Structural and biochemical alterations of Nocardia asteroides cell walls during its growth cycle

Abstract

Analysis of Nocardia asteroides 14759 cell walls were done to determine the chemical and structural composition during the growth cycle. It was found that the ultrastructural profiles of the cell wall become altered as the cultures aged. Chemical analysis revealed corresponding shifts in cell wall components as the culture went from lag to logarithmic to stationary phases of growth. The peptidoglycan from lag-phase cells (5 h) represented 15% of the total cell wall weight, and the percentage of peptidoglycan progressively increased so that, in 1-week stationary-phase cells, it represented approximately 40% of the total wall weight. In lag-phase cells it was found that 36% of the cell wall weight was lipid in nature, whereas stationary-phase cells had only 7% lipid in their wall. The overall sugar composition of the walls remained relatively constant at about 28 to 31% however, the arabinose to galactose ratio changed from approximately 1:1 in lag-phase to 2:1 in stationary-phase cells. Gas-liquid chromatography demonstrated that the fatty acids making up the cell wall lipids changed relative to one another as the cells aged. Based upon the removal of lipids by ethanol-ether, chloroform, and alkaline methanol extraction, it was shown that the classes of loosely and firmly associated lipids changed as the cells aged. Further, it was found that a carotenoid-like pigment associated to a C22 fatty acid increased in the cell wall as the culture stopped growing. Peptidolipid or lipoprotein was found to make up a significant part of the cell wall. This component increased in amount and varied in amino acid content as the culture aged. Analysis of the totally extracted basal layer of the cell wall (peptidoglycan plus arabinogalactan) showed that it too changed as the cells grew and fragmented. The data presented established that the cell wall of N. asteroides was structurally and chemically complex and that a progression of chemical and physical processes occurred within the wall as the cells developed through their growth cycle.