Lactase-phlorizin hydrolase complex from monkey small intestine: purification, properties and evidence for two catalytic sites

Abstract

Lactase-phlorizin hydrolase (EC 3.2.1.-) has been purified from the monkey small intestine by gel filtration and ion-exchange chromatographic procedures and the properties of the purified enzyme complex have been studied. Lactose was the most active substrate. Cellobiose and other synthetic hetero-β-glycosides were hydrolysed at a very much reduced rate. The rate of hydrolysis of phlorizin was about 2.5% that of lactose. Lactase and phlorizin hydrolase activities were indistinguishable by heat inactivation experiments. The purified enzyme complex also hydrolysed cerebrosides. Lactose hydrolysis was competitively inhibited by phlorizin as well as by the brain cerebroside. However, there was no mutual inhibition between phlorizin and the brain cerebroside. It is suggested that the native enzyme complex might have two catalytic sites, a phlorizin site and a cerebroside site but both hydrolysing lactose.