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Metabolic Engineering for the Production of a Variety of Biofuels and Biochemicals
Published in Kazuyuki Shimizu, Metabolic Regulation and Metabolic Engineering for Biofuel and Biochemical Production, 2017
The intermediates of the TCA cycle such as citric, fumaric, and succinic acids are localized in mitochondria in eukaryotes. These organic acids are synthesized and excreted in the medium at relatively high concentrations by filamentous fungi such as Aspergillus spp. and Rhizopus sp. (Goldberg et al. 2006). Itaconic acid is produced from aconitate in the TCA cycle in Aspergillus spp. In filamentous fungi, the citric, fumaric, and malic acids are accumulated under specific stress conditions. The overproduction of fumaric and L-malic acids is made by a separate and unique pathway such as the reductive TCA pathway, localized in the cytosol (Goldberg 1991). Citric acid is, on the other hand, produced via citrate synthase (CS), an enzyme in the mitochondrial oxidative TCA cycle, yet citosolic L-malic acid is an intermediate in citric acid synthesis.
The Citric Acid Cycle
Published in Jean-Louis Burgot, Thermodynamics in Bioenergetics, 2019
It is an union of a molecule possessing four carbons (oxaloacetate) with a group possessing two carbons, the rest acetyl, which is brought by acetyl-CoA. It is an aldol reaction in which there is a nucleophilic addition of the enolate ion derivating from the acetyl group ofacetyl-CoA, on the carbonyl of the keto group of oxaloacetate. The reaction is catalyzed by the enzyme citrate synthase. This reaction evolves spontaneously: ∆G’° = –32,2 kJ mol–1.
Effect of citrulline malate supplementation on muscle function and bioenergetics during short-term repeated bouts of fatiguing exercise
Published in Journal of Sports Sciences, 2022
Laura Meimoun, Émilie Pecchi, Christophe Vilmen, David Bendahan, Benoît Giannesini
Citrate synthase activity was assessed in another part (20–30 mg) of the freeze-clamped muscle, which was homogenised with a lysis reagent (ref. C3228; Sigma-Aldrich) and a protease inhibitor cocktail (P8340, Sigma-Aldrich). Citrate synthase activity was measured using the colorimetric Citrate Synthase Assay Kit (ref. CS0720; Sigma-Aldrich) and was normalised by the protein content measured using the colorimetric Pierce BCA Protein Assay Kit (ref. 23,225; Thermo Fisher Scientific, Waltham, MA, USA). All in vitro measurements were done on a microplate reader (Victor X3; PerkinElmer, Waltham, MA, USA).
Biochemical and transcriptional analyses of cadmium-induced mitochondrial dysfunction and oxidative stress in human osteoblasts
Published in Journal of Toxicology and Environmental Health, Part A, 2018
Cristina Monteiro, José Miguel P. Ferreira de Oliveira, Francisco Pinho, Verónica Bastos, Helena Oliveira, Francisco Peixoto, Conceição Santos
Citrate synthase specific activity was spectrophotometrically measured according to Srere (1969), with minor modifications. The assay was performed at 30ºC in a buffer, 200-mM Tris-HCl (pH 8), 2 μL 10-μM DTNB, 25 μL 1-mM oxaloacetate, and 50-μg mitochondrial. The reaction was started by the addition of 92.5 μL of 0.37-mM acetyl-CoA and monitored at 412 nm for 3 min. Citrate synthase specific activity was determined by measuring the decrease of DTNB concentration at 412 nm (ε = 13.6 mM−1cm−1) (Synergy™ HT Multi-Mode, BioTeK), and values expressed as nmol/min/mg protein.