The Brazilian Bioethanol Science and Technology Laboratory (CTBE) publications printed on 2012 and indexed by Web of Sciences (www.isiwebofknowledge.com) are listed below in chronological order.

Criteria for articles selectionFill “(CTBE OR Bioethanol OR Bioetanol) AND Brazil” in the Address Search form.

Total of CTBE Journal Articles42

42. Emergy evaluation of organic and conventional marine shrimp farms in Guaraira Lagoon, Brazil

Author(s): Lima, JSG; Rivera, EC; Focken, U | E-mail: elmer.rivera@bioetanol.org.br   Source: JOURNAL OF CLEANER PRODUCTION Volume: 35 Pages: 194-202 | DOI: 10.1016/j.jclepro.2012.05.009 | Published: NOV 2012

Abstract: Organic products are often appreciated by the consumers as being more environmentally friendly and sustainable compared to their conventional counterparts. However, in the case of aquaculture, scientific information regarding environmental impacts and costs of conventional and organic farming systems is scarce. Scientific methods able to measure the sustainability of production systems are of great relevance to identify best management practices and ensure consumers the sustainability of the production process of aquaculture products. The objective of this study was to compare organic and conventional marine shrimp farms located in Guaraira Lagoon, Brazil, using emergy analysis. The principle of emergy analysis is to express all costs of a process or product in solar energy units (sej), making it possible to compare systems with different inputs, outputs and environmental impacts. Emergy accounting was used at this study to integrate ecological and human dimensions of conventional and organic marine shrimp farms systems located at Brazilian northeast in order to better discuss and understand the multiple dimensions of the sustainability in such systems. The results showed several emergy indicators in favor of organic farming system: the renewability indicator, the emergy yield ratio indicator and the emergy investment ratio indicator. Both systems (organic and conventional) showed large flows of concentrated non-renewable emergy. Further improvements of the organic system are needed to increase the efficiency and to ensure its economic sustainability.Full Text: www.sciencedirect.com/science/article/pii/S0959652612002326

Addresses: [Goncalves Lima, Juliana Schober; Focken, Ulfert] Univ Hohenheim 480B, Inst Anim Prod Trop & Subtrop, Dept Aquaculture Syst & Anim Nutr, D-70593 Stuttgart, Germany [Rivera, Elmer Ccopa] Brazilian Bioethanol Sci & Technol Lab CTBE, BR-13083970 Campinas, SP, Brazil

41. Morphological heterogeneity of Paracoccidioides brasiliensis: relevance of the Rho-like GTPase PbCDC42

Author(s): Menino, JF; Osorio, NS; Sturme, MHJ; Barros, D; Gomes-Alves, AG; Almeida, AJ; Ludovico, P; Costa, P; Goldman, GH; Rodrigues, F | E-mail: gustavo.goldman@bioetanol.org.br

Source: MEDICAL MYCOLOGY Volume: 50 Issue: 7 Pages: 768-774 | DOI: 10.3109/13693786.2012.665614 | Published: OCT 2012

Abstract: Paracoccidioides brasiliensis budding pattern and polymorphic growth were previously shown to be closely linked to the expression of PbCDC42 and to influence the pathogenesis of the fungus. In this work we conducted a detailed morphogenetic evaluation of the yeast-forms of 11 different clinical and environmental P. brasiliensis isolates comprising four phylogenetic lineages (S1, PS2, PS3 and Pb01-like), as well as a PbCDC42 knock-down strain. High variations in the shape and size of mother and bud cells of each isolate were observed but we did not find a characteristic morphologic profile for any of the phylogenetic groups. In all isolates studied, the bud size and shape were demonstrated to be highly dependent on the mother cell. Importantly, we found strong correlations between PbCDC42 expression and both the shape of mother and bud cells and the size of the buds in all isolates and the knock-down strain. Our results suggested that PbCDC42 expression can explain approximately 80% of mother and bud cell shape and 19% of bud cell size. This data support PbCDC42 expression level as being a relevant predictor of P. brasiliensis morphology. Altogether, these findings quantitatively describe the polymorphic nature of the P. brasiliensis yeast form and provide additional support for the key role of PbCDC42 expression on yeast cell morphology.Full Text: http://informahealthcare.com/doi/abs/10.3109/13693786.2012.665614

Addresses: [Menino, Joao F.; Osorio, Nuno S.; Sturme, Mark H. J.; Barros, Diana; Gomes-Alves, Ana G.; Almeida, Agostinho J.; Ludovico, Paula; Costa, Patricio; Rodrigues, Fernando] Univ Minho, Sch Hlth Sci, Life & Hlth Sci Res Inst ICVS, P-4710057 Braga, Portugal[Menino, Joao F.; Osorio, Nuno S.; Sturme, Mark H. J.; Barros, Diana; Gomes-Alves, Ana G.; Almeida, Agostinho J.; Ludovico, Paula; Costa, Patricio; Rodrigues, Fernando] ICVS 3Bs PT Govt Associate Lab, Braga, Portugal[Goldman, Gustavo H.] Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil [Goldman, Gustavo H.] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Sao Paulo, Brazil

40. Cellulose crystals in fibrovascular bundles of sugarcane culms: orientation, size, distortion, and variability

Author(s): Driemeier, C; Santos, WD; Buckeridge, MS | E-mail: carlos.driemeier@bioetanol.org.brSource: CELLULOSE Volume: 19 Issue: 5 Pages: 1507-1515 | DOI: 10.1007/s10570-012-9743-z | Published: OCT 2012

Abstract: Cellulose crystals in fibrovascular bundles of sugarcane culms were throughly characterized by X-ray diffraction, with area-detector patterns acquired in fiber geometry. It was observed that microfibril angles are, on average, higher for bundles from pith compared to bundles from intermediate regions and rind, and higher for ratoon canes compared to plant canes. On the other hand, microfibril angles do not differ significantly among sugarcane cultivars, internode positions in the culm, or ratoon cane cut cycles. Broadening analyses of diffraction peaks yield crystal sizes (crystal width from equatorial 200 reflection and apparent crystal lengths from meridional 002 and 004 reflections) similar to other plant species (woods and bamboo). In addition, compared to reference cellulose I-beta, the 200, 110, and 110 diffraction peaks from sugarcane cellulose are notably shifted. These shifts indicate pronounced crystallite distortions, with expanded intersheet spacing d (200) , contracted d (1 <Emphasis Type="ItalicUnderline">1 0) /d (110) , and monoclinic angle gamma closer to 90A degrees. Our findings deepen the understanding of the fine structure and variability of sugarcane lignocellulose.Full Text: www.springerlink.com/content/1253471612253n70/fulltext.pdf

Addresses: [Driemeier, Carlos; Santos, Wanderley D.; Buckeridge, Marcos S.] Lab Nacl Ciencia & Tecnol Bioetanol CTBE CNPEM, BR-13083970 Campinas, SP, Brazil

39. Improving bioethanol production - Comparison between extractive and low temperature fermentation

Author(s): Dias, MOS; Junqueira, TL; Jesus, CDF; Rossell, CEV; Maciel, R; Bonomi, A | E-mail: antonio.bonomi@bioetanol.org.br Source: APPLIED ENERGY Volume: 98 Pages: 548-555 | DOI: 10.1016/j.apenergy.2012.04.030 | Published: OCT 2012

Abstract: One of the key issues that must be addressed in the biofuel production based on sugarcane industry is the energy consumption of the process. Process energy demand has direct impact on the amount of lignocellulosic material available for use as feedstock for second generation ethanol production. A significant fraction of the energy consumption in bioethanol production occurs in the purification step, since conventional fermentation systems employed in the industry require low substrate concentration and, consequently, produce wine of low (around 8.5 degrees GL) ethanol content that must be distilled in order to meet product specifications. In this study alternatives to the conventional fermentation processes employed in the industry (low temperature fermentation and vacuum extractive fermentation) were assessed, in the context of a large scale sugarcane autonomous distillery, through computer simulation. Electricity consumption and lignocellulosic material surplus on each case were evaluated. It is shown that the alternative fermentation processes allow a significant reduction on vinasse generation and increases ethanol production when compared with conventional fermentation, but increases electricity consumption (for the extractive fermentation) or steam consumption (for low temperature fermentation); when vinasse concentration is considered in the conventional process, steam consumption in the extractive fermentation is also significantly smaller. Full Text: www.sciencedirect.com/science/article/pii/S0306261912003169

Addresses: [Dias, M. O. S.; Junqueira, T. L.; Jesus, C. D. F.; Rossell, C. E. V.; Maciel Filho, R.; Bonomi, A.] CTBE CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol, BR-13083970 Campinas, SP, Brazil [Dias, M. O. S.; Junqueira, T. L.; Rossell, C. E. V.; Maciel Filho, R.] Univ Estadual Campinas, Fac Engn Quim, UNICAMP, BR-13083852 Campinas, SP, Brazil

38. Topography of funneled landscapes determines the thermodynamics and kinetics of protein folding

Author(s): Wang, J; Oliveira, RJ; Chu, XK; Whitford, PC; Chahine, J; Han, W; Wang, EK; Onuchic, JN; Leite, VBP | E-mail: ronaldo.oliveira@bioetanol.org.br Source: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Volume: 109 Issue: 39 Pages: 15763-15768 | DOI: 10.1073/pnas.1212842109 | Published: SEP 25 2012

Abstract: The energy landscape approach has played a fundamental role in advancing our understanding of protein folding. Here, we quantify protein folding energy landscapes by exploring the underlying density of states. We identify three quantities essential for characterizing landscape topography: the stabilizing energy gap between the native and nonnative ensembles delta E, the energetic roughness Delta E, and the scale of landscape measured by the entropy S. We show that the dimensionless ratio between the gap, roughness, and entropy of the system Lambda = delta E/(Delta E root 2S) accurately predicts the thermodynamics, as well as the kinetics of folding. Large Lambda implies that the energy gap (or landscape slope towards the native state) is dominant, leading to more funneled landscapes. We investigate the role of topological and energetic roughness for proteins of different sizes and for proteins of the same size, but with different structural topologies. The

landscape topography ratio Lambda is shown to be monotonically correlated with the thermodynamic stability against trapping, as characterized by the ratio of folding temperature versus trapping temperature. Furthermore, Lambda also monotonically correlates with the folding kinetic rates. These results provide the quantitative bridge between the landscape topography and experimental folding measurements.Full Text: www.pnas.org/content/109/39/15763

Addresses: [Wang, Jin; Chu, Xiakun; Wang, Erkang] Chinese Acad Sci, State Key Lab Electroanalyt Chem, Changchun Inst Appl Chem, Changchun 130012, Jilin, Peoples R China [Wang, Jin; Chu, Xiakun; Han, Wei] Jilin Univ, Coll Phys, Changchun 130021, Jilin, Peoples R China [Wang, Jin; Chu, Xiakun; Han, Wei] Jilin Univ, State Key Lab Superhard Mat, Changchun 130021, Jilin, Peoples R China [Wang, Jin] SUNY Stony Brook, Dept Chem Phys & Appl Math, Stony Brook, NY 11794 USA[Oliveira, Ronaldo J.; Chahine, Jorge; Leite, Vitor B. P.] Univ Estadual Paulista, Dept Fis, Inst Biociencias Letras & Ciencias Exatas, BR-15054000 Sao Jose Do Rio Preto, Brazil [Oliveira, Ronaldo J.] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol, BR-13083970 Campinas, SP, Brazil[Whitford, Paul C.; Onuchic, Jose N.] Rice Univ, Ctr Theoret Biol Phys, Houston, TX 77005 USA

37. Enhancement of the enzymatic digestibility of sugarcane bagasse by steam pretreatment impregnated with hydrogen peroxide

Author(s): Rabelo, SC; Rossell, CEV; Rocha, GJD; Zacchi, G | E-mail: sarita.rabelo@bioetanol.org.br Source: BIOTECHNOLOGY PROGRESS Volume: 28 Issue: 5 Pages: 1207-1217 | DOI: 10.1002/btpr.1593 | Published: SEP-OCT 2012

Abstract: Sugarcane bagasse was subjected to steam pretreatment impregnated with hydrogen peroxide. Analyses were performed using 23 factorial designs and enzymatic hydrolysis was performed at two different solid concentrations and with washed and unwashed material to evaluate the importance of this step for obtaining high cellulose conversion. Similar cellulose conversion were obtained at different conditions of pretreatment and hydrolysis. When the cellulose was hydrolyzed using the pretreated material in the most severe conditions of the experimental design (210 degrees C, 15 min and 1.0% hydrogen peroxide), and using 2% (w/w) water-insoluble solids (WIS), and 15 FPU/g WIS, the cellulose conversion was 86.9%. In contrast, at a milder pretreatment condition (190 degrees C, 15 min and 0.2% hydrogen peroxide) and industrially more realistic conditions of hydrolysis (10% WIS and 10 FPU/g WIS), the cellulose conversion reached 82.2%. The step of washing the pretreated material was very important to obtain high concentrations of fermentable sugars. Full Text: http://onlinelibrary.wiley.com/doi/10.1002/btpr.v28.5/issuetoc

Addresses: [Rabelo, Sarita Candida; Vaz Rossell, Carlos Eduardo; de Moraes Rocha, George Jackson] Lab Nacl Ciencia & Tecnol Bioetanol CTBE CNPEM, BR-13083970 Campinas, SP, Brazil[Zacchi, Guido] Lund Univ, Dept Chem Engn, SE-22100 Lund, Sweden

36. Substrate-Specific Reorganization of the Conformational Ensemble of CSK Implicates Novel Modes of Kinase Function

Author(s): Jamros, MA; Oliveira, LC; Whitford, PC; Onuchic, JN; Adams, JA; Jennings, PA | E-mail: leandro.oliveira@bioetanol.org.br Source: PLOS COMPUTATIONAL BIOLOGY Volume: 8 Issue: 9 Article Number: e1002695 | DOI:

10.1371/journal.pcbi.1002695 | Published: SEP 2012

Abstract: Protein kinases use ATP as a phosphoryl donor for the posttranslational modification of signaling targets. It is generally thought that the binding of this nucleotide induces conformational changes leading to closed, more compact forms of the kinase domain that ideally orient active-site residues for efficient catalysis. The kinase domain is oftentimes flanked by additional ligand binding domains that up- or down-regulate catalytic function. C-terminal Src kinase (Csk) is a multidomain tyrosine kinase that is up-regulated by N-terminal SH2 and SH3 domains. Although the X-ray structure of Csk suggests the enzyme is compact, X-ray scattering studies indicate that the enzyme possesses both compact and open conformational forms in solution. Here, we investigated whether interactions with the ATP analog AMP-PNP and ADP can shift the conformational ensemble of Csk in solution using a combination of small angle x-ray scattering and molecular dynamics simulations. We find that binding of AMP-PNP shifts the ensemble towards more extended rather than more compact conformations. Binding of ADP further shifts the ensemble towards extended conformations, including highly extended conformations not adopted by the apo protein, nor by the AMP-PNP bound protein. These ensembles indicate that any compaction of the kinase domain induced by nucleotide binding does not extend to the overall multi-domain architecture. Instead, assembly of an ATP-bound kinase domain generates further extended forms of Csk that may have relevance for kinase scaffolding and Src regulation in the cell.Full Text: www.ploscompbiol.org/article/info%3Adoi%2F10.1371%2Fjournal.pcbi.1002695

Addresses: [Jamros, Michael A.; Jennings, Patricia A.] Univ Calif San Diego, Dept Chem & Biochem, La Jolla, CA 92093 USA[Oliveira, Leandro C.] CTBE CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol, Campinas, SP, Brazil[Whitford, Paul C.; Onuchic, Jose N.] Rice Univ, Ctr Theoret Biol Phys, Houston, TX USA[Adams, Joseph A.] Univ Calif San Diego, Dept Pharmacol, La Jolla, CA 92093 USA

35. Aspergillus fumigatus calcineurin interacts with a nucleoside diphosphate kinase

Author(s): Dinamarco, TM; Brown, NA; de Almeida, RSC; de Castro, PA; Savoldi, M; Goldman, MHD; Goldman, GH | E-mail: gustavo.goldman@bioetanol.org.br Source: MICROBES AND INFECTION Volume: 14 Issue: 11 Pages: 922-929 | DOI: 10.1016/j.micinf.2012.05.003 | Published: SEP 2012

Abstract: The Ca2+-calcineurin pathway affects virulence and morphogenesis in filamentous fungi. Here, we identified 37 CalA-interacting proteins that interact with the catalytic subunit of calcineurin (CalA) in Aspergillus fumigatus, including the nucleoside diphosphate kinase (SwoH). The in vivo interaction between CalA and SwoH was validated by bimolecular fluorescence complementation. A. fumigatus swoH is an essential gene. Therefore, a temperature-sensitive conditional mutant strain with a point mutation in the active site, SwoH(V83F), was constructed, which demonstrated reduced growth and increased sensitivity to elevated temperatures. The SwoH(V83F) mutation did not cause a loss in virulence in the Galleria mellonella infection model. Taken together these results imply that CalA interacts with SwoH. (C) 2012 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.Full Text: www.sciencedirect.com/science/article/pii/S1286457912001219

Addresses: [Brown, Neil Andrew; de Castro, Patricia Alves; Savoldi, Marcela; Goldman, Gustavo Henrique] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Sao Paulo, Brazil [Couto de Almeida, Ricardo Sergio] Univ Londrina, Dept Microbiol, Londrina, Parana, Brazil [Dinamarco, Taisa Magnani; de Souza Goldman, Maria Helena] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Sao Paulo, BrazilFundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq), Brazil

34. Functional characterization and synergic action of fungal xylanase and arabinofuranosidase for production of xylooligosaccharides

Author(s): Goncalves, TA; Damasio, ARL; Segato, F; Alvarez, TM; Bragatto, J; Brenelli, LB; Citadini, APS; Murakami, MT; Ruller, R; Leme, AFP; Prade, RA; Squina, FM | E-mail: fabio.squina@bioetanol.org.br Source: BIORESOURCE TECHNOLOGY Volume: 119 Pages: 293-299 | DOI: 10.1016/j.biortech.2012.05.062 | Published: SEP 2012

Abstract: Plant cell wall degrading enzymes are key technological components in biomass bioconversion platforms for lignocellulosic materials transformation. Cost effective production of enzymes and identification of efficient degradation routes are two economic bottlenecks that currently limit the use of renewable feedstocks through an environmental friendly pathway. The present study describes the hypersecretion of an endo-xylanase (GH11) and an arabinofuranosidase (GH54) by a fungal expression system with potential biotechnological application, along with comprehensive characterization of both enzymes, including spectrometric analysis of thermal denaturation, biochemical characterization and mode of action description. The synergistic effect of these enzymes on natural substrates such as sugarcane bagasse, demonstrated the biotechnological potential of using GH11 and GH54 for production of probiotic xylooligosaccharides from plant biomass. Our findings shed light on enzymatic mechanisms for xylooligosaccharide production, as well as provide basis for further studies for the development of novel enzymatic routes for use in biomass-to-bioethanol applications.Full Text: www.sciencedirect.com/science/article/pii/S0960852412008061

Addresses: [Goncalves, T. A.; Damasio, A. R. L.; Segato, F.; Alvarez, T. M.; Bragatto, J.; Brenelli, L. B.; Citadini, A. P. S.; Ruller, R.; Squina, F. M.] CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil Murakami, M. T.; Paes Leme, A. F.] CNPEM, Lab Nacl Biociencias LNBio, BR-13083970 Campinas, SP, Brazil [Prade, R. A.] Oklahoma State Univ, Dept Microbiol & Mol Genet, Stillwater, OK 74078 USA

33. Shape resonance spectra of lignin subunits

Author(s): de Oliveira, EM; Sanchez, SD; Bettega, MHF; Natalense, APP; Lima, MAP; Varella, MTD | E-mail: a.natalense@bioetanol.org.br Source: PHYSICAL REVIEW A Volume: 86 Issue: 2 Article Number: 020701 | DOI: 10.1103/PhysRevA.86.020701 | Published: AUG 20 2012

Abstract: We report integral cross sections for elastic electron scattering by the lignin subunits phenol, guaiacol, and p-coumaryl alcohol. Our calculations employed the Schwinger multichannel method with pseudopotentials and indicate three to four pi* shape resonances for each of these systems, suggesting that low-energy electrons could efficiently transfer energy into the lignin matrix. We also discuss dissociation mechanisms based on the calculated cross sections, available experimental data, virtual orbital analysis, and the knowledge on electron interactions with biomolecules. Our results point out a physical- chemical basis for electron-driven biomass delignification. The latter would be an essential step for efficient biofuel production from lignocellulosic materials.Full Text: http://pra.aps.org/abstract/PRA/v86/i2/e020701

Addresses: [de Oliveira, Eliane M.; Natalense, Alexandra P. P.; Lima, Marco A. P.] Lab Nacl Ciencia & Tecnol Bioetanol CTBE CNPEM, BR-13083970 Campinas, SP, Brazil [Sanchez, Sergio d'A.; Bettega, Marcio H. F.] Univ Fed Parana, Dept Fis, BR-81531990 Curitiba, Parana, Brazil [Lima, Marco A. P.] Univ Estadual Campinas, Inst Fis Gleb Wataghin, BR-13083859 Campinas, SP, Brazil[Varella, Marcio T. do N.] Univ Sao Paulo, Inst Fis, BR-05315970 Sao Paulo, Brazil

32. Endo-xylanase GH11 activation by the fungal metabolite eugenitin

Author(s): Andrioli, WJ; Damasio, ARL; Silva, TM; da Silva, VB; Maller, A; Nanayakkara, NPD; Silva, CHTP; Polizeli, MLTM; Bastos, JK | E-mail: andre.damasio@bioetanol.org.br Source: BIOTECHNOLOGY LETTERS Volume: 34 Issue: 8 Pages: 1487-1492 | DOI: 10.1007/s10529-012-0918-3 | Published: AUG 2012

Abstract: Eugenitin, a chromone derivative and a metabolite of the endophyte Mycoleptodiscus indicus, at 5 mM activated a recombinant GH11 endo-xylanase by 40 %. The in silico prediction of ligand-binding sites on the three-dimensional structure of the endo-xylanase revealed that eugenitin interacts mainly by a hydrogen bond with a serine residue and a stacking interaction of the heterocyclic aromatic ring system with a tryptophan residue. Eugenitin improved the GH11 endo-xylanase activity on different substrates, modified the optimal pH and temperature activities and slightly affected the kinetic parameters of the enzyme.Full Text: www.springerlink.com/content/r5247550m1782473/

Addresses: [Andrioli, Willian J.; da Silva, Vinicius B.; Silva, Carlos H. T. P.; Bastos, Jairo K.] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, BR-14040903 Ribeirao Preto, SP, Brazil [Damasio, Andre R. L.; Maller, Alexandre; Polizeli, Maria L. T. M.] Univ Sao Paulo, Fac Med Ribeirao Preto, BR-14040903 Ribeirao Preto, SP, Brazil[Damasio, Andre R. L.; Polizeli, Maria L. T. M.] CNPEM Ctr Nacl Pesquisa Energia & Mat, CTBE Lab Nacl Ciencia & Tecnol Bioetanol, Campinas, SP, Brazil [Silva, Tony M.] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, BR-14040903 Ribeirao Preto, SP, Brazil [Nanayakkara, N. P. D.] Univ Mississippi, Natl Ctr Nat Prod Res, Oxford, MS 38677 USA

31. Accounting greenhouse gas emissions in the lifecycle of Brazilian sugarcane bioethanol: Methodological references in European and American regulations

Author(s): Khatiwada, D; Seabra, J; Silveira, S; Walter, A | E-mail: arnaldo.walter@bioetanol.org.br Source: ENERGY POLICY Volume: 47 Pages: 384-397 | DOI: 10.1016/j.enpol.2012.05.005 | Published: AUG 2012

Abstract: This study discusses four European and American regulatory schemes designed for accounting lifecycle GHG emissions in relation to the Brazilian sugarcane ethanol. The objective is to critically examine the methodologies and associated parameters used in existing regulatory schemes for calculating GHG emissions, and to explore methodological convergences. The issues related to direct lifecycle and indirect land use change emissions have been addressed. It is found that there are commonalities between the European Renewable Energy Directive (EU-RED) and the UK's Renewable Transport Fuels Obligation (UK-RTFO), but the US-EPA's Renewable Fuel Standard (US-EPA) and the Low Carbon Fuel Standard of the California Air Resources Board (CA-CARB) vary greatly not only among themselves, but also in relation to the European regulations. Agricultural practices (especially soil carbon and nitrogen dynamics), co-product credits from surplus electricity

and uncertainties around economic modeling approaches for indirect land use change are the major areas where methodological divergences exist. Incorporation of domestic agricultural practices, sugarcane mills operations, and realistic modeling of indirect impacts of land use change using regional models could provide more coherence in estimations of GHG emissions. Furthermore, the Brazilian trend of novelty in all phases of sugarcane bioenergy systems should be considered when projecting GHG emissions.Full Text: www.sciencedirect.com/science/article/pii/S0301421512004028

Addresses: [Khatiwada, Dilip; Silveira, Semida] KTH Royal Inst Technol, Div Energy & Climate Studies, S-10044 Stockholm, Sweden [Khatiwada, Dilip; Seabra, Joaquim; Walter, Arnaldo] Brazilian Bioethanol Sci & Technol Lab CTBE, Campinas, SP, Brazil [Seabra, Joaquim; Walter, Arnaldo] Univ Estadual Campinas, Fac Engn Mecan, BR-13083860 Campinas, SP, Brazil

30. Dynamic vapor sorption and thermoporometry to probe water in celluloses

Author(s): Driemeier, C; Mendes, FM; Oliveira, MM | E-mail: carlos.driemeier@bioetanol.org.br Source: CELLULOSE Volume: 19 Issue: 4 Pages: 1051-1063 | DOI: 10.1007/s10570-012-9727-z | Published: AUG 2012

Abstract: Dynamic vapor sorption and thermoporometry probe complementary dimensions of water interaction with cellulose. While sorption is primarily sensitive to the first hydration layers, thermoporometry is primarily sensitive to the nanometric water-filled pores. In this article, these analytical techniques are detailed and applied to model mesoporous materials and to a wide spectrum of celluloses. Correlations between techniques are explored. In dynamic vapor sorption, celluloses present a general characteristic time of desorption. On the other hand, they present highly variable characteristic times of sorption, indicating that material-specific properties may be inferred from sorption kinetics. Regarding thermoporometry, the thermodynamics of ice melting in irregular pore shapes is introduced. Moreover, in our thermoporometry analysis with differential scanning calorimeter, freezing temperature is extended to -70 A degrees C, allowing pores smaller than a few nanometers to be measured. Nevertheless, several data corrections are required for accurate thermoporometry at this condition. Comparisons between techniques show that sorption hysteresis is positively correlated with wet porosity. The presented developments and results will guide future application of these techniques to probe water in celluloses.Full Text: www.springerlink.com/content/9x45133856176170/fulltext.pdf

Addresses: [Driemeier, Carlos; Mendes, Fernanda M.; Oliveira, Marcelo M.] Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Sao Paulo, Brazil

29. High-yield secretion of multiple client proteins in Aspergillus

Author(s): Segato, F; Damasio, ARL; Goncalves, TA; de Lucas, RC; Squina, FM; Decker, SR; Prade, RA | E-mail: fabio.squina@bioetanol.org.br Source: ENZYME AND MICROBIAL TECHNOLOGY Volume: 51 Issue: 2 Pages: 100-106 | DOI: 10.1016/j.enzmictec.2012.04.008 | Published: JUL 15 2012

Abstract: Production of pure and high-yield client proteins is an important technology that addresses the need for industrial applications of enzymes as well as scientific experiments in protein chemistry and crystallization. Fungi are utilized in industrial protein production because of their ability to secrete large quantities of proteins. In this study, we engineered a high-expression-secretion vector, pEXPYR

that directs proteins towards the extracellular medium in two Aspergillii host strains, examine the effect of maltose-induced over-expression and protein secretion as well as time and pH-dependent protein stability in the medium. We describe five client proteins representing a core set of hemicellulose degrading enzymes that accumulated up to 50-100 mg/L of protein. Using a recyclable genetic marker that allows serial insertion of multiple genes, simultaneous hyper-secretion of three client proteins in a single host strain was accomplished.Full Text: www.sciencedirect.com/science/article/pii/S0141022912000580

Addresses: [Segato, Fernando; Damasio, Andre R. L.; de Lucas, Rosymar C.; Prade, Rolf A.] Oklahoma State Univ, Dept Microbiol & Mol Genet, Stillwater, OK 74078 USA [Squina, Fabio M.] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, Campinas, SP, Brazil [Goncalves, Thiago A.] Univ Fed Vicosa, Dept Bioquim & Biol Mol, Vicosa, MG, Brazil[Decker, Stephen R.] Natl Renewable Energy Lab, Golden, CO USA

28. Improving second generation ethanol production through optimization of first generation production process from sugarcane

Author(s): Dias, MOS; Junqueira, TL; Jesus, CDF; Rossell, CEV; Maciel, R; Bonomi, A | E-mail: antonio.bonomi@bioetanol.org.br Source: ENERGY Volume: 43 Issue: 1 Pages: 246-252 | DOI: 10.1016/j.energy.2012.04.034 | Published: JUL 2012

Abstract: Sugarcane bagasse and trash may be used as feedstock for second generation ethanol production. Production of second generation ethanol integrated with first generation plants processing sugarcane presents several advantages over the stand-alone second generation ethanol production process; however, bagasse is used as fuel to supply the energy demand of the first generation process, so the amount of bagasse and trash available for use as feedstock in second generation depends on the energy consumption of the integrated process. Therefore, process optimization leading to reduction in steam consumption will lead to the production of larger amounts of surplus bagasse. In this study the introduction of process improvements in the first generation autonomous distillery processing sugarcane were assessed through simulation using Aspen Plus. Second generation ethanol production was integrated to the optimized scenarios. Results show that process improvements can significantly increase the amount of lignocellulosic material available for use as feedstock for second generation ethanol production, thus increasing ethanol production. Full Text: www.sciencedirect.com/science/article/pii/S0360544212003325

Addresses: [Dias, Marina O. S.; Junqueira, Tassia L.; Jesus, Charles D. F.; Rossell, Carlos E. V.; Maciel Filho, Rubens; Bonomi, Antonio] CTBE CNPEM, Lab Natl Ciencia & Tecnol Bioetanol, BR-13083970 Campinas, SP, Brazil [Dias, Marina O. S.; Junqueira, Tassia L.; Rossell, Carlos E. V.; Maciel Filho, Rubens] Univ Estadual Campinas UNICAMP, Fac Engn Quim, BR-13083852 Campinas, SP, Brazil

27. Optical and electrical diagnostics of a spark-plug discharge in air

Author(s): Oliveira, C; Reis, JL; Souza-Correa, JA; Dal Pino, A; Amorim, J | E-mail: jayr.amorim@bioetanol.org.br Source: JOURNAL OF PHYSICS D-APPLIED PHYSICS Volume: 45 Issue: 25 Article Number: 255201 | DOI: 10.1088/0022-3727/45/25/255201 | Published: JUN 27 2012

Abstract: Optical and electrical diagnostics were used to investigate a spark discharge using a

commercial spark-plug operating in the glow phase regime. Voltage and current were measured in order to characterize the discharge. The gas temperature was estimated as a function of time and duty cycle using ro-vibrational spectra of the second positive system of nitrogen by comparison between experimental and simulated spectra. It was found that 1600 K <= T-g <= 2800 K. The reduced electric field varied between 1 and 1000 Td after the end of the current pulse. Using the line intensity method between one line from the ionic argon and the other from the neutral atom, the electronic temperature was measured and found to be between 17 000 and 20 000 K. The electron density was determined from the broadening of H-alpha line and was found to be 4.0 x 10(14) cm(-3) <= n(e) <= 1.7 x 10(15) cm(-3).Full Text: http://iopscience.iop.org/0022-3727/45/25/255201/

Addresses: [Oliveira, C.; Souza-Correa, J. A.; Amorim, J.] Lab Nacl Ciencia & Tecnol Bioetanol CTBE CNPEM, BR-13083970 Sao Paulo, Brazil [Oliveira, C.; Reis, J. L., Jr.; Dal Pino, A., Jr.; Amorim, J.] ITA, Dept Fis, BR-12228900 Sao Paulo, Brazil [Reis, J. L., Jr.] Univ Estadual Paulista, BR-12240420 Sao Paulo, Brazil

26. Electronic excitation of gas-phase furan molecules by electron impact

Author(s): da Costa, RF; Bettega, MHF; Lima, MAP; Lopes, MCA; Hargreaves, LR; Serna, G; Khakoo, MA | E-mail: maplima@bioetanol.org.br Source: PHYSICAL REVIEW A Volume: 85 Issue: 6 Article Number: 062706 | DOI: 10.1103/PhysRevA.85.062706 | Published: JUN 25 2012

Abstract: Experiments and ab initio calculations of the differential and integral cross sections for the electronic excitation from the ground state (1)A(1) to the B-3(2) and (3)A(1) states of gas-phase furan molecules by low-energy electron impact were performed. Experimental differential cross sections were measured at incident electron energies between 5 and 15 eV and for scattering angles from 10 degrees to 130 degrees. The calculated cross sections were obtained using the Schwinger multichannel method implemented with pseudopotentials. The influence of channel-coupling and polarization effects is investigated through the comparison between three different models of scattering calculations, each one considering a distinct channel-coupling scheme. The comparison of experimental and calculated cross sections for electronically inelastic electron scattering by C4H4O molecules is found to be mostly reasonable. The existing discrepancies in this combined theoretical and experimental study help to illustrate difficulties in readily establishing reliable electronic excitation cross sections of polyatomic molecules by low-energy electrons.Full Text: http://pra.aps.org/pdf/PRA/v85/i6/e062706

Addresses: [da Costa, Romarly F.] Univ Fed ABC, Ctr Ciencias Nat & Humanas, BR-09210170 Santo Andre, SP, Brazil [Bettega, Marcio H. F.] Univ Fed Parana, Dept Fis, BR-81531990 Curitiba, Parana, Brazil [Lima, Marco A. P.] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol, BR-13083970 Campinas, SP, Brazil[Lima, Marco A. P.] Univ Estadual Campinas, Inst Fis Gleb Wataghin, BR-13083970 Campinas, SP, Brazil [Lopes, Maria C. A.] Univ Fed Juiz de Fora, Dept Fis, ICE, BR-36036330 Juiz De Fora, MG, Brazil [Hargreaves, Leigh R.; Serna, Gabriela; Khakoo, Murtadha A.] Calif State Univ Fullerton, Dept Phys, Fullerton, CA 92834 USA

25. Environmental and economic assessment of sugarcane first generation biorefineries in Brazil

Author(s): Cavalett, O; Junqueira, TL; Dias, MOS; Jesus, CDF; Mantelatto, PE; Cunha, MP; Franco, HCJ; Cardoso, TF; Maciel, R; Rossell, CEV; Bonomi, A | E-mail: otavio.cavalet@bioetanol.org.br Source: CLEAN TECHNOLOGIES AND ENVIRONMENTAL POLICY Volume: 14 Issue: 3 Pages: 399-410 | DOI: 10.1007/s10098-011-0424-7 | Published: JUN 2012

Abstract: The majority of current sugarcane mills in Brazil are annexed plants, producing both ethanol and sugar; the other plants are dedicated only to ethanol production (autonomous distilleries). This study focuses on the technical, environmental, and economic impacts of these first generation sugarcane processing facilities in Brazil using an innovative framework, the so called Virtual Sugarcane Biorefinery. Results showed that optimization technologies presents potential for reducing environmental impacts and improve economic results in comparison to base scenarios for both annexed and autonomous plants. It was also observed that annexed plants that diverted more sugarcane juice for sugar production were more profitable, considering the average prices for the past 10 years in Brazil. In addition, results indicate that scenarios considering more flexibility in annexed plant are more profitable than the conventional annexed one (diverting 50% of the sugarcane juice to sugar and 50% to ethanol production) if increases in prices were to occur. This study shows quantitatively the benefits of optimization techniques and allows understanding the real benefits of the sugarcane plant flexibility in the Brazilian context.Full Text: www.springerlink.com/content/b34763660112518h/

Addresses: [Cavalett, Otavio; Junqueira, Tassia L.; Dias, Marina O. S.; Jesus, Charles D. F.; Mantelatto, Paulo E.; Cunha, Marcelo P.; Franco, Henrique C. J.; Cardoso, Terezinha F.; Maciel Filho, Rubens; Rossell, Carlos E. V.; Bonomi, Antonio] Lab Nacl Ciencia Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil [Junqueira, Tassia L.; Dias, Marina O. S.; Maciel Filho, Rubens] Univ Estadual Campinas, Fac Engn Quim, BR-13083852 Campinas, SP, Brazil

24. Carbon source pulsed feeding to attain high yield and high productivity in poly(3-hydroxybutyrate) (PHB) production from soybean oil using Cupriavidus necator

Author(s): Pradella, JGD; Ienczak, JL ; Delgado, CR; Taciro, MK | E-mail: jpradella@bioetanol.org.brSource: BIOTECHNOLOGY LETTERS Volume: 34 Issue: 6 Pages: 1003-1007 | DOI: 10.1007/s10529-012-0863-1 Published: JUN 2012

Abstract: Poly(3-hydroxybutyrate) (PHB) biosynthesis from soybean oil by Cupriavidus necator was studied using a bench scale bioreactor. The highest cell concentration (83 g l(-1)) was achieved using soybean oil at 40 g l(-1) and a pulse of the same concentration. The PHB content was 81% (w/w), PHB productivity was 2.5 g l(-1) h(-1), and the calculated Y-p/s value was 0.85 g g(-1). Growth limitation and the onset of PHB biosynthesis took place due to exhaustion of P, and probably also Cu, Ca, and Fe.Full Text: http://www.springerlink.com/content/1r81n10176656441/?MUD=MP

Addresses: [da Cruz Pradella, Jose Geraldo; Ienczak, Jaciane Lutz] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol, BR-13083970 Campinas, SP, Brazil[Delgado, Cecilia Romero] Inst Pesquisas Tecnol Estado Sao Paulo SA, Lab Biotecnol Ind, BR-05508901 Sao Paulo, Brazil[Taciro, Marilda Keico] Univ Sao Paulo, Dept Microbiol, Inst Ciencias Biomed, BR-05508900 Sao Paulo, Brazil

23. Insights on How the Activity of an Endoglucanase Is Affected by Physical Properties of

Insoluble Celluloses

Author(s): Bragatto, J; Segato, F; Cota, J; Mello, DB; Oliveira, MM; Buckeridge, MS; Squina, FM; Driemeier, C | E-mail: carlos.driemeier@bioetanol.org.br Source: JOURNAL OF PHYSICAL CHEMISTRY B Volume: 116 Issue: 21 Pages: 6128-6136 | DOI:10.1021/jp3021744 | Published: MAY 31 2012

Abstract: Cellulose physical properties like crystallinity, porosity, and particle size are known to influence cellulase activity, but knowledge is still insufficient for activity prediction from such measurable substrate characteristics. With the aim of illuminating enzyme substrate relationships, this work evaluates a purified hyperthermophilic endo-1,4-beta-glucanase (from Pyrococcus furiosus) acting on 13 celluloses characterized for crystallinity and crystal width (by X-ray diffraction), wet porosity (by thermoporometry), and particle size (by light scattering). Activities are analyzed by the Michaelis-Menten kinetic equation, which is justified by low enzyme substrate affinity. Michaelis-Menten coefficients K-m and k(cat) are reinterpreted in the context of heterogeneous cellulose hydrolysis. For a set of as-received and milled microcrystalline celluloses, activity is successfully described as a function of accessible concentration, with accessibility proportional to K-m(-1). Accessibility contribution from external particle areas, pore areas, and crystalline packing are discriminated to have comparable magnitudes, implying that activity prediction demands all these substrate properties to be considered. Results additionally suggest that looser crystalline packing increases the lengths of released cello-oligomers as well as the maximum endoglucanase specific activity (k(cat)).Full Text: http://pubs.acs.org/doi/abs/10.1021/jp3021744

Addresses: [Bragatto, Juliano; Segato, Fernando; Cota, Junio; Mello, Danilo B.; Oliveira, Marcelo M.; Buckeridge, Marcos S.; Squina, Fabio M.; Driemeier, Carlos] CTBE, Lab Nacl Ciencia & Tecnol Bioetanol, BR-13083970 Sao Paulo, Brazil

22. Functional Characterization of an Aspergillus fumigatus Calcium Transporter (PmcA) that Is Essential for Fungal Infection

Author(s): Dinamarco, TM; Freitas, FZ; Almeida, RS; Brown, NA; dos Reis, TF; Ramalho, LNZ; Savoldi, M; Goldman, MHS; Bertolini, MC; Goldman, GH | E-mail: gustavo.goldman@bioetanol.org.br Source: PLOS ONE Volume: 7 Issue: 5 Article Number: e37591 | DOI: 10.1371/journal.pone.0037591 | Published: MAY

Abstract: Aspergillus fumigatus is a primary and opportunistic pathogen, as well as a major allergen, of mammals. The Ca+2-calcineurin pathway affects virulence, morphogenesis and antifungal drug action in A. fumigatus. Here, we investigated three components of the A. fumigatus Ca+2-calcineurin pathway, pmcA,-B, and -C, which encode calcium transporters. We demonstrated that CrzA can directly control the mRNA accumulation of the pmcA-C genes by binding to their promoter regions. CrzA-binding experiments suggested that the 5'-CACAGCCAC-3' and 5'-CCCTGCCCC-3' sequences upstream of pmcA and pmcC genes, respectively, are possible calcineurin-dependent response elements (CDREs)-like consensus motifs. Null mutants were constructed for pmcA and -beta and a conditional mutant for pmcC demonstrating pmcC is an essential gene. The Delta pmcA and Delta pmcB mutants were more sensitive to calcium and resistant to manganese and cyclosporine was able to modulate the sensitivity or resistance of these mutants to these salts, supporting the interaction between calcineurin and the function of these transporters. The pmcA-C genes have decreased mRNA abundance into the alveoli in the Delta calA and Delta crzA mutant strains. However, only the A. fumigatus Delta pmcA was avirulent in the murine model of invasive pulmonary aspergillosis.Full Text: http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0037591

Addresses: [Goldman, Gustavo Henrique] Lab Nacl Ciencia & Tecnol Bioetanol CTBE, Sao Paulo, Brazil[Dinamarco, Taisa Magnani; Brown, Neil Andrew; dos Reis, Thaila Fernanda; Savoldi, Marcela; Goldman, Gustavo Henrique] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Sao Paulo, Brazil[Goldman, Maria Helena S.] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Sao Paulo, Brazil[Zambelli Ramalho, Leandra Naira] Univ Sao Paulo, Fac Med Ribeirao Preto, Sao Paulo, Brazil[Almeida, Ricardo S.] Univ Londrina, Dept Microbiol, Londrina, Parana, Brazil[Freitas, Fernanda Zanolli; Bertolini, Maria Celia] Univ Estadual Paulista, Inst Quim, Sao Paulo, Brazil

21. Configuration-Dependent Diffusion Dynamics of Downhill and Two-State Protein Folding

Author(s): Xu, WX; Lai, ZZ; Oliveira, RJ; Leite, VBP; Wang, J | E-mail: ronaldo.oliveira@bioetanol.org.br   Source: JOURNAL OF PHYSICAL CHEMISTRY B Volume: 116 Issue: 17 Pages: 5152-5159 | DOI: 10.1021/jp212132v | Published: MAY 3 2012

Abstract: Configuration-dependent diffusion (CDD) is important for protein folding kinetics with small thermodynamic barriers. CDD can be even more crucial in downhill folding without thermodynamic barriers. We explored the CDD of a downhill protein (BBL), and a two-state protein (CI2). The hidden kinetic barriers due to CDD were revealed. The increased similar to 1 k(B)T kinetic barrier is in line with experimental value based on other fast folding proteins. Compared to that of CI2, the effective free-energy profile of BBL is found to be significantly influenced by CDD, and the kinetics are totally determined by diffusion. These findings are consistent with both earlier bulk and single-molecule fluorescence measurements. In addition, we found the temperature dependence of CDD. We also found that the ratio of folding transition temperature against optimal kinetic folding temperature can provide both a quantitative measure for the underlying landscape topography and an indicator for the possible appearance of downhill folding. Our study can help for a better understanding of the role of diffusion in protein folding dynamics.Full Text: http://pubs.acs.org/doi/abs/10.1021/jp212132v

Addresses: [Xu, Weixin; Lai, Zaizhi; Wang, Jin] SUNY Stony Brook, Dept Chem, Stony Brook, NY 11794 USA[Xu, Weixin; Lai, Zaizhi; Wang, Jin] SUNY Stony Brook, Dept Phys, Stony Brook, NY 11794 USA[Xu, Weixin] E China Normal Univ, Dept Phys, State Key Lab Precis Spect, Shanghai 200062, Peoples R China[Xu, Weixin] E China Normal Univ, Inst Theoret & Computat Sci, Inst Adv Interdisciplinary Res, Shanghai 200062, Peoples R China[Oliveira, Ronaldo J.; Leite, Vitor B. P.] Univ Estadual Paulista, Inst Biociencias Letras & Ciencias Exatas, Dept Fis, BR-15054000 Sao Jose Do Rio Preto, SP, Brazil[Oliveira, Ronaldo J.] Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil[Wang, Jin] Chinese Acad Sci, Changchun Inst Appl Chem, State Key Lab Electroanalyt Chem, Changchun 130021, Jilin, Peoples R China

20. Sugarwin: A Sugarcane Insect-Induced Gene with Antipathogenic Activity

Author(s): Medeiros, AH; Franco, FP; Matos, JL; de Castro, PA; Santos-Silva, LK; Henrique-Silva, F; Goldman, GH; Moura, DS; Silva, MC. | E-mail: gustavo.goldman@bioetanol.org.brSource: MOLECULAR PLANT-MICROBE INTERACTIONS Volume: 25 Issue: 5 Pages: 613-624 DOI: 10.1094/MPMI-09-11-0254 | Published: MAY 2012

Abstract: In sugarcane fields, colonization of the stalk by opportunistic fungi usually occurs after the caterpillar Diatraea saccharalis attacks the sugarcane plant. Plants respond to insect attack by inducing and accumulating a large set of defense proteins. Two homologues of a barley wound-inducible protein (BARWIN), sugarcane wound-inducible proteins SUGARWIN1 and SUGARWIN2, have been identified in sugarcane by an in silico analysis. Antifungal properties have been described for a number of BARWIN homologues. We report that a SUGARWIN::green fluorescent protein fusion protein is located in the endoplasmic reticulum and in the extracellular space of sugarcane plants. The induction of sugarwin transcripts occurs in response to mechanical wounding, D. saccharalis damage, and methyl jasmonate treatment. The accumulation of transcripts is late induced and is restricted to the site of the wound. Although the transcripts of sugarwin genes were strongly increased following insect attack, the protein itself did not show any effect on insect development; rather, it altered fungal morphology, leading to the apoptosis of the germlings. These results suggest that, in the course of evolution, sugarwinencoding genes were recruited by sugarcane due to their antipathogenic activity. We rationalize that sugarcane is able to induce sugarwin gene expression in response to D. saccharalis feeding as a concerted plant response to the anticipated invasion by the fungi that typically penetrate the plant stalk after insect damage.Full Text: http://apsjournals.apsnet.org/doi/abs/10.1094/MPMI-09-11-0254

Addresses: [Medeiros, Ane H.; Franco, Flavia P.; Matos, Juliana L.; Silva-Filho, Marcio C.] Univ Sao Paulo, Escola Super Agr Luiz de Queiroz, Dept Genet, BR-13418900 Piracicaba, SP, Brazil[de Castro, Patricia A.; Goldman, Gustavo H.] Univ Sao Paulo, Fac Ciencias Farmaceut, BR-14049 Ribeirao Preto, SP, Brazil [Santos-Silva, Ludier K.; Henrique-Silva, Flavio] Univ Fed Sao Carlos, Dept Genet & Evolucao, BR-13560 Sao Carlos, SP, Brazil[Goldman, Gustavo H.] Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil[Moura, Daniel] Univ Sao Paulo, Dept Ciencias Biol, Escola Super Agr Luiz de Queiroz, BR-13418900 Piracicaba, SP, Brazil

19. Mass balance of pilot-scale pretreatment of sugarcane bagasse by steam explosion followed by alkaline delignification

Author(s): Rocha, GJM; Martin, C; da Silva, VFN; Gomez, EO; Goncalves, AR | E-mail: george.rocha@bioetanol.org.br Source: BIORESOURCE TECHNOLOGY Volume: 111 Pages: 447-452 | DOI: 10.1016/j.biortech.2012.02.005 | Published: MAY 2012

Abstract: Five pilot-scale steam explosion pretreatments of sugarcane bagasse followed by alkaline delignification were explored. The solubilised lignin was precipitated with 98% sulphuric acid. Most of the pentosan (82.6%), and the acetyl group fractions were solubilised during pretreatment, while 90.2% of cellulose and 87.0% lignin were recovered in the solid fraction. Approximately 91% of the lignin and 72.5% of the pentosans contained in the steam-exploded solids were solubilised by delignification, resulting in a pulp with almost 90% of cellulose. The acidification of the black liquors allowed recovery of 48.3% of the lignin contained in the raw material. Around 14% of lignin, 22% of cellulose and 26% of pentosans were lost during the process. In order to increase material recovery, major changes, such as introduction of efficient condensers and the reduction in the number of washing steps, should be done in the process setup.Full Text: www.sciencedirect.com/science/article/pii/S0960852412002210

Addresses: [Rocha, George J. M.; Gomez, Edgardo O.] Brazilian Bioethanol Sci & Technol Natl Lab CTBE, BR-13083970, Campinas, SP, Brazil[Martin, Carlos] Univ Matanzas, Dept Chem & Chem Engn, Matanzas 44740, Cuba

[Martin, Carlos] VTI Inst Wood Technol & Wood Biol, D-21031 Hamburg, Germany[da Silva, Vinicius F. N.; Goncalves, Adilson R.] Univ Sao Paulo, Escola Engn Lorena, Dept Biotecnol, BR-12602810 Lorena, SP, Brazil

18. Two structurally discrete GH7-cellobiohydrolases compete for the same cellulosic substrate fiber

Author(s): Segato, F; Damasio, ARL; Goncalves, TA; Murakami, MT; Squina, FM; Polizeli, MDTM; Mort, AJ; Prade, RA | E-mail: fabio.squina@bioetanol.org.br Source: BIOTECHNOLOGY FOR BIOFUELS Volume: 5 Article Number: 21 | DOI: 10.1186/1754-6834-5-21 | Published: APR 11 2012

Abstract: Background: Cellulose consisting of arrays of linear beta-1,4 linked glucans, is the most abundant carbon-containing polymer present in biomass. Recalcitrance of crystalline cellulose towards enzymatic degradation is widely reported and is the result of intra-and inter-molecular hydrogen bonds within and among the linear glucans. Cellobiohydrolases are enzymes that attack crystalline cellulose. Here we report on two forms of glycosyl hydrolase family 7 cellobiohydrolases common to all Aspergillii that attack Avicel, cotton cellulose and other forms of crystalline cellulose. Results: Cellobiohydrolases Cbh1 and CelD have similar catalytic domains but only Cbh1 contains a carbohydrate-binding domain (CBD) that binds to cellulose. Structural superpositioning of Cbh1 and CelD on the Talaromyces emersonii Cel7A 3-dimensional structure, identifies the typical tunnel-like catalytic active site while Cbh1 shows an additional loop that partially obstructs the substrate-fitting channel. CelD does not have a CBD and shows a four amino acid residue deletion on the tunnelobstructing loop providing a continuous opening in the absence of a CBD. Cbh1 and CelD are catalytically functional and while specific activity against Avicel is 7.7 and 0.5 U. mg prot-1, respectively specific activity on pNPC is virtually identical. Cbh1 is slightly more stable to thermal inactivation compared to CelD and is much less sensitive to glucose inhibition suggesting that an open tunnel configuration, or absence of a CBD, alters the way the catalytic domain interacts with the substrate. Cbh1 and CelD enzyme mixtures on crystalline cellulosic substrates show a strong combinatorial effort response for mixtures where Cbh1 is present in 2: 1 or 4: 1 molar excess. When CelD was overrepresented the combinatorial effort could only be partially overcome. CelD appears to bind and hydrolyze only loose cellulosic chains while Cbh1 is capable of opening new cellulosic substrate molecules away from the cellulosic fiber. Conclusion: Cellobiohydrolases both with and without a CBD occur in most fungal genomes where both enzymes are secreted, and likely participate in cellulose degradation. The fact that only Cbh1 binds to the substrate and in combination with CelD exhibits strong synergy only when Cbh1 is present in excess, suggests that Cbh1 unties enough chains from cellulose fibers, thus enabling processive access of CelD.Full Text: http://www.biotechnologyforbiofuels.com/content/5/1/21

Addresses: [Segato, Fernando; Damasio, Andre R. L.; Prade, Rolf A.] Oklahoma State Univ, Dept Microbiol & Mol Genet, Stillwater, OK 74078 USA[Segato, Fernando; Goncalves, Thiago Augusto; Squina, Fabio M.; Prade, Rolf A.] Ctr Nacl Pesquisas Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, Sao Paulo, Brazil[Mort, Andrew J.] Oklahoma State Univ, Dept Biochem & Mol Biol, Stillwater, OK 74078 USA[Murakami, Mario T.] Lab Nacl Biociencias LNBio, Sao Paulo, Brazil[Damasio, Andre R. L.] Ribeirao Preto Sch Med, Dept Biochem, Sao Paulo, Brazil[Polizeli, Maria de Lourdes T. M.] Univ Sao Paulo, FFCLRP, Biol Dept, Sao Paulo, Brazil

17. Molecular Characterization of the Putative Transcription Factor SebA Involved in Virulence in Aspergillus fumigates

Author(s): Dinamarco, TM; Almeida, RS; de Castro, PA; Brown, NA; dos Reis, TF, T; Ramalho, LNZ; Savoldi, M; Goldman, MHS; Goldman, GH | E-mail: gustavo.goldman@bioetanol.org.brSource: EUKARYOTIC CELL Volume: 11 Issue: 4 Pages: 518-531 | DOI: 10.1128/EC.00016-12 | Published: APR 2012

Abstract: Aspergillus fumigatus is a major opportunistic pathogen and allergen of mammals. Nutrient sensing and acquisition mechanisms, as well as the capability to cope with different stressing conditions, are essential for A. fumigatus virulence and survival in the mammalian host. This study characterized the A. fumigatus SebA transcription factor, which is the putative homologue of the factor encoded by Trichoderma atroviride seb1. The Delta sebA mutant demonstrated reduced growth in the presence of paraquat, hydrogen peroxide, CaCl2, and poor nutritional conditions, while viability associated with sebA was also affected by heat shock exposure. Accordingly, SebA::GFP (SebA::green fluorescent protein) was shown to accumulate in the nucleus upon exposure to oxidative stress and heat shock conditions. In addition, genes involved in either the oxidative stress or heat shock response had reduced transcription in the Delta sebA mutant. The A. fumigatus Delta sebA strain was attenuated in virulence in a murine model of invasive pulmonary aspergillosis. Furthermore, killing of the Delta sebA mutant by murine alveolar macrophages was increased compared to killing of the wild-type strain. A. fumigatus SebA plays a complex ole, contributing to several stress tolerance pathways and growth under poor nutritional conditions, and seems to be integrated into different stress responses.Full Text: http://ec.asm.org/content/11/4/518

Addresses: [Goldman, Gustavo Henrique] Lab Nacl Ciencia & Tecnol Bioetanol CTB, Sao Paulo, Brazil[Zambelli Ramalho, Leandra Naira] Univ Sao Paulo, Fac Med Ribeirao Preto, Sao Paulo, Brazil[Goldman, Maria Helena S.] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Sao Paulo, Brazil[Dinamarco, Taisa Magnani; Almeida, Ricardo S.; de Castro, Patricia Alves; Brown, Neil Andrew; dos Reis, Thaila Fernanda; Savoldi, Marcela; Goldman, Gustavo Henrique] Univ Sao Paulo, Fac Ciencias Farmaceut, Sao Paulo, Brazil

16. Metastable atomic species in the N-2 flowing afterglow

Author(s): Levaton, J ; Amorim, J | E-mail: jayr.amorim@bioetanol.org.br Source: CHEMICAL PHYSICS Volume: 397 Issue: 1 Pages: 9-17 | DOI: 10.1016/j.chemphys.2011.11.010 | Published: MAR 13 2012

Abstract: We have studied by optical emission spectroscopy the post-discharge of a pure N-2 DC flowing discharge in such experimental conditions that the pink afterglow and the Lewis-Rayleigh afterglow occur. The emission profiles originated from the N-2(B-3 Pi(g)), N-2(C-3 Pi(u)) and N-2(+)(B-2 Sigma(+)(u)) states and the N-2(B-3 Pi(g),6 <= v <= 12) and N-2(C-3 Pi(u), 0 <= v <= 4) vibrational distributions were obtained in the post-discharge region. With basis on the works of Bockel et al. [S. Bockel, A. M. Diamy, A. Ricard, Surf. Coat. Tech. 74 (1995) 474] and Amorim and Kiohara [J. Amorim, V. Kiohara, Chem. Phys. Lett. 385 (2004) 268], we have obtained the experimental N(S-4) and N(D-2) relative densities along the post-discharge. A numerical model, previously developed to describe the neutral atomic, molecular and ionic species in the afterglow, was improved to include the kinetics of N(D-2) and N(P-2) states. Several kinetic mechanisms leading to the production of N(D-2) in the post-discharge have been studied in order to explain the experimental data. We have determined that the dominant one is the reaction N-2 (X-1 Sigma(+)(g), v > 8) + N(S-4) -> N-2(X-1 Sigma(+)(g)) + N(D-2) with an estimated rate constant of 7 x 10 (14) cm(3) s (1). Also, the fit of the numerical density profiles of N-2 (C-3 Pi(u)) and N-2(+)(B-2 Sigma(+)(u)) to the experimental ones has provided the rate constant for reaction N-2 (A(3)Sigma(+)(u)) + N-2 (X-1 Sigma(+)(g), v > 18) -> N-2(C-3 Pi(u)) + N-2(X-

1 Sigma(+)(g)). Its estimated value is 4 x 10 (13) cm(3) s (1). Finally, with the new kinetic scheme, we have found that the ionization in the post-discharge region has important contribution of N(D-2) and N(P-2) species.Full Text: www.sciencedirect.com/science/article/pii/S0301010411005015

Addresses: [Levaton, J.; Amorim, J.] Lab Nacl Ciencia & Tecnol Bioetanol CTBE CNPEM, BR-13083970 Campinas, P, Brazil

15. Ammonia Losses Estimated by an Open Collector From Urea Applied to Sugarcane Straw

Author(s): Mariano, E; Trivelin, PCO; Vieira, MX; Leite, JM; Otto, R; Franco, HCJ | E-mail: henrique.franco@bioetanol.org.br Source: REVISTA BRASILEIRA DE CIENCIA DO SOLO Volume: 36 Issue: 2 Pages: 411-419 Published: MAR-APR 2012

Abstract: The quantification of ammonia (NH3) losses from sugarcane straw fertilized with urea can be performed with collectors that recover the NH3 in acid-treated absorbers. Thus, the use of an open NH3 collector with a polytetrafluoroethylene (PTFE)-wrapped absorber is an interesting option since its cost is low, handling easy and microclimatic conditions irrelevant. The aim of this study was to evaluate the efficiency of an open collector for quantifying NH3-N volatilized from urea applied over the sugarcane straw. The experiment was carried out in a sugarcane field located near Piracicaba, Sao Paulo, Brazil. The NH3-N losses were estimated using a semi-open static collector calibrated with N-15 (reference method) and an open collector with an absorber wrapped in PTFE film. Urea was applied to the soil surface in treatments corresponding to rates of 50, 100, 150 and 200 kg ha(-1) N. Applying urea-N fertilizer on sugarcane straw resulted in losses NH3-N up to 24 % of the applied rate. The amount of volatile NH3-N measured in the open and the semi-open static collector did not differ. The effectiveness of the collection system varied non-linearly, with an average value of 58.4 % for the range of 100 to 200 kg ha(-1) of urea-N. The open collector showed significant potential for use; however, further research is needed to verify the suitability of the proposed method.

Addresses: [Mariano, Eduardo; Vieira, Michele Xavier; Leite, Jose Marcos; Otto, Rafael] Univ Sao Paulo, Luiz de Queiroz Coll Agr, BR-13418900 Piracicaba, SP, Brazil[Ocheuze Trivelin, Paulo Cesar] Univ Sao Paulo, Ctr Nucl Energy Agr, BR-13416000 Piracicaba, SP, Brazil[Junqueira Franco, Henrique Coutinho] Brazilian Res Ctr Energy & Mat UNPEM, Brazilian Bioethanol Sci & Technol Lab,

14. Use of a new Trichoderma harzianum strain isolated from the Amazon rainforest with pretreated sugar cane bagasse for on-site cellulase production

Author(s): Delabona, PD; Farinas, CS; da Silva, MR; Azzoni, SF; Pradella, JGD Source: BIORESOURCE TECHNOLOGY Volume: 107 Pages: 517-521 | DOI: 10.1016/j.biortech.2011.12.048 | Published: MAR 2012 | E-mail: jpradella@bioetanol.org.br

Abstract: The on-site production of cellulases is an important strategy for the development of sustainable second-generation ethanol production processes. This study concerns the use of a specific cellulolytic enzyme complex for hydrolysis of pretreated sugar cane bagasse. Glycosyl hydrolases (FPase, xylanase, and beta-glucosidase) were produced using a new strain of Trichoderma harzianum, isolated from the Amazon rainforest and cultivated under different conditions. The influence of the carbon source was first investigated using shake-flask cultures. Selected carbon

sources were then further studied under different pH conditions using a stirred tank bioreactor. Enzymatic activities up to 121 FPU/g, 8000IU/g, and 730IU/g of delignified steam-exploded bagasse + sucrose were achieved for cellulase, xylanase and beta-glucosidase, respectively. This enzymatic complex was used to hydrolyze pretreated sugar cane bagasse. A comparative valuation, using an enzymatic extract from Trichoderma reesei RUTC30, indicated similar performance of the T. harzianum enzyme complex, being a potential candidate for on-site production of enzymes. Full Text: www.sciencedirect.com/science/article/pii/S0960852411017949

Addresses: [Delabona, Priscila da Silva; da Silva, Mateus Ribeiro; Azzoni, Sindelia Freitas; da Cruz Pradella, Jose Geraldo] Brazilian Bioethanol Sci & Technol Lab CTBE, Ind Program, BR-13083970 Campinas, SP, Brazil[Farinas, Cristiane Sanchez] Embrapa Instrumentat, BR-13560970 Sao Paulo, Brazil

13. The COP9 signalosome counteracts the accumulation of cullin SCF ubiquitin E3 RING ligases during fungal development

Author(s): Kress, MRV; Harting, R; Bayram, O; Christmann, M; Irmer, H; Valerius, O; Schinke, J; Goldman, GH; Braus, GH | E-mail: gustavo.goldman@bioetanol.org.br Source: MOLECULAR MICROBIOLOGY Volume: 83 Issue: 6 Pages: 1162-1177 | DOI: 10.1111/j.1365-2958.2012.07999.x | Published: MAR 2012

Abstract: Defects in the COP9 signalosome (CSN) impair multicellular development, including embryonic plant or animaldeath or a block in sexual development of the fungus Aspergillus nidulans. CSN deneddylates cullin-RING ligases (CRLs), which are activated by covalent linkage to ubiquitin-like NEDD8. Deneddylation allows CRL disassembly for subsequent reassembly. An attractive hypothesis is a consecutive order of CRLs for development, which demands repeated cycles of neddylation and deneddylation for reassembling CRLs. Interruption of these cycles could explain developmental blocks caused by csn mutations. This predicts an accumulation of neddylated CRLs exhibiting developmental functions when CSN is dysfunctional. We tested this hypothesis in A. nidulans, which tolerates reduced levels of neddylation for growth. We show that only genes for CRL subunits or neddylation are essential, whereas CSN is primarily required for development. We used functional tagged NEDD8, recruiting all three fungal cullins. Cullins are associated with the CSN1/CsnA subunit when deneddylation is defective. Two CRLs were identified which are specifically involved in differentiation and accumulate during the developmental block. This suggests that an active CSN complex is required to counteract the accumulation of specific CRLs during development.Full text: http://onlinelibrary.wiley.com/doi/10.1111/j.1365-2958.2012.07999.x/full

Addresses: [Kress, Marcia Regina von Zeska; Harting, Rebekka; Bayram, Ozgur; Christmann, Martin; Irmer, Henriette; Valerius, Oliver; Schinke, Josua; Braus, Gerhard H.] Univ Gottingen, Inst Mikrobiol & Genet, D-37077 Gottingen, Germany[Goldman, Gustavo H.] Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil[Goldman, Gustavo H.] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Sao Paulo, Brazil

12. Functional characterization and oligomerization of a recombinant xyloglucan-specific endo-beta-1,4-glucanase (GH12)from Aspergillus niveus

Author(s): Damasio, ARL; Ribeiro, LFC; Ribeiro, LF; Furtado, GP; Segato, F; Almeida, FBR; Crivellari, AC; Buckeridge, MS; Souza, TACB; Murakami, MT; Ward, RJ; Prade, RA ; Polizeli, MLTM | E-mail: marcos.buckeridge@bioetanol.org.br

Source: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS Volume: 1824 Issue: 3 Pages: 461-467 | DOI: 10.1016/j.bbapap.2011.12.005 | Published: MAR 2012

Abstract: Xyloglucan is a major structural polysaccharide of the primary (growing) cell wall of higher plants. It consists of a cellulosic backbone (beta-1,4-linked glucosyl residues) that is frequently substituted with side chains. This report describes Aspergillus nidulans strain A773 recombinant secretion of a dimeric xyloglucan-specific endo-beta-1,4-glucanohydrolase (XegA) cloned from Aspergillus niveus. The ORF of the A. niveus xegA gene is comprised of 714 nucleotides, and encodes a 238 amino acid protein with a calculated molecular weight of 23.5 kDa and isoelectric point of 4.38. The optimal pH and temperature were 6.0 and 60 degrees C, respectively. XegA generated a xyloglucan-oligosaccharides (XGOs) pattern similar to that observed for cellulases from family GH12, i.e., demonstrating that its mode of action includes hydrolysis of the glycosidiclinkages between glucosyl residues that are not branched with xylose. In contrast to commercial lichenase, mixed linkagebeta-glucan (lichenan) was not digested by XegA, indicating that the enzyme did not cleave glucan beta-1,3 or beta-1,6 bonds.The far-UV CD spectrum of the purified enzyme indicated a protein rich in beta-sheet structures as expected for GH12 xyloglucanases. Thermal unfolding studies displayed two transitions with mid-point temperatures of 51.3 degrees C and 81.3degrees C respectively, and dynamic light scattering studies indicated that the first transition involves a change in oligomeric state from a dimeric to a monomeric form. Since the enzyme is a predominantly a monomer at 60 degrees C. the enzymatic assays demonstrated that XegA is more active in its monomeric state.Full Text: www.sciencedirect.com/science/article/pii/S1570963911003220

Addresses: [Polizeli, Maria L. T. M.] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Biol, BR-14040901 Ribeirao Preto, SP, Brazil [Damasio, Andre R. L.; Ribeiro, Liliane F. C.; Ribeiro, Lucas F.; Furtado, Gilvan P.] Univ Sao Paulo, Dept Bioquim & Imunol, FMRP, BR-14040901 Ribeirao Preto, SP, Brazil[Segato, Fernando; Buckeridge, Marcos S.] Ctr Nacl Pesquisas Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, Campinas, SP, Brazil[Almeida, Fausto B. R.] Univ Sao Paulo, Dept Biol Celular & Mol & Bioagentes Patogen, FMRP, BR-14040901 Ribeirao Preto, SP, Brazil[Crivellari, Augusto C.; Buckeridge, Marcos S.] Univ Sao Paulo, Inst Biociencias, Dept Bot, Lab Fisiol Ecol Plantas, BR-14040901 Ribeirao Preto, SP, Brazil[Souza, Tatiana A. C. B.; Murakami, Mario T.] Ctr Nacl Pesquisas Energia & Mat, Lab Nacl Biociencias, Campinas, SP, Brazil[Ward, Richard J.] Univ Sao Paulo, Dept Quim, FFCLRP USP, BR-14040901 Ribeirao Preto, SP, Brazil[Prade, Rolf A.] Oklahoma State Univ, Dept Microbiol & Mol Genet, Stillwater, OK 74078 USA

11. Characterization of depolymerized residues from extremely low acid hydrolysis (ELA) of sugarcane bagasse cellulose: Effects of degree of polymerization, crystallinity and crystallite size on thermal decomposition

Author(s): Gurgel, LVA; Marabezi, K; Ramos, LA; Curvelo, AAD.| E-mail: karen.marabezi@bioetanol.org.br Source: INDUSTRIAL CROPS AND PRODUCTS Volume: 36 Issue: 1 Pages: 560-571 | DOI:10.1016/j.indcrop.2011.11.009 | Published: MAR 2012

Abstract: Sugarcane bagasse cellulose was subjected to the extremely low acid (ELA) hydrolysis in 0.07% H2SO4 at 90, 210 and 225 degrees C for various times. The cellulose residues from this process were characterized by TGA, XRD, PC, FIR and SEM. A kinetic study of thermal decomposition of the residues was also carried out, using the ASTM and issinger methods. The thermal studies revealed that residues of cellulose hydrolyzed at 190, 210 and 225 degrees C for

80,40 and 8 min have initial decomposition temperature and activation energy for the main decomposition step similar to those of Avicel PH-101. XRD studies confirmed this finding by showing that these cellulose residues are similar to Avicel in rystallinity index and crystallite size in relation to the 110 and 200 planes. FTIR spectra revealed no significant changes in the cellulose chemical structure and analysis of SEM micrographs demonstrated that the particle size of the cellulose residues hydrolyzed at190 and 210 degrees C were similar to that of Avicel.Full Text: http://www.sciencedirect.com/science/article/pii/S0926669011004365

Addresses: [Alves Gurgel, Leandro Vinicius; Marabezi, Karen; Ramos, Luiz Antonio; da Silva Curvelo, Antonio Aprigio] Univ Sao Paulo, Inst Quim Sao Carlos, Dept Quim Fis, Grp Fis Quim Organ, BR-13560970 Sao Carlos, SP, Brazil [da Silva Curvelo, Antonio Aprigio] Ctr Pesquisa Energia & Mat CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil

10. Ethanol from sugarcane in Brazil: a midway' strategy for increasing ethanol production while maximizing environmental benefits

Author(s): Buckeridge, M; De Souza, AP; Arundale, RA, Rebecca Teixeira, KJ ; DeLucia, E. | E-mail: marcos.buckeridge@bioetanol.org.br Source: GLOBAL CHANGE BIOLOGY BIOENERGY Volume: 4 Issue: 2 Pages: 119-126 | DOI:10.1111/j.1757-1707.2011.01122.x | Published: MAR 2012

Abstract: This article reviews the history and current state of ethanol production from sugarcane in Brazil and presents a strategy for improving ecosystem services and production. We propose that it is possible to produce ethanol from sugarcane while maintaining or even recovering some of Brazil's unique neotropical biodiversity and ecosystem climate services. This approach to the future of sustainable and responsible ethanol production is termed the midway strategy. The midway strategy involves producing the necessary biotechnology to increase productivity while synergistically protecting and regenerating rainforest. Three main areas of scientific and technological advance that are key to realizing the midway strategy are: (i) improving the quality of scientific data on sugarcane biology as pertains to its use as a bioenergy crop; (ii) developing technologies for the use of bagasse for cellulosic ethanol; and (iii) developing policies to improve the ecosystem services associated with sugarcane landscapes. This article discusses these three issues in the general context of biofuels production andhighlights examples of scientific achievements that are already leading towards the midway strategy.Full Text: www.onlinelibrary.wiley.com/doi/10.1111/j.1757-1707.2011.01122.x/full

Addresses: [Buckeridge, Marcos S.; De Souza, Amanda P.] Univ Sao Paulo, Lab Fisiol Ecol Plantas, LAFIECO, Dept Bot,Inst Biociencias, BR-05508 Sao Paulo, Brazil[Buckeridge, Marcos S.] CTBE, Brazilian Bioethanol Sci & Technol Lab, Campinas, SP, Brazil[Arundale, Rebecca A.; Anderson-Teixeira, Kristina J.; DeLucia, Evan] Univ Illinois, Dept Plant Biol, Urbana, IL 61801 USA

9. Molecular insights into substrate specificity and thermal stability of a bacterial GH5-CBM27 endo-1,4-beta-Dmannanase

Author(s): dos Santos, CR; Paiva, JH; Meza, AN; Cota, J; Alvarez, TM; Ruller, R; Prade, RA; Squina, FM; Murakami, MT | E-mail: fabio.squina@bioetanol.org.brSource: JOURNAL OF STRUCTURAL BIOLOGY Volume: 177 Issue: 2 Pages: 469-476 | DOI:10.1016/j.jsb.2011.11.021 | Published: FEB 2012

Abstract: The breakdown of beta-1,4-mannoside linkages in a variety of mannan-containing polysaccharides is of great importance in industrial processes such as kraft pulp delignification, food processing and production of second-generation biofuels, which puts a premium on studies regarding the prospection and engineering of beta-mannanases. In this work, a two-domain beta-mannanase from Thermotoga petrophila that encompasses a GH5 catalytic domain with a C-terminal CBM27 accessory domain, was functionally and structurally characterized. Kinetic and thermal denaturation experiments showed that the CBM27 domain provided thermo-protection to the catalytic domain, while no contribution on enzymatic activity was observed. The structure of the catalytic domain determined by SIRAS revealed a canonical (alpha/beta)(8)-barrel scaffold surrounded by loops and short helices that form the catalytic interface. Several structurally related ligand molecules interacting with TpMan were solved at high-resolution and resulted in a wide-range representation of the subsites forming the active-site cleft with residues W134, E198, R200, E235, H283 and W284 directly involved in glucose binding.Full Text: www.sciencedirect.com/science/article/pii/S1047847711003376

Addresses: [dos Santos, Camila Ramos; Paiva, Joice Helena; Meza, Andreia Navarro; Murakami, Mario Tyago] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Biociencias LNBio, BR-13083970 Campinas, SP, Brazil[Cota, Junio; Alvarez, Thabata Maria; Ruller, Roberto; Squina, Fabio Marcio] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil[Prade, Rolf Alexander] Oklahoma State Univ, Dept Microbiol & Mol Genet, Stillwater, OK 74078 USA

8. Molecular characterization of the Aspergillus nidulans fbxA encoding an F-box protein involved in xylanase induction

Author(s): Colabardini, AC; Humanes, AC; Gouvea, PF; Savoldi, M; Goldman, MHS; Kress, MRV; Bayram, O; Oliveira, JVD; Gomes, MD ; Braus, GH; Goldman, GH | E-mail: gustavo.goldman@bioetanol.org.brSource: FUNGAL GENETICS AND BIOLOGY Volume: 49 Issue: 2 Pages: 130-140 | DOI: 10.1016/j.fgb.2011.11.004 Published: FEB 2012 |

Abstract: The filamentous fungus Aspergillus nidulans has been used as a fungal model system to study the regulation of xylanase production. These genes are activated at transcriptional level by the master regulator the transcriptional factor XInR and repressed by carbon catabolite repression (CCR) mediated by the wide-domain repressor CreA. Here, we screened a collection of 42 A. nidulans F-box deletion mutants grown either in xylose or xylan as the single carbon source in the presence of the glucose analog 2-deoxy-D-glucose, aiming to identify mutants that have deregulated xylanase induction. We were able to recognize a null mutant in a gene (fbxA) that has decreased xylanase activity and reduced xInA and xInD mRNA accumulation. The Delta fbxA mutant interacts genetically with creAd-30, creB15, and creC27 mutants. FbxA is a novel protein containing a functional F-box domain that binds to Skp1 from the SCF-type ligase. Blastp analysis suggested that FbxA is a protein exclusive from fungi, without any apparent homologs in higher eukaryotes. Our work emphasizes the importance of the ubiquitination in the A. nidulans xylanase induction and CCR. The identification of FbxA provides another layer of complexity to xylanase induction and CCR phenomena in filamentous fungi.Full Text: http://www.sciencedirect.com/science/article/pii/S1087184511001952

Addresses: [Goldman, Gustavo Henrique] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Dept Ciencias Farmaceut, BR-14040903 Sao Paulo, Brazil [de Castro Oliveira, Juliana Velasco; Goldman, Gustavo Henrique] Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Sao Paulo, Brazil

[Humanes, Ana Carolina; Gomes, Marcelo Darnario] Univ Sao Paulo, Fac Med Ribeirao Preto, BR-14040903 Sao Paulo, Brazil [Goldman, Maria Helena S.] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Pret, BR-14040903 Sao Paulo, Brazil [Bayram, Oezguer; Braus, Gerhard H.] Univ Gottingen, Dept Mol Microbiol & Genet, Inst Microbiol & Genet, Gottingen, Germany

7. Dilute Acid Hydrolysis of Sugar Cane Bagasse at High Temperatures: A Kinetic Study of Cellulose Saccharification and Glucose Decomposition. Part I: Sulfuric Acid as the Catalyst

Author(s): Gurgel, LVA; Marabezi, K; Zanbom, MD ; Curvelo, AAD | E-mail: karen.marabezi@bioetanol.org.br

Source: INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH Volume: 51 Issue: 3 Pages: 1173-1185 | DOI: 10.1021/ie2025739 | Published: JAN 25 2012

Abstract: The kinetics of sugar cane bagasse cellulose saccharification and the decomposition of glucose under extremely low acid (ELA) conditions, (0.07%), 0.14%, and 0.28% H2SO4, and at high temperatures were investigated using batch reactors. The first-order rate constants were obtained by weight loss, remaining glucose, and fitting glucose concentration profiles determined with HPLC using the Saeman model. The maximum glucose yields reached 67.6% (200 degrees C, 0.07% H2SO4, 30 min), 69.8% (210 degrees C, 0.14% H2SO4, 10 min), and 67.3% (210 degrees C, 0.28% H2SO4, 6 min). ELA conditions produced remarkable glucose yields when applied to bagasse cellulose. The first-order rate constants were used to calculate activation energies and extrathermodynamic parameters to elucidate the reaction mechanism under ELA conditions. The effect of acid concentration on cellulose hydrolysis and glucose decomposition was also investigated. The observed activation energies and reaction orders with respect to hydronium ion for cellulose hydrolysis and glucose decomposition were 184.9 and 124.5 kJ/mol and 1.27 and 0.75, respectively.Full Text: http://pubs.acs.org/doi/abs/10.1021/ie2025739

Addresses: [Alves Gurgel, Leandro Vinicius; Marabezi, Karen; Zanbom, Marcia Dib; da Silva Curvelo, Antonio Aprigio] Univ Sao Paulo, Inst Quim Sao Carlos, Dept Fis Quim, Grp Fis Quim Organ, BR-13560970 Sao Carlos, SP, Brazil[da Silva Curvelo, Antonio Aprigio] Ctr Pesquisa Energia & Mat CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil

6. Low liquid-solid ratio (LSR) hot water pretreatment of sugarcane bagasse

Author(s): Vallejos, ME; Zambon, MD; Area, MC;Curvelo, AAD | E-mail: aprigio.curvelo@bioetanol.org.br Source: GREEN CHEMISTRY Volume: 14 Issue: 7 Pages: 1982-1989 | DOI: 10.1039/c2gc35397k | Published: 2012

Abstract: Low liquid-solid ratio (LSR) can be used to obtain high-content xylo-oligosaccharide (XOS) spend liquor by hot water pretreatment. Developing a technology based on low LSR results in more efficient water usage in the system and thus in lower capital and operating costs. Xylans from xylan rich agro-industrial waste are abundant hemicellulosic polymers with enormous potential for industrial applications. Currently, freeze-dried xylo-oligosaccharides are used as bio-based polymers and hydrolysates containing high xylose contents are converted to several chemical products. In this study, sugarcane bagasse was treated with water at low LSRs and mild temperatures in order to assess the effects of varying the pretreatment conditions on the xylo-oligosaccharide and xylose concentrations,

and use a central composite experimental design to optimize the process parameters. The pretreatments were performed in the ranges temperature: 143.3-176.7 degrees C, time: 20-70 min and LSR: 1 : 1 to 11 : 1 (g g(-1)). The maximum concentrations of xylose and xylan were 13.76 and 36.18 g L-1 (equivalent to 48.29 g L-1 of xylan), respectively, which were achieved by treating bagasse at 170 degrees C for 60 min, with LSR of 3 g g(-1). The amount of xylan removed under these conditions was almost 57%. The soluble xylan consisted mainly of xylo-oligosaccharides (74 wt% of the identified compound in the spent liquor).Full Text: http://pubs.rsc.org/en/Content/ArticleLanding/2012/GC/c2gc35397k

Addresses: [Evangelina Vallejos, Maria; Cristina Area, Maria] Univ Nacl Misiones, FCEQyN, Programa Celulosa & Papel, Posadasmisiones, Argentina[Evangelina Vallejos, Maria; Cristina Area, Maria] Consejo Nacl Invest Cient & Tecn CONICET, Buenos Aires, Argentina[Zambon, Marcia Dib; da Silva Curvelo, Antonio Aprigio] Univ Sao Paulo, Inst Quim Sao Carlos, Dept Fis Quim, Grp Fis Quim Organ, BR-13560970 Sao Paulo, Brazil[da Silva Curvelo, Antonio Aprigio] Ctr Pesquisa Energia & Mat CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil

5. Evaluation of Barros and Wolf Efficiency Correlations for Conventional and Extractive Distillation Columns in Bioethanol Production Process

Author(s): Junqueira, TL; Maciel, MRW; Maciel, R Source: SEPARATION SCIENCE AND TECHNOLOGY Volume: 47 Issue: 7 Pages: 1031-1037 | DOI: 10.1080/01496395.2011.640378 | Published: 2012

Abstract: In view of the increasing environmental concern, the use of renewable and less pollutant energy sources has been encouraged. In this context, bioethanol has been used as fuel in the anhydrous form, blended with gasoline. In this work, simulations of the extractive distillation process in anhydrous bioethanol production were carried out in Aspen Plus (R) using nonequilibrium stage model and equilibrium stage model with Barros and Wolf efficiency correlations for plate and component. Equilibrium stage model with plate efficiency determined through Barros and Wolf efficiency correlation showed a satisfactory agreement with results predicted by the nonequilibrium stage model for anhydrous bioethanol production process.Full Text: http://www.tandfonline.com/doi/abs/10.1080/01496395.2011.640378

Addresses: [Junqueira, Tassia Lopes; Wolf Maciel, Maria Regina; Maciel Filho, Rubens] Univ Estadual Campinas, Sch Chem Engn, BR-13083970 Campinas, SP, Brazil[Junqueira, Tassia Lopes; Maciel Filho, Rubens] CNPEM, Lab Nacl Ciencia & Tecnol Bioetanol CTBE, Campinas, SP, Brazil

4. Optimum Ratio of Calcium and Boron in the Nutrient Solution or in Castor Bean Shoot for Fruit Yield and Seed Oil Content

Author(s): Franco, HCJ; Lavres, J de Oliveira, A; Matias, GCS ; Cabral, CP; Malavolta, E | E-mail: henrique.franco@bioetanol.org.br   Source: JOURNAL OF PLANT NUTRITION Volume: 35 Issue: 3 Pages: 413-427 | DOI: 10.1080/01904167.2012.639921 | Published: 2012 Abstract: Calcium (Ca) and boron (B) have been reported as the major macro-and micronutrient required for castor bean plant yield. The objective of this study was to determine the Ca: B ratios (in the growth media and plant tissue) for fruit yield and shoot dry weight of the castor bean (Ricinus

communis L.), grown in a nutrient solution, and to evaluate Ca and B supply on concentration and total uptake of Ca, potassium (K), magnesium (Mg), and B, as well on the seed oil content. The treatments were arranged in a 3 x 3 factorial fashion, consisting of three rates of Ca (40, 80, and 160 mg L-1) and three of B (0.32, 0.96, and 1.60 mg L-1). Calcium and B rates increased the shoot and root dry weight and fruit yield at a Ca: B ratio in the nutrient solution of 166 and 100, respectively. Symptoms of B deficiency were observed in plants supplied with 0.32 mg B L-1, regardless of the Ca concentration in the nutrient solution. Plants which showed visual symptoms of Ca deficiency cultivated with 40 mg Ca L-1 presented concentration of Ca in plant tissue up to 10 g kg(-1). The concentration and total Ca and B uptake increased with the rates of them. Notwithstanding, the shoot Ca accumulation was improved by B rates. In addition, there were no decreases in K and Mg uptake due to Ca rates. Furthermore, addition of 80 mg L-1 of Ca and 1.60 mg L-1 of B in the growth media increased the seed oil content. The Ca: B ratio in the diagnostic leaf associated with the highest plant dry weight (shoot and root) and fruit yield, was 500 (16 to 20 g kg(-1) of Ca, and for 30 to 40 mg kg(-1) of B).Full Text: http://www.tandfonline.com/doi/abs/10.1080/01904167.2012.639921

Addresses: [Junqueira Franco, Henrique Coutinho; Lavres Junior, Jose; Cabral, Cleusa Pereira; Malavolta, Euripedes] Univ Sao Paulo, Ctr Energia Nucl Agr, Piracicaba, SP, Brazil[de Oliveira Junior, Adilson] EMBRAPA, Embrapa Soja, Londrina, Brazil[Silva Matias, Gean Carlos] Univ Sao Paulo, Escola Super Agr Luiz de Queiroz, Piracicaba, Brazil

3. Dissecting structure-function-stability relationships of a thermostable GH5-CBM3 cellulase from Bacillus subtilis 168

Author(s): Santos, CR; Paiva, JH; Sforca, ML; Neves, JL; Navarro, RZ; Cota, J; Akao, PK; Hoffmam, ZB ; Meza, AN; Smetana, JH; Nogueira, ML; Polikarpov, I; Xavier-Neto, J; Squina, FM; Ward, RJ; Ruller, R; Zeri, AC; Murakami, MT | E-mail: fabio.squina@bioetanol.org.br Source: BIOCHEMICAL JOURNAL Volume: 441 Pages: 95-104 | DOI: 10.1042/BJ20110869 Part: Part 1 | Published: JAN 1 2012

Abstract: Cellulases participate in a number of biological events, such as plant cell wall remodelling, nematode parasitism and microbial carbon uptake. Their ability to depolymerize crystalline cellulose is of great biotechnological interest for environmentally compatible production of fuels from lignocellulosic biomass. However, industrial use of cellulases is somewhat limited by both their low catalytic efficiency and stability. In the present study, we conducted a detailed functional and structural characterization of the thermostable BsCe15A (Bacillus subtilis cellulase 5A), which consists of a GH5 (glycoside hydrolase 5) catalytic domain fused to a CBM3 (family 3 carbohydrate-binding module). NMR structural analysis revealed that the Bacillus CBM3 represents a new subfamily, which lacks the classical calcium-binding motif, and variations in NMR frequencies in the presence of cellopentaose showed the importance of polar residues in the carbohydrate interaction. Together with the catalytic domain, the CBM3 forms a large planar surface for cellulose recognition, which conducts the substrate in a proper conformation to the active site and increases enzymatic efficiency. Notably, the manganese ion wasdemonstrated to have a hyper-stabilizing effect on BsCel5A, and by using deletion constructs and X-ray crystallography we determined that this effect maps to a negatively charged motif located at the opposite face of the catalytic site.Full Text: www.biochemj.org/bj/441/bj4410095.htm

Addresses: [Santos, Camila R.; Paiva, Joice H.; Sforca, Mauricio L.; Neves, Jorge L.; Navarro, Rodrigo Z.; Akao, Patricia K.; Meza, Andreia N.; Smetana, Juliana H.; Nogueira, Maria L.; Xavier-Neto, Jose; Zeri, Ana C.; Murakami, Mario T.] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Biociencias, Campinas, SP, Brazil[Cota, Junio; Hoffmam, Zaira B.; Squina, Fabio M.; Ruller, Roberto] Ctr Nacl Pesquisa Energia & Mat, Lab Nacl Ciencia & Tecnol Bioetanol, Campinas, SP, Brazil

[Polikarpov, Igor] Univ Sao Paulo, Inst Fis Sao Carlos, Sao Carlos, SP, Brazil[Ward, Richard J.] Univ Sao Paulo, Dept Quim, BR-14049 Ribeirao Preto, Brazil

2. Integrated versus stand-alone second generation ethanol production from sugarcane bagasse and trash

Author(s): Dias, MOS; Junqueira, TL; Cavalett, O; Cunha, MP; Jesus, CDF; Rossell, CEV; Maciel, R; Bonomi, A . | E-mail: carlos.rossell@bioetanol.org.brSource: BIORESOURCE TECHNOLOGY Volume: 103 Issue: 1 Pages: 152-161 | DOI:10.1016/j.biortech.2011.09.120 | Published: JAN 2012

Abstract: Ethanol production from lignocellulosic materials is often conceived considering independent, stand-alone production plants; in the Brazilian scenario, where part of the potential feedstock (sugarcane bagasse) for second generation ethanol production is already available at conventional first generation production plants, an integrated first and second generation production process seems to be the most obvious option. In this study stand-alone second generation ethanol production from surplus sugarcane bagasse and trash is compared with conventional first generation ethanol production from sugarcane and with integrated first and second generation; simulations were developed to represent the different technological scenarios, which provided data for economic and environmental analysis. Results show that the integrated first and second generation ethanol production process from sugarcane leads to better economic results when compared with the stand-alone plant, especially when advanced hydrolysis technologies and pentoses fermentation are included. Full Text: http://www.sciencedirect.com/science/article/pii/S0960852411014088

Addresses: [Dias, Marina O. S.; Junqueira, Tassia L.; Cavalett, Otavio; Cunha, Marcelo P.; Jesus, Charles D. F.; Rossell, Carlos E. V.; Maciel Filho, Rubens; Bonomi, Antonio] Lab Nacl Ciencia & Tecnol Bioetanol CTBE, BR-13083970 Campinas, SP, Brazil[Dias, Marina O. S.; Junqueira, Tassia L.; Rossell, Carlos E. V.; Maciel Filho, Rubens] Univ Estadual Campinas UNICAMP,Fac Engn Quim, BR-13083852 Campinas, SP, Brazil

1. Steam explosion pretreatment reproduction and alkaline delignification reactions performed on a pilot scale with sugarcane bagasse for bioethanol production

Author(s): Rocha, GJM; Goncalves, AR; Oliveira, BR; Olivares, EG; Rossell, CEV | E-mail: carlos.rossell@bioetanol.org.brSource: INDUSTRIAL CROPS AND PRODUCTS Volume: 35 Issue: 1 Pages: 274-279 | DOI:10.1016/j.indcrop.2011.07.010 Published: JAN 2012

Abstract: This work was focused on the steam explosion pretreatment reproduction and alkaline delignification reactions on a pilot scale for the ethanol production, through different varieties of natural sugarcane bagasse, pretreated bagasse and delignified pretreated bagasse (cellulosic pulp). The possible chemical composition differences of the various types of bagasse, as well as the chemical composition variations of the materials in the 20 processes of pretreatment and delignification on the pilot scale were verified. The analytical results of the 20 samples of most diverse varieties and origins of natural sugarcane bagasse considering planting soils, planting periods and weather; show no significant chemical differences. It is evident that only with the chemical composition it is not possible to verify the differences between the varieties of sugarcane bagasses. The research results may offer some evidences of these varieties, but it is not a reliable parameter. The pilot process of steam explosion pretreatment and the alkaline delignification process of pretreated material showed through analytical results a good capacity of reproduction, as the standard differences were below 2.7. The

average allowed in the pretreatment and alkaline delignification processes were 66.1 +/- 0.8 and 51.5 +/- 2.6 respectively, ensuring an excellent reproduction capacity of the processes obtained through chemical characterizations.Full Text: http://www.sciencedirect.com/science/article/pii/S0926669011003062

Addresses: [Rocha, G. J. M.; Oliveira, B. R.; Olivares, E. G.; Rossell, C. E. V.] Bioethanol Sci & Technol Natl Lab CTBE, BR-13083970 Campinas, SP, Brazil[Goncalves, A. R.] Univ Sao Paulo, Lorena Engn Sch, Dept Biotechnol, BR-12600970 Lorena, SP, Brazil