Atividade β-D-N-Acetilglucosaminidásica de enzimas imobilizadas extraídas da Artemia franciscana e possíveis aplicações biotecnológicas

A β-D-N-acetilglucosaminidase extracted and partially isolated from crustacean Artemia franciscana by ammonium sulfate precipitation and filtration gel chromatography Bio Gel A 1.5m. the enzyme was immobilized on ferromagnetic Dacron yielding a insoluble active derivative with 5.0 units/mg protein and 10.35% of the soluble enzyme activity. β-D-N-acetilglucosaminidase-ferromagnetic Dacron was easily removed from the reaction mixture by a magnetic field, it was reused for ten times without loss in its activity. The ferromagnetic Dacron was better activated at pH 5.0. The particles visualized at scanning electron microscope (SEM) had presented different sizes, varying between 721nm and 100µm. Infra red confirmed immobilization on support, as showed by primary amino peaks at 1640 and 1560 cm-1 . The immobilize enzyme presented Km of 2.32 ± 0.48 mM and optimum temperature of 50°C. Bought presented the same thermal stable of the soluble enzyme and larger enzymatic activity at pH 5.5. β-D-N-acetilglucosaminidase-Dacron ferromagnético showed sensible for some íons as the silver (AgNO3), with loss of activity. The β-D-N acetilglucosaminidase activity for mercury chloride (HgCl2), whom is one of the most toxic substance joined in nature, it was presented activity already diminished at 0,01mM and lost total activity at 4mM, indicating sensitivity for this type of metal. β-D-N-acetilglucosaminidase-ferromagnetic Dacron showed degradative capacity on heparan sulfate, the enzyme still demonstrated degradative capacity on heparan sulphate, suggesting a possible application to produce fractions of this glycosaminoglycan

Produção de enzimas quitosanolíticas utilizando Paenibacillus ehimensis e Paenibacillus chitinolyticus para obtenção de quitoologossacarídeos

The acquisition of oligosaccharides from chitosan has been the subject of several studies in the pharmaceutical, biochemical, food and medical due to functional properties of these compounds. This study aimed to boost its production of chitooligosaccharides (COS) through the optimization of production and characterization of chitosanolytic enzymes secreted by microorganisms Paenibacillus chitinolyticus and Paenibacillus ehimensis, and evaluating the antioxidant potential of the products obtained. In the process of optimizing the production of chitosanase were employed strategies Fractional Factorial Experimental Design and Central Composite Rotatable Design. The results identified the chitosan, peptone and yeast extract as the components that influenced the production of chitosanase by these microorganisms. With the optimization of the culture media was possible to obtain an increase of approximately 8.1 times (from 0.043 to 0.35 U.mL U.mL-1) and 7.6 times (from 0.08 U.mL-1 to 0.61 U.mL-1) in the enzymatic activity of chitosanase produced by P. chitinolyticus and P. ehimensis respectively. Enzyme complexes showed high stability in temperature ranges between 30º and 55º C and pH between 5.0 and 9.0. Has seen the share of organic solvents, divalent ions and other chemical agents on the activity of these enzymes, demonstrating high stability of these crude complexes and dependence of Mn2+. The COS generated showed the ability of DPPH radical scavenging activity, reaching a maximum rate of scavenging of 61% and 39% when they were produced with enzymes of P. ehimensis and P. chitinolyticus respectively. The use of these enzymes in raw form might facilitate its use for industrial applications

Avaliação da ação bioinseticida de SBTI e vicilina de Erythrina velutina em enzimas digestivas e membrana peritrófica de larvas de Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests

Avaliação bioquímica e hostológica de fígado de ratas wistar diabéticas e tratadas com tamoxifeno

Tamoxifen (TX), a drug used in the treatment of breast cancer, may cause hepatic changes in some patients. The consequences of its use on the liver tissues of rats with or without diabetes mellitus (DM) have not been fully explored. The purpose of this multidisciplinary study was to evaluate the correlation between plasma hepatic enzyme levels and the presence of iron overload in the hepatic tissue of female Wistar rats with or without streptozotocin-induced DM and using TX. Female rats were studied in control groups: C-0 (non-drug users), C-V (sorbitol vehicle only) and C-TX (using TX). DM (diabetic non-drug users) and DM-TX (diabetics using TX) were the test groups. Sixty days after induced DM, blood samples were collected for glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST) alkaline phosphatase (ALP) and bilirubin measures. Hepatic fragments were processed and stained with hematoxylin and eosin (H&E), Masson s trichrome, Perls. The hepatic iron content was quantified by atomic absorption spectrometry. AST, ALT and ALP levels were significantly elevated in the DM and DM-TX groups, with unchanged bilirubin levels. Liver iron overload using Perls stain and atomic absorption spectrometry were observed exclusively in groups C-TX and DM-TX. There was positive correlation between AST, ALT and ALP levels and microscopic hepatic siderosis intensity in group DM-TX. In conclusion, TX administration is associated with liver siderosis in diabetic and non-diabetic rats. In addition, TX induced liver iron overload with unaltered hepatic function in 2 non-diabetic rats and may be a useful tool for investigating the biological control of iron metabolism

Efeito do uso de probiótico sobre o desempenho e atividade de enzimas digestivas de frangos de corte

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Degradabilidade in situ e observações microscópicas de volumosos em bovinos suplementados com enzimas fibrolíticas exógenas

Avaliou-se o efeito de enzimas fibrolíticas (celulase e xilanase) sobre a degradabilidade in situ da MS, PB, FDN, FDA e hemicelulose do feno de Tifton-85 (Cynodon spp.) cortado aos 30 e 90 dias e do bagaço de cana, utilizando-se seis bovinos com cânula no rúmen. As enzimas foram extraídas dos fungos Aspergillus niger e Trichoderma longibrachiatum e fornecidas, na quantidade de 0,75 g/kgMS.dia, por meio da cânula ruminal. Os tempos de incubação ruminal foram de 0, 3, 6, 12, 24, 48, 72 e 96 horas. Os resíduos de incubação foram avaliados por meio de microscopia eletrônica de varredura (MEV). O efeito da adição de enzimas sobre a degradação da MS e PB variou em função do volumoso estudado. A degradabilidade efetiva da MS do feno Tifton cortado aos 30 e 90 dias e do bagaço de cana sem a adição de enzimas foi de 61,85; 42,35 e 28,22%, respectivamente, e de 63,51; 40,64 e 31,43%, respectivamente, com a adição das enzimas. Não houve efeito das enzimas sobre a degradação da fibra. As observações ao MEV indicaram aumento da colonização bacteriana sobre a parede celular com a suplementação enzimática. A adição de enzimas fibrolíticas na dieta de ruminantes apresentou efeito pouco expressivo sobre os parâmetros de degradação ruminal dos volumosos estudados.

Enzimas exógenas em dietas de frangos de corte: desempenho

O experimento foi conduzido para avaliar a eficiência de enzimas exógenas em dietas sobre o desempenho de frangos de corte. Um total de 1440 pintos de corte machos (Cobb 500®) foram distribuídos em um delineamento inteiramente casualizado em esquema fatorial 2x2 (duas dietas com diferentes densidades nutricionais; com e sem a adição enzimática) com 8 repetições de 45 aves em cada unidade experimental. As dietas com diferentes densidades nutricionais foram: controle positivo (níveis normais) e um controle negativo (com redução de 4,3 e 4,5% de energia metabolizável; 16,7 e 17,7% de cálcio e 35 e 42,7% de fósforo, nas fases inicial e de crescimento, respectivamente). A suplementação enzimática consistiu da combinação das enzimas fitase (100g t-1) e amilase, xilanase e protease (500g t-1). Foi avaliado o desempenho das aves nas fases de 1 a 21 e 1 a 42 dias de idade. Na fase total de criação, as aves alimentadas com a dieta controle negativo com adição enzimática tiveram consumo de ração (5,97%), peso médio (8,47%), ganho de peso (8,64%) e conversão alimentar (2,92%) melhores (P<0,05), comparados às aves alimentadas com dieta controle negativo, sem a suplementação de enzimas. Além disso, as aves alimentadas com a dieta controle negativo com adição enzimática apresentaram consumo de ração, peso vivo médio e ganho de peso similar (P>0,05) ao grupo alimentado com a dieta controle positivo com ou sem enzima. A adição de enzimas exógenas em dieta de frangos de corte com redução de energia metabolizável, cálcio e fósforo, proporciona um consumo de ração, peso vivo médio e ganho de peso similar a uma dieta com níveis nutricionais adequados.

Taxa de passagem e parâmetros ruminais em bovinos suplementados com enzimas fibrolíticas

Avaliou-se o efeito da adição de enzimas fibrolíticas (celulase e xilanase) sobre as taxas de passagem de partículas e de diluição e os parâmetros ruminais (pH e nitrogênio amoniacal) em animais consumindo dietas contendo silagem de milho e feno de tifton 85 (Cynodon spp.) como volumoso. Oito bovinos com fístulas no rúmen e no duodeno foram distribuídos em dois quadrados latinos 4 x 4, com os tratamentos distribuídos em esquema fatorial 2 x 2 (duas fontes de volumoso e adição ou não de enzimas). O complexo enzimático foi proveniente de fonte comercial e extraído dos fungos Aspergillus niger e Trichoderma longibrachiatum, sendo fornecido na proporção de 12 g/animal/dia, misturado à ração total. Para as taxas de passagem, foram utilizados como marcadores das fases sólida e líquida o cromo-mordente e o cobalto-EDTA, respectivamente. Não houve efeito das enzimas sobre o pH e as concentrações de N-NH3, que apresentaram médias de 6,5 e 22,10 mg/100 mL, respectivamente. A taxa de passagem de partículas no rúmen não foi influenciada pela suplementação enzimática, com médias de 3,4 e 3,0%/h para as dietas contendo silagem de milho e feno de tifton, respectivamente. A taxa de diluição aumentou de 10,07 para 11,84%/h com a adição de enzimas. A suplementação com enzimas fibrolíticas em dietas formuladas com silagem de milho e feno de tifton como volumoso praticamente não alterou os parâmetros avaliados.

Eficiência de síntese microbiana e atividade enzimática em bovinos submetidos à suplementação com enzimas fibrolíticas

Avaliou-se o efeito da suplementação com enzimas fibrolíticas (celulase e xilanase) sobre a eficiência de síntese microbiana e a atividade enzimática da beta-1,4-endoglucanase (celulase) em animais consumindo dietas contendo silagem de milho e feno de tifton 85 (Cynodon spp.) como volumoso. Oito bovinos com fístulas no rúmen e no duodeno foram distribuídos em dois quadrados latinos 4 x 4, em esquema fatorial 2 x 2 (duas fontes de volumoso e adição ou não de enzimas). O complexo enzimático, proveniente de fonte comercial e extraído dos fungos Aspergillus niger e Trichoderma longibrachiatum, foi fornecido misturado à ração total na proporção de 12 g/animal/dia. A adição de enzimas às dietas contendo silagem de milho e feno de tifton 85 aumentou a atividade da beta-1,4-endoglucanase no fluido ruminal. Não houve efeito das enzimas sobre a eficiência de síntese de proteína microbiana, o fluxo de nitrogênio microbiano (N-mic) e nitrogênio não-amoniacal (NNA), com valores médios de 31,8 e 33,9 g de N/kg de MODR; 0,40 e 0,41 g de N/Mcal de EDR; 78,2 e 52,5 g/dia; 89,6 e 76,3 g/dia, respectivamente, para as dietas contendo silagem de milho e feno de tifton. A adição de enzimas fibrolíticas não influenciou os parâmetros ruminais de bovinos alimentados com feno e silagem de milho.

Enzimas exógenas no desempenho e na digestibilidade ileal de nutrientes em frangos de corte

O objetivo deste trabalho foi avaliar o efeito da combinação de fitase e do complexo amilase, protease e xilanase, em dietas de milho e soja, formuladas com redução e sem redução dos níveis de energia, cálcio e fósforo sobre o desempenho e a digestibilidade ileal de nutrientes, em frangos de corte. Foi utilizado o delineamento inteiramente casualizado, com esquema fatorial 2x2, com duas dietas-controle: com redução e sem redução dos nutrientes, e duas suplementações das enzimas (sem adição e com adição de fitase e complexo enzimático), com dez repetições de 40 aves. A digesta ileal foi coletada aos 43 dias de idade, para determinação da energia digestível e dos coeficientes de digestibilidade da proteína bruta, matéria seca, cálcio e fósforo. O desempenho apresentou interação em todos os parâmetros analisados. As aves do tratamento com redução dos nutrientes mostraram pior desempenho em relação às aves da dieta sem redução dos nutrientes. Não houve efeito da matéria seca na digestibilidade e na retenção de cálcio. A adição da combinação enzimática melhorou a digestibilidade da proteína e a retenção de fósforo. Houve interação quanto à energia digestível, com efeito apenas nas dietas sem redução dos nutrientes, com maiores valores nas dietas suplementadas.

Consumo e digestibilidade aparente total em bovinos sob suplementação com enzimas fibrolíticas

Avaliou-se o efeito da suplementação com enzimas fibrolíticas (celulase e xilanase) sobre o consumo e a digestibilidade aparente total dos nutrientes de dietas compostas de silagem de milho e feno de tifton 85 (Cynodon spp.) cortado aos 90 dias. Oito bovinos (340 kg de PV) fistulados no rúmen foram distribuídos em dois quadrados latinos 4 x 4, em esquema fatorial 2 x 2 (fonte de volumoso e adição ou não de enzimas). O complexo enzimático testado foi proveniente de fonte comercial e extraído dos fungos Aspergillus niger e Trichoderma longibrachiatum, sendo fornecido na quantidade de 12 g/animal/dia, misturado à ração total. Não houve efeito da suplementação enzimática sobre o consumo de nutrientes para ambos os volumosos, porém, a adição de enzimas aumentou a digestibilidade total de FDN, FDA e CEL de 36,87; 36,21 e 46,89%, respectivamente, para 41,19; 40,01 e 50,46%, respectivamente.

Estudo da influência de pré-tratamentos de dois resíduos lignocelulósicos (bagaço do pedúnculo de caju e casca de coco) utilizados como substrato na indução à síntese de enzimas celulolíticas

Nowadays generation ethanol second, that t is obtained from fermentation of sugars of hydrolyses of cellulose, is gaining attention worldwide as a viable alternative to petroleum mainly for being a renewable resource. The increase of first generation ethanol production i.e. that obtained from sugar-cane molasses could lead to a reduction of lands sustainable for crops and food production. However, second generation ethanol needs technologic pathway for reduce the bottlenecks as production of enzymes to hydrolysis the cellulose to glucose i.e. the cellulases as well as the development of efficient biomass pretreatment and of low-cost. In this work Trichoderma reesei ATCC 2768 was cultivated under submerged fermentation to produce cellulases using as substrates waste of lignocellulosic material such as cashew apple bagasse as well as coconut bagasse with and without pretreatment. For pretreatment the bagasses were treated with 1 M NaOH and by explosion at high pressure. Enzyme production was carried out in shaker (temperature of 27ºC, 150 rpm and initial medium pH of 4.8). Results showed that T.reesei ATCC 2768 showed the higher cellulase production when the cashew apple bagasse was treated with 1M NaOH (2.160 UI/mL of CMCase and 0.215 UI/mL of FPase), in which the conversion of cellulose, in terms of total reducing sugars, was of 98.38%, when compared to pretreatment by explosion at high pressure (0.853 UI/mL of CMCase and 0.172 UI/mL of Fpase) showing a conversion of 47.39% of total reducing sugars. Cellulase production is lower for the medium containing coconut bagasse treated with 1M NaOH (0.480 UI/mL of CMcase and 0.073 UI/mL of FPase), giving a conversion of 49.5% in terms of total reducing sugars. Cashew apple bagasse without pretreatment showed cellulase activities lower (0.535 UI/mL of CMCase and 0,152 UI/mL of FPase) then pretreated bagasse while the coconut bagasse without pretreatment did not show any enzymatic activity. Maximum cell concentration was obtained using cashew nut bagasse as well as coconut shell bagasse treated with 1M NaOH, with 2.92 g/L and 1.97 g/L, respectively. These were higher than for the experiments in which the substrates were treated by explosion at high pressure, 1.93 g/L and 1.17 g/L. Cashew apple is a potential inducer for cellulolytic enzymes synthysis showing better results than coconut bagasse. Pretreatment improves the process for the cellulolytic enzyme production

Produção de enzimas extracelulares por Fusarium solani de maracujazeiro amarelo

Uma das principais doenças do maracujazeiro, na maioria dos estados produtores do Brasil, é a podridão do colo, causada por Fusarium solani. Pouco se sabe a respeito da fisiologia deste patógeno do maracujazeiro amarelo, principalmente quanto à produção de enzimas extracelulares. O objetivo do presente trabalho foi verificar, em meios de cultura individuais e apropriados, a produção das enzimas extracelulares amilase, lipase, celulase, proteases (caseinase e gelatinase), lacase (oxidase) e catalase por isolados de F. solani, provenientes de maracujazeiro amarelo. O delineamento experimental adotado foi o inteiramente casualizado, em esquema de dois fatores (nove isolados versus sete enzimas), com três repetições. Todos os isolados de F. solani produziram, de maneira semiquantitativa, as enzimas extracelulares amilase, lipase, celulase, caseinase (protease) e lacase (oxidase). No entanto, a quantidade produzida de cada enzima foi significativamente diferente entre os isolados. As enzimas extracelulares gelatinase (protease) e catalase foram produzidas em pouca quantidade e de maneira igual por todos os isolados do fungo.

Produção de enzimas por fungos em fermentação semi-sólida utilizando bagaço de coco e pedúnculo de caju como substratos

The production of enzymes by microorganisms using organic residues is important and it can be associated with several applications such as food and chemical industries and so on. The objective of this work is the production of CMCase, Xylanase, Avicelase and FPase enzymes by solid state fermentation (SSF) using as substrates: bagasse of coconut and dried cashew stem. The microorganisms employed are Penicillium chrysogenum and an isolated fungus from the coconut bark (Aspergillus fumigatus). Through the factorial design methodology and response surface analysis it was possible to study the influence of the humidity and pH. For Penicillium chrysogenum and the isolated fungus, the coconut bagasse was used as culture medium. In another fermentation, it was used the mixture of coconut bagasse and cashew stem. Fermentations were conducted using only the coconut bagasse as substrate in cultures with Penicillium chrysogenum fungus and the isolated one. A mixture with 50% of coconut and 50% of cashew stem was employed only for Penicillium chrysogenum fungus, the cultivation conditions were: 120 hours at 30 °C in BOD, changing humidity and pH values. In order to check the influence of the variables: humidity and pH, a 2 2 factorial experimental design was developed, and then two factors with two levels for each factor and three repetitions at the central point. The levels of the independent variables used in ascending order (-1, 0, +1), to humidity, 66%, 70.5% and 75% and pH 3, 5 and 7, respectively. The software STATISTICA TM (version 7.0, StatSoft, Inc.) was used to calculate the main effects of the variables and their interactions. The response surface methodology was used to optimize the conditions of the SSF. A chemical and a physic-chemical characterization of the coconut bagasse have determined the composition of cellulose (%) = 39.09; Hemicellulose (%) = 23.80, Total Lignin (%) = 36.22 and Pectin (%) = 1.64. To the characterization of cashew stem, the values were cellulose (g) = 15.91 Hemicellulose (%) = 16.77, Total Lignin (%) = 30.04 and Pectin (%) = 15.24. The results indicate the potential of the materials as substrate for semisolid fermentation enzyme production. The two microorganisms used are presented as good producers of cellulases. The results showed the potential of the fungus in the production of CMCase enzyme, with a maximum of 0.282 UI/mL and the Avicelase enzyme the maximum value ranged from 0.018 to 0.020 UI/ mL, using only coconut bagasse as substrate. The Penicillium chrysogenum fungus has showed the best results for CMCase = 0.294 UI/mL, FPase = 0.058 UI/mL, Avicelase = 0.010 UI/mL and Xylanase = 0.644 UI/ mL enzyme, using coconut bagasse and cashew stem as substrates. The Penicllium chrysogenum fungus showed enzymatic activities using only the coconut as substrate for CMCase = 0.233 UI/mL, FPase = 0.031 to 0.032 UI/ mL, Avicelase = 0.018 to 0.020 UI/mL and Xylanase = 0.735 UI/ mL. Thus, it can be concluded that the used organisms and substrates have offered potential for enzyme production processes in a semi-solid cultivation

Palma forrageira (Opuntia ficus indica e Nopalea cochenillifera) como matéria-prima para produção de etanol celulósico e enzimas celulolíticas

The need for new sources of energy and the concern about the environment have pushed the search for renewable energy sources such as ethanol. The use of lignocellulosic biomass as substrate appears as an important alternative because of the abundance of this raw material and for it does not compete with food production. However, the process still meets difficulties of implementation, including the cost for production of enzymes that degrade cellulose to fermentable sugars. The aim of this study was to evaluate the behavior of the species of cactus pear Opuntia ficus indica and Nopalea cochenillifera, commonly found in northeastern Brazil, as raw materials for the production of: 1) cellulosic ethanol by simultaneous saccharification and fermentation (SSF) process, using two different strains of Saccharomyces cerevisiae (PE-2 and LNF CA-11), and 2) cellulolytic enzymes by semi-solid state fermentation (SSSF) using the filamentous fungus Penicillium chrysogenum. Before alcoholic fermentation process, the material was conditioned and pretreated by three different strategies: alkaline hydrogen peroxide, alkaline using NaOH and acid using H2SO4 followed by alkaline delignification with NaOH. Analysis of composition, crystallinity and enzymatic digestibility were carried out with the material before and after pretreatment. In addition, scanning electron microscopy images were used to compare qualitatively the material and observe the effects of pretreatments. An experimental design 2² with triplicate at the central point was used to evaluate the influence of temperature (30, 40 and 45 °C) and the initial charge of substrate (3, 4 and 5% cellulose) in the SSF process using the material obtained through the best condition and testing both strains of S. cerevisiae, one of them flocculent (LNF CA-11). For cellulase production, the filamentous fungus P. chrysogenum was tested with N. cochenillifera in the raw condition (without pretreatment) and pretrated hydrothermically, varying the pH of the fermentative medium (3, 5 and 7). The characterization of cactus pear resulted in 31.55% cellulose, 17.12% hemicellulose and 10.25% lignin for N. cochenillifera and 34.86% cellulose, 19.97% hemicellulose and 15.72% lignin for O. ficus indica. It has also been determined, to N. cochenillifera and O. ficus indica, the content of pectin (5.44% and 5.55% of calcium pectate, respectively), extractives (26.90% and 9.69%, respectively) and ashes (5.40% and 5.95%). Pretreatment using alkaline hydrogen peroxide resulted in the best cellulose recovery results (86.16% for N. cochenillifera and 93.59% for O. ficus indica) and delignification (48.79% and 23.84% for N. cochenillifera and O. ficus indica, respectively). This pretreatment was also the only one which did not increase the crystallinity index of the samples, in the case of O. ficus indica. However, when analyzing the enzymatic digestibility of cellulose, alkali pretreatment was the one which showed the best yields and therefore it was chosen for the tests in SSF. The experiments showed higher yield of conversion of cellulose to ethanol by PE-2 strain using the pretreated N. cochenillifera (93.81%) at 40 °C using 4% initial charge of cellulose. N. cochenillifera gave better yields than O. ficus indica and PE-2 strain showed better performance than CA-11. N. cochenillifera proved to be a substrate that can be used in the SSSF for enzymes production, reaching values of 1.00 U/g of CMCase and 0.85 FPU/g. The pretreatment was not effective to increase the enzymatic activity values