Purification and Properties of Fumarate Reductase from Baker’s Yeast

Muratsubaki, Haruhiro; Katsume, Takuro

doi:10.1080/00021369.1982.10865539

Cited by 5 publications

(10 citation statements)

References 3 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For anaerobic cultivation, ergosterol (0.025 g l −1 ) and Tween 80 (1.05 g l −1 ) were added to the medium. Other conditions of cultivation were as described previously[2]. To prepare agar plates, 1.5% Difco bacto agar was added to the defined medium.…”

Section: Methodsmentioning

confidence: 99%

“…The enzymatic activity was assayed for myokinase[13], citrate synthase[14], malate dehydrogenase[15], cytochrome c oxidase[16], and kynurenine hydroxylase[17]. Fumarate reductase activity was assayed as described previously[2]. Protein was assayed according to the method of Bradford[18] using a commercial kit (Pierce) with bovine albumin as a standard.…”

Section: Methodsmentioning

confidence: 99%

“…Soluble fumarate reductase from yeast irreversibly catalyzes the conversion of fumarate to succinate and requires FADH 2 , FMNH 2 , reduced riboflavin, or reduced forms of some leuco‐pigments as electron donors [1–3]. Fumarate reductase is a soluble enzyme containing one molecule of non‐covalently bound FAD per molecule of protein.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Physiological role of soluble fumarate reductase in redox balancing during anaerobiosis inSaccharomyces cerevisiae

Enomoto

Arikawa

Muratsubaki

2002

FEMS Microbiology Letters

Self Cite

View full text Add to dashboard Cite

In Saccharomyces cerevisiae, there are two isoenzymes of fumarate reductase (FRDS1 and FRDS2), encoded by the FRDS and OSM1 genes, respectively. Simultaneous disruption of these two genes results in a growth defect of the yeast under anaerobic conditions, while disruption of the OSM1 gene causes slow growth. However, the metabolic role of these isoenzymes has been unclear until now. In the present study, we found that the anaerobic growth of the strain disrupted for both the FRDS and OSM1 genes was fully restored by adding the oxidized form of methylene blue or phenazine methosulfate, which non-enzymatically oxidize cellular NADH to NAD(+). When methylene blue was added at growth-limiting concentrations, growth was completely arrested after exhaustion of oxidized methylene blue. In the double-disrupted strain, the accumulation of succinate in the supernatant was markedly decreased during anaerobic growth in the presence of methylene blue. These results suggest that fumarate reductase isoenzymes are required for the reoxidation of intracellular NADH under anaerobic conditions, but not aerobic conditions.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Physiological role of soluble fumarate reductase in redox balancing during anaerobiosis inSaccharomyces cerevisiae

Enomoto

Arikawa

Muratsubaki

2002

FEMS Microbiology Letters

Self Cite

View full text Add to dashboard Cite

show abstract

“…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

View full text Add to dashboard Cite

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.

show abstract

“…For anaerobic cultivation, ergosterol (0.025 g l 31 ) and Tween 80 (1.05 g l 31 ) were added to the medium. Other conditions of cultivation were as described previously [2]. To prepare agar plates, 1.5% Difco bacto agar was added to the de¢ned medium.…”

Section: Media and Growth Conditionsmentioning

confidence: 99%

Physiological role of soluble fumarate reductase in redox balancing during anaerobiosis in Saccharomyces cerevisiae

Enomoto¹

2002

FEMS Microbiology Letters

View full text Add to dashboard Cite

In Saccharomyces cerevisiae, there are two isoenzymes of fumarate reductase (FRDS1 and FRDS2), encoded by the FRDS and OSM1 genes, respectively. Simultaneous disruption of these two genes results in a growth defect of the yeast under anaerobic conditions, while disruption of the OSM1 gene causes slow growth. However, the metabolic role of these isoenzymes has been unclear until now. In the present study, we found that the anaerobic growth of the strain disrupted for both the FRDS and OSM1 genes was fully restored by adding the oxidized form of methylene blue or phenazine methosulfate, which non-enzymatically oxidize cellular NADH to NAD þ . When methylene blue was added at growth-limiting concentrations, growth was completely arrested after exhaustion of oxidized methylene blue. In the double-disrupted strain, the accumulation of succinate in the supernatant was markedly decreased during anaerobic growth in the presence of methylene blue. These results suggest that fumarate reductase isoenzymes are required for the reoxidation of intracellular NADH under anaerobic conditions, but not aerobic conditions.

show abstract

Purification and Properties of Fumarate Reductase from Baker’s Yeast

Cited by 5 publications

References 3 publications

Physiological role of soluble fumarate reductase in redox balancing during anaerobiosis inSaccharomyces cerevisiae

Physiological role of soluble fumarate reductase in redox balancing during anaerobiosis inSaccharomyces cerevisiae

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

Physiological role of soluble fumarate reductase in redox balancing during anaerobiosis in Saccharomyces cerevisiae

Contact Info

Product

Resources

About