Toward an understanding of the biogenesis of the starch granule. Evidence that Chlamydomonas soluble starch synthase II controls the synthesis of intermediate size glucans of amylopectin

• Published in: The Journal of biological chemistry Vol. 268; no. 22; pp. 16223 - 16230
• Main Authors: Fontaine, T, D'Hulst, C, Maddelein, M.L, Routier, F, Pepin, T.M, Decq, A, Wieruszeski, J.M, Delrue, B, Koornhuyse, N. van den, Bossu, J.P
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 05.08.1993
• Subjects: ACTIVIDAD ENZIMATICA; ACTIVITE ENZYMATIQUE; ALMIDON; AMIDON; AMILOPECTINA; AMILOSA; Amylopectin; Amylopectin - biosynthesis; AMYLOPECTINE; AMYLOSE; Amylose - metabolism; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Carbohydrates; Chlamydomonas; Chlamydomonas - genetics; Chlamydomonas - metabolism; CHLOROPHYCEAE; Chromatography; Fundamental and applied biological sciences. Psychology; GLICOSILTRANSFERASAS; GLUCANE; GLUCANO; GLYCOSYLTRANSFERASE; Holosides; Life Sciences; Magnetic Resonance Spectroscopy; METABOLISME DES GLUCIDES; METABOLISMO DE CARBOHIDRATOS; MUTANT; MUTANTES; Mutation; Other biological molecules; Solubility; Starch Synthase; Starch Synthase - metabolism; Enzyme; Starch; Algae; Chlamydomonas reinhardtii; Chlorophycophyta; Carbohydrate; Biosynthesis; Synthase; Thallophyta
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)85409-1

Low starch mutants of Chlamydomonas reinhardtii were isolated after x-ray mutagenesis of wild-type strain 137C. The mutants accumulated 20-40% of the normal amount and displayed a 2-fold decrease of the total glycogen-primed soluble starch synthase activity. Three different mutant alleles of the st-3 gene were isolated that were characterized by similar defects and displayed a net increase in amylose content. Amylose-primed synthesis of glucan in native gels revealed a complete wipe out of one of the soluble starch synthases. Zymograms and kinetic analyses performed both in the mutant and in partially purified wild type extracts reveal at least two distinct activities that are partly analogous to higher plant soluble starch synthases I and III (SSI and II). The st-3 mutants were defective for SSII. Methylation and debranching of the purified amylopectin fraction clearly show a decrease in the number of intermediate size glucans (dp8 to 50) and an absolute and relative increase of very short glucans (dp2 to 7). These results suggest that a soluble starch synthase may be necessary for the synthesis or maintenance of intermediate size glucans that are the main component of the branched clusters of amylopectin