Journal Article

Purification and Characterization of Class-I and Class-II Fructose-1,6-bisphosphate Aldolases from the Cyanobacterium <i>Synechocystis</i> sp. PCC 6803

Ken Nakahara, Hiroshi Yamamoto, Chikahiro Miyake and Akiho Yokota

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 44, issue 3, pages 326-333
Published in print March 2003 | ISSN: 0032-0781
Published online March 2003 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcg044
Purification and Characterization of Class-I and Class-II Fructose-1,6-bisphosphate Aldolases from the Cyanobacterium Synechocystis sp. PCC 6803

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The whole genome sequence database for Synechocystis sp. PCC 6803 has revealed the presence of genes encoding class-I (CI) and class-II (CII) fructose-1,6-bisphosphate aldolases (FBAs) in this organism. Two types of FBA from Synechocystis sp. PCC 6803 were separated by chromatography on phenyl-Sepharose. The activity of the enzyme in the major peak was inhibited by the presence of 25 mM EDTA; however, the activity in the minor peak was not. Therefore, the FBA in the former fractions was designated as CII-FBA, and in the latter designated as CI-FBA. CI-FBA was functionally redundant in Synechocystis sp. PCC 6803, while no disruptant for the gene encoding CII-FBA was obtained under photoautotrophic conditions. The kinetic parameters of CI- and CII-FBAs purified from Synechocystis sp. PCC 6803 in the cleavage reaction of FBP were generally similar, except in their reactivity for SBP. The SBP/FBP activity ratio of the CII-FBA was two times higher than that of the CI-FBA.

Keywords: Keywords: Calvin cycle — Class-I aldolase — Class-II aldolase — Fructose-1,6-bisphosphate aldolase — Sedohepturose-1,7-biphosphate aldolase — Synechocystis sp. PCC6803.; Abbreviations: DHAP, dihydroxyacetone phosphate; DTT, dithiothreitol; FBA, fructose-1,6-bisphosphate aldolase; FBP, fructose-1,6-bisphosphate; FBPase, fructose-1,6-bisphosphatase; GAP, glyceraldehyde 3-phosphate; ORF, open-reading frame; PCR cycle, photosynthetic carbon reduction cycle; PMSF, phenylmethanesulphonyl fluoride; SBP, sedoheptulose-1,7-bisphosphate.

Journal Article.  5365 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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