Clavulanic acid production processes in a tower bioreactor with immobilised cells

The feasibility of using Streptomyces clavuligerus ATCC 27064 bioparticles supported on alginate gel containing alumina to produce clavulanic acid (CA) was investigated. To this end, effectiveness factors for spherical bioparticles, relating respiration rates of immobilised and free cells, were experimentally determined for various dissolved oxygen (DO) levels and bioparticle radii. Monod kinetics was assumed as representative of the oxygen consuming reaction, while internal oxygen diffusion was considered the limiting step. A comparison was made of the results from a tower bioreactor operating under batch, repeated-batch and continuous conditions with immobilised bioparticles. The theoretical curve of the effectiveness factor for the zero-order reaction model, considering an inert nucleus - the dead core model - was very well fitted to the experimental data. The results of the bioprocess indicated that the batch operation was the most efficient and productive, requiring a do concentration in the reactor above 60% of the saturation value. (C) 2007 Elsevier B.V. All rights reserved.

Optimisation of the glycerol-to-ornithine molar ratio in the feed medium for the continuous production of clavulanic acid by Streptomyces clavuligerus

Amino acids are well metabolized by Streptomyces clavuligerus during the production of clavulanic acid using glycerol as main carbon and energy source. However, only a few amino acids such as arginine and ornithine are favorable for CA biosynthesis. The aim of this work was to optimize the glycerol:ornithine molar ratio in the feed medium containing only these compounds to maximize CA production in continuous cultivation. A minimum number of experiments were performed by means of a simple two-level full-factorial central composite design to investigate the combined effect of glycerol and ornithine feeding on the CA concentration during the intermittent and continuous process in shake-flasks. Statistical analysis of the experimental data using the response surface methodology showed that a glycerol-to-ornithine molar ratio of approximately 40:1 in the feed medium resulted in the highest CA concentration when fermentation was stopped. Under these optimized conditions, in bench-scale fermentor runs, the CA concentration reached more than double the concentration obtained in shake-flasks runs. (C) 2009 Elsevier B.V. All rights reserved.

Chemical composition of Rhodocyclus gelatinosus biomass produced in poultry slaughterhouse wastewater

Rhodocyclus gelatinosus R1 foi cultivado fotoautotroficamente em águas residuais de abatedouro de aves dentro de colunas de vidro (90x670 mm) durante sete dias a 31 ± 4ºC, sob anaerobiose e intensidade luminosa fornecida pela luz do dia e por três lâmpadas incandescentes (100W) mais quatro lâmpadas fluorescentes (40W). O cultivo obtido foi centrifugado (4.500xg/20 min) e liofilizado, originando a biomassa bacteriana, que continha 7,1% de umidade. A determinação da composição centesimal indicou (base seca) 67,6% de proteína bruta, 27,6% de carboidratos totais, 0,6% de lipídeos e 4,2% de cinzas. A composição em aminoácidos da fração protéica da biomassa mostrou-se semelhante à relatada na literatura para Rhodocyclus gelatinosus e para outras bactérias fotossintetizantes. A redução na DQO do efluente após o cultivo e a retirada da biomassa foi da ordem de 90%. A valiosa composição química da biomassa de Rhodocyclus gelatinosus R1 e o alto conteúdo em aminoácidos essenciais indicam o uso potencial desse produto na suplementação de ração para aves.

Effects of organic and inorganic additives on flotation recovery of washed cells of Saccharomyces cerevisiae resuspended in water

Separation of microbial cells by flotation recovery is usually carried out in industrial reactors or wastewater treatment systems, which contain a complex mixture of microbial nutrients and excretion products. In the present study, the separation of yeast cells by flotation recovery was carried out using a simple flotation recovery systems containing washed yeast cells resuspended in water in order to elucidate the effects of additives (defined amounts of organic and inorganic acids, ethanol, surfactants and sodium chloride) on the cellular interactions at interfaces (cell/aqueous phase and cell/air bubble). When sodium chloride, organic acids (notably propionic, succinic and acetic acids) and organic surfactants (sodium dodecyl sulphate (SDS), cetyltrimethylammonium bromide (CTAB) and Nonidet P40) were added to the flotation recovery system, significant increases in the cell recovery of yeast hydrophobic cells (Saccharomyces cerevisiae, strain FLT-01) were observed. The association of ethanol to acetic acid solution (a minor by-product of alcoholic fermentation) in the flotation recovery system, containing washed cells of strain FLT-01 resuspended in water, leading to an increased flotation recovery at pH 5.5. Thus, the association among products of the cellular metabolism (e.g., ethanol and acetic acid) can improve yeast cell recovery by flotation recovery. (c) 2006 Elsevier B.V. All rights reserved.

Determination of caffeic acid in red wine by voltammetric method

The electrochemical oxidation of 3,4-dihydroxycinnamic acid, caffeic acid, leads to a stable electroactive poly(caffeic acid) thin film containing quinone moiety on a preactivated glassy polymeric carbon electrode. The properties of the deposited films as well as the stability study under different experimental conditions were investigated. Taking advantage of the electrochemical behavior, an analytical method based on differential pulse voltammetry for determination of caffeic acid in red wine was proposed.

Amperometric biosensor for ascorbic acid

Desenvolveu-se um biossensor para ácido L-ascórbico empregando ascorbato oxidase. A enzima foi extraída do mesocarpo de pepino com solução tampão fosfato 0,05 mol L-1, pH 5,8 contendo NaCl 0,5 mol L-1. Após diálise versus solução tampão fosfato 0,05 mol L-1, pH 5,8 a enzima foi imobilizada em rede de nylon através de ligação covalente com glutaraldeído. A membrana foi acoplada em eletrodo de O2 e a reação monitorada pelo consumo de oxigênio a -600 mV em análise em fluxo (solução tampão fosfato 0,05 mol L-1, pH 5,8 como carregador e vazão 0,5 mL min-1). A curva analítica apresentou-se linear entre 1,2x10-4 a 1,0x10-3 mol L-1. O tempo de vida do biossensor foi de 500 análises. Amostras de medicamentos foram analisadas com a metodologia proposta e os resultados comparados com os obtidos com HPLC.

Preparation and bactericidal effect of composites based on crosslinked copolymers containing silver nanoparticles

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Evaluation of the addition of ascorbic acid to the ration of cultivated Piaractus mesopotamicus (Characidae) on the infrapopulation of Anacanthorus penilabiatus (Monogenea)

Sixty Piaractus mesopotamicus Holmberg, 1887 (pacu) fry fed a diet containing 0, 50, 100 and 200 mg ascorbic acid/kg dry feed were studied to evaluate the effect on parasitic infestation by the monogenean Anacanthorus penilabiatus Boeger, Husak and Martins, 1995 (Monogenea: Dactylogyridae) for a period of 24 weeks. The temperature of the aquaria was measured daily and remained between 28 and 31oC. At the beginning of the experiment, fish showed 6.15 ± 0.33 cm standard length and 8.64 ± 1.62 g average body weight. A sample of fish was examined and showed 43 ± 17 monogeneans per fish. At the end of the experiment, the gills of control and vitamin C-treated fish were collected for parasite counts. Control fish had 42.5 parasites per fish, a significantly higher number (P<0.05) when compared with fish fed vitamin C, that showed 16.5 parasites per fish. Ascorbic acid fortification in the food promoted an increase in fish resistance to parasites. It is suggested that an optimum level of 139 mg/kg vitamin C supplementation either elicited better nutritional conditions by stimulating the appetite of the fish or improved the immune response.

Effect of modifiers on thermal behaviour of Se in acid digestates and slurries of vegetables by graphite furnace atomic absorption spectrometry

The influence of sample preparation strategy of vegetables on the electrothermal behaviour of Se without and with chemical modifiers such as Pd(NO3)(2), Pd(NO3)(2) + Mg(NO3)(2), Pd(NO3)(2) + Cd(NO3)(2), pre-reduced Pd, Mg(NO3)(2), and Ni(NO3)(2) was investigated. Acid digestates and slurries of vegetables (0.1% m/v in 1% m/v HNO3 + 0.005% v/v of Triton X-100) were used to prepare reference solutions or slurries. For 10 mul of each modifier tested, pyrolysis and atomization temperatures were evaluated using pyrolysis and atomization curves, respectively. Best conditions, such as thermal stability, signal profile, repeatability and sensitivity were attained using Pd(NO3)(2) as chemical modifier. The following heating program (temperature, ramp/hold time) of the graphite tube of the Varian SpectrAA-800Z atomic absorption spectrometer was used: dry step (85 degreesC, 5/0 s; 95 degreesC, 40/0 s; 120 degreesC, 10/.5 s); pyrolysis step (1400 degreesC, 10/3s); atomization step (2200 degreesC, 1/2 s); clean step (2600 degreesC, 2/0 s). This pyrolysis temperature is 800 degreesC higher than when measuring without any modifier. For 20 muL sample volume and 10 mug Pd(NO3)(2), analytical curves in the 3.0-30 mug Se 1(-1) range were obtained. The method was applied for Se determination in acid digestates and slurries of 10 vegetable samples and one standard reference material (rice flower) and results were in agreement at 95% confidence level. Recoveries varied from 89 to 95% for spiked samples. The lifetime of the graphite tube was ca. 250 firings and the relative standard deviations (n = 12) for a typical acid digestate and slurry containing 20 mug Se 1(-1) were 3.8% and 8.3%, respectively. The limits of detection were 2.0 mug Se 1(-1) and 0.6 mug Se 1(-1) Se for digestates and slurries, respectively. (C) 2002 Elsevier B.V. Ltd. All rights reserved.

Oxidation of H2S in acid solution by Thiobacillus ferrooxidans and Thiobacillus thiooxidans

Qualitative and quantitative oxidation tests of H2S in acid solution were carried out using Thiobacillus ferrooxidans and Thiobacillus thiooxidans species, Experiments were performed using solutions of H2SO4 (pH 2.0) containing H2S in initial concentrations ranging from 5 to 100 ppm. in shake flasks at 150 rpm and 30(circle)C. In these solution, this gas was not very stable and was quickly liberated. However, at low concentration (less than 5 ppm) it becomes stable and could only be removed from solution by oxidation. The results obtained indicated that the presence of either T. ferrooxidans or T. thiooxidans causes a significant reduction in H,S concentration (more than 99%) in relation to the sterile control, No differences in oxidation efficiency between these two species were detected. (C) 2001 Elsevier B.V. Ltd. All rights reserved.

Effect of iron on inhibition of acid demineralisation of bovine dental enamel in vitro

Objectives: Iron ions (Fe2+) have been shown to be cariostatic in many studies particularly by their ability to reduce bacterial metabolism. Nevertheless, the role of iron ions on dissolution of enamel is unexplored. The aim of the present study was therefore to investigate the protective effect of increasing concentrations (0-120 mmol/L) of Fe2+ on the dissolution of enamel.Design: Enamel powder was subjected to acetic acid made with increasing concentrations with respect to FeSO4 center dot 7H(2)O. In order to determine the amount of enamel dissolved, the phosphate released in the medium was analysed spectrophotometrically using the Fiske-Subarrow method. Data were tested using Kruskall-Wall and Dunn's tests (p < 0.05). The degree of protection was found to approach maximum at about 15 mmol/L Fe2+. Higher concentrations of Fe2+ did not have an extra effect on inhibition of dissolution of enamel powder. In the next step, the protective effect of 15 mmol/L Fe2+ against mineral dissolution of the bovine enamel was evaluated using a simple abiotic model system. Enamel blocks were exposed to a sequence of seven plastic vials, each containing 1 mL of 10 mmol/L acetic acid. The acid in vial 4 was made 15 mmol/L with respect to FeSO4 center dot 7H(2)O. The mineral dissolved during each challenge was thus determined by phosphate released as described above. Data were tested using two-way ANOVA (p < 0.05). Results: Lower demineralisation (around 45%) was found in vial 4 (with Fe) that continued stable until vial 7.Conclusions: Thus, our data suggest that Fe2+, can be effective on inhibition of dissolution of enamel and that this effect may be durable. (c) 2006 Elsevier Ltd. All rights reserved.

EPR investigation of the influence of side chain protecting groups on peptide-resin solvation of the Asx and Glx model containing peptides

In spite of all progressive efforts aiming to optimize SPPS, serious problems mainly affecting the assembly of aggregating sequences have persisted. Following the study intended to unravel the complex solvation phenomenon of peptide-resin beads, the XING and XAAAA model aggregating segments were labeled with a paramagnetic probe and studied via EPR spectroscopy. Low and high substituted resins were also comparatively used, with the X residue being Asx or Glx containing the main protecting groups used in the SPPS. Notably, the cyclo-hexyl group used for Asp and Glu residues in Boc-chemistry induced greater chain immobilization than its tert-butyl partner-protecting group of the Fmoc strategy. Otherwise, the most impressive peptide chain immobilization occurred when the large trytil group was used for Asn and Gln protection in Fmoc-chemistry. These surprising results thus seem to stress the possibility of the relevant influence of the amino-acid side chain protecting groups in the overall peptide synthesis yield. (C) 2007 Elsevier Ltd. All rights reserved.

EFFECT OF ASCORBIC-ACID DEFICIENCY ON THE GROWTH, GILL FILAMENT LESIONS AND BEHAVIOR OF PACU FRY (PIARACTUS-MESOPOTAMICUS HOLMBERG, 1887)

The effect of ascorbic acid deficiency was determined in Piaractus mesopotamicus Holmberg, 1887, fish (pacu) under laboratory conditions. A total of 120 fish with an average body weight of 8.64 +/- 1.62 g and measuring 6.15 +/- 0.33 cm in length at the beginning of the experiment were fed diets containing 0, 50, 100 or 200 mg palmitate-coated ascorbic acid/kg dry ration for a period of 24 weeks with measurements every 4 weeks. The experiment was conducted in 20 fiber-cement aquaria of 81-liter capacity. Each aquarium was supplied with dechlorinated water at a flow rate of 1 l/min. Water temperature was measured daily and pH, dissolved oxygen, alkalinity and water conductivity were determined weekly. A fully randomized experimental design was utilized, with 5 replicates of each treatment and 6 fish per aquarium. Ascorbic acid-supplemented fish presented significantly increased growth when compared to unsupplemented fish. Furthermore, unsupplemented fish presented a higher incidence of hyperplasia, hypertrophy and dysplasia of the bone cartilage of gill filaments. The gill lamellae of unsupplemented fish had twisted cartilage and an inflammatory infiltrate at the ends. Anorexia and increased handling stress were also observed in fish fed the unsupplemented diet. The present study suggests that 50 mg ascorbic acid/kg dry ration is sufficient to improve development of pacu fingerlings but the optimum level under aquarium conditions, determined by regression analysis, is 139 mg ascorbic acid/kg dry ration.

Treatment of 1,10-phenanthroline laboratory wastewater using the solar photo-Fenton process

The red Fe2+-phenanthroline complex is the basis of a classical spectrophotometric method for determination of iron. Due to the toxicity of this complexing agent, direct disposal of the wastewaters generated in analytical laboratories is not environmentally safe. This work evaluates the use of the solar photo-Fenton process for the treatment of laboratory wastewaters containing phenanthroline. Firstly, the degradation of phenanthroline in water was evaluated at two concentration levels (0.1 and 0.01%, w/v) and the efficiencies of degradation using ferrioxalate (FeOx) and ferric nitrate were compared. The 0.01% w/v solution presented much higher mineralization, achieving 82% after 30 min of solar irradiation with both iron sources. The solar photo-Fenton treatment of laboratory wastewater containing, in addition to phenanthroline, other organic compounds such as herbicides and 4-chlorophenol, equivalent to 4500 mg L-1 total organic carbon (TOC) resulted in total degradation of phenanthroline and 25% TOC removal after 150 min, in the presence of either FeOx or ferric nitrate. A ratio of 1: 10 dilution of the residue increased mineralization in the presence of ferrioxalate, achieving 38% TOC removal after 120 min, while use of ferric nitrate resulted in only 6% mineralization over the same period. (c) 2007 Elsevier B.V. All rights reserved.

Synthesis and some properties of hybrid gels of titanium oxide containing europium (III)

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