title: Metabolic and photosynthetic consequences of blocking starch biosynthesis in the green alga Chlamydomonas reinhardtii sta6 mutant.
abstract: Upon nutrient deprivation, microalgae partition photosynthate into starch and lipids at the expense of protein synthesis and growth. We investigated the role of starch biosynthesis with respect to photosynthetic growth and carbon partitioning in the Chlamydomonas reinhardtii starchless mutant, sta6, which lacks ADP-glucose pyrophosphorylase. This mutant is unable to convert glucose-1-phosphate to ADP-glucose, the precursor of starch biosynthesis. During nutrient-replete culturing, sta6 does not re-direct metabolism to make more proteins or lipids, and accumulates 20% less biomass. The underlying molecular basis for the decreased biomass phenotype was identified using LC-MS metabolomics studies and flux methods. Above a threshold light intensity, photosynthetic electron transport rates (water → CO2) decrease in sta6 due to attenuated rates of NADPH re-oxidation, without affecting photosystems I or II (no change in isolated photosynthetic electron transport). We observed large accumulations of carbon metabolites that are precursors for the biosynthesis of lipids, amino acids and sugars/starch, indicating system-wide consequences of slower NADPH re-oxidation. Attenuated carbon fixation resulted in imbalances in both redox and adenylate energy. The pool sizes of both pyridine and adenylate nucleotides in sta6 increased substantially to compensate for the slower rate of turnover. Mitochondrial respiration partially relieved the reductant stress; however, prolonged high-light exposure caused accelerated photoinhibition. Thus, starch biosynthesis in Chlamydomonas plays a critical role as a principal carbon sink influencing cellular energy balance however, disrupting starch biosynthesis does not redirect resources to other bioproducts (lipids or proteins) during nutrient-replete culturing, resulting in cells that are susceptible to photochemical damage caused by redox stress.
authors: Waksman Institute of Microbiology, Rutgers: The State University of New Jersey, Piscataway, NJ, 08854, USA.
authors: Anagha
authors: A
authors: Krishnan
authors: G Kenchappa
authors: GK
authors: Kumaraswamy
authors: David J
authors: DJ
authors: Vinyard
authors: Huiya
authors: H
authors: Gu
authors: Gennady
authors: G
authors: Ananyev
authors: Matthew C
authors: MC
authors: Posewitz
authors: G Charles
authors: GC
authors: Dismukes
mesh_terms: D020418:Algal Proteins / Q000235:genetics / Q000378:metabolism; D018533:Biomass; D002244:Carbon / Q000378:metabolism; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism*; D004579:Electron Transport; D051599:Glucose-1-Phosphate Adenylyltransferase / Q000235:genetics* / Q000378:metabolism; D005958:Glucosephosphates / Q000378:metabolism; D008027:Light; D055432:Metabolomics; D009154:Mutation; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D010641:Phenotype; D010788:Photosynthesis; D045331:Photosystem I Protein Complex / Q000378:metabolism; D045332:Photosystem II Protein Complex / Q000378:metabolism; D013213:Starch / Q000378:metabolism*
chemical_list: D020418:Algal Proteins; D005958:Glucosephosphates; D045331:Photosystem I Protein Complex; D045332:Photosystem II Protein Complex; D009249:NADP; D002244:Carbon; D013213:Starch; C031590:glucose-1-phosphate; D051599:Glucose-1-Phosphate Adenylyltransferase
keywords: CBB cycle; Chlamydomonas reinhardtii; NADPH re-oxidation; malonyl CoA; oxygen evolution rate; photosynthetic electron transport chain; sta6