Characterization of a Glycoside Hydrolase Family 27 α-Galactosidase from Pontibacter Reveals Its Novel Salt-Protease Tolerance and Transglycosylation Activity

J Agric Food Chem. 2016 Mar 23;64(11):2315-24. doi: 10.1021/acs.jafc.6b00255. Epub 2016 Mar 11.

Abstract

α-Galactosidases are of great interest in various applications. A glycoside hydrolase family 27 α-galactosidase was cloned from Pontibacter sp. harbored in a saline soil and expressed in Escherichia coli. The purified recombinant enzyme (rAgaAHJ8) was little or not affected by 3.5-30.0% (w/v) NaCl, 10.0-100.0 mM Pb(CH3COO)2, 10.0-60.0 mM ZnSO4, or 8.3-100.0 mg mL(-1) trypsin and by most metal ions and chemical reagents at 1.0 and 10.0 mM concentrations. The degree of synergy on enzymatic degradation of locust bean gum and guar gum by an endomannanase and rAgaAHJ8 was 1.22-1.54. In the presence of trypsin, the amount of reducing sugars released from soybean milk treated by rAgaAHJ8 was approximately 3.8-fold compared with that treated by a commercial α-galactosidase. rAgaAHJ8 showed transglycosylation activity when using sucrose, raffinose, and 3-methyl-1-butanol as the acceptors. Furthermore, potential factors for salt adaptation of the enzyme were presumed.

Keywords: glycoside hydrolase family 27; protease tolerance; salt tolerance; transglycosylation; α-galactosidase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteroidetes / enzymology*
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Gene Expression
  • Glycosylation
  • Pentanols / metabolism
  • Raffinose / metabolism
  • Recombinant Proteins
  • Sodium Chloride / pharmacology*
  • Sucrose / metabolism
  • Trypsin / metabolism
  • Trypsin / pharmacology*
  • alpha-Galactosidase / genetics
  • alpha-Galactosidase / metabolism*

Substances

  • Pentanols
  • Recombinant Proteins
  • Sodium Chloride
  • Sucrose
  • alpha-Galactosidase
  • Trypsin
  • Raffinose