Isolation, Identification, and Metabolic Role of the Sudanophilic Granules of Zoogloea ramigera1

Organisms isolated from activated sludge and identified as Zoogloea ramigera accumulated large amounts of sudanophilic granules as the cultures flocculated. The granules were extracted by chloroform and precipitated with ether from acid-hydrolyzed cells. Identification of the sudanophilic granules as poly-β-hydroxybutyric acid (PHB) was confirmed by physical, chemical, and infrared spectral analyses. The isolated polymer accounted for 12.0 to 50.5% of the dry weight of the cells. The polymer was not synthesized when the culture was grown in a growth-limiting concentration of organic substrate; it did accumulate when the culture was grown in medium enriched with carbon and energy sources. An increase in concentration of intracellular PHB was directly proportional to optical density and uptake of glucose. Aside from intracellular storage of PHB as endogenous metabolite, the accumulation of PHB is noted as a possible mechanism of flocculation.