The purification and characterization of a DNA nicking–closing enzyme from Bacillus megaterium

Abstract

Although several eucaryote DNA nicking–closing (N–C) enzymes have been characterized, only the Escherichia coli enzyme has been extensively studied amongst procaryotes. The latter enzyme is distinctly different from the eucaryotic enzymes and we have therefore purified the N–C enzyme from Bacillus megaterium to determine if procaryotes form a distinctive class of N–C enzymes. The purified B. megaterium N–C enzyme has a molecular weight of 120 000, only partly relaxes negative supercoils, does not affect positive supercoils, requires Mg2+, and is inhibited by 0.2 M KCl. The enzyme is also inhibited by 1 mM nalidixic or oxolinic acids but unaffected by novobiocin. A crude N–C enzyme preparation from Micrococcus luteus shows very similar properties.