OpenUCT :: Browsing by Subject "Kinetics"

Browsing by Subject "Kinetics"

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Open Access
A Point Mutation in the Juxtamembrane Stalk of Human Angiotensin I-converting Enzyme Invokes the Action of a Distinct Secretase
(2001) Alfalah, Marwan; Parkin, Edward T; Jacob, Ralf; Sturrock, Edward D; Mentele, Reinhard; Turner, Anthony J; HOOPER, Nigel M; Naim, Hassan Y
Angiotensin I-converting enzyme (ACE) is one of a number of integral membrane proteins that is proteolytically shed from the cell surface by a zinc metallosecretase. Mutagenesis of Asn(631) to Gln in the juxtamembrane stalk region of ACE resulted in more efficient secretion of the mutant protein (ACE(NQ)) as determined by pulse-chase analysis. In contrast to the wild-type ACE, the cleavage of ACE(NQ) was not blocked by the metallosecretase inhibitor batimastat but by the serine protease inhibitor, 1,3-dichloroisocoumarin. Incubation of the cells at 15 degrees C revealed that ACE(NQ) was cleaved in the endoplasmic reticulum, and mass spectrometric analysis of the secreted form of the protein indicated that it had been cleaved at the Asn(635)-Ser(636) bond, three residues N-terminal to the normal secretase cleavage site at Arg(638)-Ser(639). These data clearly show that a point mutation in the juxtamembrane region of an integral membrane protein can invoke the action of a mechanistically and spatially distinct secretase. In light of this observation, previous data on the effect of mutations in the juxtamembrane stalk of shed proteins being accommodated by a single secretase having a relaxed specificity need to be re-evaluated.
Open Access
Asparagine 706 and Glutamate 183 at the Catalytic Site of Sarcoplasmic Reticulum Ca 2+ -ATPase Play Critical but Distinct Roles in E 2 States
(2006) Clausen, Johannes D; McIntosh, David B; Woolley, David G; Anthonisen, Anne Nyholm; Vilsen, Bente; Andersen, Jens Peter
Mutants with alteration to Asn(706) of the highly conserved (701)TGDGVND(707) motif in domain P of sarcoplasmic reticulum Ca(2+)-ATPase were analyzed for changes in transport cycle kinetics and binding of the inhibitors vanadate, BeF, AlF, and MgF. The fluorides likely mimic the phosphoryl group/P(i) in the respective ground, transition, and product states of phosphoenzyme hydrolysis (Danko, S., Yamasaki, K., Daiho, T., and Suzuki, H. (2004) J. Biol. Chem. 279, 14991-14998). Binding of BeF, AlF, and MgF was also studied for mutant Glu(183) --> Ala, where the glutamate of the (181)TGES(184) motif in domain A is replaced. Mutations of Asn(706) and Glu(183) have in common that they dramatically impede the function of the enzyme in E2 states, but have little effect in E1. Contrary to the Glu(183) mutant, in which E2P slowly accumulates (Clausen, J. D., Vilsen, B., McIntosh, D. B., Einholm, A. P., and Andersen, J. P. (2004) Proc. Natl. Acad. Sci. U. S. A. 101, 2776-2781), E2P formation was not detectable with the Asn(706) mutants. Differential sensitivities of the mutants to inhibition by AlF, MgF, and BeF made it possible to distinguish different roles of Asn(706) and Glu(183). Hence, Asn(706) is less important than Glu(183) for gaining the transition state during E2P hydrolysis but plays critical roles in stabilization of E2P ground and E2.P(i) product states and in the major conformational changes associated with the Ca(2)E1P --> E2P and E2 --> Ca(2)E1 transitions, which seem to be facilitated by interaction of Asn(706) with domain A.
Open Access
Atomic mobility in thin solid Pa2Si films
(1985) Zingu, Edmund Charles; Comrie, Craig M
A theory for the growth kinetics of planar silicide formation in single- and bi-layer metal silicon systems has been developed on the basis that the chemical potential gradient in the growing layer is the driving force for diffusion. The predictions of the theory, when applied to single layer metal-silicon systems, is in agreement with other theories and with experimental results. Planar growth of the outer silicide layer in bilayer metal-silicon systems is predicted to proceed linearly with time, both when controlled by an interfacial reaction and when limited by diffusion through the interposed silicide layer (when this layer is sufficiently thick). In the latter case it is predicted that the growth rate of the outer silicide layer is inversely proportional to the thickness of the interposed layer.
Open Access
ATPase and Multidrug Transport Activities of the Overexpressed Yeast ABC Protein Yor1p
(1998) Decottignies, Anabelle; Grant, Althea M; Nichols, J Wylie; de Wet, Heidi; McIntosh, David B; Goffeau, André
The Saccharomyces cerevisiae genome encodes 15 full-size ATP binding cassette transporters (ABC), of which PDR5, SNQ2, and YOR1 are known to be regulated by the transcription factors Pdr1p and Pdr3p (pleiotropic drug resistance). We have identified two new ABC transporter-encoding genes, PDR10 and PDR15, which were up-regulated by the PDR1-3 mutation. These genes, as well as four other ABC transporter-encoding genes, were deleted in order to study the properties of Yor1p. The PDR1-3 gain-of-function mutant was then used to overproduce Yor1p up to 10% of the total plasma membrane proteins. Overexpressed Yor1p was photolabeled by [gamma-32P]2', 3'-O-(2,4,6-trinitrophenyl)-8-azido-ATP (K0.5 = 45 microM) and inhibited by ATP (KD = 0.3 mM) in plasma membranes. Solubilization and partial purification on sucrose gradient allowed to detect significant Yor1p ATP hydrolysis activity (approximately 100 nmol of Pi.min-1.mg-1). This activity was phospholipid-dependent and sensitive to low concentrations of vanadate (I50 = 0.3 microM) and oligomycin (I50 = 8.5 microg/ml). In vivo, we observed a correlation between the amount of Yor1p in the plasma membrane and the level of resistance to oligomycin. We also demonstrated that Yor1p drives an energy-dependent, proton uncoupler-insensitive, cellular extrusion of rhodamine B. Furthermore, cells lacking both Yor1p and Pdr5p (but not Snq2p) showed increased accumulation of the fluorescent derivative of 1-myristoyl-2-[6-(NBD)aminocaproyl]phosphatidylethanolamine. Despite their different topologies, both Yor1p and Pdr5p mediated the ATP-dependent translocation of similar drugs and phospholipids across the yeast cell membrane. Both ABC transporters exhibit ATP hydrolysis in vitro, but Pdr5p ATPase activity is about 15 times higher than that of Yor1p, which may indicate mechanistic or regulatory differences between the two enzymes.
Open Access
Decreased production of low density lipoprotein by atorvastatin after apheresis in homozygous familial hypercholesterolemia
(1997) Marais, A David; Naoumova, R P; Firth, J C; Penny, C; Neuwirth, C K Y; Thompson, G R
Apheresis only partially controls raised low density lipoprotein cholesterol levels in patients with homozygous familial hypercholesterolemia, who usually respond poorly to lipid-lowering drugs. The efficacy and mechanism of action of a new 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, atorvastatin, was therefore investigated in seven homozygotes undergoing apheresis. One receptor-negative and six receptor-defective homozygotes undergoing plasma exchange or LDL apheresis every 2 weeks were studied during 2 months each on placebo and on atorvastatin 80 mg daily. Changes in plasma lipids and mevalonic acid, an index of cholesterol synthesis, were measured and the kinetics of the rebound of low density lipoprotein cholesterol and apolipoprotein B after apheresis were analyzed. All subjects had significant improvements on atorvastatin. Mean decreases in low density lipoprotein cholesterol were 31% greater both pre- and post-apheresis on atorvastatin compared with placebo, accompanied by a 63% decrease in mevalonic acid. Percentage changes in low density lipoprotein cholesterol and mevalonic acid were closely correlated (r = 0.89, P = 0.007). The mean production rates of low density lipoprotein cholesterol and apolipoprotein B were 21% and 25% lower, respectively, on atorvastatin than on placebo (P < 0.005 and <0.02) but changes in mean fractional clearance rates were not statistically significant. We conclude that atorvastatin enhances the efficacy of plasma exchange and low density lipoprotein apheresis in patients who lack low density lipoprotein receptors. This effect appears to be due to marked inhibition of cholesterol synthesis which results in a decreased rate of production of low density lipoprotein.
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Effect of culture conditions on the competition between lactate oxidisers and fermenters in a biological sulfate reduction system
(Elsevier, 2012) Oyekola, Oluwaseun O; Harrison, Susan T L; van Hille, Robert P
Kinetic constants (μmax and Ks) describing the predominance of lactate oxidation and fermentation were determined in chemostat cultures. The kinetics of sulfate reduction and lactate utilization were determined from 0.5 to 5 d residence times at feed sulfate concentrations of 1.0–10.0 g l−1. The kinetics of lactate fermentation in the absence of sulfate were investigated at residence times of 0.5–5 d. The lactate oxidizers (LO) were characterized by a μmax of 0.2 h−1 and Ks value of 0.6 g l−1 compared with a μmax of 0.3 h−1 and Ks of 3.3 g l−1 for the lactate fermenters (LF). Using mathematical models, it was shown that LO competed more effectively for lactate at low lactate concentrations (⩽5 g l−1) and high sulfide concentrations (0.5 g l−1). Lactate fermenters outcompeted the oxidizers under conditions of excess lactate (>5 g l−1) and low sulfide (0.014–0.088 g l−1).
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Effect of culture conditions on the competitive interaction between lactate oxidizers and fermenters in a biological sulfate reduction system
(Elsevier, 2012) Oyekola, Oluwaseun O; Harrison, Susan T L; Van Hille, Robert P
Kinetic constants (lmax and Ks) describing the predominance of lactate oxidation and fermentation were determined in chemostat cultures. The kinetics of sulfate reduction and lactate utilization were determined from 0.5 to 5 d residence times at feed sulfate concentrations of 1.0–10.0 g l1 . The kinetics of lactate fermentation in the absence of sulfate were investigated at residence times of 0.5–5 d. The lactate oxidizers (LO) were characterized by a lmax of 0.2 h1 and Ks value of 0.6 g l1 compared with a lmax of 0.3 h1 and Ks of 3.3 g l1 for the lactate fermenters (LF). Using mathematical models, it was shown that LO competed more effectively for lactate at low lactate concentrations (65gl1 ) and high sulfide concentrations (0.5 g l1 ). Lactate fermenters outcompeted the oxidizers under conditions of excess lactate (>5 g l1 ) and low sulfide (0.014–0.088 g l1 ). 2011 E
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The effect of dissolved cations on microbial ferrous-iron oxidation by Leptospirillum ferriphilum in continuous culture
(Elsevier, 2008) Ojumu, Tunde V; Petersen, Jochen; Hansford, Geoffrey S
In heap bioleaching the dissolution of gangue minerals from igneous ore materials can lead to the build-up of considerable concentrations of Mg and Al sulphates in the recycled leach solution. This may interfere with microbial ferrous iron oxidation, which drives the oxidation of the target minerals. In the present study the effect of solution concentrations of Mg and Al as sulphate at individual concentrations of 0 to 10 g•dm− 3 and combined concentrations 0 to 16 g•dm− 3each (or total ionic strength from 0.2 to 1.3 M) has been investigated in continuous culture using Leptospirillum ferriphilum. Increasing the concentrations of the salts increasingly depresses the specific rate of ferrous iron oxidation and also shifts the viable range more and more into the low potential region. Aluminium significantly reduces the amount of carbon biomass maintained in the reactor, whereas magnesium actually enhances it at low concentrations. The experimental data was correlated using the Pirt equation and a simplified substrate utilisation model. The results suggest that the maximum microbial growth rate and growth yield decline significantly only at total ionic strengths above about 1 mol•dm−3. The implications of this study are that heap cultures are likely to perform sub-optimally in those operations where build-up of dissolved gangue minerals is not controlled.
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Effect of sulphate concentration on the community structure and activity of sulphate reducing bacteria
(Trans Tech Publications, 2007) Oyekola, Oluwaseun O; van Hille, Rob; Harrison, Susan T L
This study investigated the effect of sulphate concentration and residence time on the performance of anaerobic sulphate reduction by a mixed sulphate reducing bacteria (SRB) culture using lactate as the sole carbon source and electron donor. The process perforrnance is related to the population structure of the microbial consortia and dominant metabolic reactions. Laboratory scale chemostat cultures at different residence times (1-4 d) and sulphate concentrations (1.0-10.0 glL) were employed. Lactate oxidation was prevalent at feed sulphate concentrations of 1.0 to 5.0 glL. A colresponding increase in the volumetric sulphate reduction rate with increasing volumetric loading rate was also observed at this range. However, at the higher feed sulphate concentration range (10.0-15.0 glL), sulphate inhibition, lactate fermentation and an increased microbial diversity were evident. At each feed concentration of sulphate in the range 5.0 to 15.0 glL, varying dilution rates resulted in significant shifts in dominant metabolic reactions. Sulphate concentration and residence time have significant effects on both the structure of the microbial population and kinetics of biological sulphate reduction.
Open Access
Glutamate 301 of the mouse gonadotropin-releasing hormone receptor confers specificity for arginine 8 of mammalian gonadotropin-releasing hormone
(1994) Flanagan, C A; Becker, I I; Davidson, J S; Wakefield, I K; Zhou, W; Sealfon, S C; Millar, R P
The Arg residue at position 8 of mammalian GnRH is necessary for high affinity binding to mammalian GnRH receptors. This requirement has been postulated to derive from an electrostatic interaction of Arg8 with a negatively charged receptor residue. In order to identify such a residue, 8 conserved acidic residues of the mouse GnRH receptor were mutated to isosteric Asn or Gln. Mutant receptors were tested for decreased preference for Arg8-containing ligands by ligand binding and inositol phosphate production. One of the mutants, in which the Glu301 residue was mutated to Gln, exhibited a 56-fold decrease in apparent affinity for mammalian GnRH. The mutant receptor also exhibited decreased affinity for [Lys8]GnRH, but its affinity for [Gln8]GnRH was unchanged compared with the wild type receptor. The apparent affinity of the mutant receptor for the acidic analogue, [Glu8]GnRH, was increased more than 10-fold. The mutant receptor did not, therefore, distinguish mammalian GnRH from analogues with amino acid substitutions at position 8 as effectively as the wild type receptor. This loss of discrimination was specific for the residue at position 8, because the mutant receptor did distinguish mammalian GnRH from analogues with favorable substitutions at positions 5, 6, and 7. These findings show that Glu301 of the GnRH receptor plays a role in receptor recognition of Arg8 in the ligand and are consistent with an electrostatic interaction between these 2 residues.
Open Access
Importance of Conserved N-domain Residues Thr 441 , Glu 442 , Lys 515 , Arg 560 , and Leu 562 of Sarcoplasmic Reticulum Ca 2+ -ATPase for MgATP Binding and Subsequent Catalytic Steps: PLASTICITY OF THE NUCLEOTIDE-BINDING SITE
(2003) Clausen, Johannes D; McIntosh, David B; Vilsen, Bente; Woolley, David G; Andersen, Jens Peter
Nine single mutations were introduced to amino acid residues Thr441, Glu442, Lys515, Arg560, Cys561, and Leu562 located in the nucleotide-binding domain of sarcoplasmic reticulum Ca2+-ATPase, and the functional consequences were studied in a direct nucleotide binding assay, as well as by steady-state and transient kinetic measurements of the overall and partial reactions of the transport cycle. Some partial reaction steps were also examined in mutants with alterations to Phe487, Arg489, and Lys492. The results implicate all these residues, except Cys561, in high affinity nucleotide binding at the substrate site. Mutations Thr441 --> Ala, Glu442 --> Ala, and Leu562 --> Phe were more detrimental to MgATP binding than to ATP binding, thus pointing to a role for these residues in the binding of Mg2+ or to a difference between the interactions with MgATP and ATP. Subsequent catalytic steps were also selectively affected by the mutations, showing the involvement of the nucleotide-binding domain in these reactions. Mutation of Arg560 inhibited phosphoryl transfer but enhanced the E1PCa2 --> E2P conformational transition, whereas mutations Thr441 --> Ala, Glu442 --> Ala, Lys492 --> Leu, and Lys515 --> Ala inhibited the E1PCa2 --> E2P transition. Hydrolysis of the E2P phosphoenzyme intermediate was enhanced in Glu442 --> Ala, Lys492 --> Leu, Lys515 --> Ala, and Arg560 --> Glu. None of the mutations affected the low affinity activation by nucleotide of the phosphoenzyme-processing steps, indicating that modulatory nucleotide interacts differently from substrate nucleotide. Mutation Glu442 --> Ala greatly enhanced reaction of Lys515 with fluorescein isothiocyanate, indicating that the two residues form a salt link in the native protein.
Open Access
Interaction of nucleotides and cations with the (Ca2+, Mg2+)-ATPase of sarcoplasmic reticulum as determined by fluorescence changes of bound 1-anilino-8-naphthalenesulfonate
(1983) Arav, R; Aderem, A A; Berman, M C
The changes in fluorescence of 1-anilino-8-naphthalenesulfonate (ANS-) have been used to determine binding of ligands to the (Ca2+, Mg2+)-ATPase of sarcoplasmic reticulum vesicles, isolated from rabbit skeletal muscle. ANS- binds to sarcoplasmic reticulum membranes with an apparent Kd of 3.8 X 10(-5) M. The binding of ANS- had no effect on Ca2+ transport or Ca2+-dependent ATPase activity. EGTA, by binding endogenous Ca2+, increased the fluorescence intensity of bound ANS- by 10-12%. Subsequent addition of ATP, ADP, or Ca2+, in the presence or absence of Mg2+, reversed this change of fluorescence. The binding parameters, as determined by these decreases in fluorescence intensity, were as follows: for ATP, Kd = 1.0 X 10(-5) M, nH = 0.80; for ADP, Kd = 1.2 X 10(-5) M, nH = 0.89; and for Ca2+, Kd = 3.4 X 10(-7) M, nH = 1.8. The binding parameters for ITP and for the nonhydrolyzable analogue, adenyl-5'-yl-beta, gamma-methylene)diphosphate, were similar to those of ATP, but GDP, IDP, CDP, AMP, and cAMP had lower apparent affinities. Millimolar concentrations of pyrophosphate also decreased the fluorescence of bound ANS-, whereas orthophosphate caused a small (2-3%) increase in fluorescence in Ca2+-free media. Vanadate, in the presence of EGTA, decreased the fluorescence of bound ANS-with half-maximal effect at 4 X 10(-5) M. The changes of fluorescence intensity of bound ANS- appear to reflect conformational changes of the (Ca2+, Mg2+)-ATPase, consequent to ligand binding, with the low and high fluorescence intensity species corresponding to the E1 and E2 conformations, respectively. These appear to reflect similar conformational states of the (Ca2+, Mg2+)-ATPase to those reported by changes in intrinsic tryptophan fluorescence (DuPont, Y. (1976) Biochem, Biophys. Res. Commun. 71, 544-550).
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Kinetic analysis of biological sulphate reduction using lactate as carbon source and electron donor across a range of sulphate concentrations
(Elsevier, 2010) Oyekola, O O; van Hille, R P; Harrison, S T L
This study investigated the effect of feed sulphate concentration on the kinetics of anaerobic sulphate reduction by a mixed SRB culture, using lactate as the sole carbon source and electron donor. Chemostat cultures were operated across a range of residence times (0.5–5 d) and feed sulphate concentrations (1.0–10.0 g l−1). Similar phenomena were observed at feed sulphate concentrations of 1.0 and 10.0 g l−1 with the volumetric sulphate reduction rate increasing linearly with increasing volumetric sulphate loading rate. These reactors were characterised by higher specific volumetric sulphate reduction rates with maximum values of 0.24 and 0.20 g h−1 g−1. Contrastingly, the reactors fed with sulphate concentrations of 2.5 and 5.0 g l−1 showed distinctly different trends in which the volumetric sulphate reduction rate passed through a maximum at the dilution rates of 0.014 and 0.021 h−1, respectively, followed by a decline with further increase in sulphate loading rate. The maximum specific volumetric sulphate reduction rates observed were 2–6-fold lower than those observed at 1.0 and 10.0 g l−1 feed sulphate concentrations. Profiles of specific volumetric sulphate reduction rate and biomass concentration suggested a shift in lactate utilisation from oxidation to fermentation at high dilution rates, implying a change in the dominant components of the microbial consortium. The data suggest that population structure was influenced by lactate affinity and dissolved sulphide concentration. The trends observed were attributed to the greater ability of lactate oxidisers to scavenge lactate under limiting concentrations of the substrate and their greater resilience to dissolved sulphide species in comparison to lactate fermenters.
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Kinetic analysis of biological sulphate reduction using lactate as carbon source and electron donor: Effect of sulphate concentration.
(Elsevier, 2010) Oyekola, Oluwaseun O; van Hille, Robert P; Harrison, Susan T L
This study investigated the effect of feed sulphate concentration on the kinetics of anaerobic sulphate reduction by a mixed SRB culture, using lactate as the sole carbon source and electron donor. Chemostat cultures were operated across a range of residence times (0.5–5 d) and feed sulphate concentrations (1.0–10.0 g l1 ). Similar phenomena were observed at feed sulphate concentrations of 1.0 and 10.0 g l1 with the volumetric sulphate reduction rate increasing linearly with increasing volumetric sulphate loading rate. These reactors were characterised by higher specific volumetric sulphate reduction rates with maximum values of 0.24 and 0.20 g h1 g1 . Contrastingly, the reactors fed with sulphate concentrations of 2.5 and 5.0 g l1 showed distinctly different trends in which the volumetric sulphate reduction rate passed through a maximum at the dilution rates of 0.014 and 0.021 h1 , respectively, followed by a decline with further increase in sulphate loading rate. The maximum specific volumetric sulphate reduction rates observed were 2–6-fold lower than those observed at 1.0 and 10.0 g l1 feed sulphate concentrations. Profiles of specific volumetric sulphate reduction rate and biomass concentration suggested a shift in lactate utilisation from oxidation to fermentation at high dilution rates, implying a change in the dominant components of the microbial consortium. The data suggest that population structure was influenced by lactate affinity and dissolved sulphide concentration. The trends observed were attributed to the greater ability of lactate oxidisers to scavenge lactate under limiting concentrations of the substrate and their greater resilience to dissolved sulphide species in comparison to lactate fermenters.
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Kinetic measurement of biological oxidation of ferrous iron at low ferric-to-ferrous ratios in a controlled potential batch reactor
(Elsevier, 2008) Kazadi, Thierry Kamunga; Jochen, Petersen
Traditionally, the kinetics of microbial ferrous iron oxidation have been studied in continuous or in batch culture. Both methods have drawbacks: in continuous culture experiments have to be repeated at a number of dilution rates to cover the entire spectrum of ferrous to ferric ratios, which is time-consuming. Furthermore, experiments at very low ferric-to-ferrous ratios require high dilution rates which are close to or exceed those at which wash-out occurs. In batch experiments, on the other hand, the prevalent ferric to ferrous ratio rapidly changes due to substrate depletion while the microbial population continually grows, making determination of specific momentary rates difficult. The present paper describes initial work with a novel device, the Redostat™, which allows careful electrochemical control of ferric to ferrous ratio in a batch reactor. A culture of Leptospirillum ferriphilum was grown at 35 °C and 5 g dm− 3 total iron by maintaining the ferric to ferrous ratio at 0.17, 0.51 and 1.65 (corresponding to redox potentials of 419, 452 and 482 mV vs. Ag/AgCl), respectively. The correlation of data obtained from off-gas and current measurements was excellent, and fitted Monod kinetics with ferric inhibition. A hitherto unobserved effect indicates the onset of ferric iron inhibition at the low redox potentials employed here. It was also noted that the biomass concentration has an effect on the biomass specific ferrous iron consumption rate and the biomass yield on ferrous iron.
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A kinetic study on anaerobic reduction of sulphate, Part I: Effect of sulphate concentration
(Elsevier, 2002) Moosa, S; Nemati, M; Harrison, S T L
The kinetics of anaerobic reduction of sulphate was studied in continuous bioreactors. The effects of initial sulphate concentration and its volumetric loading on the kinetics of reaction and activity of sulphate-reducing bacteria were investigated. The increase in initial concentration of sulphate in the range 1.0–Full-size image (<1 K) enhanced the reaction rate from 0.007–Full-size image (<1 K). For a given initial sulphate concentration increasing the volumetric loading rate of sulphate led to a linear increase in volumetric reduction rate. The initial concentration of sulphate did not have a significant effect on maximum specific growth rate (μm), decay coefficient (kd) on bacterial yields (Yx/sulphate and Yx/acetate), with the values of these coefficients being Full-size image (<1 K) bacteria/g sulphate and Full-size image (<1 K) bacteria/g acetate, respectively. The saturation constant (Ks) was an increasing linear function of initial sulphate concentration, with the lowest and highest values being 0.027 and Full-size image (<1 K), respectively. Using the experimental data a kinetic model, incorporating terms for the effects of initial and residual concentrations of sulphate and biomass, was developed.
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The kinetics of ferrous-iron oxidation by Leptospirillum ferriphilum in continuous culture: the effect of pH
(Elsevier, 2011) Ojumu, T V; Petersen, J
The kinetics of ferrous ion oxidation by Leptospirillum ferriphilum were studied in continuous culture with a focus on the effect of solution pH (pH 0.8–2.0), assuming that the effect of pH on cell metabolism can be independently studied of reactor context and other reactions common in bioleach heaps. A simplified competitive ferric ion inhibition model and the Pirt Equation were used to analyze the experimental data. The results showed that the maximum specific activity of L. ferriphilum has a symmetrical bell-shaped curve relationship with pH. The maximum specific ferrous-iron oxidation rate,qFe2 +maxgave a highest value of 14.54 mmol Fe2+(mmol C h)− 1 at pH 1.3, and was described by a quadratic function. The steady state carbon biomass in the reactor and the apparent affinity constant, K′Fe2 +, also increased with increase in pH; however, a slight increase in the carbon biomass was observed beyond pH 1.6. The results also showed that ferric ion precipitation is significant beyond pH 1.3 and about 13% total iron from the feed was lost at pH 2.0. The maximum biomass yield increased linearly with pH, while the culture maintenance coefficient was significantly small in all experiments and was minimum at pH 1.3. The values are indicative of actively growing chemostat cultures. This study shows that microbial ferrous ion oxidation by L. ferriphilum may be sustained at pH lower than pH 0.8 as the microbial activity is much higher than reported values for common mesophilic acidophiles. This may have implications on how bioleach heap operations can be started-up to improve metal recovery.
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The kinetics of ferrous-iron oxidation by Leptospirillum ferriphilum in continuous culture: the effect of temperature
(Elsevier, 2009) Ojumu, T V; Hansford, G S; Petersen, J
A typical bioleach heap is characterized by wide variation of temperature across the heap bed, leading to oxidation of target minerals occurring at different rates. Previous studies on the effect of temperature on the microbial oxidation of ferrous-iron were limited to a narrow range of temperatures (30–40 °C) near optimum conditions and mostly toAcidithiobacillus ferrooxidans. The kinetics of ferrous-iron oxidation by Leptospirillum ferriphilum were studied in continuous culture. In this paper we focus on the effect of temperature (18–45 °C) on these kinetics. The study was based on the assumption that the effect of temperature can be studied independently of other, equally important factors such as pH, dissolved salts, etc. and independent of the reactor context. The experimental data were correlated using both, a simplified ferric-iron inhibitory model and the Pirt Equation. The results showed that the maximum specific ferrous-iron oxidation rate, increased with increasing temperature to a maximum at 42 °C. This trend can be described adequately by the Arrhenius Equation with an activation energy, Ea of 34.46 kJ mol−1 and frequency factor,K0 of 1.05 × 107 mmol Fe2+(mmolC)−1 h−1. An increase in temperature slightly reduces the steady state carbon biomass in the reactor, while the apparent affinity constant, K′Fe2+ increases. The investigation further suggests that at low temperature (18 °C) and beyond the maximum temperature (42 °C), the culture cannot be sustained in a continuous mode. The maximum biomass yield followed a linear decline with increasing temperature, while cell maintenance on ferrous-iron followed a quadratic trend, although the small values indicates that it is not significant, as would be expected in continuous culture. The results indicate that L. ferriphilum is likely to perform optimally, at warm temperatures (25–42 °C) in heap bioleach operations before being taken over by thermophiles at higher temperatures.
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Modelling of bioleach processes: connection between science and engineering
(Elsevier, 2010) Petersen, J
This paper gives a brief introduction to the modelling of bioleach processes, developed from a careful analysis of the fundamental process steps at the gas–liquid, biological and mineral interfaces, and how these interact in a given reactor environment (tanks and heaps). The insights gained from such modelling work can guide both engineers in the optimisation of processes and scientist in directing their research at areas not yet well understood. From this perspective, some future directions of the bioleaching field are discussed.
Open Access
Phosphorylated Ca 2+ -ATPase Stable Enough for Structural Studies
(2001) Henao, Fernando; Delavoie, Franck; Lacapère, Jean-Jacques; McIntosh, David B; Champeil, Philippe
The atomic structure of sarcoplasmic reticulum Ca(2+)-ATPase, in a Ca(2+)-bound conformation, has recently been elucidated (Toyoshima, C., Nakasako, M., Nomura, H. & Ogawa, H. (2000) Nature 405, 647-655). Important steps for further understanding the mechanism of ion pumps will be the atomic structural characterization of different key conformational intermediates of the transport cycle, including phosphorylated intermediates. Following our previous report (Champeil, P., Henao, F., Lacapère, J.-J. & McIntosh, D. B. (2000) J. Biol. Chem. 276, 5795-5803), we show here that it is possible to prepare a phosphorylated form of sarcoplasmic reticulum Ca(2+)-ATPase (labeled with fluorescein isothiocyanate) with a week-long stability both in membranes and in mixed lipid-detergent micelles. We show that this phosphorylated fluorescein isothiocyanate-ATPase can form two-dimensional arrays in membranes, similar to those that were used previously to reconstruct from cryoelectron microscopy images the three-dimensional structure of Ca(2+)-free unphosphorylated ATPase. The results also provide hope that crystals of phosphorylated Ca(2+)-ATPase suitable for x-ray crystallography will be achieved.