Purification and Properties of Phosphotransacetylase from Veillonella alcalescens

Abstract

Abstract Gel filtration of phosphotransacetylase purified from Veillonella alcalescens gave a molecular weight of 75,000 to 80,000. Ultracentrifugation and sedimentation in sucrose gradients showed a heterogeneous distribution of proteins. The predominant component in sucrose gradients had a sedimentation coefficient of 4.2; a minor component having a sedimentation coefficient of 2.8 could also be detected. Denaturation with urea or sodium dodecyl sulfate yielded a single polypeptide with a molecular weight of 32,000 to 40,000. Amidination with dimethyl suberimidate prior to denaturation indicated that the enzyme contained two subunits. The properties of this phosphotransacetylase were compared with those of phosphotransacetylases isolated from other bacteria.