1887

Abstract

Fumarate hydratase (EC 4.2.1.2) from the extremely thermophilic archaeobacterium has been purified to homogeneity by a rapid purification procedure using affinity chromatography and high-performance size-exclusion chromatography, and the enzyme’s physical and biochemical properties have been determined. The native enzyme has a molecular mass of 170 kDa and is composed of identical subunits with a molecular mass of 45 kDa, thus indicating a tetrameric structure similar to fumarases isolated from other organisms. The enzyme was active at temperatures ranging from 40 °C to 90 °C, with a maximum activity at 85 °C. The pH optimum for generation of fumarate was found to be pH 8·0. The enzyme showed high stability to denaturation by heat and organic solvents.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-8-1537
1990-08-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/8/mic-136-8-1537.html?itemId=/content/journal/micro/10.1099/00221287-136-8-1537&mimeType=html&fmt=ahah

References

  1. Ado Y., Kawamoto T., Masunaga I., Takayama K., Takasawa S., Kimura K. 1982; Production of l-malic acid with immobilized thermophilic bacterium, Thermusrubensnov.sp. Enzyme Engineering 6:303–304
    [Google Scholar]
  2. Beeckmans S., Kanarek L. 1982; Chicken heart fumarase: its purification and physico-chemical characterization. A comparison with the enzyme from pig heart. International Journal of Biochemistry 14:453–460
    [Google Scholar]
  3. Brock T.D., Brock K.M., Belly R.T., Weiss R.L. 1972; Sulfolobus: a new genus of sulfur-oxidizing bacteria living at low pH and high temperature. Archives of Microbiology 84:54–68
    [Google Scholar]
  4. Buonocore V., Sgambati O., De Rosa M., Esposito E., Gambacorta A. 1980; A constitutive β-galactosidase from the extreme thermoacidophilearchaebacteriumCaldariella acidophila: properties of the enzyme in the free state and in immobilized whole cells. Journal of Applied Biochemistry 2:390–397
    [Google Scholar]
  5. Chibata I., Tosa T., Sato T. 1987; Application of immobilized biocatalysts in pharmaceutical and chemical industries. In Biotechnology 7a pp. 653–684 Rehm H.-J., Reed G. Edited by Weinheim: Verlag Chemie;
    [Google Scholar]
  6. Giardina P., De Biasi M.G., De Rosa M., Gambacorta A., Buonocore V. 1986; Glucose dehydrogenase from the thermoacidophilic archaeobacterium Sulfolobus solfataricus. Biochemical Journal 239:517–522
    [Google Scholar]
  7. Görg A., Postel W., Westermeier R. 1978; Ultrathin-layer isoelectric focusing in polyacrylamide gels on cellophane. Analytical Biochemistry 86:60–70
    [Google Scholar]
  8. Kanarek L., Marler E., Bradshaw R.A., Fellows R.E., Hill R.L. 1964; The subunits of fumarase. Journal of Biological Chemistry 239:4207–4211
    [Google Scholar]
  9. Kobayashi K., Tuboi S. 1983; End-group analysis of the cytosolic and mitochondrial fumarases from rat liver. Journal of Biochemistry 94:707–713
    [Google Scholar]
  10. Kobayashi K., Yamanishi T., Tuboi S. 1981; Physicochemical, catalytic, and immunochemical properties of fumarases crystallized separately from mitochondrial and cytosolic fractions of rat liver. Journal of Biochemistry 89:1923–1931
    [Google Scholar]
  11. Merill C.R., Goldman D., Sedman S.A., Ebert M.H. 1981; Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 211:1437–1438
    [Google Scholar]
  12. Martin R.G., Ames B.N. 1961; A method for determining the sedimentation behavior of enzymes: application to protein mixtures. Journal of Biological Chemistry 236:1372–1379
    [Google Scholar]
  13. Sacchettini J.C., Meininger T., Roderick S., Banaszak L.J. 1986; Purification, crystallization, and preliminary X-ray data for porcine fumarase. Journal of Biological Chemistry 261:15183–15185
    [Google Scholar]
  14. Shibata H., Gardiner W.E., Schwartzbach S.D. 1985; Purification, characterization, and immunological properties of fumarase from Euglena gracilis var. bacillaris. Journal of Bacteriology 164:762–768
    [Google Scholar]
  15. Woese C.R., Fox G.E. 1977; Phylogenetic structure of the procaryotic domain: the primary kingdoms. Proceedings of theNational Academy of Sciences of the United States of America 74:5088–5090
    [Google Scholar]
  16. Woods S.A., Miles J.S., Roberts R.E., Guest J.R. 1986; Structural and functional relationships between fumarase and aspartase. Nucleotide sequences of the fumarase(fumC) and aspartase {aspA) genes of Escherichia coli K12. Biochemical Journal 237:547–557
    [Google Scholar]
  17. Woods S.A., Schwartzbach S.D., Guest J.R. 1988; Two biochemically distinct classes of fumarase in Escherichia coli. BiochimicaetBiophysicaActa 954:14–26
    [Google Scholar]
  18. Wu M., Tzagoloff A. 1987; Mitochondrial and cytoplasmic fumarases in Saccharomyces cerevisiae are encoded by a single nuclear gene fuml. Journal of Biological Chemistry 262:12275–12282
    [Google Scholar]
  19. Yamamoto K., Tosa T., Yamashita K., Chibata I. 1977; Kinetics and decay of fumarase activity of immobilized Brevibacterium ammoniagenes cells for continuous production of l-malic acid. Biotechnology and Bioengineering 19:1101–1114
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-8-1537
Loading
/content/journal/micro/10.1099/00221287-136-8-1537
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error