Regulation of the NADH and NADPH-ferredoxin oxidoreductases in Clostridia of the butyric group

Petitdemange, H.; Cherrier, C; Raval, G.; Gay, Robert

doi:10.1016/0304-4165(76)90300-7

Cited by 110 publications

(66 citation statements)

References 18 publications

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“…In contrast, high NADH-fd reductase activities [0*074 pmol min-' (mg protein)-'] were found for cells that exhibit a high NADH content and data on NADH-fd reductase activities showed a direct correlation between the specific activities, which increased from 0.015 to 0.074 pmol min-l (mg protein)-', and the NADH content of the cell. NADH-fd reductase activity was higher than that described for C. (Petitdemange et al, 1976;Lovitt et al, 1988). The function of pyridine nucleotide oxidoreductases depends on both the bacterial species and strain and the specific growth conditions of a given strain (Petitdemange et al, 1976;Girbal et al, 1995a).…”

Section: Enzymic Activitiesmentioning

confidence: 62%

“…NADH-fd reductase activity was higher than that described for C. (Petitdemange et al, 1976;Lovitt et al, 1988). The function of pyridine nucleotide oxidoreductases depends on both the bacterial species and strain and the specific growth conditions of a given strain (Petitdemange et al, 1976;Girbal et al, 1995a).…”

Section: Enzymic Activitiesmentioning

confidence: 62%

“…Ferredoxin (fd)-NAD(P)+ reductase [EC 1.18.1.3 (EC 1.18.1.2)] was assayed with pyruvate as the reductant of fd and under a carbon monoxide atmosphere according to Petitdemange et al (1976), modified by the addition of 1 mM purified fd. NADH-fd reductase was measured by metronidazole reduction according to the method of Blusson et al (1981).…”

Section: Enzyme Assaysmentioning

confidence: 99%

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Metabolism of cellobiose by Clostridium cellulolyticum growing in continuous culture: evidence for decreased NADH reoxidation as a factor limiting growth

et al. 1998

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Previous results indicated that molar growth yields are reduced when Clostridium cellulolyticum is cultured in media containing cellobiose concentrations greater than 1 g I-'. Continuous cultures were examined to determine the physiological basis of these poor growth yields. Acetate was the main product of C. cellulolyticum metabolism, whereas the production of reduced compounds such as ethanol or lactate was low. Such patterns of product formation were accompanied by a 12-fold increase in intracellular NADH concentration when the cellobiose flow was increased. Catabolic enzymic activities were measured in vitro. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), acetate kinase and phosphoroclastic activities were found at similar levels as in cells metabolizing higher substrate concentrations. In contrast, lactate dehydrogenase activity was low and correlated with the rate of lactate production. Furthermore, an inhibition of GAPDH activity by high NADH/NAD+ ratios was established. These results suggested that a decreased NADH reoxidation could be responsible for limiting C. cellulolyticum growth. Lactate and ethanol production were not sufficient to balance out the NADH produced in the GAPDH step of glycolysis. One consequence of poor NADH reoxidation would be an increase in intracellular concentration of NADH, which in turn could inhibit GAPDH activity.

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Section: Enzymic Activitiesmentioning

confidence: 62%

Section: Enzymic Activitiesmentioning

confidence: 62%

Section: Enzyme Assaysmentioning

confidence: 99%

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Metabolism of cellobiose by Clostridium cellulolyticum growing in continuous culture: evidence for decreased NADH reoxidation as a factor limiting growth

et al. 1998

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“…The existence of distinct ferredoxin-NAD oxidoreductases has also been shown for other clostridia. For example, when Clostridium tyrobutyricum was grown on glucose, a high NADH-ferredoxin reductase activity and a low ferredoxin-NAD reductase activity directed the electron flow from NADH to ferredoxin and H2 (34). However, when cultures were grown on pyruvate-acetate mixtures, NADH-ferredoxin reductase was not detected and only a ferredoxin-NAD reductase controlled the electron flow from reduced ferredoxin to NADH at the expense of H2 (36).…”

Section: Discussionmentioning

confidence: 99%

Regulation of carbon and electron flow in Clostridium acetobutylicum grown in chemostat culture at neutral pH on mixtures of glucose and glycerol

1994

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The metabolism of Clostridium acetobutylicum was manipulated, at neutral pH and in chemostat culture, by changing the overall degree of reduction of the substrate, using mixtures of glucose and glycerol. Cultures grown on glucose alone produced only acids, and the intracellular enzymatic pattern indicated the absence of butyraldehyde dehydrogenase activity and very low levels of coenzyme A-transferase, butanol, and ethanol dehydrogenase activities. In contrast, cultures grown on mixtures of glucose and glycerol produced mainly alcohols and low levels of hydrogen. The low production of hydrogen was not associated with a change in the hydrogenase level but was correlated with the induction of a ferredoxin-NAD reductase and a decreased level of NADH-ferredoxin reductase. The production of alcohols was related to the induction of a NAD-dependent butyraldehyde dehydrogenase and to higher expression of NAD-dependent ethanol and butanol dehydrogenases. The coenzyme A-transferase was poorly expressed, and thus no acetone was produced. These changes in the enzymatic pattern, obtained with cultures grown on a mixture of glucose and glycerol, were associated with a 7-fold increase of the intracellular level of NADH and a 2.5-fold increase of the level of ATP.The complex anaerobic metabolism of Clostridium acetobutylicum has been studied in considerable detail in recent years, but the factors involved in triggering the metabolic shift, and the physiological state associated with the transition from the acidogenic to solventogenic phase, are still not totally understood.In a number of studies, the effects of electron flow regulation, nutrient limitation, and end product accumulation on the onset and maintenance of solvent production have been investigated. In typical batch fermentations, initiation of solventogenesis is associated with a pH-acid effect. Lowering the intracellular pH and thereby increasing the concentration of undissociated butyric acid has a positive effect on the production of acetone and butanol (2,13,14,28,29,38). The alteration of the electron flow by carbon monoxide, a reversible inhibitor of the hydrogenase, can also induce the shift from an acidogenic fermentation to an alcohologenic one (9,20,25,27): alcohol (butanol and ethanol) and lactate production at very high specific rates is obtained, without acetone and little or no acetate and butyrate formation. The decrease of hydrogenase activity or concentration by iron limitation conditions (19) specifically yields butanol and ethanol as the major fermentation end products. It has also been shown, in continuous cultures at low pH, that phosphate limitation, nitrogen limitation, and glucose excess are conditions under which primarily solvents are produced by C. acetobutylicum (3,4,35).In this study, the metabolic flexibility of C. acetobutylicum was studied in chemostat culture by increasing the NAD(P)H pressure, using mixtures of glucose and a more reduced chemical like glycerol. The pH of the culture was maintained at 6.5 so that no changes could be r...

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“…Indeed, during the last two decades, research on the fermentation metabolism of Clostridium kluyveri (Hillmer & Gottschalk, 1972, 1974Lurz et al, 1979), Clostridium pasteurianum and/or Clostridium butyricum (e.g. Jungermann et al, 1973;Petitdemange et al, 1976;Mitchell et al, 1987), and certainly Clostridium acetobutylicum (see, for example, Jones & Woods, Freier & Gottschalk, 1987;Wiesenborn et al, 1989a Rao & Mutharasan, 1989), has been encouraged, because of the importance of some of these organisms for the production of solvents (ethanol, butanol and acetone) or fuels (hydrogen gas and ethanol).…”

Section: Introductionmentioning

confidence: 99%

Lactate and ethanol dehydrogenase activities in continuous cultures of Clostridium thermosaccharolyticum LMG 6564

Vancanneyt

Vos

Vennens

et al. 1990

Journal of General Microbiology

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The pattern of ethanol and lactate formation by continuous cultures of CIostridium ~~~m s a c c b o @ f k w n LMG 6564 under glucose limitation is affected by culture conditions such as pH and dilution rate. NADH-and NADPHmediated lactate dehydrogenase (LDH; EC 1.1.1.27) and alcohol (ethanol) dehydrogenase (ADH; EC 1.1.1.1 and EC 1.1.1.2) activities were measured in cell extracts from continuous cultures grown under Merent conditions. In conditions of high product formation, the NADH-mediated reaction was higher than the NADPHmediated reaction for both LDH and ADH. LDH showed an absolute requirement for fructose 1,6=bisphosphate (FBP). Both NADH-and NADPH-linked LDH reactions were cytoplasmic, not sensitive to oxygen, and had a pH optimum of 6-M.5; the temperature optimum was 55-60 OC. The reverse reaction (lactate oxidation) could not be demonstrated. ADH activity was found in the particulate fraction of the cell lysate and was sensitive (not completely, but irreversibly) to oxygen. The temperature and pH optima were 43 "C, pH 7-0 and 45 "C, pH 8.8 for the NADH-and NADPH-mediated reactions, respectively. The production of at least two different ADHs is likely. LDH and ADH seemed to be regulated at the level of enzyme synthesis (direct correlation between the in nitro activities and the lactate and ethanol yields in the culture) with a second regulation of LDH by FBP at the reaction level.

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Regulation of the NADH and NADPH-ferredoxin oxidoreductases in Clostridia of the butyric group

Cited by 110 publications

References 18 publications

Metabolism of cellobiose by Clostridium cellulolyticum growing in continuous culture: evidence for decreased NADH reoxidation as a factor limiting growth

Metabolism of cellobiose by Clostridium cellulolyticum growing in continuous culture: evidence for decreased NADH reoxidation as a factor limiting growth

Regulation of carbon and electron flow in Clostridium acetobutylicum grown in chemostat culture at neutral pH on mixtures of glucose and glycerol

Lactate and ethanol dehydrogenase activities in continuous cultures of Clostridium thermosaccharolyticum LMG 6564

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