Efficient and rapid purification of lentil <img src='/image/spc_char/alpha.gif'>-galactosidase by affinity precipitation with alginate

Abstract

<smarttagtype namespaceuri="urn:schemas-microsoft-com:office:smarttags" name="metricconverter"> <img src='/image/spc_char/alpha.gif'>-Galactosidase (<img src='/image/spc_char/alpha.gif'>-D-galactoside galactohydrolase, EC 3.2.1.22) was purified (26-fold) from the germinating seeds of lentil (<i style="">Lens culinaris</i>) by affinity precipitation with alginate. The purified enzyme gave a single band corresponding to molecular mass of 40 kDa on SDS-PAGE. The optimum temperature and pH of the enzyme were determined as 40^oC and 5.5, respectively. The enzyme was very stable at a temperature range of 4-65^oC and at a pH range of 4-7. The values of kinetic constants <i style="">K</i>_m and <i style="">V</i>_max using p-nitrophenyl-<img src='/image/spc_char/alpha.gif'>-D-galactopyranoside (PNPG) as substrate were 0.191 mM and 0.73 U, respectively. Results suggest that affinity precipitation is an attractive process for the purification of <img src='/image/spc_char/alpha.gif'>-galactosidase. </smarttagtype>