Immobilization of Aspergillus oryzae β-galactosidase in an agarose matrix functionalized by four different methods and application to the synthesis of lactulose

Bioresour Technol. 2017 May:232:53-63. doi: 10.1016/j.biortech.2017.02.003. Epub 2017 Feb 7.

Abstract

Aspergillus oryzae β-galactosidase was immobilized in monofunctional glyoxyl-agarose and heterofunctional supports (amino-glyoxyl, carboxy-glyoxyl and chelate-glyoxyl agarose), for obtaining highly active and stable catalysts for lactulose synthesis. Specific activities of the amino-glyoxyl agarose, carboxy-glyoxyl agarose and chelate-glyoxyl agarose derivatives were 3676, 430 and 454IU/g biocatalyst with half-life values at 50°C of 247, 100 and 100h respectively. Specific activities of 3490, 2559 and 1060IU/g were obtained for fine, standard and macro agarose respectively. High immobilization yield (39.4%) and specific activity of 7700IU/g was obtained with amino-glyoxyl-agarose as support. The highest yields of lactulose synthesis were obtained with monofunctional glyoxyl-agarose. Selectivity of lactulose synthesis was influenced by the support functionalization: glyoxyl-agarose and amino-glyoxyl-agarose derivatives retained the selectivity of the free enzyme, while selectivity with the carboxy-glyoxyl-agarose and chelate-glyoxyl-agarose derivatives was reduced, favoring the synthesis of transgalactosylated oligosaccharides over lactulose.

Keywords: Agarose matrix; Aspergillus oryzae; Immobilization; Lactulose; β-Galactosidase.

MeSH terms

  • Aspergillus oryzae / enzymology*
  • Biotechnology / methods*
  • Enzyme Stability / drug effects
  • Enzymes, Immobilized / metabolism*
  • Glycosylation / drug effects
  • Glyoxylates / pharmacology
  • Lactulose / biosynthesis*
  • Particle Size
  • Sepharose / pharmacology*
  • Temperature
  • beta-Galactosidase / metabolism*

Substances

  • Enzymes, Immobilized
  • Glyoxylates
  • glyoxyl agarose
  • Lactulose
  • Sepharose
  • beta-Galactosidase