1887

Abstract

An alcohol dehydrogenase of broad specificity was purified 43-fold from extracts of var. (BCG) grown on Sauton medium. It was also present in cells grown in Dubos medium with Tween 80 and bovine serum albumin. The enzyme, which appeared to be soluble, acted as an oxidoreductase in the system butan-1-ol–NADP. It was eluted from Sephadex G-200, hydroxylapatite and DEAE-cellulose in a single peak. The molecular weight, as determined by gel filtration on Sephadex G-200, was about 75 000. Results of electrophoresis in sodium dodecyl sulphate–polyacrylamide gels were compatible with the existence of two subunits each of molecular weight 37 500. The optimum pH was about 8·5 when the enzyme catalysed the oxidation of butan-1-ol, and about 8·2 for the reverse reaction. The apparent was 0·125 mm for butyraldehyde and 0·22 m for butan-1-ol. The dehydrogenase activity was maintained after heat treatment (40 min at 55 °C) in the presence of 30% (w/v) glycerol, but was abolished by heating (40 min at 55 °C) in the presence of 0·1 m-EDTA. The activity of enzyme inactivated by heat and EDTA could be fully restored at room temperature in the presence of 2 m-Zn.

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/content/journal/micro/10.1099/00221287-124-2-359
1981-06-01
2021-05-07
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References

  1. De Bruyn J., Weckx M., Beumer-Jochmans M.-P. 1981; Effect of zinc deficiency on Mycobacterium tuberculosis var.bovis (BCG). Journal of General Microbiology 124:353–357
    [Google Scholar]
  2. Dickinson J., Dickinson F. 1976; Some properties of an alcohol dehydrogenase partially purified from baker’s yeast grown without added zinc. Biochemical Journal 153:309–319
    [Google Scholar]
  3. Guerillot L., Vandecasteele J.P. 1977; Purification and characterization of two aldehyde dehydrogenases. European Journal of Biochemistry 81:185–192
    [Google Scholar]
  4. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–682
    [Google Scholar]
  5. Nason A. 1955; Extraction of soluble enzymes from higher plants. Methods in Enzymology 1:62–63
    [Google Scholar]
  6. Rudolph R., Gerschitz J., Jaenicke R. 1978; Effect of zinc (II) on the refolding and reactivation of liver alcohol dehydrogenase. European Journal of Biochemistry 87:601–606
    [Google Scholar]
  7. Spector TH. 1978; Refinement of the Coomassie blue method of protein quantitation. Analytical Biochemistry 86:142–146
    [Google Scholar]
  8. Wang L., Takayama K., Goldman D.S., Schnoes H.K. 1972; Synthesis of alcohol and wax ester by a cell-free system in Mycobacterium tuberculosis . Biochimica et biophysica acta 260:41–48
    [Google Scholar]
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