title: Cryoprotectant biosynthesis and the selective accumulation of threitol in the freeze-tolerant Alaskan beetle, Upis ceramboides.
abstract: Adult Upis ceramboides do not survive freezing in the summer but tolerate freezing to -60 degrees C in midwinter. The accumulation of two cryoprotective polyols, sorbitol and threitol, is integral to the extraordinary cold-hardiness of this beetle. U. ceramboides are the only animals known to accumulate high concentrations of threitol; however, the biosynthetic pathway has not been studied. A series of (13)C-labeled compounds was employed to investigate this biosynthetic pathway using (13)C{(1)H} NMR spectroscopy. In vivo metabolism of (13)C-labeled glucose isotopomers demonstrates that C-3-C-6 of glucose become C-1-C-4 of threitol. This labeling pattern is expected for 4-carbon saccharides arising from the pentose phosphate pathway. In vitro experiments show that threitol is synthesized from erythrose 4-phosphate, a C(4) intermediate in the PPP. Erythrose 4-phosphate is epimerized and/or isomerized to threose 4-phosphate, which is subsequently reduced by a NADPH-dependent polyol dehydrogenase and dephosphorylated by a sugar phosphatase to form threitol. Threitol 4-phosphate appears to be the preferred substrate of the sugar phosphatase(s), promoting threitol synthesis over that of erythritol. In contrast, the NADPH-dependent polyol dehydrogenase exhibits broad substrate specificity. Efficient erythritol catabolism under conditions that promote threitol synthesis, coupled with preferential threitol biosynthesis, appear to be responsible for the accumulation of high concentrations of threitol (250 mm) without concomitant accumulation of erythritol.
authors.affiliation: From the Departments of Biological Sciences, Notre Dame, Indiana 46556.
authors.forename: Kent R
authors.initials: KR
authors.lastname: Walters
authors.affiliation: Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556; Omicron Biochemicals, Inc., South Bend, Indiana 46617.
authors.forename: Qingfeng
authors.initials: Q
authors.lastname: Pan
authors.affiliation: Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556; Omicron Biochemicals, Inc., South Bend, Indiana 46617.
authors.forename: Anthony S
authors.initials: AS
authors.lastname: Serianni
authors.affiliation: From the Departments of Biological Sciences, Notre Dame, Indiana 46556. Electronic address: kwalter2@nd.edu.
authors.forename: John G
authors.initials: JG
authors.lastname: Duman
mesh_terms: D000064:Acclimatization / Q000502:physiology; D000413:Alaska; D000818:Animals; D050260:Carbohydrate Metabolism; D002247:Carbon Isotopes; D003081:Cold Climate; D001517:Coleoptera / Q000378:metabolism*; D003451:Cryoprotective Agents / Q000378:metabolism; D004896:Erythritol / Q000378:metabolism; D005615:Freezing; D006458:Hemolymph / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D008954:Models, Biological; D010427:Pentose Phosphate Pathway; D012621:Seasons; D013402:Sugar Alcohols / Q000378:metabolism*
chemical_list: D002247:Carbon Isotopes; D003451:Cryoprotective Agents; D013402:Sugar Alcohols; C003460:threitol; D004896:Erythritol