Xylooligosaccharides (XOS) are considered to be prebiotics. Prebiotics are defined as non-digestible food ingredients that benefit the host by stimulating the growth and activity of a limited number of bacteria, such as the Bifidobacterium genus, in the colon. Corn fiber separated from distillers dried grains with solubles (DDGS) could be a valuable feedstock for XOS production. The objective of the first chapter was to determine the efficacy for autohydrolysis to produce XOS using fiber separated from DDGS. Fiber was treated with deionized water in a Parr-reactor, at temperatures ranging from 140 to 220 °C to produce XOS. The maximum total yield of XOS in the solution was 18.6 wt% of the corn fiber at 180 °C. The objective of the second chapter was to evaluate and compare the prebiotic effect of XOS produced by autohydrolysis of DDGS fiber (XOS-D) with other substrates (FOS, commercial XOS (XOS-C), xylose, glucose and inulin) on intestinal bacteria, B. adolescentis, B. breve and Lactobacillus brevis. Bacterial growth on XOS-C was comparable with growth on FOS and inulin. XOS-D promoted bacterial growth more than that of control. Prebiotic potential of XOS produced from corn fiber was confirmed. The objective of third chapter is to determine the yield of XOS from corn fiber separated from ground corn flour (FC) and DDGS (FD) at different autohydrolysis temperatures and hold-times. The conditions for maximum XOS production for FD and FC were 180 °C with 20 min hold-time and 190 °C with 10 min hold-time, respectively. The fourth chapter focuses on production of XOS by enzymatic hydrolysis method for XOS production. Endo-1-4-xylanase enzyme was ineffective for corn fiber as well as corn fiber gum (CFG), despite evaluating a multitude of pretreatment methods and processing conditions. We have proposed use of Multifect Pectinase PE and Multifect Xylanase enzymes, based on work from other researchers. For commercial applications such as food industries, XOS would need to be isolated from liquor. The fifth chapter of this study focuses on literature review of purification methods used in XOS purification.

Cancer is a global risk for human wellness and health. Dietary habits could profoundly affect the risk of certain cancer, such as colorectal cancer (CRC) (Platz, 2000). CRC has listed the third leading cancer among male and female in the United States (Stewart, 2014). Surprisingly, the consumption of dietary fiber has an inverse correlation with the mortality of CRC (Song, 2018). However, most Americans do not consume enough dietary fiber to meet the recommended level of dietary fiber intake (Clemens, 2012; Lee-Kwan, 2017). Hence, it is reasonable to increase the nutrient density, i.e., dietary fiber, of current food model. Xylo-oligosaccharides (XOS), an emerging prebiotics, showed multiple advantages over fructo-oligosaccharides (FOS) and inulin. For example, Hsu et al. (2004) reported XOS are more effective than FOS on increasing Bifidobacterium level in rat cecum (Hsu, 2004). It is also suggested that dietary fiber and phenolic compounds have synergistic effect on promoting gut health (Uehara, 2001; Matsukawa, 2009). Therefore, the application of XOS into a polyphenol-dense food vehicle (strawberry puree) could be a viable way to promote gastrointestinal health and help reduce CRC risk. First, the effect of xylo-oligosaccharides (XOS) incorporation on the quality attributes of strawberry puree was investigated. Rheological properties, color and physicochemical characteristics and composition such as water activity, pH, and total soluble solids, were investigated to determine the maximum XOS content that can be incorporated into the strawberry pure without significantly altering the original properties of samples. Based on the collected data, adding xylo-oligosaccharides greater than 7.5% w/w will significantly change the quality attributes of the strawberry puree including its rheological characteristics, color profile, water activity, and total soluble solids. The addition of XOS at 2.5% and 5% w/w did not significantly alter overall quality attributes of strawberry puree. Second, the effects of heat treatment (HTST, 75℃, 15s and UHT, 121℃, 2s) and storage condition (storage time: 1, 15, and 36 days; storage temperature: 4℃ and 55℃) on the quality attributes of xylo-oligosaccharides enhanced strawberry puree was studied (texture, color water activity, total soluble solids, and pH). In addition, the physicochemical (total phenolic, tannin, flavonoids contents, and antioxidant activity) evolution was studied. A 9-point Hedonic test was conducted to evaluate the sensory properties (overall, texture, color, appearance, sweetness, aroma, flavor) of purees (with and without XOS, 5%w/w) under different thermal treatments. Briefly, all nutritional attributes were retained better under cold storage (4 ℃) as compared to high-temperature storage (55 ℃). The result revealed that thermal treatment and high storage temperature showed an inverse correlation with puree consistency. As for chemical analysis, intense thermal treatment (UHT) caused the most degradation in TPC, TFC and tannin level. However, such treatments (UHT) helped significantly increase the measurable antioxidant level. For other physicochemical properties, processing methods (HTST / UHT) and high storage temperature exhibited more significance in changing the color profiles of the specimen than XOS incorporation. Overall, the addition of XOS up to 5% w/w could increase nutritional value of strawberry puree as well as consumer preference without significantly compromising quality attributes. Based on instrumental and sensory analysis, HTST treated strawberry puree with XOS incorporation (5% w/w) meet the standard of sensorial attributes of fresh puree with improved shelf-life stability and fiber concentration.

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Submitted by Bruna Rodrigues (bruna92rodrigues@yahoo.com.br) on 2016-09-26T14:51:02Z No. of bitstreams: 1 DissMAF.pdf: 3997675 bytes, checksum: 3238bd54da33e9d0c201deb533005fa2 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-26T18:48:09Z (GMT) No. of bitstreams: 1 DissMAF.pdf: 3997675 bytes, checksum: 3238bd54da33e9d0c201deb533005fa2 (MD5) / Approved for entry into archive by Marina Freitas (marinapf@ufscar.br) on 2016-09-26T18:48:18Z (GMT) No. of bitstreams: 1 DissMAF.pdf: 3997675 bytes, checksum: 3238bd54da33e9d0c201deb533005fa2 (MD5) / Made available in DSpace on 2016-09-26T18:48:26Z (GMT). No. of bitstreams: 1 DissMAF.pdf: 3997675 bytes, checksum: 3238bd54da33e9d0c201deb533005fa2 (MD5) Previous issue date: 2015-03-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Xylooligosaccharides (XOS) are short-chain polymers of xylose (2 to 7 units) which can be produced by enzymatic hydrolysis of the xylan from the lignocellulosic feedstocks. XOS have a great potential as probiotic ingredients, and when they are incorporated in diets, they can provide many health benefits. The worldwide interest in the use of lignocellulosic residues is constantly growing, and in this scenario the soybean hull arises as a potential residue of the Brazilian agroindustry. The bioconversion of these residues to value-added products requires suitable pretreatments to deconstruct/disorganize the recalcitrant lignocellulosic complex, separating its main fractions: cellulose, hemicellulose, and lignin. In this context, this work did evaluate different biomass pretreatments aiming to produce XOS by the action of a Bacillus subtilis endoxylanase. Initially, the conditions for maximum catalytic activity of this enzyme were evaluated changing pH, buffer, and temperature. Among these parameters, 50 mM citrate buffer, pH 5.5, and 45 oC were the one that gave highest activity. The in nature soybean hull (previously chemically characterized) was hydrolyzed with soluble endoxylanase with different enzyme loads (40, 80, and 100 U/g biomass) under preestablished pH and temperature, producing around 55 mg RS/g dry biomass. This result, though little expressive, showed the viability of XOS production from soybean hull. However, this approach requires a suitable pretreatment of the lignocellulosic biomass to improve the endoxylanase accessibility to the C-5 fraction. Several pretreatments were performed in the soybean hulls, such as, enzymatic deproteinization, hydrogen peroxide/acetic acid pretreatment, and organosolv-ethanol pretreatment. For some pretreatments, reagent concentration and reaction time were evaluated, as well as, sequential pretreatment. Besides, enzymatic hydrolysis of the in nature soybean hull under microwave irradiation was also evaluated. The deproteinization of the soybean hull was not very efficient to the enzymatic hydrolysis of the remnant solid (production of 30 mg RS/dry biomass). However, this pretreatment allows the protein recovery as a high nutritional value hydrolysate. The pretreatment of the deproteinized soybean hulls with hydrogen peroxide solution (5 M, 1 h) removed 56% lignin without cellulose losses. However, this pretreatment did not contribute to an efficient action of the endoxylanase to the hemicellulose fraction (production of around 30 mg RS/g dry biomass). The organosolv-(50% v/v)ethanol pretreatment of the deproteinized soybean hulls promoted the removal of around 50% lignin, with low solubilization of hemicellulose (<17%), producing a poor substrate for the endoxylanase. The organosolv pretreatments with 50 and 70% (v/v) ethanol of the in nature soybean hull were able to solubilize around 30% hemicellulose, allowing the production of around 76 and 49 mg RS/g dry biomass, respectively, after hydrolysis with endoxylanase. Finally, the microwave action on the lignocellulosic biomass probably decreased the biomass recalcitrance, because the hydrolysis of the in nature soybean hulls catalyzed by the endoxylanase (100 IU / g of biomass) yielded approximately 100 mg of RS/g dry biomass. On the other hand, the hydrolysis performed in a reactor under conventional heating produced only 52 mg RS/g dry biomass. The results of this work did show that the combination of microwave irradiation and enzymatic hydrolysis might be a promising alternative to produce XOS. Keywords: soybean hulls; xylo-oligosaccharides; pretreatments / Xilo-oligossacarídeos (XOS) são polímeros de xilose de cadeia curta (2 a 7 unidades) que podem ser obtidos por hidrólise enzimática da xilana presente na fração de hemicelulose dos materiais lignocelulósicos. XOS possuem um grande potencial como ingredientes prebióticos, e quando incorporados na dieta, podem fornecer muitos benefícios à saúde. O interesse mundial no aproveitamento de resíduos lignocelulósicos é cada vez maior, e no cenário nacional a casca de soja se destaca como um potencial resíduo da agroindústria brasileira. Para viabilizar a bioconversão desses resíduos em produtos de interesse comercial (etanol 2G e XOS, por exemplo) são necessários pré-tratamentos, que atuam desconstituindo/desorganizando a estrutura altamente recalcitrante do complexo lignocelulósico e separando as frações principais da biomassa: celulose, hemicelulose e lignina. Neste contexto, este trabalho teve por objetivo avaliar diferentes pré-tratamentos da biomassa para produzir sequencialmente XOS por ação de uma endoxilanase de Bacillus subtilis. Inicialmente as condições de máxima atividade catalítica dessa enzima foram avaliadas variando pH, tampão e temperatura. Dentre as variáveis estudadas, as que contribuíram para uma melhor atividade da endoxilanase foram tampão citrato de sódio (50mM) pH 5,5 e 45 °C. A casca de soja in natura (previamente caracterizada quimicamente) foi hidrolisada com endoxilanase solúvel com diferentes cargas enzimáticas (40, 80 e 100 U/g casca) nas condições de pH e temperatura pré-estabelecidas, produzindo em média 55 mg de AR/g biomassa seca. Esse resultado, embora pouco expressivo, demonstrou a viabilidade da produção de XOS a partir de casca de soja, requerendo, entretanto, um pré-tratamento adequado para melhorar a acessibilidade da endoxilanase à fração C-5 da biomassa. Os pré-tratamentos avaliados foram a desproteinização enzimática da casca, prétratamento com peróxido de hidrogênio e ácido acético e pré-tratamento organossolve-etanol, variando nestes, as concentrações de solventes, tempo de reação e pré-tratamentos sequenciais. Adicionalmente, realizou-se a hidrólise enzimática da casca de soja in natura em reator micro-ondas. A desproteinização da casca de soja mostrou-se ineficiente para a hidrólise da fração sólida remanescente com endoxilanase (produção de 30 mg de AR/g biomassa seca), embora esse pré-tratamento permita a recuperação de proteínas como um hidrolisado de alto valor nutricional. O pré-tratamento com peróxido de hidrogênio (5 M, 1 h) para casca de soja desproteinizada removeu 56% de lignina sem perdas de celulose, entretanto, este pré-tratamento não contribuiu para uma eficiente atuação da endoxilanase sobre a fração hemicelulósica (produção de aproximadamente 30 mg de AR/g biomassa seca). O pré-tratamento organossolve-etanol 50% (v/v) da casca de soja desproteinizada removeu em torno de 50% de lignina com baixa solubilização de hemicelulose (< 17%), gerando, portanto, um líquido com baixa concentração de substrato para a ação da endoxilanase. Os pré-tratamentos organossolve-etanol 50 e 70% (v/v) da casca de soja in natura foram capazes de solubilizar em torno de 30% da hemicelulose, sendo possível a produção de 76 e 49 mg de AR/g de biomassa seca, respectivamente, após hidrólise com endoxilanase. Por fim, a ação das microondas sobre a biomassa lignocelulósica provavelmente reduziu a recalcitrância da biomassa, pois a hidrólise da casca in natura com endoxilanase (100 U/g de casca) produziu aproximadamente 100 mg de AR/g de biomassa seca, ao contrário da hidrólise conduzida em reator com aquecimento convencional que produziu em torno de 52 mg de AR/g de biomassa seca. Os resultados deste trabalho indicam que a combinação de irradiação micro-ondas e hidrólise enzimática pode ser uma alternativa promissora para a produção de XOS.