Solid state production of thermostable pectinases from thermophilic Thermoascus aurantiacus

Pectin lyase (Pl) and polygalacturonase (Pg) production by Thermoascus aurantiacus 179-5 was carried out by means of solid-state determination using orange bagasse and wheat bran as a carbon sources. Pg and Pl had optimum activity at pH 5.0 and 10.5 respectively. Maximal activity of the enzymes were determined at 65 °C. Pg was stable in the acidic to neutral pH range and at 60 °C for 1 h. whereas Pl was stable at acidic pH and at 60 °C for 5 h. © 2002 Elsevier Science Ltd. All rights reserved.

Heavy metal nutrients in tomato plants cultivated in soil amended with biosolid composts

The use of biosolids in horticulture could contribute to recycle residues produced by men. This study analyzed concentrations of Cu, Mn and Zn in the compost during fermentation, in the soil amended with the composts and in the tomato plant materials. Five composts were produced using sugar-cane bagasse, biosolid and cattle manure in the proportions: 75-0-25; 75-12.5-12.5; 75-25-0; 50-50- 0 and 0-100-0 (composts with 0; 12.5; 25; 50 and 100% biosolid), respectively. These composts were used in an experiment with 6 treatments (the 5 composts and a control with mineral fertilization) in a design of randomized blocks with a split plot design. The control and the treatment of 0% biosolid received inorganic nitrogen. All the treatments received the same amount of N, P and K. Two tomato plants were cultivated in each 24 L pot, in a greenhouse at the Technology Department of the Faculdade de Ciências Agrárias e Veterinárias of the Universidade Estadual Paulista in Jaboticabal County, São Paulo State, Brazil. The concentrations of Cu, Mn and Zn were evaluated in the compost 7, 27, 57, 97 and 127 days after composting began, in the soil 0 and 164 days after the compost applied, and in the plants. Compost, soil and plant samples were subjected to digestion with HNO3, H 2O2 and HCl and the metals were determined by AAS. There were positive and significant correlations between Mn in the compost and Mn uptake by the plant (0.46 p>0.05), and between Zn in the compost and Zn concentration in the plant (0.78 p>0.05). Cu, Mn and Zn concentrations increased during composting. The biosolid in the compost supplied Cu and Zn to tomato plants, and the cattle manure supplied Mn to the plants.

Purification and biochemical characterization of two xylanases produced by Aspergillus caespitosus and their potential for kraft pulp bleaching

Two extracellular xylanases produced by the thermotolerant fungus Aspergillus caespitosus grown in sugar cane bagasse were purified and characterized. Estimated molecular masses were 26.3 and 27 kDa (xyl I); 7.7 and 17.7 kDa (xyl II) for gel filtration and SDS-PAGE, respectively. Optimal temperature for both xylanases was 50-55°C. Optimal pH was 6.5-7.0 for xyl I, and 5.5-6.5 for xyl II. The thermostability (T half) at 55°C was 27.3 min (xyl I) and >90 min (xyl II). Xylanase activity was inhibited by several ions. β-mercaptoethanol activated 59 and 102% xyl I and xyl II activities, respectively. These enzymes preferentially hydrolyzed birchwood xylan, and the K m and V max values were 2.5 mg/ml and 1679 U/mg protein (xyl I), and 3.9 mg/ml and 113 U/mg protein (xyl II). The action of both xylanases mainly that of xyl II, on kraft pulp reduced kappa number and increased pulp viscosity. © 2004 Elsevier Ltd. All rights reserved.

Comparative field test between two sugar cane (Saccharum spp.) harvesters under two different operational conditions in Brazil

The sugar cane crop is one of the main products in Brazil and according to several authors can generate, besides the industrialized stalks, an amount of crop residues from the order of 15 to 30 % in weight of the aerial part of the plants, depending on the field conditions. The sugar cane area in Brazil is around 5.5×106 hectares, with an amount of 400.106 tons of stalks, with stalks yield of 72 tons.ha-1. This study took place in a sugar cane plot (Latitude 22°46'S, Longitude 47°23'W and 600m of altitude) with 3% of slope, located in São Paulo State. The sugar cane variety was SP 80-1816, in its forth cut, 11 months old and with a planted row spacing of 1.40 m. By other side, several sugar mills are bringing the crop residue to their patio to produce energy with the bagasse. One choice is to bring the crop residue at the same moment with the stalks, avoiding the next operation of baling it. The objective of this study was to analyze some operational parameters of two different sugar cane harvesters under the same field conditions, which was divided in four treatments: T1 = CAMECO CHT2500B operating normally; T2 = CAMECO CHT2500B operating without the cleaning system; T3 = CASE 7700 operating normally; T4 = CASE 7700 operating without the cleaning system. The results obtained were: Table presented CEB = Gross effective capacity; CEL = Net effective capacity. The conclusion is that under normal operation the CASE harvester worked better then CAMECO in the parameters CEL stalks and Manipulation efficiency. And without the cleaning system operating CASE also worked better in the parameters of CEB raw material, CEB stalks, CEL raw material and CEL stalks.

Sugar cane (Saccharum spp.) crop residue baling for energy purposes in Brazil

The sugar cane crop according to several authors can generate, besides the industrialized stalks, an amount of crop residues from the order of 15 to 30% in weight of the aerial part of the plants, depending on the field conditions. The sugar cane area in Brazil is around 5.5×106 hectares, with an amount of 400.106 tons of stalks, with stalks yield of 72 tons.ha-1 (Unica, 2005). This study took place in a sugar cane plot (Latitude 22°46'S, Longitude 47°23'W and 600m of altitude) with 3% of slope, located in São Paulo State. The sugar cane variety was SP 80-1816, in its forth cut, 11 months old and with a planted row spacing of 1.40m. By other side, several sugar mills are bringing the crop residue to their patio to produce energy with the bagasse. One way for that is the baling operation to bring the crop residue at the sugar mill. Some fundamental variables were obtained to define the best set of machines to work with in sugar cane crop residue removal in the baling system among the studied ones, some of the variables were: Soil Index (T1 = 0.83%, T2 = 0.46%, T3 = 0.65%, T4 = 0.57%); Energy Efficiency (T1 = 82.48%, T2 = 83.88%, T3 = 82.83% and T4 = 82.97%) of the system and Effective Cost for Equivalent Energy in US$.EBP-1 (T1 = 11.10, T2= 10.46, T3 = 11.47 and T4 = 10.57) of the baled trash delivered at the sugar mill.

Heavy metals availability and fractions in soil amended with biosolid composts

The heavy metals when linked to organic matter have a behavior in the soil that is still little known. This study aimed to evaluate the effect of sewage-sludge-based composts when incorporated in the soil, in relation to heavy metals availability. Five composts were incorporated using sugar-cane bagasse, sewage sludge and cattle manure in the respective proportions: 75-0-25, 75-12.5-12.5, 75-25-0, 50-50-0 and 0-100-0 (composts with 0, 12.5, 25, 50 and 100% sewage sludge). The experiment consisted of 6 treatments (5 composts and a control with mineral fertilization) in randomized blocks with a split-plot design. The control and the treatment of 0% sewage sludge received inorganic nitrogen (N). All the treatments received the same amount of N (8.33 g) K (5.80 g) and K (8.11 g) per pot. Tomato plants were cultivated in 24.0 L pots in a greenhouse in Jaboticabal, SP, Brazil. The concentrations of heavy metals were determined in the soil samples at day 0 after compost incorporation. The higher the sewage sludge doses, the higher heavy metal contents in the soil. Among extractants, Melhlich-1 extracted the highest amount of heavy metals, while DTPA extracted the lowest one. The residual fraction presented the highest heavy metal content, followed by Fe oxides crystalline and amorphous to Cu, Cr and Mn, and Mn oxides, and Fe amorphous to Zn, indicating strong associations to oxides and clays. There were significant positive correlations between Mn contents in the plant and Mn linked to Fe oxide amorphous and crystalline.

Environmental evaluation of cogeneration scenarios in sugar industry

The development of new techniques that allow the analysis and optimization of energy systems bearing in mind environmental issues is indispensable in a world with finite natural resources and growing demand of energy. Among the energy systems that deserve special attention, cogeneration in the sugar industry must be pointed out, because it uses efficiently a common fuel for generation of useful heat and power. Within this frame, thermoeconomical optimization - 2nd Law of Thermodynamics analysis by exergy function and economic evaluation of the thermal system - gradually is taking importance as a powerful tool to assist to the decision making process. Also, the explicit consideration of environmental issues offers a better way to explore trade-offs between different aspects to support the decisions that must be made. In this work it is used the technique of Life Cycle Analysis (LCA) which allows to consider environmental matters as an integral part of the problem, in opposite to most of the environmental approaches that only reduce residuals generation , without taking into account impacts associated to other related processes. On the other hand, the consideration of environmental issues in optimization of energy systems is a novel and promissory contribution in the state of the art of energy optimization and LCA. The system under study is a sugar plant of Tucumán (Argentina) given the particular importance that this industry had inside the regional economy of the Argentinean Northwest. Although cogeneration comes being used a while ago in sugar industry, being the main objective the generation of heat and as secondary objective the electric power generation and mechanic power to cover several needs of working machineries, to the date it is no available a versatile tool that allows to analyze economical feasible alternatives bearing in mind environmental issues. At sugar plants, steam is generated in boilers using as fuel bagasse - cellulosic fiber waste obtained crushing the sugar cane- and it is used to give useful heat and shaft work to the plant, but it can also be used to generate electricity with export opportunities to the electrical network. The great number of process alternatives outlines a serious decision making problem in order to take advantage of the resources. Although the problem turns out to be a mixed non-linear problem (MINLP), the main contribution of this work is the development of a hybrid strategy to evaluate cogeneration alternatives that combines optimization approaches with environmental indicators. This powerful tool for its versatility and robustness to analyze cogeneration systems, will be of great help in the decision making process, because of their easy implementation to analyze the kind of problems presented in the sugar industry.

Economical analysis of ethanol steam reformer for hydrogen production associated with a sugarcane industry

An expressive amount of produced hydrogen is generated by customers in-situ such as petrochemical, fertilizer and sugarcane industries. However, the most utilized feedstock is natural gas, a non-renewable and fossil fuel. The introduction of biohydrogen production process associated in a sugarcane industry is an alternative to diminish emissions and contribute to create a CO2 cycle, where the plants capture this gas by photosynthesis process and produces sucrose for ethanol production. The cost of production of ethanol has dramatically decreased (from about US$ 700/m3 in 1970s to US$ 200/m3 today), becoming this a good option at near term, inclusively for its utilization by customers localized in main regions (localized especially in regions such as Southeastern Brazil) Also in near future, it will possible the utilization of fuel cells as form of distributed generation. Its utilization could occur specially in peak hours, diminishing the cost of investments in newer transmission systems. A technical and economic analysis of steam reformer of ethanol to hydrogen production associated with sugarcane industry was recently performed. This technique will also allow the use of ethanol when its price is relatively low. This study was based on a previous R&D study (sponsored by CEMIG - State of Minas Gerais Electricity Company) where thermodynamic and economic analyses were developed, based in the development of two ethanol steam reformers prototypes.x In this study an analysis was performed considering the use of bagasse as source of heat in the steam reforming process. Its use could to diminish the costs of hydrogen production, especially at large scale, obtaining cost-competitive production and permitting that sugarcane industry produces hydrogen in large scale beyond ethylic alcohol, anhydrous alcohol (or ethanol) and sugar.

Produtividade de cana-de-açúcar sob adubação com torta de filtro enriquecida com fosfato solúvel

Phosphorus is considered an essential element for plants and it is found in small amounts in Brazilian soils. The filter cake residue, composed of a mixture of bagasse and decanting sludge, has high levels of organic matter, phosphorus and calcium. The phosphorus present in the filter cake is organic, and its release, as it happens to the nitrogen, occurs gradually by mineralization and by microorganisms attack in the soil. This study aimed to evaluate sugarcane vegetative growth and yield under fertilization with filter cake enriched with soluble phosphate. The experiment was carried out in Presidente Prudente, São Paulo State, Brazil, by using a randomized complete block design, in a 5x4 factorial scheme, where the first factor consisted of filter cake doses (0 t ha-1, 0.5 t ha-1, 1.0 t ha-1, 2.0 t ha-1, and 4.0 t ha-1) and the second of phosphorus fertilizer doses (0 kg ha-1, 50 kg ha -1, 100 kg ha-1, and 200 kg ha-1 of P 2O5), with 4 repetitions, totalizing 80 plots. The experiment evaluated the tiller number, at 30, 60, 90, and 120 days after planting, oBrix, and yield. The stalk yield and tillering were influenced by the filter cake rates applied to the soil. Filter cake doses and their combination with phosphate did not change the juice quality (Brix) at harvest.

Production of pectate lyase by penicillium viridicatum RFC3 in solid-state and submerged fermentation

Pectate lyase (PL) was produced by the filamentous fungus Penicillium viridicatum RFC3 in solid-state cultures of a mixture of orange bagasse and wheat bran (1 : 1 w/w), or orange bagasse, wheat bran and sugarcane bagasse (1 : 1 : 0.5 w/w), and in a submerged liquid culture with orange bagasse and wheat bran (3%) as the carbon source. PL production was highest (1,500U mL -1 or 300Ug -1 of substrate) in solid-state fermentation (SSF) on wheat bran and orange bagasse at 96 hours. PL production in submerged fermentation (SmF) was influenced by the initial pH of the medium. With the initial pH adjusted to 4.5, 5.0, and 5.5, the peak activity was observed after 72, 48, and 24 hours of fermentation, respectively, when the pH of the medium reached the value 5.0. PL from SSF and SmF were loaded on Sephadex-G75 columns and six activity peaks were obtained from crude enzyme fromSSF and designated PL I, II, III, IV, V, andVI, while five peaks were obtained fromcrude enzyme fromSmF and labeled PL I', II', III', IV', and VII'. Crude enzyme and fraction III from each fermentative process were tested further. The optimum pH for crude PL from either process was 5.5, while that for PL III was 8.0. The maximum activity of enzymes from SSF was observed at 35°C, but crude enzyme was more thermotolerant than PL III, maintaining its maximum activity up to 45°C. Crude enzyme from SmF and PL III' showed thermophilic profiles of activity, with maximum activity at 60 and 55°C, respectively. In the absence of substrate, the crude enzyme from SSF was stable over the pH range 3.0-10.0 and PL III was most stable in the pH range 4.0-7.0. Crude enzyme from SmF retained 70%-80% of its maximum activity in the acid-neutral pH range (4.0-7.0), but PIII showed high stability at alkaline pH (7.5-9.5). PL from SSF was more thermolabile than that from SmF. The latter maintained 60% of its initial activity after 1 h at 55°C. The differing behavior of the enzymes with respect to pH and temperature suggests that they are different isozymes. Copyright © 2010 Viviani Ferreira et al.

Selecting the most adequate bedding material for broiler production in Brazil

Broiler chicken production is widely dispersed across the globe, and one important issue for growers is the selection of adequate bedding material, as the availability and price of substrates varies among countries and regions within a same country. This study aimed at applying a multiple criteria analysis approach for the selection of the most appropriate bedding material for broiler production. Based on field research data and growers' experience, the most desirable characteristics of a litter material were chosen as the main criteria. The selected materials were wood shavings, rice husks, chopped Napier grass (Pennisetum pupureum), 50% sugar cane bagasse (Saccharum L.) plus 50% wood shavings, 50% sugar cane bagasse (Saccharum L.) plus 50% rice husks, and pure sugar cane bagasse (Saccharum L.). The analytical hierarchy process (AHP) was applied for selecting the most suitable bedding material. Validation was performed using data from previous studies carried out in central-western Brazil on the effects of different types of bedding material on broiler carcass quality. Considering the selected criteria, several bedding materials were tested and ranked, and the results showed that wood-shavings litter was the best option (weight = 0.28), followed by rice husks (weight = 0.24). All other tested alternatives presented lower scores and were, therefore, not considered for use. The AHP approach was found to be an efficient tool to select the most appropriate litter material under specific scenarios.

Xylo-oligosaccharides from lignocellulosic materials: Chemical structure, health benefits and production by chemical and enzymatic hydrolysis

Currently, there is worldwide interest in the technological use of agro-industrial residues as a renewable source of food and biofuels. Lignocellulosic materials (LCMs) are a rich source of cellulose and hemicellulose. Hemicellulose is rich in xylan, a polysaccharide used to develop technology for producing alcohol, xylose, xylitol and xylo-oligosaccharides (XOSs). The XOSs are unusual oligosaccharides whose main constituent is xylose linked by β 1-4 bonds. The XOS applications described in this paper highlight that they are considered soluble dietary fibers that have prebiotic activity, favoring the improvement of bowel functions and immune function and having antimicrobial and other health benefits. These effects open a new perspective on potential applications for animal production and human consumption. The raw materials that are rich in hemicellulose include sugar cane bagasse, corncobs, rice husks, olive pits, barley straw, tobacco stalk, cotton stalk, sunflower stalk and wheat straw. The XOS-yielding treatments that have been studied include acid hydrolysis, alkaline hydrolysis, auto-hydrolysis and enzymatic hydrolysis, but the breaking of bonds present in these compounds is relatively difficult and costly, thus limiting the production of XOS. To obviate this limitation, a thorough evaluation of the most convenient methods and the opportunities for innovation in this area is needed. Another challenge is the screening and taxonomy of microorganisms that produce the xylanolytic complex and enzymes and reaction mechanisms involved. Among the standing out microorganisms involved in lignocellulose degradation are Trichoderma harzianum, Cellulosimicrobium cellulans, Penicillium janczewskii, Penicillium echinulatu, Trichoderma reesei and Aspergillus awamori. The enzyme complex predominantly comprises endoxylanase and enzymes that remove hemicellulose side groups such as the acetyl group. The complex has low β-xylosidase activities because β-xylosidase stimulates the production of xylose instead of XOS; xylose, in turn, inhibits the enzymes that produce XOS. The enzymatic conversion of xylan in XOS is the preferred route for the food industries because of problems associated with chemical technologies (e.g., acid hydrolysis) due to the release of toxic and undesired products, such as furfural. The improvement of the bioprocess for XOS production and its benefits for several applications are discussed in this study. © 2012 Elsevier Ltd.

Fontes de carbono de baixo custo para a produção de xilanase termoestável por aspergillus niger

A strain of the flamentous fungus Aspergillus niger was isolated and shown to possess extracellular xylanolytic activity. These enzymes have biotechnological potential and can be employed in various industries. This fungus produced its highest xylanase activity in a medium made up of 0.1% CaCO3, 0.5% NaCl, 0.1% NH4Cl, 0.5% corn steep liquor and 1% carbon source, at pH 8.0. A low-cost hemicellulose residue (powdered corncob) proved to be an excellent inducer of the A. niger xylanolytic complex. Filtration of the crude culture medium with suspended kaolin was ideal for to clarify the extract and led to partial purifcation of the xylanolytic activity. The apparent molecular mass of the xylanase was about 32.3 kDa. Maximum enzyme activity occurred at pH 5.0 and 55-60oC. Apparent Km was 10.41 ± 0.282 mg/mL and Vmax was 3.32 ± 0.053 U/mg protein, with birchwood xylan as the substrate. Activation energy was 4.55 kcal/mol and half-life of the crude enzyme at 60oC was 30 minutes. Addition of 2% glucose to the culture medium supplemented with xylan repressed xylanase production, but in the presence of xylose the enzyme production was not affected.

Comparing submerged and solid-state fermentation of agro-industrial residues for the production and characterization of lipase by Trichoderma harzianum

Lipase production by Trichoderma harzianum was evaluated in submerged fermentation (SF) and solid-state fermentation (SSF) using a variety of agro-industrial residues. Cultures in SF showed the highest activity (1.4 U/mL) in medium containing 0.5 % (w/v) yeast extract, 1 % (v/v) olive oil and 2.5 C:N ratio. This paper is the first to report lipase production by T. harzianum in SSF. A 1:2 mixture of castor oil cake and sugarcane bagasse supplemented with 1 % (v/w) olive oil showed the best results among the cultures in SSF (4 U/g ds). Lipolytic activity was stable in a slightly acidic to neutral pH, maintaining 50 % activity after 30 min at 50 C. Eighty percent of the activity remained after 1 h in 25 % (v/v) methanol, ethanol, isopropanol or acetone. Activity was observed with vegetable oils (olive, soybean, corn and sunflower) and long-chain triacylglycerols (triolein), confirming the presence of a true lipase. The results of this study are promising because they demonstrate an enzyme with interesting properties for application in catalysis produced by fermentation at low cost. © 2012 Springer-Verlag and the University of Milan.

Development and Biotechnological Application of a Novel Endoxylanase Family GH10 Identified from Sugarcane Soil Metagenome

Metagenomics has been widely employed for discovery of new enzymes and pathways to conversion of lignocellulosic biomass to fuels and chemicals. In this context, the present study reports the isolation, recombinant expression, biochemical and structural characterization of a novel endoxylanase family GH10 (SCXyl) identified from sugarcane soil metagenome. The recombinant SCXyl was highly active against xylan from beechwood and showed optimal enzyme activity at pH 6,0 and 45°C. The crystal structure was solved at 2.75 Å resolution, revealing the classical (β/α)8-barrel fold with a conserved active-site pocket and an inherent flexibility of the Trp281-Arg291 loop that can adopt distinct conformational states depending on substrate binding. The capillary electrophoresis analysis of degradation products evidenced that the enzyme displays unusual capacity to degrade small xylooligosaccharides, such as xylotriose, which is consistent to the hydrophobic contacts at the +1 subsite and low-binding energies of subsites that are distant from the site of hydrolysis. The main reaction products from xylan polymers and phosphoric acid-pretreated sugarcane bagasse (PASB) were xylooligosaccharides, but, after a longer incubation time, xylobiose and xylose were also formed. Moreover, the use of SCXyl as pre-treatment step of PASB, prior to the addition of commercial cellulolytic cocktail, significantly enhanced the saccharification process. All these characteristics demonstrate the advantageous application of this enzyme in several biotechnological processes in food and feed industry and also in the enzymatic pretreatment of biomass for feedstock and ethanol production. © 2013 Alvarez et al.