram J.D., Burr I.D., Elmore M.J., Minton N.P. 1993; Cloning and sequence analysis of the genes encoding phosphotransbutyrylase and butyrate kinase from clostridium acetobutylicum ncimb 8052. Gene 131:107–112
    [Google Scholar]
  34. Rose I.A., Grunberg-Manago M., Korey R.S., Ochoa S. 1954; Enzymatic phosphorylation of acetate. J Biol Chem 211:737–756
    [Google Scholar]
  35. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  36. Schweizer H.P., Datta P. 1991; The complete nucleotide sequence of the tdc region of. Nucleic Acids Res 17:3994
    [Google Scholar]
  37. Singh-Wissmann K., Ferry J.G. 1995; Transcriptional regulation of the phosphotransacetylase-encoding and acetateencoding genes (pta and ack) from. J Bacteriol 177:1699–1702
    [Google Scholar]
  38. Walter K.A., Nair R.V., Cary J.W., Bennett G.N., Papoutsakis E.T. 1993; Sequence and arrangement of two genes of the butyrate-synthesis pathway of clostridium acetobutylicum atcc 824. Gene 134:107–111
    [Google Scholar]
  39. Wiesenborn D.P., Rudolph F.B., Papoutsakis E.T. 1989; Phosphotransbutyrylase from clostridium acetobutylicum atcc 824 and its role in acidogenesis. Appl Environ Microbiol 55:317–322
    [Google Scholar]
  40. Wilkinson S.R., Young M., Goodacre R., Morris J.G., Farrow J.A.E., Collins M.D. 1995; Phenotypic and genotypic differences between certain strains of clostridium acetobutylicum . FEMS Microbiol Lett 125:199–204
    [Google Scholar]
  41. Winzer K. 1992 Reinigung und Charakterisierung der Acetat Kinase aus Acetobacterium woodii Diploma thesis University of Gottingen;
    [Google Scholar]
  42. Yanisch-Perron C., Vieira J., Messing J. 1985; Improved m13 phage cloning vectors and host strains: Nucleotide sequences of the m13mpl8 and puc19 vectors. Gene 33:103–119
    [Google Scholar]
  43. Yoshimura F. 1978; Purification and characterization of acetate kinase from veillonella alcalescens atcc 17748. Arch Biochem Biophys 189:424–432
    [Google Scholar]
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