Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1981 Jan;145(1):164–170. doi: 10.1128/jb.145.1.164-170.1981

Genetic and biochemical characterization of D-arabinose dehydrogenase from Neurospora crassa.

A Carrasco, G Pincheira, T Ureta
PMCID: PMC217257  PMID: 6450742

Abstract

D-Arabinose dehydrogenase has been purified to homogeneity from wild-type Neurospora crassa 74-A (FGSC 262) and from two colonial mutants, col-15a (FGSC 1391) and col-16a (FGSC 1349), found to contain more of the enzyme. The enzymes were characterized by measurement of several kinetic and physicochemical parameters. The enzymes were the same in all characteristics studied thus far. Immunological studied performed with enzyme preparations from the three strains showed antigenic identity and indicated that those colonial strains contain more normal enzyme, rather than the usual amount of an altered "improved" enzyme. Quantitation of the enzyme in crude extracts, performed by single radial immunodiffusion, showed that the colonial strains have twice the level of enzyme as the wild-type strain. Genetic characterization, performed by analysis of meiotic products, heterokaryosis, and reversions, indicated that the difference in D-arabinose dehydrogenase activity detected among the three strains is probably determined by one gene. The genetic control, structural or regulatory of this enzyme activity is different from that determining the morphological alterations exhibited by mutant strains carrying the col-15 or col-16 gene.

Full text

PDF
164

Images in this article

166Image
on p.166
167Image
on p.167
168Image
on p.168
168Image
on p.168

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cline A. L., Hu A. S. Enzymatic characterization and comparison of three sugar dehydrogenases from a pseudomonad. J Biol Chem. 1965 Nov;240(11):4493–4497. [PubMed] [Google Scholar]
  2. Cline A. L., Hu A. S. The isolation of three sugar dehydrogenases from a psuedomonad. J Biol Chem. 1965 Nov;240(11):4488–4492. [PubMed] [Google Scholar]
  3. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  4. Drysdale J. W., Righetti P., Bunn H. F. The separation of human and animal hemoglobins by isoelectric focusing in polyacrylamide gel. Biochim Biophys Acta. 1971 Jan 19;229(1):42–50. doi: 10.1016/0005-2795(71)90315-1. [DOI] [PubMed] [Google Scholar]
  5. Fleischer S., Kervina M. Subcellular fractionation of rat liver. Methods Enzymol. 1974;31:6–41. doi: 10.1016/0076-6879(74)31005-1. [DOI] [PubMed] [Google Scholar]
  6. Hedrick J. L., Smith A. J. Size and charge isomer separation and estimation of molecular weights of proteins by disc gel electrophoresis. Arch Biochem Biophys. 1968 Jul;126(1):155–164. doi: 10.1016/0003-9861(68)90569-9. [DOI] [PubMed] [Google Scholar]
  7. Heston W. E., Hoffman H. A., Rechcigl M., Jr Genetic analysis of liver catalase activity in two substrains of C57BL mice. Genet Res. 1965 Nov;6(3):387–397. doi: 10.1017/s0016672300004274. [DOI] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Maijub A. G., Pecht M. A., Miller G. R., Carper W. R. Arabinose (fucose) dehydrogenase from pig liver. I. Isolation and characterization. Biochim Biophys Acta. 1973 Jul 5;315(1):37–42. doi: 10.1016/0005-2744(73)90126-5. [DOI] [PubMed] [Google Scholar]
  10. Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
  11. Metzger R. P., Wick A. N. Partial purification of rat liver D-arabinose dehydrogenase. Biochem Biophys Res Commun. 1967 Mar 21;26(6):742–747. doi: 10.1016/s0006-291x(67)80136-0. [DOI] [PubMed] [Google Scholar]
  12. Mobley P. W., Metzger R. P. The physical properties NAD-dependent L-fucose dehydrogenase from sheep liver. Arch Biochem Biophys. 1978 Feb;186(1):184–188. doi: 10.1016/0003-9861(78)90478-2. [DOI] [PubMed] [Google Scholar]
  13. Mobley P. W., Metzger R. P., Wick A. N. NAD-dependent L-fucose dehydrogenase from sheep liver. Arch Biochem Biophys. 1970 Jul;139(1):83–86. doi: 10.1016/0003-9861(70)90047-0. [DOI] [PubMed] [Google Scholar]
  14. Pincheira G., Leon G., Ureta T. Aldosugar dehydrogenases from Neurospora crassa Partial purification and characterization of D-arabinose: NAD dehydrogenase. FEBS Lett. 1973 Feb 15;30(1):111–114. doi: 10.1016/0014-5793(73)80630-1. [DOI] [PubMed] [Google Scholar]
  15. Rechcigl M., Jr, Heston W. E. Genetic regulation of enzyme activity in mammalian system by the alteration of the rates of enzyme degradation. Biochem Biophys Res Commun. 1967 Apr 20;27(2):119–124. doi: 10.1016/s0006-291x(67)80049-4. [DOI] [PubMed] [Google Scholar]
  16. Schachter H., Sarney J., McGuire E. J., Roseman S. Isolation of diphosphopyridine nucleotide-dependent L-fucose dehydrogenase from pork liver. J Biol Chem. 1969 Sep 10;244(17):4785–4792. [PubMed] [Google Scholar]
  17. Schacterle G. R., Pollack R. L. A simplified method for the quantitative assay of small amounts of protein in biologic material. Anal Biochem. 1973 Feb;51(2):654–655. doi: 10.1016/0003-2697(73)90523-x. [DOI] [PubMed] [Google Scholar]
  18. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]
  19. Weber K., Pringle J. R., Osborn M. Measurement of molecular weights by electrophoresis on SDS-acrylamide gel. Methods Enzymol. 1972;26:3–27. doi: 10.1016/s0076-6879(72)26003-7. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES