A novel heterofunctional epoxy-amino sepabeads for a new enzyme immobilization protocol: immobilization-stabilization of beta-galactosidase from Aspergillus oryzae.

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  • Additional Information
    • Source:
      Publisher: Wiley-Blackwell Country of Publication: United States NLM ID: 8506292 Publication Model: Print Cited Medium: Print ISSN: 8756-7938 (Print) Linking ISSN: 15206033 NLM ISO Abbreviation: Biotechnol Prog Subsets: MEDLINE
    • Publication Information:
      Publication: <2010-> : Hoboken, NJ : Wiley-Blackwell
      Original Publication: [New York, N.Y. : American Institute of Chemical Engineers, c1985-
    • Subject Terms:
      Aspergillus oryzae/*chemistry ; Aspergillus oryzae/*enzymology ; Enzymes, Immobilized/*biosynthesis ; Enzymes, Immobilized/*chemistry ; Epoxy Resins/*chemistry ; Ethylenediamines/*chemistry ; beta-Galactosidase/*biosynthesis ; beta-Galactosidase/*chemistry; Amino Acids/chemistry ; Coated Materials, Biocompatible/chemical synthesis ; Coated Materials, Biocompatible/chemistry ; Enzyme Activation ; Enzyme Stability ; Microspheres
    • Abstract:
      The properties of a new and commercially available amino-epoxy support (amino-epoxy-Sepabeads) have been compared to conventional epoxy supports to immobilize enzymes, using the beta-galactosidase from Aspergillus oryzae as a model enzyme. The new support has a layer of epoxy groups over a layer of ethylenediamine that is covalently bound to the support. This support has both a great anionic exchanger strength and a high density of epoxy groups. Epoxy supports require the physical adsorption of the proteins onto the support before the covalent binding of the enzyme to the epoxy groups. Using conventional supports the immobilization rate is slow, because the adsorption is of hydrophobic nature, and immobilization must be performed using high ionic strength (over 0.5 M sodium phosphate) and a support with a fairly hydrophobic nature. Using the new support, immobilization may be performed at moderately low ionic strength, it occurs very rapidly, and it is not necessary to use a hydrophobic support. Therefore, this support should be specially recommended for immobilization of enzymes that cannot be submitted to high ionic strength. Also, both supports may be expected to yield different orientations of the proteins on the support, and that may result in some advantages in specific cases. For example, the model enzyme became almost fully inactivated when using the conventional support, while it exhibited an almost intact activity after immobilization on the new support. Furthermore, enzyme stability was significantly improved by the immobilization on this support (by more than a 12-fold factor), suggesting the promotion of some multipoint covalent attachment between the enzyme and the support (in fact the enzyme adsorbed on an equivalent cationic support without epoxy groups was even slightly less stable than the soluble enzyme).
    • Accession Number:
      0 (Amino Acids)
      0 (Coated Materials, Biocompatible)
      0 (Enzymes, Immobilized)
      0 (Epoxy Resins)
      0 (Ethylenediamines)
      60V9STC53F (ethylenediamine)
      EC 3.2.1.23 (beta-Galactosidase)
    • Publication Date:
      Date Created: 20030607 Date Completed: 20040524 Latest Revision: 20191210
    • Publication Date:
      20221213
    • Accession Number:
      10.1021/bp025771g
    • Accession Number:
      12790680