Purification and properties of fumarate reductase from baker's yeast.

Muratsubaki, Haruhiro; Katsume, Takuro

doi:10.1271/bbb1961.46.2909

Cited by 14 publications

(16 citation statements)

References 2 publications

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“…Where necessary, membrane samples were first clarified by using deoxycholate (final concentration 0.05%). Enzyme preparations obtained by the procedures quoted or as gifts from colleagues included: submitochondrial particles (ETP) from beef heart [4], rat liver [5], human placenta [6] and Ascaris suum (from Dr. R. Komuniecki, University of Toledo, OH); aerobic cytoplasmic membranes of E. coli (Dr. R. Gennis, University of Illinois) and R subtilis (Dr. L. Hederstedt, University of Lund, Sweden); purified FRD complex of E. coli (Dr. G. Cecchini, University of California, San Francisco); isolated complex (Complex II) [7] and soluble SDH [8] of beef heart; and the yeast cytoplasmic FRD isolated essentially according to the procedure of Muratsubaki and Katsume [9]. Protein was measured by the biuret method, after precipitation with trichloroacetic acid and, in the case of Complex II, an additional pre-wash with acetone/HC1 [10].…”

Section: Methodsmentioning

confidence: 99%

Classification of fumarate reductases and succinate dehydrogenases based upon their contrasting behaviour in the reduced benzylviologen/fumarate assay

et al. 1993

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Reduction of fumarate by soluble beef heart succinate dehydrogenase has been shown previously by voltammetry to become increasingly retarded as the potential is lowered below a threshold potential of -80 mV at pH 7.5. The behaviour resembles that of a tunnel diode, an electronic device exhibiting the property of negative resistance. The enzyme thus acts to oppose fumarate reduction under conditions of high thermodynamic driving force. We now provide independent evidence for this phenomenon from spectrophotometric kinetic assays. With reduced benzylviologen as electron donor, we have studied the reduction of fumarate catalysed by various enzymes classified either as succinate dehydrogenases or fumarate reductases.For succinate dehydrogenases, the rate increases as the concentration of reduced dye (driving force) decreases during the reaction. In contrast, authentic fumarate reductases of anaerobic cells (and 'succinate dehydrogenase' from Bacillus subtilis) neither exhibit the electrochemical effect nor deviate from simple kinetic behaviour in the cuvette assay. The 'tunnel-diode' effect may thus represent an evolutionary adaptation to aerobic metabolism.

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Section: Methodsmentioning

confidence: 99%

Classification of fumarate reductases and succinate dehydrogenases based upon their contrasting behaviour in the reduced benzylviologen/fumarate assay

et al. 1993

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“…In contrast, the fumarate reductase enzyme of Escherichia coli [2] and Vibrio succinogenes [3] are membrane-bound enzymes bound covalently with FAD, that exhibit weak succinate oxidation activity. Yeasts contain two fumarate reductase isoenzymes that can be fractionated into the absorbed (FRDS1; fumarate reductase soluble 1) and non-absorbed (FRDS2; fumarate reductase soluble 2) fractions on a DE-52 column [4]. Recently, we determined that FRDS1, located in the cytosol, is encoded by the FRDS gene [5] and FRDS2, located in promitochondria, is encoded by the OSM1 gene [6].…”

Section: Introductionmentioning

confidence: 99%

Soluble fumarate reductase isoenzymes from Saccharomyces cerevisiae are required for anaerobic growth

Arikawa

1998

FEMS Microbiology Letters

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In Saccharomyces cerevisiae, the cytosolic and promitochondrial isoenzymes of fumarate reductase are encoded by the FRDS and OSM1 genes, respectively. The product of the OSM1 gene is reported to be required for growth in hypertonic medium. Simultaneous disruption of the FRDS and OSM1 genes resulted in the inability of the yeasts to grow anaerobically on glucose as a carbon source, and disruption of the OSM1 gene caused poor growth under anaerobic conditions. However, the disruption of both the FRDS and/or OSM1 genes had no effect on aerobic growth or growth under hypertonic conditions. These results suggest that the fumarate reductase isoenzymes in Saccharomyces cerevisiae are essential for anaerobic growth but not for growth under hypertonic conditions. z

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“…Similarly, the FRD activity in the presence of 40 M NADH was estimated to be 70% of that with 0.2 mM NADH (data not shown). In other organisms, the K m values for fumarate were reported to vary from 0.005 to 3 mM (10,19,29,33,36,38). Although the K m values for fumarate and NADH were too low to be determined accurately, the affinities for fumarate (K m Ͻ 50 M) and NADH (K m Ͻ 40 M) were considered to be high.…”

Section: Purification Of Frd From H Thermophilusmentioning

confidence: 99%

A Soluble NADH-Dependent Fumarate Reductase in the Reductive Tricarboxylic Acid Cycle of Hydrogenobacter thermophilus TK-6

et al. 2008

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Fumarate reductase (FRD) is an enzyme that reduces fumarate to succinate. In many organisms, it is bound to the membrane and uses electron donors such as quinol. In this study, an FRD from a thermophilic chemolithoautotrophic bacterium, Hydrogenobacter thermophilus TK-6, was purified and characterized. FRD activity using NADH as an electron donor was not detected in the membrane fraction but was found in the soluble fraction. The purified enzyme was demonstrated to be a novel type of FRD, consisting of five subunits. One subunit showed high sequence identity to the catalytic subunits of known FRDs. Although the genes of typical FRDs are assembled in a cluster, the five genes encoding the H. thermophilus FRD were distant from each other in the genome. Furthermore, phylogenetic analysis showed that the H. thermophilus FRD was located in a distinct position from those of known soluble FRDs. This is the first report of a soluble NADH-dependent FRD in Bacteria and of the purification of a FRD that operates in the reductive tricarboxylic acid cycle.

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Purification and properties of fumarate reductase from baker's yeast.

Cited by 14 publications

References 2 publications

Classification of fumarate reductases and succinate dehydrogenases based upon their contrasting behaviour in the reduced benzylviologen/fumarate assay

Classification of fumarate reductases and succinate dehydrogenases based upon their contrasting behaviour in the reduced benzylviologen/fumarate assay

Soluble fumarate reductase isoenzymes from Saccharomyces cerevisiae are required for anaerobic growth

A Soluble NADH-Dependent Fumarate Reductase in the Reductive Tricarboxylic Acid Cycle of Hydrogenobacter thermophilus TK-6

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