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1.
β-Thalassemia is the most common single gene disorder in Iran and more than 25,000 affected individuals have been reported. It has been reported that in patients with β-thalassemia in the presence of Xmn1 polymorphic site the level of Hb F and Gγ: Aγ ratio is increased. The prevalence of Xmn1 polymorphic site, Gγ: Aγ ratio and Hb F in 197 β-thalassemia major patients from the Kermanshah Province of Iran were studied. The Xmn1 polymorphic site was determined by PCR-RFLP procedure. The levels of Gγ and Aγ chains were detected by HPLC. The percent of Hb F was determined using electrophoresis method. In β-thalassemia major patients the frequency of presence Xmn1 was 0.39. The mean of Gγ: Aγ ratio was found to be 2.5. In the present study it was found that in the presence of Xmn1 polymorphic site Gγ percent and Gγ: Aγ ratio were significantly increased (P = 0.01) and the clinical features such as splenomegaly and bone marrow expansion were significantly improved (P = 0.01). We found that in the presence of Xmn1 polymorphic site on both chromosomes (+/+) the level of Hb F tended to be increased compared to the absence of Xmn1 (−/−). The present investigation has studied the frequency of Xmn1 polymorphic site in β-thalassemia major patients from Western Iran and has revealed that the presence of this polymorphic site caused a positive influence on Hb F production and the Gγ percent which could improve the clinical symptoms of β-thalassemia patients.  相似文献   

2.
5′-O-β-d-Galactosyl-floxuridine, a potential novel prodrug, was synthesized with a yield of 75% through β-galactosidase-catalyzed transgalactosylation. This enzyme displayed absolute regioselectivity toward the 5′-position of floxuridine. For the reaction, the optimal conditions were pH 6.5 at 45°C for 60 h with floxuridine to o-nitrophenyl-β-d-galactoside at 2:1 (mol/mol). Under these conditions, the initial reaction rate and the maximum yield were 0.28 mM h−1 and 75%, respectively.  相似文献   

3.
4.
The recombinant β-carotene 15,15′-monooxygenase from chicken liver was purified as a single 60 kDa band by His-Trap HP and Resource Q chromatography. It had a molecular mass of 240 kDa by gel filtration indicating the native form to be tetramer. The enzyme converted β-carotene under maximal conditions (pH 8.0 and 37°C) with a k cat of 1.65 min−1 and a K m of 26 μM and its conversion yield of β-carotene to retinal was 120% (mol mol−1). The enzyme displayed catalytic efficiency and conversion yield for β-carotene, β-cryptoxanthin, β-apo-8′-carotenal, β-apo-4′-carotenal, α-carotene and γ-carotene in decreasing order but not for zeaxanthin, lutein, β-apo-12′-carotenal and lycopene, suggesting that the presence of one unsubstituted β-ionone ring in a substrate with a molecular weight greater than C30 seems to be essential for enzyme activity.  相似文献   

5.
GABAA receptors composed of α, β and γ subunits display a significantly higher single-channel conductance than receptors comprised of only α and β subunits. The pore of GABAA receptors is lined by the second transmembrane region from each of its five subunits and includes conserved threonines at the 6′, 10′ and 13′ positions. At the 2′ position, however, a polar residue is present in the γ subunit but not the α or β subunits. As residues at the 2′, 6′ and 10′ positions are exposed in the open channel and as such polar channel-lining residues may interact with permeant ions by substituting for water interactions, we compared both the single-channel conductance and the kinetic properties of wild-type α1β1 and α1β1γ2S receptors with two mutant receptors, αβγ(S2′A) and αβγ(S2′V). We found that the single-channel conductance of both mutant αβγ receptors was significantly decreased with respect to wild-type αβγ, with the presence of the larger valine side chain having the greatest effect. However, the conductance of the mutant αβγ receptors remained larger than wild-type αβ channels. This reduction in the conductance of mutant αβγ receptors was observed at depolarized potentials only (ECl = −1.8 mV), which revealed an asymmetry in the ion conduction pathway mediated by the γ2′ residue. The substitutions at the γ2′ serine residue also altered the gating properties of the channel in addition to the effects on the conductance with the open probability of the mutant channels being decreased while the mean open time increased. The data presented in this study show that residues at the 2′ position in M2 of the γ subunit affects both single-channel conductance and receptor kinetics.  相似文献   

6.
Propionibacterium shermanii CDB 10014 is able to grow even at high oxygen transfer rates (24.0 mmol O2 l−1 h−1), in contrast to reports in the specialised literature, where all Propionibacteria are considered oxygen-sensitive microorganisms. Propionic acid is the main product in anaerobiosis. The presence of oxygen in the system leads to an inhibition of propionic acid production while acetic acid formation is enhanced. At high oxygen supply rates no propionic acid is produced and acetic acid is the main product. Lactic acid is also produced in reasonable quantities (2.7 g l−1). The growth rate (μmax) is higher in anaerobiosis (0.19 h−1) than in aerobiosis (0.12–0.15 h−1). The cell yield is higher in aerobiosis (0.18–0.22 g g−1) than in anaerobiosis (0.14 g g−1) suggesting the oxidative metabolism of glucose by Propionibacterium shermanii CDB 10014. No corrinoid production was detected at oxygen transfer rates of more than 13.6 mmol l−1 h−1. Received: 10 September 1997 / Received revision: 6 January 1998 / Accepted: 9 January 1998  相似文献   

7.
To produce recombinant β-carotene in vitro, synthetic operons encoding genes governing its biosynthesis were engineered into Escherichia coli. Constructs harboring these operons were introduced into either a high-copy or low-copy cloning vector. β-Carotene production from these recombinant E. coli cells was either constitutive or inducible depending upon plasmid copy number. The most efficient β-carotene production was with the low-copy based vector. The process was increased incrementally from a 5 l to a 50 l fermentor and finally into a 300 l fermentor. The maximal β-carotene yields achieved using the 50 l and 300 l fermentor were 390 mg l−1 and 240 mg l−1, respectively, with overall productivities of 7.8 mg l−1 h−1 and 4.8 mg l−1 h−1.  相似文献   

8.
In this work, we characterized an ecto-ATPase activity in intact mycelial forms of Fonsecaea pedrosoi, the primary causative agent of chromoblastomycosis. In the presence of 1 mM EDTA, fungal cells hydrolyzed adenosine-5′-triphosphate (ATP) at a rate of 84.6 ± 11.3 nmol Pi h−1 mg−1 mycelial dry weight. The ecto-ATPase activity was increased at about five times (498.3 ± 27.6 nmol Pi h−1 mg−1) in the presence of 5 mM MgCl2, with values of V max and apparent K m for Mg-ATP2−corresponding to 541.9 ± 48.6 nmol Pi h−1 mg−1 cellular dry weight and 1.9 ± 0.2 mM, respectively. The Mg2+-stimulated ecto-ATPase activity was insensitive to inhibitors of intracellular ATPases such as vanadate (P-ATPases), bafilomycin A1 (V-ATPases), and oligomycin (F-ATPases). Inhibitors of acid phosphatases (molybdate, vanadate, and fluoride) or alkaline phosphatases (levamizole) had no effect on the ecto-ATPase activity. The surface of the Mg2+-stimulated ATPase in F. pedrosoi was confirmed by assays in which 4,4′-diisothiocyanostylbene-2,2′-disulfonic acid (DIDS), a membrane impermeant inhibitor, and suramin, an inhibitor of ecto-ATPase and antagonist of P2 purinoreceptors. Based on the differential expression of ecto-ATPases in the different morphological stages of F. pedrosoi, the putative role of this enzyme in fungal biology is discussed.  相似文献   

9.
Kogawa K  Kato N  Kazuma K  Noda N  Suzuki M 《Planta》2007,226(6):1501-1509
A UDP-glucose: anthocyanin 3′,5′-O-glucosyltransferase (UA3′5′GT) (EC 2.4.1.-) was purified from the petals of Clitoria ternatea L. (Phaseoleae), which accumulate polyacylated anthocyanins named ternatins. In the biosynthesis of ternatins, delphinidin 3-O-(6″-O-malonyl)-β-glucoside (1) is first converted to delphinidin 3-O-(6″-O-malonyl)-β-glucoside-3′-O-β-glucoside (2). Then 2 is converted to ternatin C5 (3), which is delphinidin 3-O-(6″-O-malonyl)-β-glucoside-3′,5′-di-O-β-glucoside. UA3′5′GT is responsible for these two steps by transferring two glucosyl groups in a stepwise manner. Its substrate specificity revealed the regioselectivity to the anthocyanin′s 3′- or 5′-OH groups. Its kinetic properties showed comparable k cat values for 1 and 2, suggesting the subequality of these anthocyanins as substrates. However, the apparent K m value for 1 (3.89 × 10−5 M), which is lower than that for 2 (1.38 × 10−4 M), renders the k cat/K m value for 1 smaller, making 1 catalytically more efficient than 2. Although the apparent K m value for UDP-glucose (6.18 × 10−3 M) with saturated 2 is larger than that for UDP-glucose (1.49 × 10−3 M) with saturated 1, the k cat values are almost the same, suggesting the UDP-glucose binding inhibition by 2 as a product. UA3′5′GT turns the product 2 into a substrate possibly by reversing the B-ring of 2 along the C2-C1′ single bond axis so that the 5′-OH group of 2 can point toward the catalytic center. K. Kogawa, N. Kato, K. Kazuma, and N. Noda contributed equally to this work.  相似文献   

10.
Yarrowia lipolytica converts methyl ricinoleate to γ-decalactone, a high-value fruity aroma compound. The highest amount of 3-hydroxy-γ-decalactone produced by the yeast (263 mg l-1) occurred by increasing the kLa up to 120 h−1 at atmospheric pressure; above it, its concentration decreased, suggesting a predominance of the activity of 3-hydroxyacyl-CoA dehydrogenase. Cultures were grown under high-pressure, i.e., under increased O2 solubility, but, although growth was accelerated, γ-decalactone production decreased. However, by applying 0.5 MPa during growth and biotransformation gave increased concentrations of dec−2-en-4-olide and dec-3-en-4-olide (70 mg l−1).  相似文献   

11.
An isolated, perfused salmon tail preparation showed oxyconformance at low oxygen delivery rates. Addition of pig red blood cells to the perfusing solution at a haematocrit of 5 or 10% allowed the tail tissues to oxyregulate. Below ca. 60 ml O2 kg−1 h−1 of oxygen delivery (DO2), VO2 was delivery dependent. Above this value additional oxygen delivery did not increase VO2 of resting muscle above ca. 35 ml O2 kg−1 h−1. Following electrical stimulation, VO2 increased to ca. 65 ml O2 kg−1 h−1, with a critical DO2 of ca. 150 ml O2 kg−1 h−1. Dorsal aortic pressure fell to 69% of the pre-stimulation value after 5 min of stimulation and to 54% after 10 min. Microspheres were used to determine blood flow distribution (BFD) to red (RM) and white muscle (WM) within the perfused myotome. Mass specific BFD ratio at rest was found to be 4.03 ± 0.49 (RM:WM). After 5 min of electrical stimulation the ratio did not change. Perfusion with saline containing the tetrazolium salt 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) revealed significantly more mitochondrial activity in RM. Formazan production from MTT was directly proportional to time of perfusion in both red and WM. The mitochondrial activity ratio (RM:WM) did not change over 90 min of perfusion.  相似文献   

12.
Ko BS  Rhee CH  Kim JH 《Biotechnology letters》2006,28(15):1159-1162
The effects of glycerol and the oxygen transfer rate on the xylitol production rate by a xylitol dehydrogenase gene (XYL2)-disrupted mutant of Candida tropicalis were investigated. The mutant produced xylitol near the almost yield of 100% from d-xylose using glycerol as a co-substrate for cell growth and NADPH regeneration: 50 g d-xylose l−1 was completely converted into xylitol when at least 20 g glycerol l−1 was used as a co-substrate. The xylitol production rate increased with the O2 transfer rate until saturation and it was not necessary to control the dissolved O2 tension precisely. Under the optimum conditions, the volumetric productivity and xylitol yield were 3.2 g l−1 h−1 and 97% (w/w), respectively.  相似文献   

13.
Sequential batch and continuous operation of a rotating biological contacting (RBC) reactor and the effects of dissolved oxygen on the decoloration of amaranth by Trametes versicolor were evaluated. Amaranth belongs to the group of azo dyes which are potential carcinogens and/or mutagens that can be transformed into toxic aryl amines under anaerobic conditions. Cultivation of T. versicolor in a stirred tank reactor was found to be unsuitable for amaranth decoloration due to significant biomass fouling and increase in medium viscosity. Assuming that decoloration follows first-order kinetics, amaranth was decolorized more rapidly when T. versicolor was immobilized on jute twine in a RBC reactor operated either in a sequential batch (k=0.25 h–1) or in a continuous (0.051 h−1) mode compared to a stirred tank reactor (0.015 h−1). Oxygen was found to be essential for decoloration with the highest decoloration rates occurring at oxygen saturation. Although longer retention times resulted in more decoloration when the RBC was operated in the continuous mode (about 33% amaranth decoloration), sequential batch operation gave better results (>95%) under similar nutrient conditions. Our data indicate that the fastest decoloration should occur in the RBC using nitrogen-free Kirk’s medium with 1 g/l glucose in sequential batch operation at rotational speeds and/or aeration rates which maintain oxygen saturation in the liquid phase.  相似文献   

14.
The freshwater microalga Chlorella vulgaris was grown heterotrophically in fed-batch 50–600-L fermenters at 36°C, on aerated and mixed nutrient solution with urea as a nitrogen and glucose as a carbon and energy source. Cell density increased from the initial value 6.25 to 117.18 g DW L−1 in 32 h in the fermenter 50 L at a mean growth rate 3.52 g DW L−1 h−1. The DW increase in the fermenter 200 L was from 7.25 to 94.82 g DW L−1 in 26.5 h at a mean growth rate 3.37 g DW L−1 h−1. Mean specific growth rate μ was about 0.1 h−1 in the both fermenters, if nutrients and oxygen were adequately supplied. The DW increase in the fermenter 600 L was from 0.8 to 81.6 g DW L−1 in 66.5 h at a mean growth rate 1.22 g DW L−1 h−1 and μ = 0.07 h−1. A limitation of the cell growth rate in 600 L fermenter caused by a low dissolved oxygen concentration above cell densities higher than 10 g DW L−1) occurred. Specific growth rate decreased approximately linearly with increasing glucose concentration (25–80 g glucose L−1) at the beginning of cultivation and decreased with the time of cultivation. The cell yield was 0.55–0.69 g DW (g glucose)−1. The content of proteins, β-carotene, and chlorophylls in the cells steadily increased and starch content decreased, by keeping aerated and mixed culture another 12 h in fermenter after the cell growth was stopped due to glucose deficiency.  相似文献   

15.
The production of compound K and aglycon protopanaxadiol (APPD) from ginsenoside Rd and ginseng root extract was performed using a recombinant β-glycosidase from Pyrococcus furiosus. The activity for Rd was optimal at pH 5.5 and 95°C with a half-life of 68 h at 95°C. β-Glycosidase converted Rb1, Rb2, Rc, and Rd to APPD via compound K. With increases in the enzyme activity, the productivities of compound K and APPD increased. The substrate concentration was optimal at 4.0 mM Rd or 10% (w/v) ginseng root extract; 4 mM of Rd was converted to 3.3 mM compound K with a yield of 82.5% (mol/mol) and a productivity of 2,010 mg l−1 h−1 at 1 h and was hydrolyzed completely to APPD with 364 mg l−1 h−1 after 5 h. Rb1, Rb2, Rc, and Rd at 3.9 mM in 10% ginseng root extract were converted to 3.1 mM compound K with 79.5% and 1,610 mg l−1 h−1 at 1.2 h and were hydrolyzed completely to APPD with 300 mg l−1 h−1 after 6 h. The concentrations and productivities of compound K and APPD in the present study are the highest ever reported.  相似文献   

16.
The biodegradation of phenol by a pure culture of Pseudomonas putida was investigated in a continuously fed stirred-tank reactor, under aerobic conditions. The dilution rate was varied between 0.0174 h−1 and 0.278 h−1, covering a wide range of dissolved oxygen and the inhibition region of phenol. Through non-linear analysis of the data, a dual-substrate growth kinetics, Haldane kinetics for phenol and Monod kinetics for oxygen, was derived with high correlation coefficients. Respective biokinetic parameters were evaluated as μm = 0.569 h−1, K p = 18.539 mg/l, K i = 99.374 mg/l, K o = 0.048 mg/l, Y x/p = 0.521 g microorganism/g phenol and Y x/o = 0.338 g microorganism/g oxygen, being in good agreement with other studies in the literature. Maintenance factors for both phenol and oxygen were calculated for the first time for P. putida while the saturation coefficient for oxygen, K o, was genuinely evaluated from the constructed model, not imported or adapted from other studies as reported in the literature. All pertinent biokinetic parameters for P. putida have been calculated from continuous system data, which are most appropriate for use in continuous bioprocess applications. Received: 29 July 1996 / Received revision: 18 November 1996 / Accepted: 23 November 1996  相似文献   

17.
Songbirds are widely studied to investigate the hormonal control of behavior. However, little is known about the effects of steroids on neurotransmission in these birds. We used electrophysiological and pharmacological techniques to characterize γ-aminobutyric acid (GABA) type A receptors (GABAA) of primary cultured telencephalic and hippocampal neurons from developing zebra finches. Additionally, their modulation by 17β-estradiol(E2), 5α- and 5β-dihydrotestosterone (DHT), 5α- and 5β-pregnan-3α-ol-20-one, and corticosterone was examined. Whole-cell GABA-evoked currents were inhibited by picrotoxin (10 μmol l−1) and bicuculline methiodide (10 μmol l−1) and potentiated by pentobarbital (100 μmol l−1) and propofol (3 μmol l−1). Loreclezole (10 μmol l−1) potentiated GABA-evoked currents, suggesting the presence of β2, β3 and/or β4 subunits. Diazepam (1 μmol l−1) potentiated currents, while Zn2+ (1 μmol l−1) caused no inhibition, indicating the presence of γ subunits. 5α- and 5β-Pregnan-3α-ol-20-one (100 nmol l−1) potentiated currents, whereas E2 (1 μmol l−1), 5α- and 5β-DHT (1 μmol l−1), and corticosterone (10 μmol l−1) had no detectable effect. We conclude that zebra finch telencephalic and hippocampal GABAA receptors include α, β, and γ subunits and are similar to their mammalian counterparts in both their biophysical and pharmacological properties. Additionally, GABA-evoked currents are greatly potentiated by 5α- and 5β-pregnan-3α-ol-20-one but show little or no acute modulation by sex steroids or corticosterone. Accepted: 12 November 1997  相似文献   

18.
Two reactors, initially operated at 14 and 23±1°C (RA and RB, respectively), were inoculated with a bacterial consortium enriched and acclimatized to the respective temperatures over 4 months. The biofilms, formed in the reactors, were studied using scanning electron microscopy, cultivation of the biofilm microflora, and physiological analysis of the isolates. Two bacteria able to mineralize chlorophenols under a large range of temperature (10–30°C) were isolated from the biofilm communities of reactors RA and RB and characterized as Alcaligenaceae bacterium R14C4 and Cupriavidus basilensis R25C6, respectively. When temperature was decreased by 10°C, the chlorophenols removal capacity was reduced from 51.6 to 22.8 mg l−1 h−1 in bioreactor RA (from 14 to 4°C) and from 59.3 to 34.7 mg l−1 h−1 in bioreactor RB (from 23±1 to 14°C). Fluorescence in situ hybridization (FISH) of the biofilm communities showed that, in all temperatures tested, the β-proteobacteria were the major bacterial community (35–47%) followed by the γ-proteobacteria (12.0–6.5%). When the temperature was decreased by 10°C, the proportions of γ-proteobacteria and Pseudomonas species increased significantly in both microbial communities.  相似文献   

19.
Air artificially contaminated with increasing concentrations of benzene was treated in a laboratory scale compost-packed biofilter for 240 days with a removal efficiency of 81–100%. The bacterial community in the packing material (PM) at different heights of the biofilter was analysed every 60 days. Bacterial plate counts and ribosomal intergenic spacer analysis (RISA) of the isolated strains showed that the number of cultivable aerobic heterotrophic bacteria and the species diversity increased with benzene availability. Identification of the isolated species and the main bands in denaturing gradient gel electrophoresis (DGGE) profiles from total compost DNA during the treatment revealed that, at a relatively low volumetric benzene load (1.2≤VBL≤6.4 g m−3 PM h−1), besides low G+C Gram positive bacteria, originally present in the packing compost, bacteroidetes and β- and γ-proteobacteria became detectable in the colonising population. At the VBL value (24.8 g m−3 PM h−1) ensuring the maximum elimination capacity of the biofilter (20.1 g m−3 PM h−1), strains affiliated to the genus Rhodococcus dominated the microflora, followed by β-proteobacteria comprising the genera Bordetella and Neisseria. Under these conditions, more than 35% of the isolated strains were able to grow on benzene as the sole carbon source. Comparison of DGGE and automated RISA profiles of the total community and isolated strains showed that a complex bacterial succession occurred in the reactor in response to the increasing concentrations of the pollutant and that cultivable bacteria played a major role in benzene degradation under the adopted conditions.  相似文献   

20.
The change of dilution rate (D) on both Methylophilus methylotrophus NCIMB11348 and Methylobacterium sp. RXM CCMI908 growing in trimethylamine (TMA) chemostat cultures was studied in order to assess their ability to remove odours in fish processing plants. M. methylotrophus NCIMB11348 was grown at dilution rates of 0.012–0.084 h−1 and the biomass level slightly increased up to values of D around 0.07 h−1. The maximum cell production rate was obtained at 0.07 h−1 corresponding to a maximum conversion of carbon into cell mass (35%). The highest rate of TMA consumption was 3.04 mM h−1 occurring at D=0.076 h−1. Methylobacterium sp. RXM CCMI908 was grown under similar conditions. The biomass increased in a more steep manner up to values of D around 0.06 h−1. The maximum cell production rate (0.058 g l−1h−1) was obtained in the region close to 0.06 h−1 where a maximum conversion of the carbon into cell mass (40%) was observed. The maximum TMA consumption was 2.33 mM h−1 at D=0.075 h−1. The flux of carbon from TMA towards cell synthesis and carbon dioxide in both strains indicates that the cell is not excreting products but directing most of the carbon source to growth. Carbon recovery levels of approximately 100% show that the cultures are carbon-limited. Values for theoretical maximum yields and maintenance coefficients are presented along with a kinetic assessment based on the determination of the substrate saturation constant and maximum growth rate for each organism. Received: 25 February 1999 / Received revision: 14 May 1999 / Accepted: 17 May 1999  相似文献   

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