Characterization and biotechnological application of an acid alpha-galactosidase from Tachigali multijuga Benth. seeds

Phytochemistry. 2008 Oct;69(14):2579-85. doi: 10.1016/j.phytochem.2008.08.017. Epub 2008 Oct 2.

Abstract

Tachigali multijuga Benth. seeds were found to contain protein (364 mg g(-1)dwt), lipids (24 mg g(-1)dwt), ash (35 mg g(-1)dwt), and carbohydrates (577 mg g(-1)dwt). Sucrose, raffinose, and stachyose concentrations were 8.3, 3.0, and 11.6 mg g(-1)dwt, respectively. alpha-Galactosidase activity increased during seed germination and reached a maximum level at 108 h after seed imbibition. The alpha-galactosidase purified from germinating seeds had an M(r) of 38,000 and maximal activity at pH 5.0-5.5 and 50 degrees C. The enzyme was stable at 35 degrees C and 40 degrees C, but lost 79% of its activity after 30 min at 50 degrees C. The activation energy (E(a)) values for p-nitrophenyl-alpha-d-galactopyranoside (pNPGal) and raffinose were 13.86 and 4.75 kcal mol(-1), respectively. The K(m) values for pNPGal, melibiose, raffinose, and stachyose were 0.45, 5.37, 39.62 and 48.80 mM, respectively. The enzyme was sensitive to inhibition by HgCl(2), SDS, AgNO(3), CuSO(4), and melibiose. d-Galactose was a competitive inhibitor (K(i)=2.74 mM). In addition to its ability to hydrolyze raffinose and stachyose, the enzyme also hydrolyzed galactomannan.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fabaceae / enzymology*
  • Germination
  • Hydrogen-Ion Concentration
  • Molecular Weight
  • Oligosaccharides / metabolism
  • Seeds / enzymology*
  • Substrate Specificity
  • Temperature
  • alpha-Galactosidase / isolation & purification*
  • alpha-Galactosidase / metabolism*

Substances

  • Oligosaccharides
  • alpha-Galactosidase