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. 1968 Oct;96(4):1181–1186. doi: 10.1128/jb.96.4.1181-1186.1968

Purification of β-Glucosidase from Saccharomyces lactis Strain Y-1231

George L Marchin a,2, Jacob D Duerksen a
PMCID: PMC252432  PMID: 4176642

Abstract

Saccharomyces lactis strain Y-123, a constitutive high producer of β-glucosidase (Bh), was grown in an enriched medium. β-Glucosidase, extracted most easily by cell autolysis, was purified by successive ammonium sulfate precipitation, ethyl alcohol precipitation, gel filtration, calcium phosphate gel adsorption-elution, and hydroxyapatite column chromatography. The specific activity of the enzyme increased 200-fold during the purification. The electrophoretic and catalytic properties of the enzyme did not change during the procedure. Polyacrylamide gel disc electrophoresis of the partially purified enzyme revealed one major and several minor protein-staining bands. β-Glucosidase activity in the polyacrylamide gel columns was measured directly by assaying sections of columns frozen and sliced immediately after electrophoresis. Most of the activity coincided with the major protein-staining band. Prolonged assay produced minor activity coinciding with the less intense protein bands. Properties of the enzyme suggest that it is a single, unconjugated, intracellular, high molecular weight protein. The purification procedure is applicable to strains of S. lactis which possess alleles of the B locus for β-glucosidase synthesis.

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Selected References

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

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