BIOMIMETIC, AMORPHOUS GRANULES OF POLYHYDROXYALKANOATES - COMPOSITION, MOBILITY, AND STABILIZATION IN-VITRO BY PROTEINS

It is a remarkable feature of poly(3-hydroxybutyrate) (PHB) that although the isolated polymer is highly crystalline, native PHB storage granules in cells are only found in an amorphous, mobile state. It has recently been proposed that the failure of PHB granules to crystallize is simply the result of slow nucleation kinetics that are operative for small, isolated particles. In support of this new model, we present here a straightforward procedure by which pure crystalline PHB can be reconstituted into submicron-size, detergent-coated artificial granules. The artificial granules are amorphous and stable in suspension, and they are essentially indistinguishable from their native counterparts in terms of size, morphology, molecular mobility, and density. Furthermore, when the surfactant coating is removed from the artificial granules by dialysis, the granules crystallize, verifying the nucleation hypothesis. In vivo, the PHB granule surface is likely to consist of both polypeptide and lipid; in vitro it is possible to prepare amorphous PHB granules that are stabilized solely by phospholipids or by any of several common proteins (serum albumin, casein, or ovalbumin). Artificial amorphous granules may be prepared from a variety of different bacterial PHAs and from blends of incompatible polyesters.