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1.
Adsorption of Hg2+ and methylmercury (MeHg) to multi-walled carbon nanotubes (MWCNTs) modified, respectively, with hydroxyl, amine and carboxyl groups was studied. The effect of various factors like the initial pH, natural organic matter (NOM), Cl- and adsorbent dose on the sorption efficiency were evaluated. It was found that amine-modified MWCNTs showed a strong adsorption capacity to Hg2+ and MeHg, and the removal efficiency could reach up to 92% (0.5 g/L MWCNTs, and 100 μg/L Hg2+ and MeHg) which is independent of pH. NOM had complex effects on the adsorption of Hg2+ and MeHg to MWCNTs. Cl- inhibited the adsorption of Hg2+ and MeHg to MWCNTs. The adsorption of Hg2+ and MeHg was found to be inhomogeneous and homogeneous chemisorption, respectively. Our results suggested that MWCNTs modified with different functional groups can efficiently adsorb both Hg2+ and MeHg in aqueous environment. 相似文献
2.
The selective interaction between polyelectrolyte multilayers (PEM) consecutively adsorbed from poly(ethyleneimine) (PEI) and poly(acrylic acid) (PAC) and a binary mixture containing concanavalin A (COA) and lysozyme (LYZ) based on electrostatic interaction is reported. The composition and structure of the PEM and the uptake of proteins were analyzed by in situ attenuated total reflection (ATR) Fourier transform infrared (FTIR) spectroscopy, and the morphology and thickness were characterized by atomic force microscopy (AFM) and ellipsometry. The PEM dissociation degree and charge state and the protein adsorption were shown to be highly dependent on the outermost layer type and the pH in solution. High protein uptake was obtained under electrostatically attractive conditions. This was used to bind selectively one protein from a binary mixture of LYZ/COA. In detail it could be demonstrated that six-layered PEM-6 at pH = 7.3 showed a preferential sorption of positively charged LYZ, while at PEM-5 and pH = 7.3 negatively charged COA could be selectively bound. No protein sorption from the binary mixture was observed at pH = 4.0 for both PEM, when COA, LYZ, and the outermost PEI layer of PEM-5 were positively charged or the outermost PAC layer of PEM-6 was neutral. Furthermore, from factor analysis of the spectral data the higher selectivity was found for PEM-5 compared to PEM-6. Increasing the ionic strength revealed a drastic decrease in the selectivity of both PEM. Evidence was found that the proteins were predominantly bound at the surface and to a minor extent in the bulk phase of PEM. These results suggest possible working regimes and application fields of PEI/PAC multilayer assemblies related to the preparative separation of binary and multicomponent protein mixtures (biofluids, food) as well as to the design of selective protein-resistant surfaces. 相似文献
3.
Structural determinants of the affinity of saxitoxin for neuronal sodium channels. Electrophysiological studies on frog peripheral nerve 总被引:5,自引:4,他引:1
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G Strichartz 《The Journal of general physiology》1984,84(2):281-305
The potencies of saxitoxin (STX) and of five structurally related toxins were determined by their ability to block impulses at equilibrium in frog sciatic nerve. The order of potency, with values relative to STX potency in parentheses, was: neo-STX (4.5) greater than gonyautoxin (GTX) III (1.4) greater than STX (1.0) greater than GTXII (0.22) greater than 12 alpha-dihydroSTX (0.050) greater than 12 beta-dihydroSTX (0.0014). When equipotent solutions of STX and neo-STX were exchanged, impulses in the treated nerve were transiently overblocked or underblocked, thus kinetically distinguishing neo-STX from STX. Similar phenomena occurred with exchanges of STX and GTXIII. No consistent evidence was found for any blocking activity of STX molecules that were not protonated at the C8 guanidinium, but the pH dependence of STX potency cannot be described simply by the titration of this guanidinium group. The effects of pH and of various substituents on STX potency are accounted for by changes in the molecular forms of STX and by alterations in specific electrical charges on STX and at the receptor. The results support a model in which toxin molecules bind in two steps; initial binding of the C8 guanidinium to an anionic group induces the loss of water from the normally hydrated ketone (at carbon 12), which then forms a weak covalent bond with a nucleophilic group on the receptor. 相似文献
4.
Y Hirata H Yoshimi F Marumo T X Watanabe S Kumagaye K Nakajima T Kimura S Sakakibara 《Biochemical and biophysical research communications》1989,162(1):441-447
The effects of synthetic analogs of sarafotoxin (STX) S6b, a snake venom peptide with a high sequence homology to the endothelium-derived vasoconstrictor endothelin (ET), on ET receptor binding activity and cytosolic free Ca2+ concentration [( Ca2+]i) were studied in cultured rat vascular smooth muscle cells. Binding studies revealed that [Cys1-15, Cys3-11] STX competed with 125I-ET for the binding to its vascular receptors with lower affinity than that of ET, but was far more effective than [Cys1-11, Cys3-15]STX in inhibiting the binding. [Cys1-15, Cys3-11]STX had a less potent effect on increasing [Ca2+]i than ET, whereas [Cys1-11, Cys3-15]STX was inactive. These data suggest that there may exist heterogenous subpopulations of the vascular ET/STX receptors, and that the proper double cyclic structure of STX is essential for interacting with its putative receptors to induce the [Ca2+]i response. 相似文献
5.
The sorption of Cd(II) from aqueous solution by rice husk, a surplus agricultural byproduct was investigated. Some simple and low-cost chemical modifications resulted in increasing the sorption capacity of raw rice husk (RRH) from 8.58 mg/g to 11.12, 20.24, 16.18 mg/g and reducing the equilibrium time from 10 h of RRH to 2, 4 and 1 h for epichlorohydrin treated rice husk (ERH), NaOH treated rice husk (NRH), sodium bicarbonate treated rice husk (NCRH), respectively. The effect of pH, sorption kinetics and isotherms were studied in batch experiments. Good correlation coefficient was obtained for pseudo second-order kinetic model, which agreed with chemisorption as the rate-limiting mechanism. Sorption isotherm test showed that equilibrium sorption data were better represented by Langmuir model than the Freundlich model. The highly efficient low cost and the rapid uptake of Cd(II) by NCRH indicated that it could be an excellent alternative for the removal of heavy metal by sorption process. 相似文献
6.
7.
Sharaf EF 《Acta microbiologica Polonica》2002,51(3):293-306
The third oxidation pond at 10th of Ramadan desert receives a number of industrial waste water effluents contaminated with the heavy metal ions Zn, Cd, Cu and Ni. The species diversity and fungal community structure of seven different sites at the onshore sediments and offshore were studied. Mycological analysis resulted in isolation of 3912 fungal colonies, 11.7% of this count were recovered from the onshore sediment sites (4 sites) whereas 88.3% were from the offshore sites (3 sites), in the desert. Fungal counts and species diversity at the onshore sites tend to increase with increasing distance far from the waste water input. A complete accordance was observed among the total fungal counts and species variabilities with organic matter content at each sampling site. This relationship was reversed in case of heavy metal contents with both counts and diversity. Seventeen fungal species belonging to seven genera were isolated from all sites under study. Aspergillus spp. constituted the majority of the isolates (51.7% of the total isolates), followed by Curvularia, Cephalosporium, and Humicola. Of nine isolated Aspergillus spp., A. humicola was the most dominant (37.4% of the total catch) and appeared at all polluted sites. Therefore, A. humicola was chosen to investigate its potential for heavy metals sorption from the contaminated waste water effluent. Four days old biomass pellets could sorb a large amount of heavy metals according to the following sequence: Zn>Cd>Cu>Ni ions. Agitation significantly increased Zn and Cd sorption, but not Cu and Ni. Heavy metals sorption took place at a wide pH range and particularly increased at alkaline pH (8-9). 相似文献
8.
J. Bernhard Wehr F. Pax C. Blamey Peter M. Kopittke Neal W. Menzies 《Plant and Soil》2010,332(1-2):319-330
Pectin, which is an important component of plant cell walls, strongly binds Al and this may play a role in expression of Al toxicity. Sorption of aluminium (Al) and lanthanum (La) from aqueous solutions onto pectic acid, Ca-pectate and plant cell wall material was pH dependent. For Al at pH 3, sorption was less than the available sorption sites (i.e., the cation exchange capacity) on all three sorbents, whereas at pH 4, sorption of Al was in excess of available sorption capacity. By contrast, sorption of the trivalent Al analogue La corresponded to the available sorption capacity on all three sorbents at pH 5. This indicates, therefore, that Al hydrolyses at ≥ pH 4, and hydrolysis increases with Al concentration in solution. Further, it is proposed that the sorption of Al to pectin leads to deprotonation of the galacturonic acid (GalA) residues. Sorption of Al to pectin limits hydrolysis of Al, thereby shifting the pH of hydrolysis to a higher value. Hydrolysis of Al results in its sorption in excess of the stoichiometric equivalent (assuming the free Al3+ ion), leading to oversaturation of the pectin with Al. Staining of the metal-pectate complexes with the metachromatic dye eosin showed that with increasing Al saturation (but not La saturation), the complex developed a positive net charge, due to formation of some positively charged Al-complexes. The development of a positive charge on the Al-pectate complex may have major effects on cellular transmembrane potential and nutrient acquisition by plant roots. This is the first report of Al binding in excess of binding sites and development of a net positive charge on Al-pectate. 相似文献
9.
J S D'Arrigo 《The Journal of membrane biology》1976,29(3):231-242
The effects of uranyl ion (UO22+; at low concentrations binds specifically to phosphate groups) and the cationic dye methylene blue (MB+; binds strongly to carboxyl groups) on saxitoxin (STX) potency in crayfish axon has been studied by means of intracellular microelectrodes. At pH 6.00 +/- 0.05 and 13.5 mM Ca2+, addition of 10.0 muM UO22+ + 5.0 nM STX had only slightly, if any, less effect on the spike's maximum rate of rise [0.79 +/- 0.04 (viz., mean +/- SEM) of control value] than did addition of 5.0 nM STX alone (0.72 +/- 0.05). Under the same conditions of pH and Ca2+ concentration, 1.0 mM MB+ had approximately the same effect: 1.0 mM MB+ + 5.0 nM STX, 0.76 +/- 0.03; 5.0 nM STX alone, 0.70 +/- 0.04. However, at pH 7.00 +/- 0.05 and lower Ca2+ concentrations, 1.0 mM MB+ significantly reduced STX potency. Using 6.0 mM Ca2+: 1.0 mM MB+ + 5.0 nM STX, 0.92 +/- 0.01; 5.0 nM STX alone, 0.68 +/- 0.08. Using 3.0 mM Ca2+, the corresponding values were 0.94 +/- 0.03 and 0.67 +/- 0.04. It is concluded that: (1) In accord with previous suggestions, the ionized acidic group known to exist in the Na channel (and to which a guanidinium group of STX appears to bind) is very likely a carboxyl group and not a phosphate group. (2) The accessible part of the Na channel mouth serving as the saxitoxin receptor probably does not include phospholipid in its structure proper. 相似文献
10.
This paper reports biosorption of Zn(II), Cu(II) and Co(II) onto O. angustissima biomass from single, binary and ternary metal solutions, as a function of pH and metal concentrations via Central Composite Design generated by statistical software package Design Expert 6.0. The experimental design revealed that metal interactions could be best studied at lower pH range i.e. 4.0-5.0, which facilitates adequate availability of all the metal ions. The sorption capacities for single metal decreased in the order Zn(II)>Co(II)>Cu(II). In absence of any interfering metals, at pH 4.0 and an initial metal concentration of 0.5 mM in the solution, the adsorption capacities were 0.33 mmol/g Zn(II), 0.26 mmol/g Co(II) and 0.12 mmol/g Cu(II). In a binary system, copper inhibited both Zn(II) and Co(II) sorption but the extent of inhibition of former was greater than the latter; sorption values being 0.14 mmol/g Zn(II) and 0.27 mmol/g Co(II) at initial Zn(II) and Co(II) concentration of 1.5 mM each, pH 4.0 and 1mM Cu(II) as the interfering metal. Zn(II) and Co(II) were equally antagonistic to each others sorption; Zn(II) and Co(II) sorption being 0.23 and 0.24 mmol/g, respectively, at initial metal concentration of 1.5 mM each, pH 4.0 and 1mM interfering metal concentration. In contrast, Cu(II) sorption remained almost unaffected at lower concentrations of the competing metals. Thus, in binary system inhibition dominance observed was Cu(II)>Zn(II), Cu(II)>Co(II) and Zn(II) approximately Co(II), due to this the biosorbent exhibited net preference/affinity for Cu(II) sorption over Zn(II) or Co(II). Hence, the affinity series showed a trend of Cu(II)>Co(II)>Zn(II). In a ternary system, increasing Co(II) concentration exhibited protection against the inhibitory effect of Cu(II) on Zn(II) sorption. On the other hand, the inhibitory effect of Zn(II) and Cu(II) on Co(II) sorption was additive. The model equation for metal interactions was found to be valid within the design space. 相似文献
11.
HPC-1/syntaxin 1A (STX1A) is abundantly expressed in neurons. STX1A is believed to regulate exocytosis in synaptic vesicles.
In our recent studies, STX1A knockout (KO) mice showed normal development, and basal synaptic transmission in cultured hippocampal neurons appeared to
be normal. However, behavioral abnormalities were observed in STX1A KO mice. In the normal rodent retina, the STX1A protein is expressed in two synaptic layers (plexiform layers). Here, to
evaluate the effects of the loss of STX1A on retinal structure, we examined the retinal layer structure in STX1A KO mice using hematoxylin staining and immunostaining. We found that the general layer structures in the retina were preserved
in all genotypes. However, the outer plexiform layer (OPL) was significantly thicker in KO and heterozygous mutant (HT) mice
compared with that in wild-type (WT) mice. No significant differences were observed in the thicknesses of the other layers.
Immunostaining for protein kinase C α showed that the alignment of rod bipolar cell bodies in the inner nuclear layer (INL)
was slightly disrupted in HT and KO retinas. Furthermore, the dendrites of these cells in the OPL of KO mice were sparse,
compared to those in WT mice. Our results show that STX1A KO mice have increased thickness of the OPL and changes in the morphology of the INL that may contribute to the change in
OPL thickness. We suggest that STX1A may play a role in the structural formation of the INL and OPL in the retina. 相似文献
12.
Extracellular polysaccharide (EPS) produced by a Pseudomonas aeruginosa strain BU2 was characterized for its ability to remove uranium from aqueous solution. The EPS was acidic in nature and found as a potent biosorbent for uranium (U), showing pH dependence and fast saturating metal sorption, being maximum (985 mg U g? 1 EPS) at pH 5.0. The polymer showed enhanced uranium sorption capacity and affinity with increasing solution pH, suggesting a preferential sorption of monovalent uranyl hydroxide ions over the nonhydroxylated divalent species. Pseudo-first-order and pseudo-second-order kinetic models were applied to the experimental data, assuming that the external mass transfer limitations in the system can be neglected and biosorption is sorption controlled. Equilibrium metal binding showing conformity to the Freundlich model suggested a multilayer sorption involving specific binding sites with affinity distribution. The presence of two types of metal binding sites corresponding to strong and weak binding affinity was interpreted from the Scatchard model equation. Uranium sorption by EPS was unaffected or only slightly affected in the presence of several interfering cations and anions, except iron and thorium. Fourier transform infrared (FTIR) spectroscopy ascertained the strong binding of uranium with the carboxylic groups of uronic acids of bacterial EPS at pH 5.0, whereas at lower pH, amino and hydroxyl groups played a major role in metal binding. 相似文献
13.
Francesco Pomati Brett A. Neilan Takahiro Suzuki Gianluca Manarolla Carlo Rossetti 《Journal of phycology》2003,39(3):535-542
The metabolic effect of three different concentrations of lidocaine hydrochloride (0.01, 0.1, and 1 μM) on growth and saxitoxin (STX) production of the freshwater cyanobacterium Cylindrospermopsis raciborskii (Wolosznska) T3 was analyzed. Lidocaine hydrochloride increased both the growth rate and the final growth yield in the toxic cyanobacterium, with a maximum of 25% and 18% for a 1‐μM dose, respectively. Moreover, C. raciborskii T3 samples harvested at the end of the growth phase and analyzed for STX content by HPLC showed an increase in STX intracellular concentration of 14.3% and 49.3% after exposure to 0.01 and 0.1 μM lidocaine hydrochloride, respectively, whereas 1 μM lidocaine hydrochloride resulted in a 114% incremental change in STX content. The time course of the 1‐μM lidocaine hydrochloride effect showed the highest rate of increase in mean STX intracellular concentration (298%) within the first 2 h after induction. The increase in STX content induced by lidocaine hydrochloride in C. raciborskii T3 was dependent on the concentration of Na+ ions in the culture medium and alkaline pH. The results suggest a possible action of lidocaine hydrochloride on membrane ion fluxes and the hypothesis of a potential linkage between cyanobacterial homeostasis and STX regulation. 相似文献
14.
Specific neosaxitoxin interactions with the Na+ channel outer vestibule determined by mutant cycle analysis
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The voltage-gated Na+ channel alpha-subunit consists of four homologous domains arranged circumferentially to form the pore. Several neurotoxins, including saxitoxin (STX), block the pore by binding to the outer vestibule of this permeation pathway, which is composed of four pore-forming loops (P-loops), one from each domain. Neosaxitoxin (neoSTX) is a variant of STX that differs only by having an additional hydroxyl group at the N1 position of the 1,2,3 guanidinium (N1-OH). We used this structural variant in mutant cycle experiments to determine interactions of the N1-OH and its guanidinium with the outer vestibule. NeoSTX had a higher affinity for the adult rat skeletal muscle Na+ channel (muI or Scn4a) than for STX (DeltaG approximately = 1.3 kcal/mol). Mutant cycle analysis identified groups that potentially interacted with each other. The N1 toxin site interacted most strongly with muI Asp-400 and Tyr-401. The interaction between the N1-OH of neoSTX and Tyr-401 was attractive (DeltaDeltaG = -1.3 +/- 0.1 kcal/mol), probably with formation of a hydrogen bond. A second possible attractive interaction to Asp-1532 was identified. There was repulsion between Asp-400 and the N1-OH (DeltaDeltaG = 1.4 +/- 0.1 kcal/mol), and kinetic analysis further suggested that the N1-OH was interacting negatively with Asp-400 at the transition state. Changes in pH altered the affinity of neoSTX, as would be expected if the N1-OH site were partially deprotonated. These interactions offer an explanation for most of the difference in blocking efficacy between neoSTX and STX and for the sensitivity of neoSTX to pH. Kinetic analysis suggested significant differences in coupling energies between the transition and the equilibrium, bound states. This is the first report to identify points of interaction between a channel and a non-peptide toxin. This interaction pattern was consistent with previous proposals describing the interactions of STX with the outer vestibule (Lipkind, G. M., and H. A. Fozzard. 1994. Biophys. J. 66:1-13; Penzotti, J. L., G. Lipkind, H. A. Fozzard, and S. C. Dudley, Jr. 1998. Biophys. J. 75:2647-2657). 相似文献
15.
Equilibrium isotherm studies for the uptake of cadmium and lead ions onto sugar beet pulp 总被引:2,自引:0,他引:2
The adsorption of Cd2+ and Pb2+ on sugar beet pulp (SBP), a low-cost material, has been studied. In the present work, the abilities of native (SBP) to remove cadmium (Cd2+) and lead (Pb2+) ions from aqueous solutions were compared. The (SBP) an industrial by product and solid waste of sugar industry were used for the removal of Cd2+ and Pb2+ ions from aqueous water. Batch adsorption studies were carried out to examine the influence of various parameters such as initial pH, adsorbent dose, initial metal ion concentration, and time on uptake. The sorption process was relatively fast and equilibrium was reached after about 70 min of contact. As much as 70-75% removal of Cd2+ and Pb2+ ions for (SBP) are possible in about 70 min, respectively, under the batch test conditions. Uptake of Cd2+ and Pb2+ ions on (SBP) showed a pH-dependent profile. The overall uptake for the (SBP) is at a maximum at pH 5.3 and gives up to 46.1 mg g(-1) for Cd2+ and at pH 5.0 and gives 43.5 mg g(-1) for Pb2+ for (SBP), which seems to be removed exclusively by ion exchange, physical sorption and chelation. A dose of 8 gL(-1) was sufficient for the optimum removal of both the metal ions. The Freundlich represented the sorption data for (SBP). In the presence of 0.1M NaNO3 the level of metal ion uptake was found to reach its maximum value very rapidly with the speed increasing both with the (SPB) concentration and with increasing initial pH of the suspension. The reversibility of the process was investigated. The desorption of Cd2+ and Pb2+ ions which were previously deposited on the (SBP) back into the deionised water was observed only in acidic pH values during one day study period and was generally rather low. The extent of adsorption for both metals increased along with an increase of the (SBP) dosage. (SBP), which is cheap and highly selective, therefore seems to be a promising substrate to entrap heavy metals in aqueous solutions. 相似文献
16.
Ariff A.B. Mel M. Hasan M.A. Karim M.I.A. 《World journal of microbiology & biotechnology》1999,15(2):291-298
The kinetics and mechanism of lead biosorption by powderized Rhizopus oligosporus were studied using shake flask experiment. The optimum biomass concentration and initial solution pH for lead sorption at initial lead concentrations ranging from 50–200 mg/l was obtained at 0.5 g/l and pH5, respectively. In term of the ratio of initial lead concentration to biomass concentration ratio, the highest lead adsorption was obtained at 750 mg/g which gave the maximum lead uptake capacity of 126 mg/g. The experimental data of lead sorption by R.oligosporus fitted well to the Langmuir sorption isotherm model, indicating that the sorption was similar to that for an ion-exchange resin. This means that the sorption is a single layer metal adsorption that occurred as a molecular surface coverage. This assumption was confirmed by the examination of lead sorption using transmission electron microscope and energy dispersive X-ray analysis, which showed that during sorption most of the lead was adsorbed on the surface of cell. 相似文献
17.
Katherine C. Young Patricia A. Maurice Larry E. Hersman 《Geomicrobiology journal》2013,30(3-4):183-188
Iron (Fe) is an essential nutrient to most microorganisms. Aerobic microorganisms exhibit various strategies for acquiring Fe at near-neutral pH conditions, where Fe oxyhydroxides are insoluble. Although much research has focused on microbial acquisition of Fe from minerals, little is known about Fe acquisition from natural organic matter (NOM). Yet, in surface waters, soils and shallow sediments, Fe is often associated with natural organic matter (NOM), and this NOM-associated Fe could represent an important pool of Fe for microorganisms. Here, we investigated the growth of aerobic Pseudomonas mendocina on soil and surface water NOM samples containing Fe, under Fe-limited conditions. In the presence of NOM, bacteria grew to population sizes greater than in no-Fe-added controls, indicating that the bacteria were able to access Fe associated with NOM. Maximum population size correlated with the NOM-associated Fe concentration. In an additional experiment, Pseudomonas putida was able to acquire Fe from an NOM sample, demonstrating that this ability is not limited to P. mendocina. When Fe was added as 30 μ M FeEDTA plus NOM, together in the same reaction flasks, P. mendocina and P. putida growth was less than in the presence of 30 μM FeEDTA alone. The fact that Fe sources are not simply additive and that the presence of NOM inhibits growth in FeEDTA suggests that further study on the responses of bacteria to a combination of Fe sources is needed to understand the complexities of bacterial Fe acquisition in the subsurface. 相似文献
18.
Post-repolarization block of cloned sodium channels by saxitoxin: the contribution of pore-region amino acids. 总被引:2,自引:1,他引:1
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Sodium channels expressed in oocytes exhibited isoform differences in phasic block by saxitoxin (STX). Neuronal channels (rat IIa co-expressed with beta 1 subunit, Br2a + beta 1) had slower kinetics of phasic block for pulse trains than cardiac channels (RHI). After the membrane was repolarized from a single brief depolarizing step, a test pulse at increasing intervals showed first a decrease in current (post-repolarization block) then eventual recovery in the presence of STX. This block/unblock process for Br2a + beta 1 was 10-fold slower than that for RHI. A model accounting for these results predicts a faster toxin dissociation rate and a slower association rate for the cardiac isoform, and it also predicts a shorter dwell time in a putative high STX affinity conformation for the cardiac isoform. The RHI mutation (Cys374-->Phe), which was previously shown to be neuronal-like with respect to high affinity tonic toxin block, was also neuronal-like with respect to the kinetics of post-repolarization block, suggesting that this single amino acid is important for conferring isoform-specific transition rates determining post-repolarization block. Because the same mutation determines both sensitivity for tonic STX block and the kinetics of phasic STX block, the mechanisms accounting for tonic block and phasic block share the same toxin binding site. We conclude that the residue at position 374, in the putative pore-forming region, confers isoform-specific channel kinetics that underlie phasic toxin block. 相似文献
19.
High-temperature-oriented production of bacterial penicillin acylase (PAC), which is usually expressed at low temperatures (less than 30 degrees C), was demonstrated in this study via heterologous expression of the Providencia rettgeri (P. rettgeri) pac gene in Escherichia coli (E. coli). While it is possible to produce PAC at a temperature as high as 37 degrees C, the environmental condition (specifically, culture pH) critically affected culture performance. Production of PAC at 37 degrees C was feasible only when culture pH was close to neutral (i.e., 6.5-7.5). Outside this pH range, cell physiology for the host/vector system was seriously affected, resulting in poor culture performance. In acidic culture environments, temperature significantly affected the pac expression level and specific PAC activity decreased with an increase in culture temperature. In basic culture environments, cell growth was seriously inhibited though the pac expression level was minimally affected by temperature. Such unusual types of pH and temperature effects on pac expression were never reported for bacterial PACs. The results suggest that culture pH should be precisely controlled for the current host/vector systems being applied on the overproduction of P. rettgeri PAC in E. coli at high temperatures. 相似文献