Obtenção e caracterização de queratinase de bacillus sp. P45 a partir de coprodutos e aplicação na produção de queijos cremosos enriquecidos com chia e quinoa

As proteases constituem 60-65% do mercado global das enzimas industriais e são utilizadas na indústria de alimentos no processo de amaciamento de carne, na síntese de peptídeos, preparo de fórmulas infantis, panificação, cervejarias, produtos farmacêuticos, diagnósticos médicos, como aditivos na indústria de detergentes e na indústria têxtil no processo de depilação e transformação do couro. Proteases específicas produzidas por micro-organismos queratinolíticos são chamadas de queratinases e distinguem-se de outras proteases pela maior capacidade de degradação de substratos compactos e insolúveis como a queratina. Atualmente, processos que apontem o uso total das matérias-primas e que não resultem em impactos negativos ao meio ambiente tem ganhado destaque. Dentro desta temática, destacam-se a reutilização da farinha de penas residual durante o cultivo do Bacillus sp. P45 para produção de proteases e a biomassa residual de levedura, ambas com elevados teores de proteínas, podendo ser utilizadas no cultivo do Bacillus sp. P45 para obtenção de proteases. O objetivo deste trabalho foi obter a enzima queratinase purificada em grandes quantidades, sua caracterização, bem como a sua aplicação em processos de coagulação enzimática do leite para o desenvolvimento de um queijo cremoso enriquecido com farinha de chia e quinoa. Além disso, aplicar diferentes coprodutos para produção de enzimas proteolíticas e queratinolíticas. A presente tese foi dividida em quatro artigos: no primeiro foi realizado a obtenção da queratinase purificada em maiores quantidades e a determinação dos parâmetros de estabilidade térmica e a influência de componentes químicos na atividade enzimática. A obtenção da enzima em maiores quantidades alcançou fatores de purificação de 2,6, 6,7 e 4,0 vezes, paras 1º SAB, 2º SAB e diafiltração, respectivamente. A recuperação enzimática alcançou valores de 75,3% para o 1º SAB, 75,1% no 2º sistema e 84,3% na diafiltração. A temperatura de 55ºC e o pH 7,5 foram determinados como ótimos para atividade da enzima queratinase. O valor da energia de desativação (Ed) médio foi de 118,0 kJ/mol e os valores de z e D variaram de 13,6 a 18,8ºC, e 6,9 a 237,3 min, respectivamente. Além disso a adição de sais (CaCl2, CaO, C8H5KO4 e MgSO4) elevou a atividade da enzima na presença destes compostos. O segundo artigo apresenta a aplicação da queratinase como coagulante de leite bovino e sua aplicação na obtenção de queijo cremoso enriquecido com chia e quinoa. A enzima mostrou atividade de coagulação semelhante ao coagulante comercial, na concentração de 30mg/mL. A enzima purificada foi empregada de forma eficiente na fabricação do queijo cremoso, que apresentou valores de pH de 5,3 e acidez de 0,06 a 0,1 mol/L, com elevação durante os 25 dias de armazenamento. O terceiro artigo apresenta o perfil do queijo cremoso enriquecido com farinha de chia e quinoa, o qual apresentou alto índice de retenção de água (>99,0%) e baixos valores de sinérese (<0,72%). Elevados teores de fibras foi verificado (3,0 a 5,0%), sugerindo seu consumo como fonte de fibras. As análises microbiológicas foram de acordo com a legislação vigente. Na análise sensorial foi verificado altos valores de suavidade ao paladar e verificado maiores valores de consistência e untabilidade nas amostras com maiores concentrações de nata e quinoa. O quarto artigo traz a extração de β-galactosidase por ultrassom e o uso da biomassa residual da levedura, bem como o uso de farinha de penas residuais como substrato para obtenção de proteases. O ultrassom foi eficiente para ruptura celular e extração de β-galactosidase, apresentando alta atividade (35,0 U/mL) e rendimento (876,0 U/g de biomassa). A maior atividade proteolítica (1300 U/mL em 32 h) e queratinolítica (89,2 U/mL) verificadas ocorreram utilizando-se a biomassa e a farinha de penas residuais, respectivamente. Maior produtividade proteolítica (40,8 U/mL/h) foi verificado no meio utilizando biomassa residual como substrato. Já a maior produtividade queratinolítica (2,8 U/mL/h) foi alcançada utilizando farinha de penas reutilizada.

Life cycle assessment of residual forestry biomass chips at a power plant: a Portuguese case study

The residual forest biomass (RFB) sector has been experiencing strong development at European level and particularly in Portugal mainly due to the increase of energy production from renewable sources. The aim of this study is to assess the environmental impacts of eucalyptus RFB chips production chain in Portugal. The environmental and economic impact comparison of the processes included in the production chain is presented as well. The environmental impacts were calculated by the life cycle assessment approach described in the ISO 14040 series of standards. The production chain assessed included all processes from eucalyptus forest until the delivery of RFB chips at the power plant. The main conclusion of this study is that eucalyptus wood production is the process that presents the greatest environmental impact through the product life cycle.

Estudi de la viabilitat de l’aprofitament de biomassa residual amb implantació d’una planta de cogeneració al sector del Pla de Baix de Domeny

L’estudi pretén incorporar el residu de fusta i poda com a combustible en una planta de cogeneració al sector del Pla de Baix de Domeny, per obtenir un benefici econòmic amb la venda d’energia elèctrica generada i utilitzar l’energia tèrmica residual per a la calefacció i l’aigua calenta sanitària dels habitatges. S’han analitzat els habitatges, els residus de biomassa i els diferents tipus de cogeneració. I s’ha proposat una cogeneració amb el cicle orgànic de Rankine, utilitzant l’estella de residus de fusta i poda per a 1219 habitatges del Pla de Baix de Domeny i s’ha estudiat la viabilitat econòmica

Estimating the spatial scale of herbicide and soil interactions by nested sampling, hierarchical analysis of variance and residual maximum likelihood

An unbalanced nested sampling design was used to investigate the spatial scale of soil and herbicide interactions at the field scale. A hierarchical analysis of variance based on residual maximum likelihood (REML) was used to analyse the data and provide a first estimate of the variogram. Soil samples were taken at 108 locations at a range of separating distances in a 9 ha field to explore small and medium scale spatial variation. Soil organic matter content, pH, particle size distribution, microbial biomass and the degradation and sorption of the herbicide, isoproturon, were determined for each soil sample. A large proportion of the spatial variation in isoproturon degradation and sorption occurred at sampling intervals less than 60 m, however, the sampling design did not resolve the variation present at scales greater than this. A sampling interval of 20-25 m should ensure that the main spatial structures are identified for isoproturon degradation rate and sorption without too great a loss of information in this field.

Lead distribution in organic matter of loamy oxisol treated with manure and its effect on microbial biomass

This paper reports a study regarding the distribution of lead in the organic matter fractions of a loamy oxisol treated with stable manure, and its effect on the soil microbial biomass. The experimental design was a completely randomized factorial with three replicates. Treatments were four lead (Pb) rates (0, 200, 400, and 600 mg kg(-1) soil), two levels of manure (0 and 30 Mg ha(-1)) and two sampling times (30 and 60 days after lead application). Total and soil lead soluble in Mehlich 1 extractant, and Ph in the different fractions of the organic matter (fulvic acid, humic acid, and residual) were evaluated along with the soil microbial biomass. Data showed that lead, applied as PbCl2, was concentrated in the residual fraction. The application of manure at the rate of 30 Mg ha(-1) did not affect lead distribution in the fractions until 60 days after incorporation. Lead extracted by Mehlich 1 increased as function of the quantity applied. Manure stimulated the growth of the soil microbial biomass which was reduced by the 200 mg kg(-1) doses of lead, with manure application, increasing, however, with larger rates. Mehlich 1 extracted only part of the lead present in the residual fraction.

Efeito residual de fontes de fósforo nos atributos químicos do solo, nutrição e produtividade de biomassa da cana-de-açúcar

In tropical regions, the residual effect of phosphate fertilizer source is of crucial importance. This study aimed to evaluate the effects of application of P sources in chemical properties of soil, nutrition and productivity of sugarcane plant and during the first and second ratoon. The sources of P applied in furrows at planting were, triple superphosphate, Arad phosphate, bone meal (100 kg ha-1 P2O5 total) and a control without P, the varieties of sugarcane studied were IAC86-2480 and SP79-1011. A randomized block design in a 4 × 2 factorial was used, with four replications. The study was conducted in the municipality of Alta Floresta-MT, Brazil, in Typic Hapludox. After each cutting, in soil P content, pH and Ca were evaluated. In the plant, the production of green and dry matter of shoots, the concentration of P in the biomass harvested, the export of P and agronomic efficiency of the sources of P were determined. The bone meal had a higher agronomic efficiency resulted in greater accumulation and export of P by crop, higher content of available Ca and P and increase soil pH. The Arad reactive phosphate increased its agronomic efficiency between crop cycles.

Metodologia de análise de risco de investimento em projetos de eficiência energética com aplicação no aproveitamento da biomassa residual do dendê na Amazônia

A promoção das energias renováveis, como estratégias para mitigar as alterações climáticas é um grande desafio mundial, principalmente para países em desenvolvimento como Brasil que busca diversificar sua matriz energética a partir de fontes renováveis, mas possui restrições de custo de investimentos. Neste contexto, são necessárias ferramentas que mensurem os riscos de investimento em projetos de fontes de energia renovável. Este trabalho contribui para o desenvolvimento sustentável da Amazônia através do desenvolvimento de uma metodologia de análise de risco de investimento em projetos de eficiência energética aplicada no aproveitamento da biomassa do dendê. A crescente produção de dendê na região amazônica para geração de produtos alimentícios e biodiesel está gerando um grande volume de biomassa. Verificou-se que a utilização do resíduo da biomassa de dendê para a geração de energia e comercialização no mercado aumenta o seu valor econômico, para tanto se identificou os Critérios e Indicadores (C&I) de sustentabilidade da cadeia do dendê agrupando-os em: econômicos, ambientais, sociais e técnicos. Qualificando e quantificando os riscos que afetam os indicadores identificados. A metodologia desenvolvida foi analisada em três cenários futuros de crescimento econômico para o Brasil que foram construídos balizados nos cenários contidos no Plano Nacional de Energia 2030.

Leaching potential and residual effect of amicarbazone in soils of contrasting texture

The infestation of weeds is a major biotic factor in the agroecosystem of cane sugar that may interfere in development and crop productivity. This study aimed to evaluate the potential for leaching and residual effects of the herbicide amicarbazone in contrasting soils. Samples were Quartzarenic Neosol (NR - sandy texture) and Red Latosol (LR - clay texture). For the leaching potential, after application of herbicide amicarbazone (NR 1.05 kg ha(-1) and LR 1.40 kg ha(-1)), layers of 0, 20, 40, 60, 80 and 100 mm of water were applied to soil columns. We evaluated the residual effect after the permanence of the herbicide in soil of clay texture and sandy for periods of 0, 25, 50, 75 and 100 days after application (DAA) of amicarbazone (0, 1.05, 1.40 kg ha(-1)) treatments. The amicarbazone started showing high leaching from the 60 mm layer of water in sandy texture soils, evidencing a shorter residual effect. In clay soil, slides from 20 to 80 mm of water reduced the biomass until a depth of 5-10 cm, with the use of this herbicide. Based on these results, we conclude that the amicarbazone showed higher leaching and lower residual effects in sandy soil. The residual effect of amicarbazone was prolonged as the content of clay and organic matter present in the soil increased.

Mycelial biomass and biochemical properties of proteases produced by Lentinus citrinus DPUA 1535 (higher basidiomycetes) in submerged cultivation

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influential factors in determination of biomass termal susceptibility by termal analysis

Las biomasas almacenadas en silos son potencialmente capaces de absorber oxígeno produciendo reacciones exotérmicas de oxidación. Si el calor producido en estas reacciones no se dispersa adecuadamente, provoca un auto-calentamiento de la materia orgánica que puede ser causa de descomposición e inflamación. En el ámbito de la posible auto-combustión en el almacenamiento y manipulación de las biomasas existen diversos factores que influyen en la susceptibilidad térmica de las biomasas, es decir, en su tendencia a la oxidación y posterior inflamación de la materia. Con el fin de analizar este comportamiento se han estudiado diferentes tipos de biomasas. En este estudio se ha trabajado con biomasas de origen agrícola, forestal y residual con distinta composición química. Las muestras de estudio se han acondicionado mediante distintos tratamientos físicos para valorar la influencia de diferentes factores en su auto-combustión.

Characterization of sugar cane waste biomass derived chars from pressurized gasification

There is interest in the use of sugar cane waste biomass for electricity cogeneration, by integrated gasification combined cycle (IGCC) processes. This paper describes one aspect of an overall investigation into the reactivity of cane wastes under pressurized IGGC conditions, for input into process design. There is currently a gap in understanding the morphological transformations experienced by cane waste biomass undergoing conversion to char during pressurized gasification, which is addressed by this work. Char residuals remaining after pressurized pyrolysis and carbon dioxide gasification were analysed by optical microscope, nitrogen (BET) adsorption analysis, SEM/EDS, TEM/EDS and XPS techniques. The amorphous cane plant silica structures were found to remain physically intact during entrained flow gasification, but chemically altered in the presence of other inorganic species. The resulting crystalline silicates were mesoporous (with surface areas of the order of 20 m(2) g(-1)) and contributed to much of the otherwise limited pore volume present in the residual chars. Coke deposition and intimate blending of the carbonaceous and inorganic species was identified. Progressive sintering of the silicates appeared to trap coke deposits in the pore network. As a result ash residuals showed significant organic contents, even after extensive additional oxidation in air. The implications of the findings are that full conversion of cane trash materials under pressurized IGCC conditions may be significantly hampered by the silica structures inherent in these biomass materials and that further research of the contributing phenomena is recommended.

Production of renewable phenolic resins by thermochemical conversion of biomass: a review

This review covers the production and utilisation of liquids from the thermal processing of biomass and related materials to substitute for synthetic phenol and formaldehyde in phenol formaldehyde resins. These resins are primarily employed in the manufacture of wood panels such as plywood, MDF, particle-board and OSB. The most important thermal conversion methods for this purpose are fast pyrolysis and vacuum pyrolysis, pressure liquefaction and phenolysis. Many feedstocks have been tested for their suitability as sources of phenolics including hard and softwoods, bark and residual lignins. Resins have been prepared utilising either the whole liquid product, or a phenolics enriched fraction obtained after fractional condensation or further processing, such as solvent extraction. None of the phenolics production and fractionation techniques covered in this review are believed to allow substitution of 100% of the phenol content of the resin without impacting its effectiveness compared to commercial formulations based on petroleum derived phenol. This survey shows that considerable progress has been made towards reaching the goal of a price competitive renewable resin, but that further research is required to meet the twin challenges of low renewable resin cost and satisfactory quality requirements. Particular areas of concern are wood panel press times, variability of renewable resin properties, odour, lack of reactive sites compared to phenol and potential for increased emissions of volatile organic compounds.

Intermediate pyrolysis studies of aquatic biomass and potential applications in the BtVB-process

Aquatic biomass is seen as one of the major feedstocks to overcome difficulties associated with 1st generation biofuels, such as competition with food production, change of land use and further environmental issues. Although, this finding is widely accepted only little work has been carried out to investigate thermo-chemical conversion of algal specimen to produce biofuels, power and heat. This work aims at contributing fundamental knowledge for thermo-chemical processing of aquatic biomass via intermediate pyrolysis. Therefore, it was necessary to install and commission an analytical pyrolysis apparatus which facilitates intermediate pyrolysis process conditions as well as subsequent separation and detection of pyrolysates (Py- GC/MS). In addition, a methodology was established to analyse aquatic biomass under intermediate conditions by Thermo-Gravimetric Analysis (TGA). Several microalgae (e.g. Chlamydomonas reinhardtii, Chlorella vulgaris) and macroalgae specimen (e.g. Fucus vesiculosus) from main algal divisions and various natural habitats (fresh and saline water, temperate and polar climates) were chosen and their thermal degradation under intermediate pyrolysis conditions was studied. In addition, it was of interest to examine the contribution of biochemical constituents of algal biomass onto the chemical compounds contained in pyrolysates. Therefore, lipid and protein fractions were extracted from microalgae biomass and analysed separately. Furthermore, investigations of residual algal materials obtained by extraction of high valuable compounds (e.g. lipids, proteins, enzymes) were included to evaluate their potential for intermediate pyrolysis processing. On basis of these thermal degradation studies, possible applications of algal biomass and from there derived materials in the Bio-thermal Valorisation of Biomass-process (BtVB-process) are presented. It was of interest to evaluate the combination of the production of high valuable products and bioenergy generation derived by micro- and macro algal biomass.