Utilization of pineapple stem juice to enhance enzyme-hydrolytic efficiency for sugarcane bagasse after an optimized pre-treatment with alkaline peroxide

The enzymatic hydrolysis of sugarcane bagasse was investigated by treating a peroxide-alkaline bagasse with a pineapple stem juice, xylanase and cellulase. Pre-treatment procedures of sugarcane bagasse with alkaline hydrogen peroxide were evaluated and compared. Analyses were performed using 2(4) factorial designs, with pre-treatment time, temperature, magnesium sulfate and hydrogen peroxide concentration as factors. The responses evaluated were the yield of cellobiose and glucose released from pretreated bagasse after enzymatic hydrolysis. The results show that the highest enzymatic conversion was obtained for bagasse using 2% hydrogen peroxide at 60 degrees C for 16 h in the presence of 0.5% magnesium sulfate. Bagasse (5%) was treated with pineapple stem extract, which contains mixtures of protease and esterase, in combination with xylanase and cellulase. It was observed that the amount of glucose and cellobiose released from bagasse increased with the mixture of enzymes. It is believed that the enzymes present in pineapple extracts are capable of hydrolyze specific linkages that would facilitate the action of digesting plant cell walls enzymes. This increases the amount of glucose and other hexoses that are released during the enzymatic treatment and also reduces the amount of cellulase necessary in a typical hydrolysis. (C) 2010 Elsevier Ltd. All rights reserved.

Purification and Biochemical Properties of a Glucose-Stimulated beta-D-Glucosidase Produced by Humicola grisea var. thermoidea Grown on Sugarcane Bagasse

The effect of several carbon sources on the production of mycelial-bound beta-glucosidase by Humicola grisea var. thermoidea in submerged fermentation was investigated. Maximum production occurred when cellulose was present in the culture medium, but higher specific activities were achieved with cellobiose or sugarcane bagasse. Xylose or glucose (1%) in the reaction medium stimulated beta-glucosidase activity by about 2-fold in crude extracts from mycelia grown in sugarcane bagasse. The enzyme was purified by ammonium sulfate precipitation, followed by Sephadex G-200 and DEAE-cellulose chromatography, showing a single band in PAGE and SDS-PAGE. The beta-glucosidase had a carbohydrate content of 43% and showed apparent molecular masses of 57 and 60 kDa, as estimated by SDS-PAGE and gel filtration, respectively. The optimal pH and temperature were 6.0 and 50 degrees C, respectively. The purified enzyme was thermostable up to 60 min in water at 55 degrees C and showed half-lives of 7 and 14 min when incubated in the absence or presence of 50 mM glucose, respectively, at 60 degrees C. The enzyme hydrolyzed p-nitrophenyl-beta-D-glucopyranoside, p-nitrophenyl-beta-D-galactopyranoside, p-nitrophenyl-beta-D-fucopyranoside, p-nitrophenyl-beta-D-xylopyranoside, o-nitrophenyl-beta-D-galactopyranoside, lactose, and cellobiose. The best synthetic and natural substrates were p-nitrophenyl-beta-D-fucopyranoside and cellobiose, respectively. Purified enzyme activity was stimulated up to 2-fold by glucose or xylose at concentrations from 25 to 200 mM. The addition of purified or crude beta-glucosidase to a reaction medium containing Trichoderma reesei cellulases increased the saccharification of sugarcane bagasse by about 50%. These findings suggest that H. grisea var. thermoidea beta-glucosidase has a potential for biotechnological applications in the bioconversion of lignocellulosic materials.

In vitro evaluation of fibrolytic enzymes as additives for maize (Zea mays L.) silage - II. Effects on rate of acidification, fibre degradation during ensiling and rumen fermentation

A study was carried out to determine the influence of fibrolytic enzymes derived from mesophilic or thermophilic fungal sources, added at ensiling, on time-course fermentation characteristics and in vitro rumen degradation of maize silage. The mesophilic enzyme was a commercial product derived from Trichodenna reesei (L), whereas the thermophilic enzyme was a crude extract produced from Thermoascus aurantiacus (Ta) in this laboratory. The fungus was cultured using maize cobs as a carbon source. The resulting fermentation extract was deionised to remove sugars and characterised for its protein concentration, main and side enzymic activities, optimal pH, protein molecular mass and isoelectric point. In an additional study, both enzymes were added to maize forage (333.5 g DM/kg, 70.0, 469.8, 227.1 and 307.5 g/kg DM of CP, NDF, ADF and starch, respectively) at two levels each, normalized according to xylanase activity, and ensiled in 0.5 kg capacity laboratory minisilos. Duplicate silos were opened at 2, 4, 8, 15, and 60 days after ensiling, and analysed for chemical characteristics. Silages from 60 days were bulked and in vitro gas production (GP) and organic matter degradability (OMD) profiles evaluated using the Reading Pressure Technique (RPT), in a completely randomised design. The crude enzyme extract contained mainly xylanase and endoglucanase activities, with very low levels of exoglucanase, which probably limited hydrolysis of filter paper. The extract contained three major protein bands of between 29 and 55 kDa, with mainly acidic isoelectric points. Ensiling maize with enzymes lowered (P < 0.05) the final silage pH, with this effect being observed throughout the ensiling process. All enzyme treatments reduced (P < 0.05) ADF contents. Treatments including Ta produced more gas (P < 0.05) than the controls after 24 h incubation in vitro, whereas end point gas production at 96 h was not affected. Addition of Ta increased (P < 0.01) OMD after 12 h (410 and 416 g/kg versus 373 g/kg), whereas both L and Ta increased (P < 0.05) OMD after 24 h. Addition of enzymes from mesophilic or thermophilic sources to maize forage at ensiling increased the rate of acidification of the silages and improved in vitro degradation kinetics, suggesting an improvement in the nutritive quality. (C) 2003 Elsevier B.V All rights reserved.

Protease production by different thermophilic fungi

A comparative study was carried out to evaluate protease production in solid-state fermentation (SSF) and submerged fermentation (SmF) by nine different thermophilic fungi - Thermoascus aurantiacus Miehe, Thermomyces lanuginosus, T. lanuginosus TO.03, Aspergillus flavus 1.2, Aspergillus sp. 13.33, Aspergillus sp. 13.34, Aspergillus sp. 13.35, Rhizomucor pusillus 13.36 and Rhizomucor sp. 13.37 - using substrates containing proteins to induce enzyme secretion. Soybean extract (soybean milk), soybean flour, milk powder, rice, and wheat bran were tested. The most satisfactory results were obtained when using wheat bran in SSF. The fungi that stood out in SSF were T. lanuginosus, T. lanuginosus TO.03, Aspergillus sp. 13.34, Aspergillus sp. 13.35, and Rhizomucor sp. 13.37, and those in SmF were T. aurantiacus, T. lanuginosus TO.03, and 13.37. In both fermentation systems, A. flavus 1.2 and R. pusillus 13.36 presented the lowest levels of proteolytic activity.

Isolamento de leveduras fermentadoras de pentoses e suas aplicações na produção de xilitol e etanol a partir do licor negro proveniente do processo Kraft de extração da celulose

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Aplicação de extratos enzimáticos fúngicos na ensilagem do milho

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Análise dos compostos flavorizantes da cana-de-açúcar e otimização da aplicação de extratos ricos em β-glicosidases para liberação de aroma na produção de aguardente de cana

Pós-graduação em Química - IBILCE

Aplicação de xilanase e/ou ciclodextria glicotransferase (CGTase) na produção de pães

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Aplicações de coquetéis de celulases para a sacarificação enzimática do bagaço de cana-de-açúcar

Pós-graduação em Química - IBILCE

Production of a xylose-stimulated beta-glucosidase and a cellulase-free thermostable xylanase by the thermophilic fungus Humicola brevis var. thermoidea under solid state fermentation

Humicola brevis var. thermoidea cultivated under solid state fermentation in wheat bran and water (1:2 w/v) was a good producer of beta-glucosidase and xylanase. After optimization using response surface methodology the level of xylanase reached 5,791.2 +/- A 411.2 U g(-1), while beta-glucosidase production was increased about 2.6-fold, reaching 20.7 +/- A 1.5 U g(-1). Cellulase levels were negligible. Biochemical characterization of H. brevis beta-glucosidase and xylanase activities showed that they were stable in a wide pH range. Optimum pH for beta-glucosidase and xylanase activities were 5.0 and 5.5, respectively, but the xylanase showed 80 % of maximal activity when assayed at pH 8.0. Both enzymes presented high thermal stability. The beta-glucosidase maintained about 95 % of its activity after 26 h in water at 55 A degrees C, with half-lives of 15.7 h at 60 A degrees C and 5.1 h at 65 A degrees C. The presence of xylose during heat treatment at 65 A degrees C protected beta-glucosidase against thermal inactivation. Xylanase maintained about 80 % of its activity after 200 h in water at 60 A degrees C. Xylose stimulated beta-glucosidase activity up to 1.7-fold, at 200 mmol L-1. The notable features of both xylanase and beta-glucosidase suggest that H. brevis crude culture extract may be useful to compose efficient enzymatic cocktails for lignocellulosic materials treatment or paper pulp biobleaching.

Cantharellus aurantiacus, der orangfarbene Pfefferling, ist ein giftiger Pilz

Pfuhl

Cantharellus aurantiacus Wulf. (Falscher Eierpilz)

B. Studer

Phylogenetic heterogeneity within the genus Herpetosiphon: Transfer of the marine species Herpetosiphon cohaerens, Herpetosiphon nigricans and Herpetosiphon persicus to the genus Lewinella gen, nov. in the Flexibacter-Bacteroides-Cytophaga phylum

Analysis of the 16S rDNA sequences of species currently assigned to the genus Herpetosiphon revealed intrageneric phylogenetic heterogeneity. The thermotolerant freshwater species Herpetosiphon geysericola is most closely related to the type species Herpetosiphon aurantiacus in the Chloroflexus Subdivision of the green non-sulfur bacteria, The marine species Herpetosiphon cohaerens, Herpetosiphon nigricans and Herpetosiphon persicus, on the other hand, were found to form a cluster with the sheathed bacterium Haliscomenobacter hydrossis in the Saprospira group of the Flexibacter-Bacteroides-Cytophaga (FBC) phylum. A proposal is made to transfer these marine species to the genus Lewinella gen. nov. as Lewinella cohaerens comb, nov., Lewinella nigricans comb. nov, and Lewinella persica comb. nov. The marine sheathed gliding bacterium Flexithrix dorotheae was also found to be a member of the FBC phylum but on a separate phylogenetic line to the marine herpetosiphons now assigned to the genus Lewinella.

Taxonomia da subfamília Corinninae (Araneae, Corinnidae) nas regiões neotropical e neártica

The subfamily Corinninae is characterized and diagnosed. Two synapomorphies are hypothesized for the subfamily, both regarding the male palpal reservoir, which is primarily coiled and presents a sclerotized distal sector. Seventeen genera are recognized, six of which are new: Abapeba (type species Corinna lacertosa Simon), Erendira (type species Corinna pallidoguttata Simon), Septentrinna (type species Corinna bicalcarata Simon), Simonestus (type species Diestus validus Simon), Tapixaua (type species T. callida sp. nov.) and Tupirinna (type species T. rosae sp. nov.). The genera Creugas Thorell, Falconina Brignoli and Paradiestus Mello-Leitão are revalidated. Diestus Simon and Lausus Simon are newly synonymized with Corinna C. L. Koch. Chemmis Simon is included in the synonymy of Megalostrata Karsch. Hypsinotus L. Koch is removed from the synonymy of Corinna and included in the synonymy of Creugas. Thirteen new species are described: Septentrinna yucatan and S. potosi from Mexico; Tupirinna rosae from Venezuela and Brazil; Tapixaua callida from Brazil and Peru; Abapeba hoeferi, A. rioclaro, A. taruma, Corinna ducke, C. colombo, C. mourai, C. recurva and Parachemmis manauara from Brazil; Creugas lisei from Brazil, Argentina and Uruguay. Twenty seven species are redescribed. Fifty eight new combinations are presented: from Chemmis, Septentrinna steckleri (Gertsch); from Corinna, Abapeba abalosi (Mello-Leitão), A. cleonei (Petrunkevitch), A. echinus (Simon), A. grassima (Chickering), A. guanicae (Petrunkevitch), A. lacertosa (Simon), A. luctuosa (F. O. Pickard-Cambridge), A. lugubris (Schenkel), A. pennata (Caporiacco), A. kochi (Petrunkevitch), A. saga (F. O. Pickard-Cambridge), A. wheeleri (Petrunkevitch), Creugas annamae (Gertsch & Davis), C. apophysarius (Caporiacco), C. bajulus (Gertsch), C. bellator (L. Koch), C. bicuspis (F.O. Pickard-Cambridge), C. epicureanus (Chamberlin), C. falculus (F. O. Pickard-Cambridge), C. mucronatus (F. O. Pickard-Cambridge), C. navus (F. O. Pickard-Cambridge), C. nigricans (C. L. Koch), C. plumatus (L. Koch), C. praeceps (F. O. Pickard-Cambridge), C. silvaticus (Chickering), C. uncatus (F. O. Pickard-Cambridge), Erendira luteomaculatta (Petrunkevitch), E. pallidoguttata (Simon), E. subsignata (Simon), Falconina albomaculosa (Schmidt), F. crassipalpis (Chickering), F. gracilis (Keyserling), Megalostrata raptrix (L. Koch), Paradiestus egregius (Simon), P. giganteus (Karsch), P. penicillatus (Mello-Leitão), P. vitiosus (Keyserling), Septentrinna bicalcarata (Simon), S. paradoxa (F. O. Pickard-Cambridge), S. retusa (F. O. Pickard-Cambridge), Simonestus pseudobulbolus (Caporiacco), S. robustus (Chickering), S. semiluna (F.O. Pickard-Cambridge), Stethorrhagus maculatus (L. Koch) and Xeropigo smedigari (Caporiacco); from Diestus, Corinna alticeps (Keyserling), C. kochi (Simon), Simonestus occidentalis (Schenkel), S. separatus (Schmidt) and S. validus (Simon); from Lausus, Corinna grandis (Simon) and Abapeba sicarioides (Mello-Leitão); from Medmassa, Corinna andina (Simon) and C. venezuelica (Caporiacco); from Megalostrata, Erendira atrox (Caporiacco) and Erendira pictitorax (Caporiacco); from Parachemmis, Tupirinna trilineata (Chickering). Five combinations are restaured: Corinna aenea Simon, Creugas cinnamius Simon, Creugas gulosus Thorell, Falconina melloi (Schenkel), Paradiestus aurantiacus Mello-Leitão. Twenty five new synonymies are proposed: Diestus altifrons Mello-Leitão with Corinna nitens (Keyserling); Corinna tomentosa Simon, C. tridentina Mello-Leitão, Hypsinotus flavipes Keyserling, H. humilis Keyserling and Xeropigo scutulatus Simon with Xeropigo tridentiger (O. Pickard-Cambridge); Corinna cribosa Mello-Leitão and C. stigmatica Simon with Falconina gracilis (Keyserling); Corinna casueta Chickering with SIMONestus separatus (Schmidt); Corinna abnormis Petrunkevitch, C. antillana BRYANT, C. consobrina Simon, C. inornata Kraus, C. nervosa F. O. Pickard-Cambridge, C. wolleboeki Banks, Creugas cetratus Simon, C. senegalensis Simon and Hypsinotus gracilipes Keyserling with Creugas gulosus Thorell; Chemmis frederici Simon, Delozeugma formidabile O. Pickard-Cambridge, D. mordicans O. Pickard-Cambridge, Megalostrata sperata Kraus and M. venifica KARSCH with Megalostrata raptrix (L. Koch); Megalostrata lohmanderi Caporiacco with Erendira atrox (Caporiacco); Corinna tenubra Chickering with Parachemmis fuscus Chickering. One new name, Creugas berlandi, is erected for Corinna bellatrix Schmidt. Males of Creugas cinnamius, Corinna kochi, Methesis semirufa Simon, Paradiestus aurantiacus, Septentrinna steckleri and Xeropigo smedigari, the females of Paradiestus giganteus, Septentrinna bicalcarata and the adult female of S. steckleri are described for the first time.

A new species of Trypanosyllis (Polychaeta : Syllidae) from Brazil, with a redescription of Brazilian material of Trypanosyllis zebra

A new species of Trypanosyllis was found in a collection of polychaetes living on algae, sponges, ascidians and sabelariid reef; at the intertidal zone of a rocky shore, at Praia do Guarau, south-eastern Brazil. Trypanosyllis aurantiacus sp. nov., is characterized by having an orange body in life, with dark red antennae and cirri throughout, falcigers with short, sub-bidentate blades, and parapodia with thick, distally sharp, protruding aciculae, two to three aciculae on each anterior parapodium, two aciculae on midbody segments, single acicula per parapodium on posteriormost chaetigers. Trypanosyllis aurantiacus sp. nov., is compared with the most similar congeners and a redescription of Trypanosyllis zebra, based on Brazilian specimens collected from similar environments at nearby beaches, is given.