As dietas de vacas em lactação são compostas principalmente por carboidratos que não estão totalmente disponíveis para fermentação microbiana no rúmen, o que pode ser um fator crítico para a obtenção de energia em ruminantes. O objetivo do presente estudo foi avaliar o efeito da adição de enzima fibrolítica (Fibrozyme®, Alltech Inc., Springfield, KY) e amilolítica (Amaize, Alltech Inc., Springfield, KY) nas dietas sobre o consumo de nutrientes, índice de seleção de partículas, digestibilidade aparente total, fermentação ruminal, metabolismo de nutrientes, produção e composição do leite de vacas em lactação. Para o experimento, 32 vacas multíparas da raça Holandesa com 181.3 ± 35.3 (média ± SD) dias em lactação (DEL), 571 ± 72.7 kg de peso vivo (PV) e 29.6 ± 5.24 kg/d de produção de leite (PL), foram distribuídas em 8 quadrados Latinos 4×4. Os tratamentos foram obtidos por esquema fatorial 2×2 pela combinação de enzima fibrolítica e amilolítica, como segue: 1) Controle (CONT): dieta basal sem adição de enzimas exógenas; 2) Enzima fibrolítica (FIB): dieta basal com adição de 1 g/kg de Fibrozyme± no concentrado da dieta (51 UI de atividade xilanase/kg de matéria seca (MS) da dieta; Alltech, Nicholasville, KY, USA; batch#: 417990-2); 3) Enzima amilolítica (AMI): dieta basal com adição de 0,66 g/kg de Amaize no concentrado da dieta (203 FAU/kg MS da dieta; Alltech Nicholasville, KY, USA; batch: 432715-1); e 4) FIB+AMI: adição de 1 e 0,66 g/kg de Fibrozyme® e Amaize, respectivamente. As enzimas foram aplicadas para atender um consumo de 12 g/dia de Fibrozyme® e 8 g/dia de Amaize, de acordo com as recomendações do fabricante. A enzima foi adicionada ao concentrado durante a preparação na fábrica de ração (toda semana). Não foram observados efeitos das enzimas sobre o consumo e digestibilidade dos nutrientes. Contudo, houve efeito de interação entre FIB e AMI para o consumo de partículas entre 19 e 8 mm. A enzima amilolítica aumentou o consumo de partículas entre 19 e 8 mm nos animais que não recebiam FIB. Além disso, AMI reduziu o consumo de partículas maiores que 19 mm. Outro efeito observado foi a interação entre FIB e AMI sobre a concentração de butirato ruminal. A enzima amilolítica aumentou a concentração de butirato apenas nos animais tratados com FIB. Houve tendência de interação entre enzimas sobre a excreção de nitrogênio (N) no leite, em que FIB reduziu a excreção de N no leite apenas nos animais não tratados com AMI. Ainda, AMI reduziu a excreção de N na urina. Ainda, houve interação entre os efeitos de enzima fibrolítica e amilolítica sobre a concentração de colesterol e a atividade enzimática da aspartato aminotransferase (AST) e gama glutamiltransferase (GGT). A enzima fibrolítica reduziu a concentração de colesterol e aumentou a atividade da enzima GGT nos animais não tratados com enzima amilolítica. No entanto, a enzima amilolítica aumentou a concentração de colesterol e a atividade da AST nos animais alimentados com enzima fibrolítica. Houve interação entre enzimas exógenas sobre a produção de proteína e lactose no leite. A enzima fibrolítica reduziu a produção de proteína e lactose nos animais não alimentados com AMI. Não houve mudanças na produção de leite com a suplementação de enzimas exógenas. Em nosso estudo foram encontrados efeitos no consumo de partículas, fermentação ruminal, excreção de N e mudanças na composição do leite, contudo não foram observadas alterações no desempenho produtivo dos animais. / Lactating cows diets are comprised mostly of carbohydrates which are not fully available for microbial fermentation in the rumen, critical factor for to obtain energy in ruminants. The aimed of this study was to evaluate the effect of fibrolytic enzyme (Fibrozyme®, Alltech Inc., Springfield, KY) on nutrient intake, sorting index, total tract digestion, ruminal fermentation, nitrogen utilization, metabolic profile, milk yield and composition in diets with or without amylolytic enzyme (Amaize, Alltech Inc., Springfield, KY) of mid-lactating dairy cows. Thirty-two multiparous Hostein cows with 181.3 ± 35.3 (mean ± SD) days in milk (DIM), 571 ± 72.7 kg of body weight (BW) and 29.6 ± 5.24 kg/d of milk yield, were blocked and randomly allocated to a sequence of treatments in a 4 × 4 Latin square experimental design. Treatments were obtained in a 2 × 2 factorial arrangement, as follows: 1) Control (CONT), basal diet without exogenous enzymes; 2) Fibrolytic enzyme (FIB), provision of Fibrozyme® (batch#: 417990-2; Alltech, Nichollasvile, KY) at 1 g/kg of concentrate (51 IU of xylanase activity/kg diet DM); 3) Amylolytic enzyme (AMY), provision of AmaizeTM (batch#: 432715-1; Alltech) at 0.66 g/kg of concentrate (203 FAU/kg diet DM); and 4) Fibrolytic enzyme plus amylolytic enzyme (FIB+AMY), enzymes added at the same rate in FIB and AMY treatments. The enzymes were applied to meet the intake of 12 and 8 g/cow/day of Fibrozyme® and AmaizeTM, respectively. Enzymes products were added to concentrate during its preparation (once a week). Enzymes no effects on intake and digestibility of nutrients. However, there was FIB and AMY interaction effect on selection index of particle size between 19 and 8 mm. Amylolytic enzyme increase selection index of particles with 19 and 8 mm, only treatments without FIB. Furthermore, AMY decreased the sorting for feed with particle size greater than 19 mm. There was FIB and AMY interaction effect on butyric acid concentration. Amylolytic enzyme increased on butyrate concentrations in cows treated with fibrolytic enzyme. Fibrolytic and amylolytic enzyme interaction effect tended on N excreted in milk, in which FIB decrease N excreted in milk only on animals non-treated with AMY. Whereas AMY reduced urinary N excretion. There was interaction effects of fibrolytic and amylolytic enzymes on cholesterol concentration and the enzymatic activity of AST and GGT. The fibrolytic enzyme reduced cholesterol concentration and increased GGT enzyme activity in animals not treated with amylolytic enzyme. There was FIB and AMY interaction effect on lactose and protein production. Fibrolytic enzyme decreased lactose and protein production, only on animals non-treated with AMY. Exogenous enzymes had no impact on milk production of dairy cows. Enzymes exogenous affected on particle size greater, ruminal fermentation and milk composition. This study did not show evidences that fibrolytic and amylolytic enzymes can alter total tract nutrient digestibility and performance of mid-lactating cows.

Made available in DSpace on 2014-06-11T19:31:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-04-08Bitstream added on 2014-06-13T19:40:50Z : No. of bitstreams: 1 oliveira_ds_dr_sjrp.pdf: 1933738 bytes, checksum: baca389347279bb3f5a0eea7681d038d (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O desenvolvimento da tecnologia de pães é um fenômeno de grande impacto na indústria de alimentos. Ao longo dos anos vários aditivos foram incorporados à tecnologia de panificação, elevando a qualidade destes produtos e fazendo crescer a sua aceitação pela população em geral. O objetivo deste trabalho foi estudar os efeitos sobre a qualidade do pão e sobre o processo de envelhecimento, da adição de enzimas na massa. As enzimas usadas foram xilanase produzida pelo fungo Thermoascus aurantiacus CBMAI 756 nas concentrações de 20, 35 e 50U/100 g de farinha de trigo e/ou da ciclodextrina glicosiltransferase (CGTase) produzida pela bactéria Bacillus clausii E16 nas concentrações de 15 e 30U/100 g de farinha de trigo, parcialmente purificadas. O mecanismo de ação dessas enzimas sobre os seus respectivos substratos arabinoxilana e amido, respectivamente, também foram avaliadas. Os pães adicionados de xilanase, CGTase e xilanase/CGTase foram produzidos em três etapas distintas. Para o estudo do envelhecimento os pães foram mantidos a temperatura de 4ºC durante 10 dias, e foram avaliados quanto à perda de água, a textura e a retrogradação da amilopectina. Os produtos obtidos pela ação da xilanase e CGTase sobre as arabinoxilanas e o amido, respectivamente, isolados da farinha de trigo, foram analisados por HPAEC-PAD e HPLC. O fungo T. aurantiacus exibiu uma variação no perfil enzimático de acordo com cada substrato usado no seu cultivo. O substrato que resultou no melhor perfil enzimático para o uso em panificação foi o sabugo de milho, porque esse extrato enzimático exibiu alta atividade xilanolítica e baixa atividade amilolítica e proteolítica. A adição de xilanase, CGTase e xilanase/CGTase aumentou o volume da massa e o volume específico dos pães. Quanto ao... / The development of bread technology is a phenomenon of great impact on food industry. For years, many additives were used on bread technology, increasing the bread quality and improving the acceptance of general population. The aim of this work was to study the effects on the bread quality and on the staling process, by adding enzymes to the dough. The enzymes used were xylanase from the fungus Thermoascus aurantiacus CBMAI 756 in the concentrations 20, 35 and 50U/100 g wheat flour and/or cyclodextryn glycosiltransferase (CGTase) from the bacteria Bacillus clausii E16 in the concentration 15 and 30U/100 g wheat flour, partially purified. The action mechanisms of these enzymes under the substrates arabinoxylan and starch, respectively, were also evaluated. The breads added of xylanase, CGTase and xylanase/CGTase were produced in three distinct stages. For bread staling study, the breads were stored at 4ºC for 10 days, and they were analyzed regarding to the moisture content, texture and amylopectin retrogadation. The products obtained by the action of xylanase and CGTase on arabinoxylans and starch, respectively, isolated from wheat flour, were analyzed by using HPAEC-PAD and HPLC. The fungus T. aurantiacus exhibited a variation on the enzymatic profile according to each substrate used on its cultivation. The substrate which resulted in a better enzymatic profile for using on breadmaking was corncob, since this enzymatic extract exhibited high xylanolytic activity and low amylolytic and proteolytic activities. The addition of xylanase, CGTase and xylanase/CGTase increased the dough volume and the specific volume of the breads. Regarding to the staling study, the enzymes added separated or together reduced amylopectin retrogradation and firmness of the crumb during storage, when compared... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:31:04Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-06-26Bitstream added on 2014-06-13T18:41:37Z : No. of bitstreams: 1 benedetti_acep_dr_arafcf.pdf: 555249 bytes, checksum: c34c45268021bed2af4612d36cdf1d73 (MD5) / Fundação para o Desenvolvimento da UNESP (FUNDUNESP) / Universidade Estadual Paulista (UNESP) / A descoberta de resíduos agrícolas, como fonte de energia renovável tem colaborado para o desenvolvimento industrial e preservação do meio ambiente. Desta maneira, a degradação da parede celular destes resíduos tem grande importância na fabricação de pães, alimentos, bebidas, têxtil, papel e celulose entre outros produtos. A degradação enzimática destes polímeros esta se tornando uma alternativa mais atraente do que a utilização de substâncias químicas e processos mecânicos. Alguns fungos termófilos participam desta degradação enzimática, como Humicola grisea var. thermoidea, isolado do solo, é conhecido como bom produtor do complexo xilanolítico. Além deste, um fungo termófilo isolado da polpa do fruto cupuaçuzeiro em decomposição foi estudado e produziu xilanase. Esses fungos foram mantidos em meios sólidos contendo ágar e farinha de aveia. O fungo isolado da polpa do fruto cresceu em meio liquido empregando diferentes resíduos agrícolas como fonte de carbono. Dentre os vários resíduos empregados para se otimizar a produção de xilanase, o melhor foi o sabugo de milho. Esse foi utilizado nos experimentos na proporção de 1,0% (m/v), em meio líquido contendo 0,1% CaCO3, 0,5% NaCl, 0,1% NH4Cl; 0,5% água de maceração de milho. O pH ótimo foi 5,0 e a temperatura ótima de 60ºC. A xilanase presente no extrato clarificado em caulim apresentou cinética Michaeliana com Km de 10,41 ± 0,282 mgmL-1 e Vmax 3,32 ± 0,053 U (mg protein)-1. Para acompanhar o processo de clarificação da enzima solúvel realizou-se SDS-PAGE verificando a presença de uma banda protéica com atividade xilanolítica de massa molecular de aproximadamente 30kDa e a Cromatografia Sílica Gel P60 para verificar os produtos da hidrólise da xilana em função do tempo e da associação de xilosidase do fungo termófilo Humicola com a xilanase do fungo isolado do fruto... / The researches of agricultural residues as renewable energy sources have been contributing for the industrial development and the environment conservation. Thus, the degradation of these residues’ cell walls has great importance in fabrication of bread, food, drinks, textile, paper, cellulose, and other products. The enzymatic degradation of such polymers is becoming an attractive option when compared to the use of chemical substances and mechanical processes. Some thermophilic fungi take role in this enzymatic degradation, such as Humicola grisea var. thermoidea, isolated from soil, and is known to be a good producer of the xylanolytic complex. Besides it, another thermophilic fungus, isolated from the Theobroma grandiflorum decomposing fruit pulp, was studied and also produced xylanases. These fungi were kept in solid media containing agar and oat bran. The fungus isolated from the fruit pulp grew in liquid media in wich different kinds of agricultural residues were employed as carbon sources. Among the various types of residues that were used to optimize the xylanase production, corn cobs proved to be the best one. It was used in assays in a proportion of 1,0% (m/v), in liquid media containing 0.1% of CaCO3, 0.5% of NaCl, 0.1% of NH4Cl, and 0.5% of corn steep liquor. The pH optima was 5.0 and the temperature optima was 60ºC. The xylanase present in the kaolin clarified extract showed a Km of 10.41 ± 0.282 mg.mL-1, and Vmax 3.32 ± 0.053 U (mg.protein)-1. In order to follow the soluble enzyme clarification process, SDS-PAGE was run, verifying the presence of a proteic band with xylanolytic activity of, approximately, 30kDa of molecular mass. Furthermore, Silica Gel 60 Thin Layer Chromatography (TLC) was used to verify the xylan hydrolysis products in function of time and the association of Humicola sp. thermophylic fungus xylosidase with the xylanase produced by the fruit pulp... (Complete abstract click electronic access below)

O presente trabalho teve como objetivo a caracterização da xilanase presente no extrato bruto de Bacillus pumilus, com a finalidade de usá-la na extração de xilana do bagaço de cana-de-açúcar. Foram testados dois meios de cultura em pHs 8,5 e 9,5 com diferentes substratos. As maiores atividades de xilanase foram produzidas em xilana comercial de oat spelts (228 U/mL) e em farelo de trigo (220 U/mL), no pH 8,5. Independentemente das condições de cultivo empregada, não foi detectada atividades de &szlig;-xilosidase, &szlig;-glicosidase e arabinofuranosidase e a produção de endoglucanase foi baixa (< 0,7 U/mL). O perfil de proteínas em eletroforese foi amplo, e a banda com 23 kDa correspondeu a uma xilanase. A xilanase apresentou pH ótimo na faixa de 7-8, temperatura ótima entre 45-50 ºC e se apresentou mais estável a 40 ºC (pH8,0). A hidrólise de xilanas comerciais produziu xilotriose, xilotetraose, xilopentaose e xilo-oligossacarídeos (XOS) com massas maiores, que não foram identificados por cromatografia em camada fina (TLC). A influência de vários fatores, tais como a carga de xilanase, tipo de substrato, temperatura e a adição de vários compostos foi avaliada no rendimento de extração de xilanas desde o bagaço de cana-de-açúcar prétratado. O extrato bruto de B. pumilus, rico em xilanases, foi aplicado em um bagaço pré-tratado com sulfito/álcali, e em um bagaço deslignificado por clorito em meio ácido, evidenciando que a menor quantidade de lignina residual como principal fator para aumentar a extração de xilana, aumentando a extração de 4,2 para 42,5%. A lavagem extensiva do material pré-tratado com sulfito alcalino também auxiliou no aumento do rendimento de xilana, de 18,5 para 25,1 %. Foram testadas outras alternativas para aumentar o rendimento de hidrólise da xilana do bagaço pré-tratado, sem a necessidade da lavagem do material. A reutilização do licor sulfito alcalino, a adição de tween 80, polietileno glicol, albumina não favoreceram a extração de xilanas, enquanto que a adição de MgSO4 teve um pequeno efeito positivo na extração da xilana (de 18% para 20,9 %). A extração alcalina a frio, realizada após a extração enzimática, influenciou positivamente o rendimento de extração da xilana do bagaço pré-tratado com 10% Na2SO3/5%NaOH. A extração de xilana obtido pela aplicação de xilanase (20 U/g) seguida da extração com 20% de NaOH produziu o mesmo resultado que utilizando apenas NaOH (70%), de 24,1 e 24,9 %, respectivamente. A extração enzimática da xilana permitiu a obtenção de um resíduo com características favoráveis para a completa sacarificação, com Cellic CTec2 na carga de 10 FPU/g de material. / The present work aimed to characterize the xylanase present in the crude extract of Bacillus pumilus, with the purpose of using it in the xylan extraction of the sugarcane bagasse. Two culture media were tested at pHs 8.5 and 9.5 with different substrates. The highest xylanase activities were produced in commercial xylan oat spelled (228 U / mL) and wheat bran (220 U / mL) at pH 8.5. Regardless of the culture conditions employed, &szlig;xylosidase, &szlig;-glycosidase and arabinofuranosidase activities were not detected and endoglucanase production was low (<0.7 U/mL). The protein profile on electrophoresis was extensive, and the 23 kDa band corresponded to a xylanase. The xylanase presented optimum pH in the range of 7-8, optimal temperature between 45-50ºC and was more stable at 40 ºC (pH8.0). Hydrolysis of commercial xylanes produced xylotriose, xylotetraose, xylopentaose and xylooligosaccharides (XOS) with larger masses, which were not identified by thin layer chromatography (TLC). The influence of various factors, such as xylanase loading, substrate type, temperature and the addition of various compounds was evaluated in the xylan extraction yield from the pretreated sugarcane bagasse. The crude extract of B. pumilus, rich in xylanases, was applied to a bagasse pretreated with sulfite/alkali, and to a bagasse delignified by chlorite in acid medium, showing that the lower amount of residual lignin as the main factor to increase the extracting xylan, increasing the extraction from 4.2 to 42.5%. Extensive washing of the alkaline sulfite pretreated material also assisted in increasing xylan yield, from 18.5 to 25.1%. Further alternatives were tested to increase the hydrolysis yield of pretreated bagasse xylan without the need for material washing. The reuse of the alkali sulfite liquor, the addition of tween 80, polyethylene glycol, albumin did not favor xylan extraction, while addition of MgSO4 had a small positive effect on xylan extraction (from 18% to 20.9%). The cold alkaline extraction, after enzymatic extraction, positively influenced the xylan extraction yield of the pretreated bagasse with 10% Na2SO3/5% NaOH. Extraction of xylan obtained by applying xylanase (20 U / g) followed by extraction with 20% NaOH produced the same result as using only NaOH (70%), 24.1 and 24.9%, respectively. The enzymatic extraction of xylan allowed to obtain a residue with favorable characteristics for the complete saccharification, with Cellic CTec2 in the load of 10 FPU / g of material.

The objective of this thesis was to evaluate the effect of exogenous enzyme supplementation, phytase and xylanase-glucanase, on AME value of barley in poultry and swine. In the first study, 280 broilers were assigned 1 of 8 treatments. Barley inclusion in the diet resulted in decreased (P < 0.05) performance. There was a treatment × phytase × xylanase-glucanase interaction for dry matter retention with birds fed the corn-SBM-barley diet supplemented with phytase and xylanase-glucanase having higher (P < 0.05) DM retention compared to birds fed corn-SBM-based diet with only xylanase-glucanase supplementation. AME and AMEn of corn-SBM-based diets were greater (P < 0.05) than the corn-SBM-barley-based diets. Energy metabolizability and AMEn of barley significantly increased with xylanase-glucanase supplementation. In the second study, 24 pigs (12 pigs/phase) were assigned to 1 of 4 treatments with xylanase-glucanase and phytase. After a 7-d adaption period, urine and feces were quantitatively collected for 5 d. DE of the barley-based diet supplemented with xylanase-glucanase (3,578 kcal/kg) and phytase and xylanase-glucanase in combination (3,617 kcal/kg) were significantly different. Compared to control diets, exogenous enzymes either significantly improved or had a tendency to improve AME and AMEn value of barley in broilers, but not in growing pigs.

Des travaux de recherche en fermentation liquide ont montré que P. canescens est une souche hyperproductrice de xylanases non contaminées par des activités cellulolytiques et amylolytiques. Selon les scientifiques, lintérêt de lutilisation industrielle de ces hémicellulases dans différents secteurs (particulièrement dans la formulation daliments pour le bétail, en industries des jus de fruits et brassicoles, en amidonnerie, en industrie du papier, en pharmacie, dans les textiles et dans la production du bioéthanol) va croître significativement. Mais le développement de ces enzymes est fréquemment limité par le coût de production. Ce travail sest intéressé à loptimisation de la production des xylanases de P. canescens à partir de matières premières peu coûteuses telles les résidus agro-industriels par fermentation solide, une technique traditionnellement utilisée dans la fermentation des aliments en Asie. Létude a démontré que le tourteau de soja est un bon inducteur de la production des xylanases. La teneur initiale en eau, le pH initial, la température de la culture et laération active influencent la synthèse de l'enzyme. Compte tenu des résultats obtenus à léchelle du laboratoire, la transposition à léchelle industrielle serait facilitée naturellement par de fines épaisseurs de cultures statiques, ce qui réduit de moitié le coût de production comparativement à la fermentation liquide. Les expérimentations ont confirmé que la production de xylanases par P. canescens répondait à des phénomènes dinduction et de répression dépendant du substrat et des conditions physico-chimiques de croissance, et non pas à des phénomènes de régulation de type quorum sensing. Lenzyme sous forme liquide concentrée présente une bonne stabilité pendant six mois sans protection préalable (stérilisation, stabilisation ou inhibition de protéases).

The overall objective of this research is to identify and optimize enzymatic applications that have the potential to degrade middle lamella lignin, so as to decrease economic and environmental costs associated with the production of mechanical pulp. Non-productive binding of enzyme to lignin in lignocellulosic biomass reduces enzyme availability and efficiency. The elucidation of non-productive binding behavior between hydrolytic enzymes and lignocellulosic substrates could significantly improve the efficiency of corresponding industrial bioprocesses. The first part of this report presents a study that characterizes non-catalytic interactions between enzymes and fibre. The second part of this report presents the biochemical and mutational studies of a novel, small laccase SCO6712 from Streptomyces coelicolor. The findings from this research support the design, control, and optimization of enzymatic treatments of lignocellulosic fibres in the pulp and biofuel industries.

The overall objective of this research is to identify and optimize enzymatic applications that have the potential to degrade middle lamella lignin, so as to decrease economic and environmental costs associated with the production of mechanical pulp. Non-productive binding of enzyme to lignin in lignocellulosic biomass reduces enzyme availability and efficiency. The elucidation of non-productive binding behavior between hydrolytic enzymes and lignocellulosic substrates could significantly improve the efficiency of corresponding industrial bioprocesses. The first part of this report presents a study that characterizes non-catalytic interactions between enzymes and fibre. The second part of this report presents the biochemical and mutational studies of a novel, small laccase SCO6712 from Streptomyces coelicolor. The findings from this research support the design, control, and optimization of enzymatic treatments of lignocellulosic fibres in the pulp and biofuel industries.

Endo-b-1,4 xylanases (EC. 3.2.1.8) are typically produced as a mixture of different hydrolytic enzymes such as b-1,4-xylosidase (EC. 3.2.1.37) , a-Larabinofuranosidases (EC. 3.2.1.55), and feruloyl esterase (EC 3.1.1.73) that hydrolyze xylan molecule, which constitutes 20-30% of the weight of wood and agricultural wastes. Thus, xylan, a renewable biomass, can be utilized as a substrate for the preparation of many products such as fuels, solvents and pharmaceuticals. Besides, xylanolytic enzymes themselves are also used in food,feed, textile industries and pre-bleaching of kraft. In the first part of the study, xylanolytic systems of a soil isolate Bacillus pumilus SB-M13 and a thermophilic fungus Scytalidium thermophilum were investigated. Production rate and type of xylanolytic changed depending on the carbon source and the microorganism. However, xylanolytic enzyme production was found to be sequential, in synergy and under the control of carbon catabolite repression for both microorganisms. In the second part, B. pumilus SB-M13 b-1,4 xylanase was purified and biochemically characterized. The enzyme was stable at alkaline pHs and highest activity was observed at 60&deg / C and pH 7.5. Enzyme Km and kcat values were determined as 1.87 mg/ml and 43,000 U/mg, respectively. B. pumilus SB-M13 and S .thermophilum a-L-arabinofuranosidases were also purified and biochemically characterized. Although produced from a mesophilic microorganism, B. pumilus SB-M13 arabinofuranosidase was quite thermostable. Moreover, unlike other fungi, S. thermophilum produced alkaline stable arabinofuranosidases. Both enzymes were multimeric, alkaline stable and most active at 70&deg / C and pH 7.0. However, when compared to S. thermophilum, catalytic power of B. pumilus SB-M13 arabinofuranosidase was higher.

In this study, the aim was enzymatic xylooligosaccharide production from cotton and sunflower stalks, two of main agricultural residues in Turkey. In first two parts of the study, alkali extracted xylan from both of the stalks was hydrolyzed by commercial xylanases Veron and Shearzyme. The effect of temperature, pH, enzyme and substrate concentrations were investigated to determine optimum enzymatic hydrolysis conditions of xylan. Sunflower and cotton stalk xylans were hydrolyzed by Shearzyme more efficiently than Veron under the conditions studied. Shearzyme produced different product profiles containing xylobiose (X2), xylotriose (X3), xylotetrose (X4) and xylopentose (X5) from cotton and sunflower stalk xylan. On the other hand, Veron hydrolyzed both xylan types to produce X2, X3, X5, X6 and larger xylooligosaccharides without any change in product profiles. In the third part of the study, home produced xylanase from Bacillus pumilus SB-M13, was also investigated for the production of xylooligosaccharides from both cotton and sunflower stalk xylan. The main products obtained by hydrolysis of both substrates by pure B. pumilus xylanase were X5 and X6, while crude B. pumilus xylanase generated X4 and X5 as the main products. Xylooligosaccharide production from pretreated cotton stalk without alkali extraction of xylan was the final part of the study. Three different pretreatment methods including biomass pretreatment by Phanerochaete chrysosporium fermentation, cellulase pretreatment and hydrothermal pretreatment were investigated to break down complex lignocellulosic structure of cotton stalk to improve the subsequent enzymatic hydrolysis of xylan in pretreated cotton stalk for xylooligosaccharide production. However, xylooligosaccharide was not effectively produced from pretreated cotton stalk. Shearzyme inhibiton was observed after all the pretreatment methods during further hydrolysis of pretreated cotton stalk probably due to production of inhibitory compounds of the enzyme.