Modeling and simulation of fructooligosaccharides synthesis in a batch basket reactor.

Fructooligosaccharides (FOS) production was carried out in a batch basket reactor with immobilized fructosyltransferase from Rhodotorula sp. from 500×10(3) g m(-3) of sucrose in 50 mM sodium acetate buffer at pH 6.0, 48 °C at 85 rpm and with an activity of 22.44×10(3) U m(-3). The experimental data were well adjusted to the mathematical model for FOS production using SIMULINK(®) (MATLAB(®)). The highest regression coefficient (R(2)>90%) and the lowest percentual residual standard deviation (%RSD<4.0) and chi-square (χ(2) <1.0) were obtained for sucrose (GF), kestose (GF2) and total FOS. The mass transfer coefficient (kL) was determined as 5.6×10(-5) m h(-1) and the diffusivity (DS) was 2.11×10(-11) m(2) s(-1). The best predicted FOS yield (after 96 h) was 60.62%, with an equivalent productivity of 3.16×10(3) g m(-3) h(-1). These results reaffirm the good potential of this enzyme for industrial application and, in addition, are in conformation to other studies conducted with the same enzyme from the same and different microbial sources.

Enzymatic biodiesel synthesis from yeast oil using immobilized recombinant Rhizopus oryzae lipase.

The recombinant Rhizopus oryzae lipase (1-3 positional selective), immobilized on Relizyme OD403, has been applied to the production of biodiesel using single cell oil from Candida sp. LEB-M3 growing on glycerol from biodiesel process. The composition of microbial oil is quite similar in terms of saponifiable lipids than olive oil, although with a higher amount of saturated fatty acids. The reaction was carried out in a solvent system, and n-hexane showed the best performance in terms of yield and easy recovery. The strategy selected for acyl acceptor addition was a stepwise methanol addition using crude and neutralized single cell oil, olive oil and oleic acid as substrates. A FAMEs yield of 40.6% was obtained with microbial oils lower than olive oil 54.3%. Finally in terms of stability, only a lost about 30% after 6 reutilizations were achieved.

Cultivation of Candida sp. LEB-M3 in glycerol: lipid accumulation and prediction of biodiesel quality parameters.

The quality of biodiesel from lipids produced by the yeast Candida sp. LEB-M3 was predicted, by the use of mathematical models for parameters that specify quality as a function of the fatty acid profile. The lipid production was studied according to the experimental design methodology, for different cultivation conditions for agitation and aeration. Lipid compositions were affected by the cultivation conditions, and the agitation presented a positive effect for the formation of monounsaturated fatty acids and negative effect for saturated fatty acids. Aeration had a positive effect on the formation of polyunsaturated fatty acids. According to the predictions by the mathematical models, the cetane number varied from 61 to 67, the oxidative stability from 11 to 17h, the iodine index from 55 to 75gI2/100g, density from 852 to 868kg/m(3). All cultivation conditions led to lipid compositions, whose predicted bioparameter values indicate that biodiesel from this lipid source should present current standard quality.

Exploration of Brazilian biodiversity and selection of a new oleaginous yeast strain cultivated in raw glycerol.

The objective of this study was to use glycerol generated from the synthesis of biodiesel to study the oleaginous potential of wild yeasts. An initial selection was performed via a rapid and qualitative technique by staining with Sudan Black B. Initially 129 yeasts were present, from which 5 were selected and cultivated in liquid medium containing pure or raw glycerol. The yeast LEB-M3, isolated from the Pantanal, presented lipid content of 20.46% and 56.58% for cultivation in pure and raw glycerol, respectively. This strain was genotypically identified as Candida sp. The fatty acid profile showed predominance of oleic acid (C18:1), 57.35% for cultivation in pure glycerol, and in raw glycerol linoleic acid (C18:2) was predominant (46.0%). It was possible to select a yeast with high lipid concentrations 9.14 g/L and fatty acid profile similar to vegetable oils commonly used in the synthesis of biodiesel.

Modeling of ion exchange expanded-bed chromatography for the purification of C-phycocyanin.

This work is focused on the experimental evaluation and mathematical modeling of ion exchange expanded-bed chromatography for the purification of C-phycocyanin from crude fermentative broth containing Spirulina platensis cells. Experiments were carried out in different expansion degree to evaluate the process performance. The experimental breakthrough curves were used to estimate the mass transfer and kinetics parameters of the proposed model, using the Particle Swarm Optimization algorithm (PSO). The proposed model satisfactorily fitted the experimental data. The results from the model application pointed out that the increase in the initial bed height does not influence the process efficiency, however enables the operation of expanded-bed column at high volumetric flow rates, improving the productivity. It was also shown that the use of mathematical modeling was a good and promising tool for the optimization of chromatographic processes.

Characterization of a commercial cellulase for hydrolysis of agroindustrial substrates.

This work is focused on the characterization of a commercial cellulase in terms of optimum pH and temperature, stability to pH and temperature and affinity of this enzyme to several substrates, determining the Michaelis-Menten parameters. Maximum activity of cellulase was obtained for the temperature range from 40 to 50 °C and pH from 5.2 to 5.5. Enzyme activity decreased only 15% after 150 h of reaction at temperatures between 30 and 50 °C. No loss of activity was observed at pH 5.0 and 5.5. The cellulase showed satisfactory results in the hydrolysis of agroindustrial substrates, since similar activity was verified on filter paper and other agroindustrial substrates.

Mathematical modeling of Kluyveromyces marxianus growth in solid-state fermentation using a packed-bed bioreactor.

This work investigated the growth of Kluyveromyces marxianus NRRL Y-7571 in solid-state fermentation in a medium composed of sugarcane bagasse, molasses, corn steep liquor and soybean meal within a packed-bed bioreactor. Seven experimental runs were carried out to evaluate the effects of flow rate and inlet air temperature on the following microbial rates: cell mass production, total reducing sugar and oxygen consumption, carbon dioxide and ethanol production, metabolic heat and water generation. A mathematical model based on an artificial neural network was developed to predict the above-mentioned microbial rates as a function of the fermentation time, initial total reducing sugar concentration, inlet and outlet air temperatures. The results showed that the microbial rates were temperature dependent for the range 27-50 degrees C. The proposed model efficiently predicted the microbial rates, indicating that the neural network approach could be used to simulate the microbial growth in SSF.

Production of an acidic and thermostable lipase of the mesophilic fungus Penicillium simplicissimum by solid-state fermentation.

The production of a lipase by a wild-type Brazilian strain of Penicillium simplicissimum in solid-state fermentation of babassu cake, an abundant residue of the oil industry, was studied. The enzyme production reached about 90 U/g in 72 h, with a specific activity of 4.5 U/mg of total proteins. The crude lipase showed high activities at 35-60 degrees C and pH 4.0-6.0, with a maximum activity at 50 degrees C and pH 4.0-5.0. Enzyme stability was enhanced at pH 5.0 and 6.0, with a maximum half-life of 5.02 h at 50 degrees C and pH 5.0. Thus, this lipase shows a thermophilic and thermostable behavior, what is not common among lipases from mesophilic filamentous fungi. The crude enzyme catalysed the hydrolysis of triglycerides and p-nitrophenyl esters (C4:0-C18:0), preferably acting on substrates with medium-chain fatty acids. This non-purified lipase in addition to interesting properties showed a reduced production cost making feasible its applicability in many fields.

Use of a sequential strategy of experimental design to optimize the inulinase production in a batch bioreactor.

A potential application of inulinase in the food industry is the production of fructoligosaccharides (FOS) by the transfructosilation of sucrose. The FOSs present many interesting functional properties besides their ability to increase the shelf-life and flavor of many products. The use of an industrial medium represents a good alternative to producing inulinase at low cost, since the activity may improve, or at least remain the same, as that obtained using a synthetic medium. This work was an optimization study of the inulinase production by Kluyveromyces marxianus NRRL Y-7571 using industrial pre-treated culture medium in a bioreactor employing a sequential strategy of experimental design. Initially, a Plackett-Burman (Screening Design) design was used, where the studied variables were molasses, corn steep liquor, yeast extract concentration, and agitation and aeration rates. After the analysis of the effects, a central composite rotational design (CCRD) was carried out. The optimized condition for the inulinase production was: 250 g/l of molasses, 80 g/l of corn steep liquor, 6 g/l of yeast extract, 300 rpm of agitation and 1.5 vvm aeration rate, which resulted in an enzymatic activity of 1,317 +/- 65 U/ml.

Purification and characterisation of a fructosyltransferase from Rhodotorula sp.

The present work was devoted to investigations concerning the purification and characterisation of the fructooligosaccharide (FOS)-producing extracellular enzyme of Rhodotorula sp. LEB-V10. FOS are functional food ingredients showing prebiotic properties, meaning that it could stimulate selectively the growth and/or activity of probiotic bacteria in the gut. The purification of the enzyme was carried out according to the following sequential procedure: cell separation by centrifugation, recovering by ethanol precipitation and purification by anion exchange chromatography. The molecular weight was estimated to be 170 kDa by preparative gel filtration and 77 kDa by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, signifying that the native enzyme exists as a dimer. With sucrose as substrate, the data failed to fit the Michaelis-Menten behaviour, rather showing a sigmoid shape similar to that of the allosteric enzymes (cooperative behaviour), requiring high sucrose concentrations to obtain high reaction rates. The enzyme showed both fructofuranosidase (FA) and fructosyl-transferase (FTA) activities. The optimum pH and temperature for FA activity were found to be around 4.0 and 72-75 degrees C, respectively, while FTA showed optimum activity at pH 4.5 and 65-70 degrees C. Both activities were very stable at temperatures below 66 degrees C, while for FA, the enzyme was more stable at pH 4.0 and for FTA at pH 5.0.