Lidia Maria Melo Santa Anna

Petróleo Brasileiro S.A., Rio de Janeiro, Rio de Janeiro, Brazil

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Publications (15)23.08 Total impact

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    ABSTRACT: Increasing interest in the production of second-generation ethanol necessitates the low-cost production of enzymes from the cellulolytic complex (endoglucanases, exoglucanases, and β-glucosidases), which act synergistically in cellulose breakdown. The present work aimed to optimise a bioprocess to produce these biocatalysts from the fungus Penicillium funiculosum ATCC11797. A statistical full factorial design (FFD) was employed to determine the optimal conditions for cellulase production. The optimal composition of culture media using Avicel (10 g·L(-1)) as carbon source was determined to include urea (1.2 g·L(-1)), yeast extract (1.0 g·L(-1)), KH2PO4 (6.0 g·L(-1)), and MgSO4 ·7H2O (1.2 g·L(-1)). The growth process was performed in batches in a bioreactor. Using a different FFD strategy, the optimised bioreactor operational conditions of an agitation speed of 220 rpm and aeration rate of 0.6 vvm allowed the obtainment of an enzyme pool with activities of 508 U·L(-1) for FPase, 9,204 U·L(-1) for endoglucanase, and 2,395 U·L(-1) for β-glucosidase. The sequential optimisation strategy was effective and afforded increased cellulase production in the order from 3.6 to 9.5 times higher than production using nonoptimised conditions.
    Enzyme research. 01/2014; 2014:703291.
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    Electronic Journal of Biotechnology 09/2013; 16(5):1-1. · 0.83 Impact Factor
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    ABSTRACT: This study aimed to produce a cellulase blend and to evaluate its application in a simultaneous saccharification and fermentation process (SSF) for second generation ethanol production from sugar cane bagasse. The sugar cane bagasse was subjected to pretreatments (diluted acid and alkaline), as for disorganizing the ligocellulosic complex, and making the cellulose component more amenable to enzymatic hydrolysis. The residual solid fraction was named sugar cane bagasse partially delignified cellulignin (PDC), and was used for enzyme production and ethanol fermentation. The enzyme production was performed in a bioreactor with two inoculum concentrations (5 and 10% v/v). The fermentation inoculated with higher inoculum size reduced the time for maximum enzyme production (from 72 to 48). The enzyme extract was concentrated using tangential ultrafiltration in hollow fiber membranes, and the produced cellulase blend was evaluated for its stability at 37°C, operation temperature of the simultaneous saccharification and fermentation process (SSF), and at 50°C, optimum temperature of cellulase blend activity. The cellulolytic preparation was stable for at least 300h at both 37°C and 50°C. The ethanol production was carried out by PDC fed-batch SSF process, using the onsite cellulase blend. The feeding strategy circumvented the classic problems of diffusion limitations by diminishing the presence of a high solid:liquid ratio at any time, resulting in high ethanol concentration at the end of the process (100g/L), which corresponded to a fermentation efficiency of 78% of the maximum obtainable theoretically. The experimental results led to the ratio of 380 L of ethanol per ton of sugar cane bagasse PDC.
    Journal of Biotechnology 10/2012; · 3.18 Impact Factor
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    ABSTRACT: Dentre as diversas matérias-primas renováveis disponíveis para a produção de etanol, o sorgo sacarino apresenta-se como uma das opções mais promissoras devido à sua ampla adaptabilidade em diferentes tipos de clima e solo. Além disso, é a única cultura que fornece colmos e grãos que podem ser usados para a produção de etanol por via enzimática, e a biomassa excedente (resíduo ligno-celulósico) pode ser utilizada tanto na cogeração de energia, como na produção de etanol de segunda geração. O objetivo deste tra-balho foi avaliar a produção de bioetanol a partir das frações sacarínea, amilácea e lignocelulósica do sorgo sacarino. A conversão da fração amilácea contida nos grãos atingiu a produção de 87g.L -1 de etanol, a do caldo sacarino resultou em 72g.L -1 , e da fração lignoce-lulósica do sorgo sacarino foi possível produzir 30g.L -1 e 84,4g.L -1 de etanol das frações hemicelulósicas e celulósicas, respectivamente. Com este processo foi possível atingir a relação de aproximadamente 160L de etanol/ton de sorgo sacarino como um todo, o que corresponde a 13.610L de etanol/ha, aproveitando 79,1% do potencial teórico das frações de açúcares do sorgo. Estes resultados indicam a possibilidade de usar esta cultura excepcional como principal matéria-prima para a produção de etanol e outros bioprodutos de alto valor agregado em regiões com condições de solo e clima desfavoráveis ao cultivo de cana-de-açúcar e como cultura comple-mentar na entressafra do cultivo de cana. palavras-chave: sorgo sacarino fermentação alcoólica pré-tratamentos materiais lignocelulósicos bioetanol
    Boletim Tecnico da PETROBRAS 12/2011; 54(3):29-46.
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    ABSTRACT: In this study, we investigated the enzymatic hydrolysis of pretreated sugarcane bagasse using eight different enzymatic blends obtained from concentrated crude enzyme extracts produced by Penicillium funiculosum and Trichoderma harzianum as well as from the extracts in combination with a commercial enzymatic cocktail. The influence of different levels of biomass delignification, degree of crystallinity of lignicellulose, composition of enzymatic activities and BSA on enzymatic hydrolysis yields (HYs) was evaluated. Our X-ray diffraction studies showed that crystallinity of lignocellulose is not a key determinant of its recalcitrance toward enzymatic hydrolysis. In fact, under the experimental conditions of our study, an increase in crystallinity of lignocellulosic samples resulted in increased glucose release by enzymatic hydrolysis. Furthermore, under the same conditions, the addition of BSA had no significant effect on enzymatic hydrolysis. The most efficient enzyme blends were obtained by mixing a commercial enzymatic cocktail with P. funiculosum or T. harzianum cellulase preparations (HYs above 97%) followed by the concentrated extract of P. funiculosum alone (HY = 88.5%). Increased hydrolytic efficiencies appeared to correlate with having an adequate level of both β-glucosidase and xylanase activities in the blends.
    Process Biochemistry. 01/2011; 46(5):1196-1201.
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    ABSTRACT: The PA1 strain of Pseudomonas aeruginosa isolated from oil waste produces rhamnolipid, a biodegradable surfactant with applications in several industrial and environmental fields. The metabolic pathway and genetic regulation of rhamnolipid production in P. aeruginosa are poorly understood. Herein, several proteins directly or indirectly related to rhamnolipid production and their genetic regulations were identified by comparative proteomic. We compared the proteome of P. aeruginosa PA1 after fermentation in two different conditions of carbon and nitrogen sources: condition A allowed rhamnolipid production and condition B prevented it. Protein extracts from cellular pellets were compared using 2D-PAGE stained with colloidal Coomassie followed by MALDI-TOF/TOF mass spectrometry. We identified 21 differentially expressed proteins, including those involved in secretion, quorum sensing, oxidative response and metabolism.
    Process Biochemistry 09/2010; · 2.44 Impact Factor
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    ABSTRACT: The present study aimed at maximizing cellulase production by Penicillium funiculosum using sequential experimental design methodology for optimizing the concentrations of nitrogen sources. Three sequential experimental designs were performed. The first and the second series of experiments consisted of a 2(4) and a 2(3) factorial designs, respectively, and in the third one, a central composite rotational design was used for better visualizing the optimum conditions. The following nitrogen sources were evaluated: urea, ammonium sulfate, peptone, and yeast extract. Peptone and ammonium sulfate were removed from the medium optimization since they did not present significant statistical effect on cellulase production. The optimal concentrations of urea and yeast extract predicted by the model were 0.97 and 0.36 g/L, respectively, which were validated experimentally. By the use of the desirability function, it was possible to maximize the three main enzyme activities simultaneously, which resulted in values for FPase of 227 U/L, for CMCase of 6,917 U/L, and for beta-glucosidase of 1,375 U/L. These values corresponded to increases of 3.3-, 3.2-, and 6.7-folds, respectively, when compared to those obtained in the first experimental design. The results showed that the use of sequential experimental designs associated to the use of the desirability function can be used satisfactorily to maximize cellulase production by P. funiculosum.
    Applied biochemistry and biotechnology 12/2009; 161(1-8):411-22. · 1.94 Impact Factor
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    ABSTRACT: Rhamnolipids have been pointed out as promising biosurfactants. The most studied microorganisms for the aerobic production of these molecules are the bacteria of the genus Pseudomonas. The aim of this work was to produce a rhamnolipid-type biosurfactant in a bench-scale bioreactor by one strain of Pseudomonas aeruginosa isolated from oil environments. To study the microorganism growth and production dependency on oxygen, a nondispersive oxygenation device was developed, and a programmable logic controller (PLC) was used to set the dissolved oxygen (DO) concentration. Using the data stored in a computer and the predetermined characteristics of the oxygenation device, it was possible to evaluate the oxygen uptake rate (OUR) and the specific OUR (SOUR) of this microorganism. These rates, obtained for some different DO concentrations, were then compared to the bacterial growth, to the carbon source consumption, and to the rhamnolipid and other virulence factors production. The SOUR presented an initial value of about 60.0 mgO(2)/g(DW) h. Then, when the exponential growth phase begins, there is a rise in this rate. After that, the SOUR reduces to about 20.0 mgO(2)/g(DW) h. The carbon source consumption is linear during the whole process.
    Applied biochemistry and biotechnology 04/2008; 147(1-3):33-45. · 1.94 Impact Factor
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    ABSTRACT: Rhamnolipids have been pointed out as promising biosurfactants. The most studied microorganisms for the aerobic production of these molecules are the bacteria of the genus Pseudomonas. The aim of this work was to produce a rhamnolipid-type biosurfactant in a bench-scale bioreactor by one strain of Pseudomonas aeruginosa isolated from oil environments. To study the microorganism growth and production dependency on oxygen, a nondispersive oxygenation device was developed, and a programmable logic controller (PLC) was used to set the dissolved oxygen (DO) concentration. Using the data stored in a computer and the predetermined characteristics of the oxygenation device, it was possible to evaluate the oxygen uptake rate (OUR) and the specific OUR (SOUR) of this microorganism. These rates, obtained for some different DO concentrations, were then compared to the bacterial growth, to the carbon source consumption, and to the rhamnolipid and other virulence factors production. The SOUR presented an initial value of about 60.0 mg02/gdw h. Then, when the exponential growth phase begins, there is a rise in this rate. After that, the SOUR reduces to about 20.0 mg02/gdw h. The carbon source consumption is linear during the whole process. KeywordsPseudomonas aeruginosa-Biosurfactant-Oxygenation-Rhamnolipid-Bioreactor
    12/2007: pages 401-413;
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    ABSTRACT: The castor bean cake is rich in starch (48 ± 0.53%) and bears a problem linked to the occurrence of a toxic protein (ricin). The chemical hydrolysis (ratio solid:liquid = 1:6; H2SO4= 0.1 mol L-1; 120 °C; 40 min) generated a medium with 27 g L-1 of reducing sugars (hydrolysis efficiency= 32%). The hydrolyzed product was fermented and produced 11 g L-1 of ethanol (volumetric productivity=1.38 g L-1 h-1 and ethanol yield on substrate consumed=0.45 g g-1). In vivo experiments (DL50) revealed a reduction of roughly 240 times in the CBC toxicity (2.11 µg g-1).
    Química Nova 12/2007; 31(5):1104-1106. · 0.74 Impact Factor
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    ABSTRACT: The effectiveness of cell-free rhamnolipid biosurfactant, derived from the culture medium at the end of fermentation was investigated for the removal of two different kinds of oil from contaminated sandy soils. The crude cultivation medium, containing 13.2 g L−1 of rhamnolipids, had a surface tension, interfacial tension and critical micellar concentration of 30 mN m−1, 2 mN m−1 and 60 mg L−1, respectively. The evaluation of biosurfactant in the culture medium (BM) and oil concentrations in the removal of oil from different contaminated sandy soil was performed using a statistical experimental design tool. Oil in sandy soil, containing predominantly aromatic or paraffinic hydrocarbons (5 to 10% w/w), was removed by as much as 91 and 78%, respectively, in the presence of reduced amounts of BM (6.3 to 7.9 g L−1). The progress of oil removal was monitored for 101 days and results indicated that removal efficiency in sandy soil with aromatic characteristics was relatively stable over the entire period. Based on these studies, it is concluded that use of a BM was effective in reducing oil concentrations in contaminated sandy soil. Copyright © 2007 Society of Chemical Industry
    Journal of Chemical Technology & Biotechnology 07/2007; 82(7):687 - 691. · 2.50 Impact Factor
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    ABSTRACT: The production of biosurfactant by Rhodococcus erythropolis during the growth on glycerol was investigated. The process was carried out at 28 degrees C in a 1.5-L bioreactor using glycerol as carbon source. The bioprocess was monitored through measurements of biosurfactant concentration and glycerol consumption. After 51 h of cultivation, 1.7 g/L of biosurfactant, surface, and interfacial tensions values (with n-hexadecane) of 43 and 15 mN/m, respectively, 67% of Emulsifying Index (E (24)), and 94% of oil removal were obtained. The use of glycerol rather than what happens with hydrophobic carbon source allowed the release of the biosurfactant, originally associated to the cell wall.
    Applied biochemistry and biotechnology 04/2006; 131(1-3):880-6. · 1.94 Impact Factor
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    ABSTRACT: The production of biosurfactant by Rhodococcus erythropolis during the growth on glycerol was investigated. The process was carried out at 28°C in a 1.5-L bioreactor using glycerol as carbon source. The bioprocess was monitored through measurements of biosurfactant concentration and glycerol consumption. After 51 h of cultivation, 1.7 g/L of biosurfactant, surface, and interfacial tensions values (with n-hexadecane) of 43 and 15 mN/m, respectively, 67% of Emulsifying Index (E24), and 94% of oil removal were obtained. The use of glycerol rather than what happens with hydrophobic carbon source allowed the release of the biosurfactant, originally associated to the cell wall.
    Applied Biochemistry and Biotechnology 12/2005; 131(1-3):880-886. · 1.89 Impact Factor
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    ABSTRACT: Culture conditions involving variations in carbon and nitrogen sources and different C:N ratios were examined with the aim of increasing productivity in the process of rhamnolipid synthesis by Pseudomonas aeruginosa. In addition to the differences in productivity, the use of different carbon sources resulted in several proportions related to the types of rhamnolipids synthesized (monorhamnolipids and dirhamnolipids). Furthermore, the variation in nutrients, mainly the nitrogen source, resulted in different amounts of virulence factors, as phenazines and extracellular proteins. The data point out a new concern in the choice of substrate to be used for rhamnolipid production by P. aeruginosa: toxic byproducts.
    Applied Biochemistry and Biotechnology 02/2002; 98-100:1025-35. · 1.89 Impact Factor
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    ABSTRACT: The Pseudomonas aeruginosa PA1 strain, isolated from the water of oil production in Sergipe, Northeast Brazil, was evaluated as a potential rhamnolipid type of biosurfactant producer. The production of biosurfactants was investigated using different carbon sources (n-hexadecane, paraffin oil, glycerol, and babassu oil) and inoculum concentrations (0.0016-0.008 g/L). The best results were obtained with glycerol as the substrate and an initial cell concentration of 0.004 g/L. A C:N ratio of 22.8 led to the greatest production of rhamnolipids (1700 mg/L) and efficiency (1.18 g of rhamnolipid/g of dry wt).
    Applied Biochemistry and Biotechnology 02/2001; 91-93:459-67. · 1.89 Impact Factor