Abstract
β-d-glucosidase purified from commercial preparations of clarified culture broth ofAspergillus niger (Novo SP188) was shown to elute as two distinct species during analytical anion-exchange chromatography (AEC). However, the two enzyme forms behaved identically on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE), high-performance size-exclusion chromatography (HPSEC), and isoelectric focusing. Also, the N-terminal amino acid sequence, amino acid composition, fingerprint of tryptic-digest peptides, circular dichroism spectra, and reaction kinetics appear identical for these forms. This feature of theA. niger enzyme is distinctly different from β-d-glucosidase isozymes reported from other sources, where multiple forms tend to differ in molecular weight and/or isoelectric pH. Michaelis-Menten kinetic analysis also gave comparable results for the two forms. The distinct behavior on AEC was explained by considering the differences in N-linked carbohydrates liberated from both species following treatment with endoglycosidase H or F.
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Himmel, M.E., Adney, W.S., Fox, J.W. et al. Isolation and characterization of two forms of β-d-glucosidase fromAspergillus niger . Appl Biochem Biotechnol 39, 213–225 (1993). https://doi.org/10.1007/BF02918991
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DOI: https://doi.org/10.1007/BF02918991