title: Pyruvate is a by-product of catalysis by ribulosebisphosphate carboxylase/oxygenase.
abstract: Pyruvate is a minor product of the reaction catalyzed by ribulosebisphosphate carboxylase/oxygenase from spinach leaves. Labeled pyruvate was detected, in addition to the major labeled product, 3-phosphoglycerate, when 14CO2 was the substrate. Pyruvate production was also measured spectrophotometrically in the presence of lactate dehydrogenase and NADH. The Km for CO2 of the pyruvate-producing activity was 12.5 microM, similar to the CO2 affinity of the 3-phosphoglycerate-producing activity. No pyruvate was detected by the coupled assay when ribulose 1,5-bisphosphate was replaced by 3-phosphoglycerate or when the carboxylase was inhibited by the reaction-intermediate analog, 2'-carboxyarabinitol 1,5-bisphosphate. Therefore, pyruvate was not being produced from 3-phosphoglycerate by contaminant enzymes. The ratio of pyruvate produced to ribulose bisphosphate consumed at 25 degrees C was 0.7%, and this ratio was not altered by varying pH or CO2 concentration or by substituting Mn2+ for Mg2+ as the catalytically essential metal. The ratio increased with increasing temperature. Ribulose-bisphosphate carboxylases from the cyanobacterium Synechococcus PCC 6301 and the bacterium Rhodospirillum rubrum also catalyzed pyruvate formation and to the same extent as the spinach enzyme. When the reaction was carried out in 2H2O, the spinach carboxylase increased the proportion of its product partitioned to pyruvate to 2.2%. These observations provide evidence that the C-2 carbanion form of 3-phosphoglycerate is an intermediate in the catalytic sequence of ribulose-bisphosphate carboxylase. Pyruvate is formed by beta elimination of a phosphate ion from a small portion of this intermediate.
authors.affiliation: Plant Environmental Biology Group, Research School of Biological Sciences, Australian National University, Canberra.
authors.forename: T J
authors.initials: TJ
authors.lastname: Andrews
authors.forename: H J
authors.initials: HJ
authors.lastname: Kane
mesh_terms: D002384:Catalysis; D002853:Chromatography, Liquid; D000458:Cyanobacteria / Q000201:enzymology; D010944:Plants / Q000201:enzymology; D011773:Pyruvates / Q000737:chemistry / Q000378:metabolism*; D019289:Pyruvic Acid; D012247:Rhodospirillum rubrum / Q000201:enzymology; D012273:Ribulose-Bisphosphate Carboxylase / Q000737:chemistry / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature
chemical_list: D011773:Pyruvates; D019289:Pyruvic Acid; D012273:Ribulose-Bisphosphate Carboxylase