Organization of Enzymes in the Polyaromatic Synthetic Pathway: Separability in Bacteria

Abstract

Ultracentrifugation in sucrose density gradients was employed to estimate the molecular weights and to determine possible physical aggregation of the five enzymes catalyzing steps two to six in the prechorismic acid portion of the polyaromatic synthetic pathway in six species of bacteria: Escherichia coli, Salmonella typhimurium, Aerobacter aerogenes, Bacillus subtilis, Pseudomonas aeruginosa , and Streptomyces coelicolor . The five enzymes were not aggregated in extracts of any of the species examined, nor are the genes encoding these enzymes clustered in those bacterial species for which genetic evidence exists. (An initial examination of the blue-green alga Anabaena variabilis indicates nonaggregation in this species also.) This situation in bacteria is in marked contrast to that found in Neurospora crassa and other fungi in which the same five enzymes are associated as an aggregate encoded (at least in the case of N. crassa ) by a cluster of five genes. In addition, also in contrast to N. crassa , no evidence was obtained for more than one kind of dehydroquinase activity in any of the bacteria examined. These comparative results are discussed in relation to the origin, evolution, and functional significance of the gene-enzyme relationships existing in the early steps of aromatic biosynthesis in bacteria and fungi.