Continuous ethanol production in a nonconventional five-stage system operating with yeast cell recycling at elevated temperatures

Three ranges of increasing temperatures (35-43, 37-45, 39-47degreesC) were sequentially applied to a five-stage system continuously operated with cell recycling so that differences of 2degreesC (between one reactor to the next) and 8degreesC (between the first reactor at the highest temperature and the fifth at the lowest temperature) were kept among the reactors for each temperature range. The entire system was fed through the first reactor. The lowest values of biomass and viability were obtained for reactor R-3 located in the middle of the system. The highest yield of biomass was obtained in the effluent when the system was operated at 35-43degreesC. This nonconventional system was set up to simulate the local fluctuations in temperature and nutrient concentrations that occur in different regions of the medium in an industrial bioreactor for fuel ethanol production mainly in tropical climates. Minimized cell death and continuous sugar utilization were observed at temperatures normally considered too high for Saccharomyces cerevisiae fermentations.

A continuously differentiable upwinding scheme for the simulation of fluid flow problems

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Continuously deformable topological structure

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Influence of stationary and bioreactor cultivation on Lentinula edodes (Berk) pegler lignocellulolitic activity

This work aimed to study the stationary and periodically mixed culture of L. edodes to the production of lignocellulolitic enzymes activity. LE 95/17, LE 96/22 and Leax strains were incubated in 25 g of eucalyptus sawdust substrate in Erlenmeyer flasks in stationary culture at 25 degrees C and in a bioreactor with four complete rotations daily at 25 degrees C and 3% CO2. The samples were collected at 8, 11, 14, 17 and 20 days after the incubation. Oxidative and hydrolytic enzymes analyses were performed. Lignin peroxidase enzyme was not found in the lignolytic systernfor LE 95/17, LE 96/22 and Leax strains in the different incubation methods. The use of bioreactor could be a practicable system to induce the laccase activity for L22 and Leax and MnP activity for L17 and L22. The activity of the hydrolytic enzymes was higher in the stationary system in comparison to periodically mixed system in the bioreactor.

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.

Genetic and physiological alterations occurring in a yeast population continuously propagated at increasing temperatures with cell recycling

This work investigated the effects of increasing temperature from 30 degrees C to 47 degrees C on the physiological and genetic characteristics of Saccharomyces cerevisiae strain 63M after continuous fermentation with cell recycling in a system of five reactors in series. Steady state was attained at 30 degrees C, and then the temperature of the system was raised so it ranged from 35 degrees C in the last reactor to 43 degrees C in the first reactor or feeding reactor with a 2 degrees C difference between reactors. After 15 days at steady state, the temperature was raised from 37 degrees C to 45 degrees C for 25 days at steady state, then from 39 degrees C to 47 degrees C for 20 days at steady state. Starter strain 63M was a hybrid strain constructed to have a MAT a/alpha, LYS/lys, URA/ura genotype. This hybrid yeast showed vigorous growth on plates at 40 degrees C, weak growth at 41 degrees C, positive assimilation of melibiose, positive fermentation of galactose, raffinose and sucrose. of 156 isolates obtained from this system at the end of the fermentation process, only 17.3% showed the same characteristics as starter strain 63M. Alterations in mating type reaction and in utilization of raffinose, melibiose, and sucrose were identified. Only 1.9% of the isolates lost the ability to grow at 40 degrees C. Isolates showing requirements for lysine and uracil were also obtained. In addition, cell survival was observed at 39-47 degrees C, but no isolates showing growth above 41 degrees C were obtained.

Modelling and optimization of the cephalosporin C production bioprocess in a fed-batch bioreactor with invert sugar as substrate

Cephalosporin C production process optimization was studied based on four experiments carried out in an agitated and aerated tank fermenter operated as a fed-batch reactor. The microorganism Cephalosporium acremonium ATCC 48272 (C-10) was cultivated in a synthetic medium containing glucose as major carbon and energy source. The additional medium contained a hydrolyzed sucrose solution as the main carbon and energy source and it was added after the glucose depletion. By manipulating the supplementary feed rate, it was possible to increase antibiotic production. A mathematical model to represent the fed-batch production process was developed. It was observed that the model was applicable under different operation conditions, showing that optimization studies can be made based on this model. (C) 1999 Elsevier B.V. Ltd. All rights reserved.

Cephalosporin C production by immobilized Cephalosporium acremonium cells in a repeated batch tower bioreactor

The industrial production of antibiotics with filamentous fungi is usually carried out in conventional aerated and agitated tank fermentors. Highly viscous non-Newtonian broths are produced and a compromise must be found between convenient shear stress and adequate oxygen transfer. In this work, cephalosporin C production by bioparticles of immobilized cells of Cephalosporium acremonium ATCC 48272 was studied in a repeated batch tower bioreactor as an alternative to the conventional process. Also, gas-liquid oxygen transfer volumetric coefficients, k(L)a, were determined at various air flow-rates and alumina contents in the bioparticle. The bioparticles were composed of calcium alginate (2.0% w/w), alumina (<44 micra), cells, and water. A model describing the cell growth, cephalosporin C production, oxygen, glucose, and sucrose consumption was proposed. To describe the radial variation of oxygen concentration within the pellet, the reaction-diffusion model forecasting a dead core bioparticle was adopted. The k(L)a measurements with gel beads prepared with 0.0, 1.0, 1.5, and 2.0% alumina showed that a higher k(L)a value is attained with 1.5 and 2.0%. An expression relating this coefficient to particle density, liquid density, and air velocity was obtained and further utilized in the simulation of the proposed model. Batch, followed by repeated batch experiments, were accomplished by draining the spent medium, washing with saline solution, and pouring fresh medium into the bioreactor. Results showed that glucose is consumed very quickly, within 24 h, followed by sucrose consumption and cephalosporin C production. Higher productivities were attained during the second batch, as cell concentration was already high, resulting in rapid glucose consumption and an early derepression of cephalosporin C synthesizing enzymes. The model incorporated this improvement predicting higher cephalosporin C productivity. (C) 2004 Wiley Periodicals, Inc.

Immobilized cells of Acidithiobacillus ferrooxidans in PVC strands and sultite removal in a pilot-scale bioreactor

The biooxidation of ferrous ion into ferric ion by Acidithiobacillus ferrooxidans can be potentially used for the removal of H2S from industrial gases. In this work, Fe3+ ions were obtained through the oxidation of Fe2+ using the LR strain of At. ferrooxidans immobilized in PVC stands in a pilot-scale bioreactor, while H2S was removed in an absorption tower equipped with Rasching rings. At. ferrooxidans LR strain cells were immobilized by inoculating the bacterium in a Fe2+-mineral medium and percolating it through the support. After complete Fe2+ oxidation, which took around 90 h, the reactor was washed several times with sulfuric acid (pH 1.7) before a new cycle was started. Four additional cycles using fresh Fe2+ mineral medium were then run. During these colonization cycles, the time required for complete iron oxidation decreased, dropping to about 60 h in the last cycle. The batch experiments in the H2S gas removal trials resulted in a gas removal rate of about 98-99% under the operational conditions employed. In the continuous experiments with the bioreactor coupled to the gas absorption column, a gas removal efficiency of almost 100% was reached after 500 min. Precipitate containing mainly sulfur formed during the experimental trial was identified by EDX. (c) 2005 Elsevier B.V. All rights reserved.

Studies on the respiration rate of free and immobilized cells of Cephalosporium acremonium in cephalosporin C production

Bioprocesses using filamentous fungi immobilized in inert supports present many advantages when compared to conventional free cell processes. However, assessment of the real advantages of the unconventional process demands a rigorous study of the limitations to diffusional mass transfer of the reagents, especially concerning oxygen. In this work, a comparative study was carried out on the cephalosporin C production process in defined medium containing glucose and sucrose as main carbon and energy sources, by free and immobilized cells of Cephalosporium acremonium ATCC 48272 in calcium alginate gel beads containing alumina. The effective diffusivity of oxygen through the gel beads and the effectiveness factors related to the respiration rate of the microorganism were determined experimentally. By applying Monod kinetics, the respiration kinetics parameters were experimentally determined in independent experiments in a complete production medium. The effectiveness factor experimental values presented good agreement with the theoretical values of the approximated zero-order effectiveness factor, considering the dead core model. Furthermore, experimental results obtained with immobilized cells in a 1.7-L tower bioreactor were compared with those obtained in 5-L conventional fermenter with free cells. It could be concluded that it is possible to attain rather high production rates working with relatively large diameter gel beads (ca. 2.5 mm) and sucrose consumption-based productivity was remarkably higher with immobilized cells, i.e., 0.33 gCPC/kg sucrose/h against 0.24 gCPC/kg sucrose/h in the aerated stirred tank bioreactor process. (C) 1999 John Wiley & Sons, Inc.

Use of sorghum seed tissue as a biocatalyst in a stirred reactor for oxalic acid determination

The enzyme oxalate oxidase, E.C. 1.2.3.4 from Sorghum vulgare seeds (variety BR303) was used to develop a new sensor for oxalate determination without any purification. The sorghum seeds were conditioned in a 0.10 mol I-1 KCl solution. Then, these seeds were put in a stirring bar type enzymic reactor and coupled with an electrode for CO2. This device was introduced into a cell containing 10.0 ml of a 0.10 mol I-1 KCl solution saturated with oxygen. This sensor showed a linear response between 1.0 and 4.0 × 10-3 mol I-1 with a slope of 30 mV per decade of oxalate concentration at 25.0°C. The sensor was stable for one month or 200 determinations. The response time was about 60 s. The Michaelis-Menten constant determined for this enzyme was 1.5 × 10-3 mol I-1.

Immobilization of lipases and assay in continuous fixed bed reactor

Lipases are versatile enzymes regarding the range of reactions they catalyse and substrates on which they act. They are as well important as catalyst in organic synthesis. Their immobilization on appropriate supports confer them greater stability besides the possibility of operating in continuous reactors. In order to explore these abilities, the reactions involving hydrolysis of p-nitrophenyl acetate (PNPA) and transesterification of PNPA with n-butanol were chosen. Lipases from two different sources were assayed, namely: microbial (Candida rugosa, CRL, Sigma Type VII) and pancreatic (PPL, Sigma, Type 11). Two immobilization methods were also used, namely: 1) adsorption, using as support the following silica derivatives (150-300μm e 450μ): phenyl, epoxy, amino and without derivation, and 2) covalent binding, using glutaraldehyde as binding agent and silica amino as support. This later method led to better results. Hydrolytic activity was 6.1 U/gsupport for CRL and 0.97U/gsupport for PPL, and of transesterification, 2,8U/gsupport for CRL and 1,9U/gsupport for PPL. Stability of the immobilized enzyme as a function of temperature was evaluated for CRL at 40°C and 50°C and for PPL at 32°C and 40°C. The assays were initially carried out batchwise, both for soluble and immobilized enzymes, aiming to the obtention of parameters for the continues reactor. Lipases immobilized by covalent binding were used in the assays of operacional stability in continuos reactors. For PPL in aqueous medium, at 32°C, and CRL in organic medium at 40°C, both operating continuously, no significant loss of activity was detected along the analysis period of 17 days. In the case of CRL in aqueous medium at 40°C there was a loss of activity around 40% after 18 days. For PPL in organic medium at 40°C the loss was 33% after 20 days. Compairing both sources with each other, very different results were obtained. Higher activitiy was found for CRL, both for hydrolysis and for transesterification reactions, with higher stability in organic medium. PPL showed lower activity as well as higher stability in aqueous medium. The immobilization method by covalent binding showed to be the most appropriate. Immobilized lipases are therefore relatively stable both in aqueous and organic medium.

Simultaneous effects of total solids content, milk base, heat treatment temperature and sample temperature on the rheological properties of plain stirred yogurt

Response surface methodology was used to establish a relationship between total solids content, milk base, heat treatment temperature, and sample temperature, and consistency index, flow behaviour index, and apparent viscosity of plain stirred yogurts. Statistical treatments resulted in developments of mathematical models. All samples presented shear thinning fluid behaviour. The increase of the content of total solids (9.3-22.7 %) and milk base heat treatment temperature (81.6-98.4°C) resulted in a significant increase in consistency index and a decrease in flow behaviour index. Increase in the sample temperature (1.6-18.4°C) caused a decrease in consistency index and increase in flow behaviour index. Apparent viscosity was directly related to the content of total solids. Rheological properties of yogurt were highly dependent on the content of total solids in milk.

Otimização do cultivo de Lasiodiplodia theobromae para obtenção de EPS e caracterização física e química dos polissacarídeos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The effect of biomass immobilization support material and bed porosity on hydrogen production in an upflow anaerobic packed-bed bioreactor

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)