scholarly journals A comparison of mitochondria from Torulopsis utilis grown in continous culture with glycerol, iron, ammonium, magnesium or phosphate as the growth-limiting nutrient

1971 ◽  
Vol 124 (1) ◽  
pp. 123-134 ◽  
Author(s):  
P. A Light ◽  
P. B. Garland
Keyword(s):  
Energy Conservation ◽  
Electron Flow ◽  
Respiratory Control ◽  
Indirect Evidence ◽  
Iron Limitation ◽  
Limited Growth ◽  
Respiratory Inhibitor ◽  
Limiting Nutrient ◽  
Haem Iron

1. Mitochondria prepared from Torulopsis utilis grown in a chemostat with iron-limited growth were found to lack energy conservation but not electron flow in that segment of the respiratory chain leading from intramitochondrial NADH to the cytochromes [i.e. the site 1 segment (Lehninger, 1964)]. 2. Site 1 energy conservation was present in mitochondria prepared from cells grown under conditions of limitation by glycerol, ammonium and magnesium. Phosphate-limited growth resulted in mitochondrial preparations without respiratory control. 3. Mitochondria from cells grown under conditions of iron limitation were insensitive to the respiratory inhibitor piericidin A, whereas sensitivity was present in mitochondria prepared from glycerol-, ammonium-, magnesium- or phosphate-limited cells. 4. These observations are considered to provide indirect evidence for a role of non-haem iron in the mechanism of energy conservation and also piericidin A sensitivity in T. utilis mitochondria. 5. A readily constructed and inexpensive pH-measuring and -controlling circuit is described for use with continuous-culture apparatus.

scholarly journals Non-haem iron and the dissociation of piericidin A sensitivity from site 1 energy conservation in mitochondria from Torulopsis utilis

1971 ◽  
Vol 124 (1) ◽  
pp. 135-151 ◽  
Author(s):  
R. A. Clegg ◽  
P. B. Garland
Keyword(s):  
Energy Conservation ◽  
Electron Flow ◽  
Iron Concentration ◽  
Normal Function ◽  
Aerobic Incubation ◽  
Submitochondrial Particles ◽  
Limited Growth ◽  
Iron Deficient ◽  
Haem Iron ◽  
Acid Labile

1. The aerobic incubation of iron-deficient Torulopsis utilis cells for 12h under non-growing conditions results in the recovery by mitochondria of the previously absent site 1 energy conservation and sensitivity to piericidin A. 2. The recovery of piericidin A sensitivity but not site 1 is prevented by the presence of cycloheximide (100μg/ml) in the medium used for aerobic incubation of the cells. Rotenone sensitivity behaved similarly. 3. Chloramphenicol, erythromycin and tetracycline were without effect on the recovery of site 1 and piericidin A sensitivity. 4. Inclusion of 59Fe in the growth medium can be used as the basis for a highly sensitive assay for non-haem iron. 5. Iron-limited growth of T. utilis lowers the concentration of both non-haem iron and acid-labile sulphide of submitochondrial particles by over 20-fold compared with the ‘normal’ situation with iron-supplemented glycerol-limited growth. 6. Increases in the non-haem iron and acid-labile sulphide concentrations of submitochondrial particles occur when site 1 and piericidin A sensitivity are recovered. The increase is approximately halved by the presence of cycloheximide. 7. The non-haem iron of T. utilis submitochondrial particles does not exchange with added iron. 8. Continuous culture of T. utilis at the transition between glycerol- and iron-limitation results in cells where mitochondria possess site 1 energy conservation but lack piericidin A sensitivity. 8. It is concluded, in contrast with widely held views to the opposite, that energy conservation at site 1 does not require electron flow to proceed through a piericidin A- or rotenone-sensitive route. 9. Restriction of the iron supplied to growing T. utilis to a concentration just above that required for growth limitation demonstrates that a 10- to 20-fold decrease of the ‘normal’ non-haem iron concentration of both cells and mitochondria is without effect on the growth yield per unit of carbon source. Submitochondrial particles prepared from such iron-restricted but otherwise functionally normal cells have a non-haem iron concentration of about 0.5–0.8ng-atom/mg of protein. It is concluded that the concentration of iron–sulphur protein required for normal function by the respiratory chain is close to the concentrations of cytochromes and flavoproteins.

scholarly journals Effect of sulphate-limited growth on mitochondrial electron transfer and energy conservation between reduced nicotinamide–adenine dinucleotide and the cytochromes in Torulopsis utilis

1971 ◽  
Vol 124 (1) ◽  
pp. 155-170 ◽  
Author(s):  
B. A. Haddock ◽  
P. B. Garland
Keyword(s):  
Energy Conservation ◽  
Cytochrome B ◽  
Antimycin A ◽  
Alternative Route ◽  
Aerobic Incubation ◽  
Submitochondrial Particles ◽  
Limited Growth ◽  
Sulphide Concentration ◽  
Haem Iron ◽  
Acid Labile

1. Conditions have been established for the sulphate-limited growth of Torulopsis utilis in continuous culture. 2. Mitochondria prepared from sulphate-limited cells lack both piericidin A sensitivity and the first energy-conservation site (site 1). Sensitivity to antimycin A or cyanide and the second and third energy-conservation sites were apparently unaffected by sulphate-limited growth. 3. Aerobic incubation for 8h of sulphate-limited cells with a low concentration of sulphate (50μm or less) resulted in the recovery of mitochondrial piericidin A sensitivity and site 1. The use of higher concentrations of sulphate (250μm or more) still resulted in the recovery of mitochondrial piericidin A sensitivity and site 1, but also resulted in the appearance of a non-phosphorylating oxidase, which mediated oxidation of the respiratory chain at about the level of cytochrome b in an antimycin A- and cyanide-insensitive manner. Both this alternative route and the conventional normal route of respiration were shown to coexist and to intercommunicate at the level of cytochrome b. 4. Low-temperature spectroscopy failed to identify any new respiratory component to explain the alternative route. 5. The apparent affinity of the alternative route for oxygen was similar to that for the conventional route through cytochrome oxidase, namely half-maximal activity at 0.1μm-oxygen or less. 6. The non-haem iron concentration of submitochondrial particles was unaffected by sulphate limitation, whereas the acid-labile sulphide concentration was lowered tenfold. Marked increases (between four- and 30-fold) in the acid-labile sulphide concentration of submitochondrial particles were observed in sulphate-limited cells after aerobic incubation with various concentrations of sulphate. The lowest increase (fourfold) was observed without added sulphate, the highest (30-fold) with 1.0mm added sulphate. 7. The ratio of non-haem iron to acid-labile sulphide in submitochondrial particles varied with different growth conditions from a maximum of 15.0 to a minimum of 0.72. It is suggested that analytical measurements of non-haem iron are an inadequate guide to the concentration of iron–sulphur protein in complex systems. 8. The effects of sulphate-limited growth on site 1 and piericidin sensitivity are interpreted to indicate a role for iron–sulphur protein in these properties. 9. The aerobic incubation of sulphate-limited cells with cycloheximide resulted in the recovery by mitochondria of site 1 but not of piericidin sensitivity. 10. The appearance of the alternative route for cyanide- and antimycin-A (but not piericidin A-) insensitive respiration on incubating sulphate-limited cells with sulphate concentrations higher than 250μm indicates that the alternative route involves an iron–sulphur protein.

Relationship between iron-limited growth and energy limitation during phased cultivation of Candida utilis

1978 ◽  
Vol 24 (4) ◽  
pp. 440-447 ◽  
Author(s):  
K. C. Thomas ◽  
P. S. S. Dawson
Keyword(s):  
Ethyl Acetate ◽  
Candida Utilis ◽  
Energy Charge ◽  
Tca Cycle ◽  
Iron Limitation ◽  
Limited Growth ◽  
Acetyl Coa ◽  
Limiting Nutrient ◽  
Energy Limitation

The yeast Candida utilis was continuously synchronized by the phasing technique (6 h doubling time) with either iron or nitrogen as the limiting nutrient. Iron limitation resulted in decreased molar growth yields with respect to the carbon substrates and ammonia and in increased specific rates of oxygen uptake. Relatively low energy-charge values were maintained by the iron-limited culture. All these taken together seemed to indicate that the growth of the yeast under iron limitation was also limited by metabolically available energy.Considerable amounts of ethyl acetate were produced by the yeast under phased cultivation when the growth was limited by iron but not by nitrogen. In vitro studies using cell-free extracts showed that the substrates for ethyl acetate synthesis were acetyl coenzyme A (acetyl CoA) and ethanol. Under iron-limited growth acetyl CoA seemed to be diverted to ethyl acetate formation rather than being oxidized through the tricarboxylic acid (TCA) cycle. The possibility of energy limitation under iron-limited growth being brought about by the reduced capacity of the yeast to oxidize acetyl CoA through the TCA cycle is considered.

Plastoquinones in photosynthesis

1978 ◽  
Vol 284 (1002) ◽  
pp. 591-599 ◽  
Keyword(s):  
Electron Transport ◽  
Photosystem Ii ◽  
Energy Conservation ◽  
Thylakoid Membrane ◽  
Electron Flow ◽  
Proton Translocation ◽  
Photosynthetic Electron Transport ◽  
Atp Formation ◽  
Electron Transport Chains

Several plastoquinones with different or modified side chains have been characterized in plant material: they are localized in the inner thylakoid membrane of the chloroplast. So far only plastoquinone-45 (PQ-45) has been identified as an obligatory functional component of the photosynthetic electron transport chain in chloroplasts between photosystem II and photosystem I. A special form (semiquinone) of PQ-45 acts as primary acceptor Q of photosystem II, a large pool of PQ-45 as electron buffer, interconnecting several electron transport chains. The rôle of PQ, in energy conservation (ATP formation) is of particular current interest. Owing to vectorial electron flow across the thylakoid membrane, plastoquinone is thought to be reduced on the outside and plastohydroquinone to be oxidized on the inside of the membrane. This results in a proton translocation across the membrane and a build-up of a proton motive force which drives ATP formation. Old and new plastoquinone antagonists are described and the relevance of inhibitor studies on the rôle of plastoquinone in electron flow and photophosphorylation is discussed. Open questions and current problems of the mechanism of plastoquinone/plastoquinol transport across the membrane - and of proton translocation connected to it - relevant for the mechanism of energy conservation in photosynthesis, are pointed out.

Physiological Role of Cyclic Electron Flow in Higher Plants

2012 ◽  
Vol 30 (1) ◽  
pp. 100
Author(s):  
Wei HUANG ◽  
Shi-Bao ZHANG ◽  
Kun-Fang CAO
Keyword(s):  
Higher Plants ◽  
Electron Flow ◽  
Physiological Role ◽  
Cyclic Electron Flow

Role of pulmonary reflexes in ventilation and respiratory control of acutely obstructed dogs during assisted ventilation

Lung ◽  
1980 ◽  
Vol 158 (1) ◽  
pp. 1-8
Author(s):  
Zab Mohsenifar ◽  
David Campisi ◽  
Daniel H. Simmons
Keyword(s):  
Respiratory Control ◽  
Assisted Ventilation
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