Immobilization of mutated xylanase from Neocallimastix patriciarum in E coli and application for kraft pulp biobleaching / Imobilização de xilanase mutada de Neocallimastix patriciarum em E coli e aplicação para polpa kraft

As an important enzyme, xylanase is widely used in the food, pulp, and textile industry. Different applications of xylanase warrant specific conditions including temperature and pH. This study aimed to carry out sodium alginate beads as carrier to immobilize previous reported mutated xylanase from Neocallimastix patriciarum which expressed in E. coli, the activity of immobilization of mutated xylanase was elevated about 4% at pH 6 and 13% at 62 °C. Moreover, the immobilized mutated xylanase retained a greater proportion of its activity than the wide type in thermostability. These properties suggested that the immobilization of mutated xylanase has potential to apply in biobleaching industry.

Como importante enzima, a xilanase é amplamente utilizada na indústria alimentícia, de celulose e têxtil. Diferentes aplicações de xilanase garantem condições específicas, incluindo temperatura e pH. Este estudo teve como objetivo realizar grânulos de alginato de sódio como carreador para imobilizar xilanase mutada relatada anteriormente de Neocallimastix patriciarum que expressa em E. coli, a atividade de imobilização da xilanase mutada foi elevada em cerca de 4% em pH 6 e 13% a 62 °C. Além disso, a xilanase mutada imobilizada reteve uma proporção maior de sua atividade do que o tipo amplo em termoestabilidade. Essas propriedades sugerem que a imobilização da xilanase mutada tem potencial para aplicação na indústria de biobranqueamento.

Modulating the production of xylanase by Bacillus pumilus BS131 through optimization using waste fiber sludge / Modulando a produção de xilanase por Bacillus pumilus BS131 por meio da otimização usando lodo de fibra residual

In recent days, cheapest alternative carbon source for fermentation purpose is desirable to minimize production cost. Xylanases have become attractive enzymes as their potential in bio-bleaching of pulp and paper industry. The objective of the present study was to identify the potential ability on the xylanase production by locally isolated Bacillus pumilus BS131 by using waste fiber sludge and wheat bran media under submerged fermentation. Culture growth conditions were optimized to obtain significant amount of xylanase. Maximum xylanase production was recorded after 72 hours of incubation at 30 °C and 7 pH with 4.0% substrate concentration. In the nutshell, the production of xylanase using inexpensive waste fiber sludge and wheat-bran as an alternative in place of expensive xylan substrate was more cost effective and environment friendly.

Nos últimos dias, a fonte alternativa de carbono mais barata para fins de fermentação é desejável para minimizar o custo de produção. As xilanases têm se tornado enzimas atraentes como seu potencial no biobranqueamento da indústria de papel e celulose. O objetivo do presente estudo foi identificar a capacidade potencial na produção de xilanase por Bacillus pumilus BS131 isolado localmente usando lodo de fibra residual e farelo de trigo em meio de fermentação submersa. As condições de crescimento da cultura foram otimizadas para obter uma quantidade significativa de xilanase. A produção máxima de xilanase foi registrada após 72 horas de incubação a 30 °C e pH 7 com concentração de substrato de 4,0%. Resumindo, a produção de xilanase usando lodo de fibra residual de baixo custo e farelo de trigo como uma alternativa no lugar do substrato de xilano caro foi mais econômica e ecológica.

Modulating the production of xylanase by Bacillus pumilus BS131 through optimization using waste fiber sludge / Modulando a produção de xilanase por Bacillus pumilus BS131 por meio da otimização usando lodo de fibra residual

In recent days, cheapest alternative carbon source for fermentation purpose is desirable to minimize production cost. Xylanases have become attractive enzymes as their potential in bio-bleaching of pulp and paper industry. The objective of the present study was to identify the potential ability on the xylanase production by locally isolated Bacillus pumilus BS131 by using waste fiber sludge and wheat bran media under submerged fermentation. Culture growth conditions were optimized to obtain significant amount of xylanase. Maximum xylanase production was recorded after 72 hours of incubation at 30 °C and 7 pH with 4.0% substrate concentration. In the nutshell, the production of xylanase using inexpensive waste fiber sludge and wheat-bran as an alternative in place of expensive xylan substrate was more cost effective and environment friendly.(AU)

Nos últimos dias, a fonte alternativa de carbono mais barata para fins de fermentação é desejável para minimizar o custo de produção. As xilanases têm se tornado enzimas atraentes como seu potencial no biobranqueamento da indústria de papel e celulose. O objetivo do presente estudo foi identificar a capacidade potencial na produção de xilanase por Bacillus pumilus BS131 isolado localmente usando lodo de fibra residual e farelo de trigo em meio de fermentação submersa. As condições de crescimento da cultura foram otimizadas para obter uma quantidade significativa de xilanase. A produção máxima de xilanase foi registrada após 72 horas de incubação a 30 °C e pH 7 com concentração de substrato de 4,0%. Resumindo, a produção de xilanase usando lodo de fibra residual de baixo custo e farelo de trigo como uma alternativa no lugar do substrato de xilano caro foi mais econômica e ecológica.(AU)

Immobilization of mutated xylanase from Neocallimastix patriciarum in E coli and application for kraft pulp biobleaching / Imobilização de xilanase mutada de Neocallimastix patriciarum em E coli e aplicação para polpa kraft

As an important enzyme, xylanase is widely used in the food, pulp, and textile industry. Different applications of xylanase warrant specific conditions including temperature and pH. This study aimed to carry out sodium alginate beads as carrier to immobilize previous reported mutated xylanase from Neocallimastix patriciarum which expressed in E. coli, the activity of immobilization of mutated xylanase was elevated about 4% at pH 6 and 13% at 62 °C. Moreover, the immobilized mutated xylanase retained a greater proportion of its activity than the wide type in thermostability. These properties suggested that the immobilization of mutated xylanase has potential to apply in biobleaching industry.(AU)

Como importante enzima, a xilanase é amplamente utilizada na indústria alimentícia, de celulose e têxtil. Diferentes aplicações de xilanase garantem condições específicas, incluindo temperatura e pH. Este estudo teve como objetivo realizar grânulos de alginato de sódio como carreador para imobilizar xilanase mutada relatada anteriormente de Neocallimastix patriciarum que expressa em E. coli, a atividade de imobilização da xilanase mutada foi elevada em cerca de 4% em pH 6 e 13% a 62 °C. Além disso, a xilanase mutada imobilizada reteve uma proporção maior de sua atividade do que o tipo amplo em termoestabilidade. Essas propriedades sugerem que a imobilização da xilanase mutada tem potencial para aplicação na indústria de biobranqueamento.(AU)

Beta-glucanases e xilanases na nutrição de não ruminantes / Beta-glucanases and xylanases in non-ruminant nutrition / Beta glucanasas y xilanasas en la nutrición de no rumiantes

A busca por custo x benefício na avicultura envolve compreender como melhorar o desempenho animal através da alimentação ao mesmo tempo em que haja diminuição dos custos. Tendo em vista o crescente aumento monetário da matéria prima da ração, várias pesquisas se voltam para novas fontes energéticas derivados de plantas, que contem quantidades significativas de fibras e agem como fatores antinutricionais, interferindo no desempenho zootécnico das aves. Os principais componentes das fibras, as beta glucanas e xilanas, podem ser quebradas e melhor aproveitadas quando no uso de enzimas exógenas chamadas de carboidrases, sendo a beta glucanase e xilase as respectivas enzimas que fazem a lise desses componentes. É necessário, portanto, melhor compreensão da forma de ação de cada enzima, assim como a fonte nutritiva das bases alimentares alternativas. Essa obra tem por objetivo abordar e revisar as duas principais fontes de fibras, beta glucanas e xilanas, e suas respectivas enzimas, beta glucanase e xilase.(AU)

The pursuit for cost-benefit in poultry farming involves understanding how to improve animal performance through feeding while reducing costs. In view of the growing monetary increase in feed raw material, several researches are focused on new energy sources derived from plants, which contain significant amounts of fiber and act as anti-nutritional factors, interfering with the zootechnical performance of birds. The main components of fiber, beta glucans and xylans, can be broken down and better utilized when using exogenous enzymes called carbohydrases, with beta glucanase and xylase being the respective enzymes that lyse these components. It is therefore necessary to better understand the mode of action of each enzyme, as well as the nutritive source of alternative food bases. This work aims to address and review the two main sources of fiber, beta glucans and xylans, and their respective enzymes, beta glucanase and xylase.(AU)

La búsqueda de la relación costo-beneficio en la avicultura implica comprender cómo mejorar el rendimiento animal a través de la alimentación y reducir los costos. Ante el creciente aumento monetario de la materia prima de los alimentos, varias investigaciones se enfocan en nuevas fuentes de energía derivadas de las plantas, que contienen cantidades significativas de fibra y actúan como factores antinutricionales, interfiriendo en el desempeño zootécnico de las aves. Los principales componentes de la fibra, los betaglucanos y los xilanos, se pueden descomponer y utilizar mejor cuando se usan enzimas exógenas llamadas carbohidrasas, siendo la betaglucanasa y la xilasa las enzimas respectivas que lisan estos componentes. Por lo tanto, es necesario comprender mejor el modo de acción de cada enzima, así como la fuente nutritiva de las bases alimenticias alternativas. Este trabajo tiene como objetivo abordar y revisar las dos principales fuentes de fibra, beta glucanos y xilanos, y sus respectivas enzimas, beta glucanasa y xilasa.(AU)

Cell wall remodeling under salt stress: Insights into changes in polysaccharides, feruloylation, lignification, and phenolic metabolism in maize.

Although cell wall polymers play important roles in the tolerance of plants to abiotic stress, the effects of salinity on cell wall composition and metabolism in grasses remain largely unexplored. Here, we conducted an in-depth study of changes in cell wall composition and phenolic metabolism induced upon salinity in maize seedlings and plants. Cell wall characterization revealed that salt stress modulated the deposition of cellulose, matrix polysaccharides and lignin in seedling roots, plant roots and stems. The extraction and analysis of arabinoxylans by size-exclusion chromatography, 2D-NMR spectroscopy and carbohydrate gel electrophoresis showed a reduction of arabinoxylan content in salt-stressed roots. Saponification and mild acid hydrolysis revealed that salinity also reduced the feruloylation of arabinoxylans in roots of seedlings and plants. Determination of lignin content and composition by nitrobenzene oxidation and 2D-NMR confirmed the increased incorporation of syringyl units in lignin of maize roots. Salt stress also induced the expression of genes and the activity of enzymes enrolled in phenylpropanoid biosynthesis. The UHPLC-MS-based metabolite profiling confirmed the modulation of phenolic profiling by salinity and the accumulation of ferulate and its derivatives 3- and 4-O-feruloyl quinate. In conclusion, we present a model for explaining cell wall remodeling in response to salinity.

Feruloylated Arabinoxylans from Nixtamalized Maize Bran Byproduct: A Functional Ingredient in Frankfurter Sausages.

Feruloylated arabinoxylans obtained from nixtamalized maize bran were evaluated in terms of physicochemical characteristics and antioxidant capacity when incorporated in frankfurter sausages. Concentrations of 0.15% and 0.30% of feruloylated arabinoxylans were incorporated in frankfurter sausages formulations and a control without feruloylated arabinoxylans was also prepared. Shear force, hardness, color measurement, proximate analysis, pH, titratable acidity, water-holding capacity, total phenols, and antioxidant capacity were evaluated. Phenolic content and antioxidant capacity were significantly higher (P < 0.0001) in all treatments, sausages containing feruloylated arabinoxylans compared to the control. The results showed that there was a significant difference (P < 0.0001) in total phenolic content and antioxidant capacity with all feruloylated arabinoxylans sausages treatments higher than control. Additionally, significant differences (P < 0.0001) were obtained in the physicochemical parameters.

Fiber source and xylanase on performance, egg quality, and gastrointestinal tract of laying hens

The objective of this study was to ascertain the influence of different dietary fiber sources and the usage of xylanase on diet of commercial layers and their influence on productive performance, egg quality, and digestive organ biometry. A total of 864 Lohmann® White hens was fed diets with three different fiber sources (wheat bran, soybean hull, or coffee husk) with or without xylanase inclusion (concentration of 160,000 BXU/g) in a 3×2 factorial arrangement, with six replicates of 24 birds each, from 25 to 44 weeks of age. There were no interactions between dietary fiber and xylanase inclusion. The enzyme supplementation did not influence any parameters evaluated. There were dietary fiber effects on body weight gain, viability, egg weight, eggshell quality, yolk pigmentation, and liver and gizzard relative weights. Wheat bran, soybean hull, and coffee husk can be used in laying hen diets without detrimental effect on productive performance. The enzyme used had no effect on the performance and eggshell quality of laying hens.(AU)

Structural diversity of alkali-soluble polysaccharides from the fruit cell walls of tucumã (Astrocaryum aculeatum), a commelinid monocotyledon from the family Arecaceae.

The polysaccharide compositions of primary and secondary cell walls of members of the family Arecaceae in the commelinid clade of monocotyledonous plants have previously been found to be distinguishable from other commelinid families, and to be more similar to those of non-commelinids. However, few studies have been conducted. We aimed to extract and characterize the main cell-wall polysaccharides in the fruit pulp of tucumã (Astrocaryum aculeatum), a member of Arecaceae family. Hemicellulosic polysaccharides extracted by alkali from the fruit pulp were present in greater proportions (6.4% yield) than water-extracted ones (3.0% yield). Thus, the former was analyzed using monosaccharide composition, methylation, molecular weight determination and 13C-NMR data. The tucumã alkaline extract presented a highly ramified acidic galactoarabinoxylan (53.7%), a linear (1 â†’ 5)-linked α-L-arabinan (27.8%), a low branched glucuronoxylan (14.1%) and small portions of a xyloglucan (4.4%). The major polysaccharide found in A. aculeatum (acidic galactoarabinoxylan) is similar to those found in other commelinid plants such as grasses and cereals.

Reuse of wastewater from pulp industry for the optimization of fungal xylanase production / Reutilização de efluentes da indústria de celulose para otimização da produção de xilanase fúngica

The production of enzymes using agro-industrial waste is a low cost alternative for the reuse of byproducts, with the subsequent impact decrease on the environment. Current analysis produced xylanase using fungus Aspergillus niger, with two types of wastewater generated during the pulp chemical bleaching phase as inducers. Xylanase was produced by submerged liquid fermentation and factorial design optimized parameters that influence production (concentration of wastewater and production period). Initial culture conditions (pH, temperature and agitation) were optimized independently. Alkaline wastewater was more effective than acidic wastewater for the induction of xylanase in optimized conditions: 50% of culture medium, 7-day production, 30C, pH 6.0 and agitation at 160 rpm. Despite different results, acidic and alkaline wastewaters induced xylanase production by A. niger when employed in concentrations lower than or equal to 50% of culture medium and in the most optimal conditions described above. Alkaline wastewater is highlighted as the most efficient for such production. 

A produção de enzimas, a partir de resíduos agroindustriais, é uma alternativa para reutilização destes subprodutos como substrato de baixo custo reduzindo seu impacto causado pelo descarte no meio ambiente. Diante disso, o objetivo deste estudo foi a produção de xilanase por Aspergillus niger, utilizando dois efluentes gerados nas fases de branqueamento químico de polpa de celulose como indutores. A produção de xilanase foi realizada por fermentação líquida submersa, e utilizou-se planejamento fatorial para otimização dos parâmetros influentes de produção (concentração de efluentes e período de produção) e as condições iniciais de cultivo (pH, temperatura e agitação) foram otimizadas de forma independente. O efluente alcalino se mostrou mais efetivo do que o efluente com característica ácida, na indução de xilanase em condições otimizadas: 50% em relação ao meio de cultura, sete dias de produção, 30°C, pH 6,0 e agitação de 160 rpm. Conclui-se que ambos os efluentes, ácido e alcalino, apesar de levarem a diferentes resultados, são capazes de induzir a produção de xilanase por A. niger quando empregues em concentrações menores ou iguais a 50% em relação ao meio de cultura e nas condições ótimas descritas acima, destacando-se o efluente alcalino como mais eficiente para produção de tal enzima.(AU)

Isolation and characterization of a xylan with industrial and biomedical applications from edible açaí berries (Euterpe oleraceae).

The chemical features of xylan largely determine its physical and biological properties and its use in the industry. In this work, we describe the occurrence, purification and partial characterization of a xylan in edible açaí berries (Euterpe oleraceae), using a fairly simple and inexpensive method of purification from alkaline açaí extract. A mainly linear (1→4)-ß-d-xylan was found as the majority (70%) of alkali extract and 4.2% of the dry matter açaí pulp. This represents the biggest source of xylan found so far in a fruit pulp and could be suitable for applications in the industry and biomedical field.

Anatomy, nutritional value and cell wall chemical analysis of foliage leaves of Guadua chacoensis (Poaceae, Bambusoideae, Bambuseae), a promising source of forage.

BACKGROUND: The present study combines morphological and anatomical studies, cell wall chemical composition analysis, as well as assessment of the nutritional value of Guadua chacoensis foliage leaves. RESULTS: Foliage leaves of G. chacoensis are a promising source of forage because: (a) as a native woody bamboo, it is adapted to and helps maintain environmental conditions in America; (b) leaf anatomical studies exhibit discontinuous sclerenchyma, scarcely developed, while pilose indumentum, silica cells, prickles and hooks are also scarce; (c) it has a high protein content, similar to that of Medicago sativa, while other nutritional parameters are similar to those of common forages; and (d) glucuronoarabinoxylan, the major extracted polysaccharide, has one-third of the 4-linked ß-d-xylopyranosyl units of the backbone substituted mainly with α-l-arabinofuranose as single stubs or non-reducing end of short chains, but also 5-linked α-l-arabinofuranose units, terminal ß-d-xylopyranose and d-galactopyranose units, as well as α-d-glucuronic acid residues and small amounts of its 4-O-methylated derivative. CONCLUSION: These results constitute the first report on this species, and as culms are utilized in constructions and crafts, the remaining leaves, when used as forage, constitute a byproduct that allows an additional income opportunity. © 2016 Society of Chemical Industry.

Ultrasound-assisted extraction of hemicelluloses from grape pomace using response surface methodology.

An ultrasound-assisted procedure was applied to the extraction of hemicelluloses from grape pomace at a mild temperature (20°C). A Central composite design (CCD) was employed to optimize the ultrasound-assisted extraction (UAE) of hemicelluloses from grape pomace with the aim to maximize their extraction yield, and, also, the obtention of the main polymers forming this fraction: Xyloglucans (XLG), Mannans (MAN) and Xylans (XN). Extraction time (X1), solid:liquid ratio (X2) and KOH concentration (X3) were the variables used to optimize the process. The conditions that maximize (1) the extraction yield of hemicelluloses and the contents of (2) XLG, (3) MAN and (4) XN, were: (1) X1=2.6h; X2=1:48 (w/v); X3=0.4M, (2) X1=2.9h; X2=1:57 (w/v); X3=2.25M, (3) X1=2.7h; X2=1:58(w/v);X3=2.2M, and (4) X1=3h; X2=1:60 (w/v); X3=2.3M, respectively. Under these conditions, the maximum extraction yield of hemicelluloses, XLG, MAN and XN contents were: ∼7.9±0.2%, ∼3.6±0.02%, ∼1.1±0.04% and ∼1.2±0.02%, respectively. Close agreement between experimental and predicted values was found. The results suggest that the ultrasound-assisted extraction could be a good option for the extraction of hemicellulosic polysaccharides from grape pomace at industrial level.

Interaction between xylanase, glucose oxidase and ascorbic acid on the technological quality of whole wheat bread / Interação entre xilanase, glicose oxidase e ácido ascórbico na qualidade tecnológica de pão elaborado com farinha do trigo integral

This study aimed to verify the performance of xylanase and its interaction with oxidants agents (glucose oxidase and ascorbic acid) on the quality of whole wheat bread. The experiment was based on a central composite rotational design and the Response Surface Methodology was used to analyze the results. None of the xylanase, glucose oxidase or ascorbic acid concentrations within the studied range led to a significant difference in the specific volume. The highest moisture content and the lowest firmness values were reported in the bread with lower and intermediate levels of xylanase and larger amounts of glucose oxidase and ascorbic acid. This effect was observed mainly at the end of the storage period. A minimum amount of xylanase (from 33 to 63 EDX kg-¹ flour) showed to be essential for obtaining best results. Levels of ascorbic acid above 63mg kg-¹ and glucose oxidase above 91 SRU kg-¹ proved to be necessary to offer the beneficial effect of xylanase.(AU)

O objetivo deste estudo foi verificar o desempenho da xilanase e sua interação com agentes oxidantes (glicose oxidase e ácido ascórbico) na qualidade de pão elaborado com farinha do trigo integral. O experimento foi baseado em um delineamento composto central rotacional (DCCR) e a Metodologia de Superfície de Resposta foi utilizada para analisar os resultados. Nenhuma das concentrações de xilanase, glicose oxidase e ácido ascórbico, dentro da faixa estudada, levaram a uma diferença significativa no volume específico. Mais alto conteúdo de umidade e menor firmeza foram encontrados nos pães com concentrações menores e intermediárias de xilanase e concentrações maiores de glicose oxidase e ácido ascórbico. Este efeito foi observado principalmente no final do período de estocagem. Uma quantidade mínima de xilanase (de 33 a 63 EDX kg-¹ farinha) mostra ser essencial na obtenção dos melhores resultados. Concentrações de ácido ascórbico acima de 63mg kg-¹ e glicose oxidase acima de 91 SRU kg-¹ mostraram ser necessárias para que o efeito benéfico da xilanase fosse observado.(AU)

Characterization of water extractable arabinoxylans from a spring wheat flour: rheological properties and microstructure.

In the present study water extractable arabinoxylans (WEAX) from a Mexican spring wheat flour (cv. Tacupeto F2001) were isolated, characterized and gelled and the gel rheological properties and microstructure were investigated. These WEAX presented an arabinose to xylose ratio of 0.66, a ferulic acid and diferulic acid content of 0.526 and 0.036 µg/mg WEAX, respectively and a Fourier Transform Infra-Red (FT-IR) spectrum typical of arabinoxylans. The intrinsic viscosity and viscosimetric molecular weight values for WEAX were 3.5 dL/g and 504 kDa, respectively. WEAX solution at 2% (w/v) formed gels induced by a laccase as cross-linking agent. Cured WEAX gels registered storage (G') and loss (G'') modulus values of 31 and 5 Pa, respectively and a diferulic acid content of 0.12 µg/mg WEAX, only traces of triferulic acid were detected. Scanning electron microscopy analysis of the lyophilized WEAX gels showed that this material resembles that of an imperfect honeycomb.

Comparison of the impact of ionic liquid pretreatment on recalcitrance of agave bagasse and switchgrass.

Lignocellulose represents a sustainable source of carbon for transformation into biofuels. Effective biomass to sugar conversion strategies are needed to lower processing cost without degradation of polysaccharides. Since ionic liquids (ILs) are excellent solvents for pretreatment/dissolution of biomass, IL pretreatment was carried out on agave bagasse (AGB-byproduct of tequila industry) and digestibility and sugar yield was compared with that obtained with switchgrass (SWG). The IL pretreatment was conducted using ([C2mim][OAc]) at 120 and 160 °C for 3h and 15% biomass loading. While pretreatment using [C2mim][OAc] was very effective in improving the digestibility of both feedstocks, IL pretreatment at 160 °C resulted in higher delignification for AGB (45.5%) than for SWG (38.4%) when compared to 120 °C (AGB-16.6%, SWG-8.2%), formation of a highly amorphous cellulose structure and a significant enhancement of enzyme kinetics. These results highlight the potential of AGB as a biofuel feedstock that can produce high sugar yields with IL pretreatment.

Lignocellulolytic enzyme production of Pleurotus ostreatus growth in agroindustrial wastes

The mushroom Pleurotus ostreatus has nutritional and medicinal characteristics that depend on the growth substrate. In nature, this fungus grows on dead wood, but it can be artificially cultivated on agricultural wastes (coffee husks, eucalyptus sawdust, corncobs and sugar cane bagasse). The degradation of agricultural wastes involves some enzyme complexes made up of oxidative (laccase, manganese peroxidase and lignin peroxidase) and hydrolytic enzymes (cellulases, xylanases and tanases). Understanding how these enzymes work will help to improve the productivity of mushroom cultures and decrease the potential pollution that can be caused by inadequate discharge of the agroindustrial residues. The objective of this work was to assess the activity of the lignocellulolytic enzymes produced by two P. ostreatus strains (PLO 2 and PLO 6). These strains were used to inoculate samples of coffee husks, eucalyptus sawdust or eucalyptus bark add with or without 20 % rice bran. Every five days after substrate inoculation, the enzyme activity and soluble protein concentration were evaluated. The maximum activity of oxidative enzymes was observed at day 10 after inoculation, and the activity of the hydrolytic enzymes increased during the entire period of the experiment. The results show that substrate composition and colonization time influenced the activity of the lignocellulolytic enzymes.

Endoglucanase production with the newly isolated Myceliophtora sp. i-1d3b in a packed bed solid state fermentor

This work is aimed to produce endoglucanase through solid state fermentation in a packed bed bioreactor with the use of the fungus Myceliophtora sp. I-1D3busing a mixture of wheat bran (WB) and sugar cane bagasse (SCB) as culture medium. Preliminary tests were performed in polypropylene plastic bags, controlling the variables temperature (40, 45, and 50ºC), initial moisture content (75, 80, and 85%, w.b.), and weight proportion SCB/WB (1:1, 7:3, and 9:1). The highest enzyme activities in plastic bags were obtained using the substrate proportion of 7:3, 50ºC temperature, and 80% initial moisture content (878 U/grams of dry solid). High activities of filter-paper cellulase and xylanase were also obtained in plastic bags and some results are reported. For the packed bed experiments, the temperature (45 and 50ºC) and the air flow rate (80, 100 and 120L/h) were the controlled variables. Activity of endoglucanase was similar to plastic bag tests. A longitudinal gradient of moisture content, was observed increasing from the bottom to the top of the reactor, even though the longitudinal enzyme activity profile was flat for almost the whole bed. Air flow rate did not affect enzyme activity, while experiments carried out at 50ºC showed higher enzyme activities. The maximum temperature peak observed was at about 6ºC above the process temperature.

Utilization of Agro-industrial Wastes for the Simultaneous Production of Amylase and Xylanase by Thermophilic Actinomycetes

Agro-industrial wastes such as sugarcane bagasse, wheat bran, rice bran, corn cob and wheat straw are cheapest and abundantly available natural carbon sources. The present study was aimed to production of amylase and xylanase simultaneously using agro-industrial waste as the sole carbon source. Seven thermophilic strains of actinomycete were isolated from the mushroom compost. Among of these, strain designated MSC702 having high potential to utilize agro-industrial wastes for the production of amylase and xylanase. Strain MSC702 was identified as novel species of Streptomyces through morphological characterization and 16S rRNA gene sequence. Enzyme production was determined using 1% (w/v) of various agro-industrial waste in production medium containing (g/100mL): K2HPO4(0.1), (NH4)2SO4(0.1), NaCl (0.1), MgSO4(0.1) at pH 7.0 after incubation of 48 h at 50°C. The amylase activity (373.89 IU/mL) and xylanase activity (30.15 IU/mL) was maximum in rice bran. The decreasing order of amylase and xylanase activity in different type of agro-industrial wastes were found rice bran (RB) > corn cob (CC) > wheat bran (WB) > wheat straw (WS) > sugarcane bagasse (SB) and rice bran (RB) > wheat bran (WB) > wheat straw (WS) > sugarcane bagasse (SB) > corn cob (CC), respectively. Mixed effect of different agro-industrial wastes was examined in different ratios. Enzyme yield of amylase and xylanase was ~1.3 and ~2.0 fold higher with RB: WB in 1:2 ratio.

Selection of thermophilic and thermotolerant fungi for the production of cellulases and xylanases under solid-state fermentation

Twenty-seven thermophilic and thermotolerant fungal strains were isolated from soil, decaying organic matter and sugarcane piles based on their ability to grow at 45ºC on medium containing corn straw and cardboard as carbon sources. These fungi were identified in the genera Aspergillus, Thermomyces, Myceliophthora, Thermomucor and Candida. The majority of the isolated strains produced xylanase and cellulases under solid state fermentation (SSF). The highest cellulase and xylanase productions were obtained by the cultivation of the strains identified as Aspergillus fumigatus M.7.1 and Myceliophthora thermophila M.7.7. The enzymes from these strains exhibited maximum activity at pH 5.0 and at 60 and 70ºC. The endo-glucanase from A. fumigatus was stable from 40ºC to 65ºC and both endo-glucanase and xylanase from M. thermophila were stable in this temperature range when in absence of substrate. The enzymes were stable from pH 4.0 to 9.0.