title: Alginate formation in Azotobacter vinelandii UWD during stationary phase and the turnover of poly-beta-hydroxybutyrate.
abstract: Azotobacter vinelandii UWD is a mutant of strain UW that is defective in the respiratory oxidation of NADH. This mutation causes an overproduction of polyhydroxyalkanoates (PHAs), as polyester synthesis is used as an alternative electron sink. Since PHAs have potential for use as natural, biodegradable plastics, studies of physiology related to their production are of interest. Alginate production by this strain is limited to < 11 microg (mg cell protein)(-1), which permits high efficiency conversion of carbon source into PHA. However, < or = 400 microg (mg cell protein)(-1) was formed when UWD cells were oxygen-limited and in the stationary phase of growth. Alginate formation was fuelled by PHA turnover, which was coincident with the synthesis of alkyl resorcinols, under conditions of exogenous glucose limitation. However, alginate production was a phenotypic and reversible change. Alginate production was stopped by interruption of algD with Tn5lacZ. LacZ activity in UWD was shown to increase in stationary phase, while LacZ activity in a similarly constructed mutant of strain UW did not. Transcription of algD in strain UWD started from a previously identified RpoD promoter and not from the AlgU (RpoE) promoter. This is because strain UWD has a natural insertion element in algU. Differences between strain UW and UWD may reside in the defective respiratory oxidation of NADH, where the NADH surplus in strain UWD may act as a signal of stationary phase. Indeed, a backcross of UW DNA into UWD generated NADH-oxidase-proficient cells that failed to form alginate in stationary phase. Evidence is also presented to show that the RpoD promoter may be recognized by the stationary phase sigma factor (RpoS), which may mediate alginate production in strain UWD.
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, CanadaT6G 2E91.
authors.forename: William J
authors.initials: WJ
authors.lastname: Page
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, CanadaT6G 2E91.
authors.forename: Anne
authors.initials: A
authors.lastname: Tindale
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, CanadaT6G 2E91.
authors.forename: Mark
authors.initials: M
authors.lastname: Chandra
authors.affiliation: Department of Biological Sciences, University of Alberta, Edmonton, Alberta, CanadaT6G 2E91.
authors.forename: Elena
authors.initials: E
authors.lastname: Kwon
mesh_terms: D000464:Alginates / Q000378:metabolism*; D016948:Azotobacter vinelandii / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D002237:Carbohydrate Dehydrogenases / Q000235:genetics / Q000378:metabolism; D003470:Culture Media; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006885:Hydroxybutyrates / Q000378:metabolism*; D008969:Molecular Sequence Data; D010100:Oxygen / Q000494:pharmacology; D011091:Polyesters / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012808:Sigma Factor / Q000378:metabolism; D014158:Transcription, Genetic; D000094122:Polyhydroxybutyrates
chemical_list: D000464:Alginates; D001426:Bacterial Proteins; D002237:Carbohydrate Dehydrogenases; D003470:Culture Media; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006885:Hydroxybutyrates; D010100:Oxygen; D011091:Polyesters; D012808:Sigma Factor; C079619:AlgU protein, Pseudomonas aeruginosa; C069567:sigma factor KatF protein, Bacteria; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C020562:GDPmannose dehydrogenase