Enolase from spores and cells of Bacillus megaterium: two step purification of the enzyme and some of its properties

Abstract

A simple 2-step procedure for purification of enolase from germinated spores or vegetative cells of B. megaterium is described. The procedure resulted in a 1,200-fold purification with production of homogeneous enzyme in ~75% yield; the enzymes from spores and cells seemed identical. The molecular weight of the native enzyme was 335,000, with a subunit molecular weight of 42,000. The enzyme required Mg2+ and was inhibited by ethylenediaminetetraacetic acid and fluoride ions. The Michaelis constants for 2-phosphoglyceric acid and Mg2+ were 7.1 x 10-4 and 4.7 x 10-4 M, respectively.