Glycerol synthesis in freeze-resistant rainbow smelt: towards the characterization of a key enzyme glycerol-3-phosphatase.

ABSTRACT

Rainbow smelt (Osmerus mordax) synthesize high amounts of glycerol in winter as a cryoprotectant through the direct dephosphorylation of glycerol-3-phosphate by a phosphatase, glycerol-3-phosphatase (G3Pase). Such a protein is well described in a few species including fungi, bacteria and plants but never studied beyond tissue homogenates in any animal species. Purification, identification and characterization of this enzyme is thus crucial for a better comprehension of the biochemical adaptation in rainbow smelt in response to low temperature and more generally of the biochemical mechanisms involved in glycerol synthesis in animals. This work presents the first attempt to purify G3Pase from smelt liver, the main site of glycerol synthesis for the whole animal. A partial purification was performed, and some characteristics of the protein determined, including optimal pH, K(m) and cation requirements. Smelt G3Pase is most likely a low molecular weight, Mg⁺-dependent and cytosolic phosphatase.