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1.
Lim YR  Yeom SJ  Kim YS  Oh DK 《Bioresource technology》2011,102(5):4277-4280
The optimum conditions for the production of l-arabinose from debranched arabinan were determined to be pH 6.5, 75 °C, 20 g l−1 debranched arabinan, 42 U ml−1 endo-1,5-α-l-arabinanase, and 14 U ml−1 α-l-arabinofuranosidase from Caldicellulosiruptor saccharolyticus and the conditions for sugar beet arabinan were pH 6.0, 75 °C, 20 g l−1 sugar beet arabinan, 3 U ml−1 endo-1,5-α-l-arabinanase, and 24 U ml−1 α-l-arabinofuranosidase. Under the optimum conditions, 16 g l−1l-arabinose was obtained from 20 g l−1 debranched arabinan or sugar beet arabinan after 120 min, with a hydrolysis yield of 80% and a productivity of 8 g l−1 h−1. This is the first reported trial for the production of l-arabinose from the hemicellulose arabinan by the combined use of endo- and exo-arabinanases.  相似文献   

2.
Olive-mill wastewater (OMW) was investigated for its suitability to serve as a medium for lipase production by Candida cylindracea NRRL Y-17506. The OMW that best supported enzyme production was characterized by low COD and low total sugars content. In shake flask batch cultures, OMW supplementation with 2.4 g l−1 NH4Cl and 3 g l−1 olive oil led to an enzyme activity of about 10 U ml−1. The addition of glucose or malt extract and supplements containing organic N (e.g., peptone, yeast extract) either depressed or did not affect the enzyme production. Further experiments were then performed in a 3-l stirred tank reactor to assess the impact of medium pH and stirring speed on the yeast enzyme activity. The lipase activity was low (1.8 U ml−1) when the pH was held constant at 6.5, significantly increased (18.7 U ml−1) with uncontrolled pH and was maximum (20.4 U ml−1) when the pH was let free to vary below 6.5. A stirring regime, that varied depending on the dissolved oxygen concentration in the medium, both prevented the occurrence of anoxic conditions during the exponential growth phase and enabled good lipase production (i.e., 21.6 U ml−1) and mean volumetric productivity (i.e., 123.5 U l−1 h−1).  相似文献   

3.
A new polymer bearing aldehyde groups was designed and synthesized by grafting 4-pyridinecarboxaldehyde onto poly(epichlorohydrin). Antibodies can be directly immobilized on the surface of the polymer film through the covalent bonding of aldehyde groups of the film with amino groups of antibodies. In this study, human immunoglobulin G (IgG) was used as a model analyte for the fabrication of an electrochemical impedance immunosensor. Using the proposed immunosensor, IgG in the range from 0.1 to 80 ng ml−1 was detected with a detection limit of 0.07 ng ml−1 (signal/noise [S/N] = 3). In addition, the electrochemical impedance immunosensor displays good stability and reproducibility.  相似文献   

4.
In this study, interactions between Cry1Ac, a toxic crystal protein produced by Bacillus thuringiensis (Berliner), and Beauveria bassiana on the mortality and survival of Ostrinia furnacalis was evaluated in the laboratory. The results showed that Cry1Ac is toxic to O. furnacalis. Not only were larval growth and development delayed, but pupation, pupal weight and adult emergency also decreased when larvae were fed on artificial diet containing purified Cry1Ac toxin. When third instars O. furnacalis were exposed to combination of B. bassiana (1.8 × 105, 1.8 × 106 or 1.8 × 107 conidia ml−1) and Cry1Ac, (0.2 or 0.8 μg g−1), the effect on mortality was additive, however, the combinations of sublethal concentrations showed antagonism between Cry1Ac (3.2 or 13 μg g−1) and B. bassiana (1.8 × 105 or 1.8 × 106 conidia ml−1). When neonates were reared on sublethal concentrations of Cry1AC until the third instar, and survivors exposed B. bassiana conidial suspension, such treatments showed additive effect on mortality of O. furnacalis except for the combination of Cry1Ac (0.2 μg g−1) and B. bassiana (1.8 × 106 conidia ml−1) that showed antagonism.  相似文献   

5.
In this study, a novel sensitive electrochemiluminescence (ECL) immunosensor was constructed by carboxyl graphene (GR) for enhancing luminol–O2 system emission. Here, carboxyl GR was used to enhance the ECL intensity of luminol that had excellent electron transfer ability and good solubility. The sensing platform was constructed by depositing carboxyl GR on electrodes and immobilizing antibodies on the surface of carboxyl GR through amidation. The specific immunoreaction between α-fetoprotein (AFP) and antibodies resulted in a decrease of ECL intensity, and the intensity decreased linearly with AFP concentrations in the range of 5 pg ml−1 to 14 ng ml−1 with a detection limit of 2.0 pg ml−1. The proposed immunosensor exhibits high specificity, good reproducibility, and longtime stability. It may become a promising technique for protein detection.  相似文献   

6.
Batch cultivations were performed to evaluate the influence of phosphate concentrations (0.25, 0.5, 0.75, and 1.0 g L−1) for pH regimes (9.5, 10.0, and 10.5) on the biomass production by Spirulina platensis. The best condition for cell growth (3.099 g L−1) was found at 0.5 g L−1 phosphate and pH value of 10.0. Cultivation time, phosphate, and pH caused to increase significantly (p < 0.01) in biomass production by S. platensis. Lag time was observed up to 4 h. After then, biomass production increased sharply (p < 0.01) from 0.020 g L−1 to 2.063, 2.213, 1.532, and 0.797 g L−1 at 0.25, 0.5, 0.75, and 1.0 g L−1 phosphate values, respectively. Modified Gompertz model could be regarded as sufficient to describe the biomass production by S. platensis with high determination coefficients and low sum of square value indicated that. Biological parameters for biomass production were successfully predicted by modified Gompertz model.  相似文献   

7.
Colurella dicentra clones isolated from bay water in the Mississippi Gulf Coast were cultured with artificial seawater. Experiments were conducted to determine the effects of six algae species (Nannochloropsis oculata, Tetraselmis chuii, Chaetoceros gracilis, Rhodomonas salina, Isochrysis galbana, and Prorocentrum micans), six C. gracilis densities, and six N. oculata densities (25,000, 50,000, 100,000, 250,000, 500,000, and 1,000,000 cells ml− 1) on C. dicentra population growth. Algae type influenced rotifer production (p < 0.0001). C. gracilis treatment (9120 ± 3351SD) produced the highest number of rotifers followed by N. oculata (5760 ±2232SD). P. micans had the lowest number of rotifers, although not significantly different from numbers in T. chuii, R. salina, and I. galbana treatments (p > 0.05).The population growth rate (r) varied with algae species treatment. The highest values were recorded for C. gracilis treatment (0.22 to 0.26 d− 1), followed by N. oculata (0.21 to 0.24 d− 1), and the lowest for P. micans (− 0.19 to 0.14 d− 1). C. gracilis and N. oculata densities had significant effects (p < 0.0001) on C. dicentra population growth. The highest rotifer production was recorded at a C. gracilis density of 100,000 cells ml− 1, followed by 250,000 cells ml− 1 and 50,000 cells ml− 1. Algae densities of 500,000 cells ml− 1 and above produced the lowest rotifer numbers. Population growth rate (r) varied with C. gracilis densities. The highest values were observed for C. gracilis concentrations of 100,000 cells ml− 1 (0.17 to 0.19 d− 1), and the lowest for concentrations of 500,000 cells ml− 1 and above (− 0.19 to 0.09 d− 1). The 100,000 cells ml− 1N. oculata density gave the highest rotifer production followed by 50,000, 250,000, 25,000, and 500,000 cells ml− 1. Algae densities of 1,000,000 cells ml− 1 produced the lowest rotifer numbers. Population growth rate (r) varied with N. oculata densities, with the highest values obtained for algae densities of 100,000 cells ml− 1 (0.35 to 0.40 d− 1), and the lowest for concentrations of 1,000,000 cells ml− 1 (0.05 to 0.012 d− 1). This is the first report of C. dicentra in Mississippi Coastal waters, and perhaps the smallest marine rotifer species (93 by 49 μm) ever cultured successfully.  相似文献   

8.
In this study, interactions on the mortality and debilitating effects between Cry1Ac, a toxic protein produced by Bacillus thuringiensis (Berliner) and HaCPV (Chinese strain) on first and third instars larvae of Helicoverpa armigera were evaluated in laboratory. When first instar was exposed to combination of Bt cotton leaf discs containing HaCPV (6 × 106, 1 × 107, and 3 × 107 PIB ml−1) the effect on mortality was additive, when such instar larvae exposed to combination of Cry1Ac (0.9, 2.7, or 8.1 μg g−1) and the same concentrations of HaCPV the effect on mortality was additive except for the combination of Cry1Ac (0.3 μg g−1) and HaCPV concentrations that showed synergism. When third instars of H. armigera were infected using a suspension containing both HaCPV and Cry1Ac, most combinations of them showed additive effect except for the combination of Cry1Ac (0.3 μg g−1) and HaCPV (3 × 107 PIB ml−1) that showed synergism. However, when they exposed to Bt cotton leaf discs and HaCPV the effect on mortality was synergism except combination of Bt cotton leaf discs and HaCPV (6 × 106 PIB ml−1) that showed additive. Most of the combinations are showed additive effect in the toxicity and in combinations of Cry1Ac at lowest and HaCPV at highest concentrations synergism is observed. Not only were larval growth and development delayed, but pupation and pupal weight also decreased when larvae were fed on artificial diet containing Cry1Ac and HaCPV or transgenic Bt cotton leaf discs specially in first instar.  相似文献   

9.
Electroreduction and adsorption of cefixime was studied in phosphate buffer by cyclic voltammetry (CV), differential pulse cathodic adsorptive stripping voltammetry (DPCAdSV), and square-wave cathodic adsorptive stripping voltammetry (SWCAdSV) at hanging mercury drop electrode (HMDE). These fully validated sensitive and reproducible cathodic adsorptive stripping voltammetric procedures were applied for the trace determination of the bulk drug in pharmaceutical formulations and in human urine. The optimal experimental parameters were as follows: accumulation potential = −0.1 V (vs. Ag/AgCl, 3 M KCl), accumulation time = 50 s, frequency = 140 Hz, pulse amplitude = 0.07 V, and scan increment = 10 mV in phosphate buffer (pH 2.6). The first peak current showed a linear dependence with the drug concentration over the range of 50 ng ml−1 to 25.6 μg ml−1. The achieved limit of detection and limit of quantitation were 3.99 and 13.3 ng ml−1 by SWCAdSV and 7.98 and 26.6 ng ml−1 by DPCAdSV, respectively. The procedure was applied to assay the drug in tablets. Applicability was also tested in urine samples. Peak current was linear with the drug concentration in the range of 1 to 60 μg ml−1 of the urine, and minimum detectability was found to be 12.6 ng ml−1 by SWCAdSV and 58.4 ng ml−1 by DPCAdSV.  相似文献   

10.
Immortal cell lines have not yet been reported from Penaeus monodon, which delimits the prospects of investigating the associated viral pathogens especially white spot syndrome virus (WSSV). In this context, a method of developing primary hemocyte culture from this crustacean has been standardized by employing modified double strength Leibovitz-15 (L-15) growth medium supplemented with 2% glucose, MEM vitamins (1×), tryptose phosphate broth (2.95 g l−1), 20% FBS, N-phenylthiourea (0.2 mM), 0.06 μg ml−1 chloramphenicol, 100 μg ml−1 streptomycin and 100 IU ml−1 penicillin and hemolymph drawn from shrimp grown under a bio-secured recirculating aquaculture system (RAS). In this medium the hemocytes remained viable up to 8 days. 5-Bromo-2′-deoxyuridine (BrdU) labeling assay revealed its incorporation in 22 ± 7% of cells at 24 h. Susceptibility of the cells to WSSV was confirmed by immunofluoresence assay using a monoclonal antibody against 28 kDa envelope protein of WSSV. A convenient method for determining virus titer as MTT50/ml was standardized employing the primary hemocyte culture. Expression of viral genes and cellular immune genes were also investigated. The cell culture could be demonstrated for determining toxicity of a management chemical (benzalkonium chloride) by determining its IC50. The primary hemocyte culture could serve as a model for WSSV titration and viral and cellular immune related gene expression and also for investigations on cytotoxicity of aquaculture drugs and chemicals.  相似文献   

11.
3-Phenyllactic acid (PLA), which is produced by some strains of lactic acid bacteria (LAB), is a known antimicrobial agent with a broad spectrum. Batch and fed-batch fermentation by the strain Lactobacillus sp. SK007 for PLA production have been reported. With batch fermentation without pH-control, PLA production yield was 2.42 g L−1. When fed-batch fermentation by Lactobacillus sp. SK007 was conducted in 3 L initial volume with pH-control at 6.0 and intermittent feeding, which was developed after fermentation for 12 h and every 2 h with 120 mL 100 g L−1 PPA phenylpyruvic acid (PPA) and 50 mL 500 g L−1 glucose each time, PLA production yield reached 17.38 g L−1. The final conversion ratio of PPA to PLA was 51.1%, and the PLA production rate was 0.241 g L−1 h−1. This indicated that PPA was the ideal substrate for PLA fermentation production, and fed-batch fermentation with intermittent PPA feeding and pH-control was an effective approach to improve PLA production yield.  相似文献   

12.
Copolymer particles for removal of endotoxins (lipopolysaccharides, LPSs) were prepared by suspension copolymerization of γ-cyclodextrin (CyD) and 1,6-hexamethylenediisocyanate. The LPS-removing activity of the copolymer particles was compared with that of poly(ε-lysine)-immobilized Cellufine (cationic adsorbent) or polystyrene particles (hydrophobic adsorbent) by a batch method. When DNA was present in solution with LPSs under physiological conditions (pH 6.0, ionic strength of μ = 0.05–0.8), LPS-removing activity of the cationic or hydrophobic adsorbent was unsatisfactory because both the DNA and the LPSs were adsorbed onto each adsorbent. By contrast, the copolymer particles with γ-CyD cavity (CyD content: 14–20 mol%) could selectively remove LPSs from a DNA solution (50 μg ml−1, pH 6.0, and μ = 0.05–0.2) containing LPSs (15 EU ml−1) without the adsorption of DNA. The residual concentration of LPSs in the treated DNA solution was below 0.1 EU ml−1, and the recovery of DNA was 99%.  相似文献   

13.
Prostate-specific antigen (PSA), as the specificity of prostate cancer markers, has been widely used in prostate cancer diagnosis and screening. In this study, we fabricated an electrochemical immunosensor for PSA detection using the amino-functionalized graphene sheet–ferrocenecarboxaldehyde composite materials (NH2-GS@FCA) and silver hybridized mesoporous silica nanoparticles (Ag@NH2-MCM48). Under optimal conditions, the fabricated immunosensor showed a wide linear range with PSA concentration (0.01–10.0 ng·ml−1). Low detection limit (2 pg·ml−1) proved the high sensitivity. In addition, the immunosensor possessed good stability and reproducibility. Moreover, the application to PSA analysis in serum samples yielded satisfactory results.  相似文献   

14.
Batch fermentative production of 2,3-butanediol by Klebsiella oxytoca was investigated using various oxygen supply methods though varying agitation speed. Based on the analysis of three kinetic parameters including specific cell growth rate (μ), specific glucose consumption rate (qs) and specific 2,3-butanediol formation rate (qp), a two-stage agitation speed control strategy, aimed at achieving high concentration, high yield and high productivity of 2,3-butanediol, was proposed. At the first 15 h, agitation speed was controlled at 300 rpm to obtain high μ for cell growth, subsequently agitation speed was controlled at 200 rpm to maintain high qp for high 2,3-butanediol accumulation. Finally, the maximum concentration of 2,3-butanediol reached 95.5 g l−1 with the yield of 0.478 g g−1 and the productivity of 1.71 g l−1 h−1, which were 6.23%, 6.22% and 22.14% over the best results controlled by constant agitation speeds.  相似文献   

15.
An electrochemical immunosensor for quantitative detection of α-fetoprotein (AFP) in human serum was developed using graphene sheets (GS) and thionine (TH) as electrode materials and mesoporous silica nanoparticles (MSNs) loaded with ferroferric oxide (Fe3O4) nanoparticles and horseradish peroxidase (HRP) as labels for signal amplification. In this study, the compound of GS and TH (GS–TH) was used as a substrate for promoting electron transfer and immobilization of primary antibody of AFP (Ab1). MSNs were used as a carrier for immobilization of secondary antibody of AFP (Ab2), Fe3O4, and HRP. The synergistic effect occurred between Fe3O4 and HRP and greatly improved the sensitivity of the immunosensor. This method could detect AFP over a wide concentration range from 0.01 to 25 ng ml−1 with a detection limit of 4 pg ml−1. This strategy may find wide potential application in clinical analysis or detection of other tumor markers.  相似文献   

16.
A novel immunosensor for α-1-fetoprotein based on dialdehyde cellulose/ionic liquid composite film as a matrix has been developed. Microcrystalline cellulose was activated by sodium metaperiodate to produce dialdehyde cellulose. Antibodies can be immobilized on the electrode by a one-step method through covalent bonding of the aldehyde groups of dialdehyde cellulose with the amino groups of antibodies, in which no additional chemical cross-linking step is required. Moreover, ionic liquid added can improve the conductivity of the sensing interface and, therefore, can enhance the electrochemical signal. In this work, α-1-fetoprotein was detected within the range from 0.1 to 60 ng ml−1 with a detection limit of 0.07 ng ml−1 (signal/noise = 3). The proposed immunosensor had good specificity and reproducibility. It was used to determine real samples with satisfactory results.  相似文献   

17.
Short antimicrobial peptides with nine and eleven residues were developed against several clinically important bacterial and fungal pathogens (specifically Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and Fusarium solani). Twelve analogues of previously reported peptides BP76 (KKLFKKILKFL) and Pac-525 (KWRRWVRWI) were designed, synthesized, and tested for their antimicrobial activities. Two of our eleven amino acid peptides, P11-5 (GKLFKKILKIL) and P11-6 (KKLIKKILKIL), have very low MICs of 3.1-12.5 μg ml−1 against all five pathogens. The MICs of these two peptides against S. aureus, C. albicans and F. solani are four to ten times lower than the corresponding MICs of the reference peptide BP76. P9-4 (KWRRWIRWL), our newly designed nine-amino acid analogue, also has particularly low MICs of 3.1-6.2 μg ml−1 against four of the tested pathogens; these MICs are two to eight times lower than those reported for Pac-525 (6.2-50 μg ml−1).These new peptides (P11-5, P11-6 and P9-4) also exhibit improved stability in the presence of salts, and have low cytotoxicity as shown by the hemolysis and MTT assays. From the results of field-emission scanning electron microscopy, membrane depolarization and dye-leakage assays, we propose that these peptides exert their action by disrupting membrane lipids. Molecular dynamics simulation studies confirm that P11-6 peptide maintains relatively stable helical structure and exerts more perturbation action on the order of acyl tail of lipid bilayer.  相似文献   

18.
Glycerol was utilized by Cupriavidus necator DSM 545 for production of poly-3-hydroxybutyrate (PHB) in fed-batch fermentation. Maximal specific growth rates (0.12 and 0.3 h−1) and maximal specific non-growth PHB production rate (0.16 g g−1 h−1) were determined from two experiments (inocula from exponential and stationary phase). Saturation constants for nitrogen (0.107 and 0.016 g L−1), glycerol (0.05 g L−1), non-growth related PHB synthesis (0.011 g L−1) and nitrogen/PHB related inhibition constant (0.405 g L−1), were estimated. Five relations for specific growth rate were tested using mathematical models. In silico performed optimization procedures (varied glycerol/nitrogen ratio and feeding) has resulted in a PHB content of 70.9%, shorter cultivation time (23 h) and better PHB yield (0.347 g g−1). Initial concentration of biomass 16.8 g L−1 and glycerol concentration in broth between 3 and 5 g L−1 were decisive factors for increasing of productivity.  相似文献   

19.
Lu H  Conneely G  Pravda M  Guilbault GG 《Steroids》2006,71(9):760-767
Electrochemical based immunosensors for the detection of boldenone and methylboldenone in bovine urine were described in this paper. The immunosensors were fabricated by immobilizing boldenone-bovine serum albumin conjugate on the surface of screen-printed electrodes (SPEs), and followed by the competition between the free analyte and coating conjugate with corresponding antibodies. The use of anti-species IgG-horseradish peroxidase conjugate determined the degree of competition. The electrochemical technique chosen was chronoamperometry, performed at a potential of +100 mV whereby the product of the catalysis of 3,3′,5,5′-tetramethylbenzidine undergoes reduction produced by the enzyme label. The limits of detection of assay were 30.9 ± 4.3 pg ml−1 for boldenone and 120.2 ± 8.2 pg ml−1 for methylboldenone, respectively. Results of repeated analysis of each androgen carried out using three different batches of electrodes indicate suitable repeatability (EC50 = 1.0 ± 0.3 ng ml−1 (n = 3, N = 3), R2 = 0.969, R.S.D. = 9.6% for boldenone and 1.5 ± 0.3 ng ml−1, 0.971, 10.5% for methylboldenone, respectively). Urine samples were determined directly after a single dilution step, omitting extraction and hydrolysis. This method offers the advantage to pick up both boldenone and its major metabolites in an efficient manner due to the high cross-reactivity pattern of α-boldenone with this antibody. The concentration of methylboldenone in urine detected by developed methods does indicate methylboldenone administration to heifers. Gas chromatography coupled to mass spectrometry analysis was performed to quantitate the individual metabolites present in urine samples, and results were validated with both ELISA and immunosensor data.  相似文献   

20.
Sulfated polysaccharides potently inhibit the infectivity of herpes simplex virus (HSV) in cultured cells. In this study, we have analyzed sulfated xylogalactofucan and alginic acid containing fractions generated from Laminaria angustata, a marine alga. The xylogalactofucan that has apparent molecular mass of 56 ± 5 kDa and unusually low sulfate content contains, inter alia, 1,3-, 1,4- and 1,2-linked fucopyranosyl residues. The algin (molecular mass: 32 ± 5 kDa) contains gulo- (55.5%) and mannuronic (44.5%) acid residues. Introduction of sulfate groups enhanced the macromolecules capability to inhibit the infection of cells by HSV-1. The 50% inhibitory concentration (IC50) values of these macromolecules against HSV-1 were in the range of 0.2-25 μg ml−1 and they lacked cytotoxicity at concentrations up to 1000 μg ml−1. The sulfate content appeared to be an important hallmark of anti-HSV-1 activity. Our results suggest the feasibility of inhibiting HSV attachment to cells by direct interaction of polysaccharides with viral particles.  相似文献   

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