Properties of alpha-glucosidase from Lactobacillus acidophilus NCTC 1723.

Lactobacillus acidophilus NCTC 1723 produced intracellular and extracellular alpha-glucosidase (alpha-D-glucoside glucohydrolase, EC 3.2.1.20). The alpha-glucosidase was partially purified by ammonium sulphate fractionation and DEAE-cellulose column chromatography, and attained a 10.3-fold purification. The Km for alpha-PNPG was 2.9 mM and the Vmax for alpha-PNPG hydrolysis was 6.45 mumole ml-1 min-1. The enzyme was stable only at pH 6.0-7.5 while incubated at 25 degrees C. At pH 6.5, a 100% activity was retained at 15 degrees-37 degrees C. However, the enzyme was easily destroyed at 50 degrees C. The pH optimum for stability of the enzyme at low temperature (2 degrees C) was between 5 and 6. It was found that addition of Mn++, Ba++ and EDTA, to the medium stimulated alpha-glucosidase activity, while the presence of Hg++, Cu++, Co++, Ni++, Zn++, L-histidine, arabitol, erythritol, sorbitol and glycerol inhibited enzyme activity. Although isomaltase activity was found in the partially purified alpha-glucosidase, it was not known whether this activity was an intrinsic capability of the enzyme. Transglucosylase and weak glucoamylase activities were also found to associate with the partially purified alpha-glucosidase. Since only the alpha-1,6 linked isomaltose was detected as the transferase product, it was thought that the alpha-glucosidase was capable of glucosyl transfer via alpha-1,6-glucosidic bonds.