Dissertations / Theses on the topic 'Brewers grains'

Master of Science
Department of Animal Sciences and Industry
Micheal J. Brouk
An evaluation of short-term replacement of corn silage (CS) and soybean meal with a blend of wet brewers grains (BG) and cracked corn on milk production and DMI was completed utilizing 8 primiparous (192 DIM) and 4 multiparous (191 DIM) mid-lactation Holstein cows. Milk production, composition, DMI, production efficiency, fatty acid composition and diet digestibility were evaluated. Cows were allotted to a 4 x 4 Latin Square with 3 replications blocked by parity, DIM and energy corrected milk (ECM). Crude protein and starch levels were balanced between diets by varying the levels of cracked corn and soybean meal in four diets; 0 BG (0% wet BG and 24% CS of diet DM), 12 BG (12% wet BG and 12% CS), 18 BG (18% wet BG and 6% CS), and 24 BG (24% wet BG and 0% CS). Fifteen day periods were used, d11-15 designated for collection. Orts were collected daily and TMRs were fed at 5 to 10% of previous day’s intake. Cows were milked 3x/day and individual milk weights recorded at every milking. Milk samples, body weights and BCS were taken -2 and -1d pre-trial to obtain baseline data and d14 and 15 of each period. During collection, samples of TMR and orts were taken d1, 3 and 5. Fecal grab samples were taken d12-15 at 8 hr intervals and advanced 2 hrs every 24 hr period to account for diurnal variation. Dry matter intake was similar (P=0.33) among treatments (20.3, 20.8, 20.9 and 21.2 kg/cow) for 0 BG, 12 BG, 18 BG AND 24 BG respectively, however CP intake of 24 BG tended to be greater (P=0.05) than 0 BG. NDF intake was lower for 0 BG compared to all other treatments and 24 BG was higher than 12 BG (P=0.0007). Dietary fat intake was different (P<0.001) across all treatments, increasing with greater BG inclusion. Inclusion of BG had no effect (P=0.37) on milk production (30.5, 31.5, 31.6 and 32.1 kg/cow), fat percent or amount, protein percent, SNF, lactose or SCC, but protein yield (P=0.04) was lower and MUN (P=0.05)tended to be lower with 0 BG compared to 18 BG and 24 BG. Efficiency of milk production did not differ (P=0.93) among treatments. Milk fatty acid profiles were different among treatments, with general increases of individual fatty acids as BG inclusion increased. No differences were found in DM, CP or ADF digestibility across treatments. Results suggest wet BG fed simultaneously with grass hay can be utilized as a short-term replacement for CS in mid-lactation dairy cow diets.

Over the last several decades, there have been a tremendous developments and greatinnovations in photocatalysis process along with the development of efficient nanosized catalysts for simple approach and economic viability. In this study, magnetic core@doubleshell nanomaterials were investigated and synthesized in lab with three-step innovative approach where Fe3O4 nanoparticles (NPs) were produced first to act as cores without using any surfactants. The magnetite/silica core–shell structure was then prepared by hydrolysis of tetraethoxysilane (TEOS) in the presence of core particles under alkaline conditions. And the outermost shell, the α-Fe2O3/TiO2 nanoparticles, were grown over magnetic core of Fe3O4@SiO2 using coprecipitation and calcination method. Furthermore, the Fe3O4@SiO2@α-Fe2O3/TiO2 NPs were then loaded on the reduced graphene oxide (r-GO) using hydrothermal method and are also mixed by kneading with the layered double hydroxides (LDH) of Mg2+ and Al3+. These nanoparticles were characterized with scanning electron microscope (SEM), transmission electron microscope (TEM), dynamic light scattering (DLS), and energy dispersive X-ray spectroscopy (EDS). Different model compounds like microcrystalline cellulose (90 μm), D-xylose, and sodium lignosulfonate representing cellulose, hemicellulose, and lignin, respectively, were converted to valuable chemicals with different NPs under visible light for different time periods. For example, valeric acid (VA) and vanillylmandelic acid (VMA) were produced when cellulose was used for the conversion with core-double shell NPS which were quantified using high performance liquid chromatography (HPLC). Similar approach was adopted for the conversion of brewers’ spent grain (BSG), a lignocellulosic biomass, without oxygen under visible light, which yielded ethanol as the main product along with other sugars and acids of very low concentrations. The magnetic property of the nanomaterials made it easy for recycle and reuse. From a sustainability point of view, this study will fill a large need in the biomass photocatalysis field by developing core-shell multi-functional photocatalysts for direct transformation of lignocellulose into valuable chemicals under low temperatures, atmospheric pressure, and visible light from the sun.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico
Two experiments were conducted, the first to determine the chemical and digestible energy of dried brewers grain (DBG) and the second to evaluate the effect of the inclusion of this ingredient in the feeding of growing rabbits on performance, carcass characteristics, meat quality and economic viability. For the digestibility assay a total of 28 males rabbits were used, White New Zealand x California, with 50 days of age with an initial weight of 1.400Â0.100 kg, distributed on a completely randomized block design, with two treatments and fourteen replications, using the total of feces method. The treatments were: basal diet (BD) and ration test (RT) comprised 60% of basal diet and 40% dried brewers grain. The dried brewers grain showed 90.43% of dry matter, 37.94% of crude protein, 51.72% of neutral detergent fiber, 22.08% of acid detergent fiber, 8.06% of ether extract, 5.30% of mineral matters and 3371.24 kcal of digestible energy/kg. To test the performance, a total of eighty rabbits (40 males and 40 females), White New Zealand x California, in the period from 40 to 90 days of age, allocated in a completely randomized block design in a 5x2 factorial arrangement, considering five levels of inclusion of the DBG (0, 7, 14, 21 e 28%), and sex (male and female), with four replicates of two animals. No effects were observed on the performance, however, it was observed that the carcass weight decreased linearly, while the pÃncreas weight increased, due to the higher level of inclusion of the DBG. The weight of the liver of animals fed with DBG over 14% was lower than in animals that received feed without this ingredient. There was no effect of treatments on quality characteristics of meat rabbits. No effects were observed on relation meat/bone and qualitative meat characteristics. Considering the lower cost value of feed and better economic efficiency and cost indexes, the DBG may be included up to 28% in growing rabbits diets.
Foram realizados dois ensaios, sendo o primeiro para determinar a composiÃÃo quÃmica e a energia digestÃvel do resÃduo desidratado de cervejaria (RDC) e o segundo para avaliar o efeito da inclusÃo deste ingrediente na dieta de coelhos em crescimento sobre o desempenho, caracterÃsticas da carcaÃa, qualidade da carne e viabilidade econÃmica. Para o ensaio de digestibilidade foram utilizados 28 coelhos machos com 50 dias de idade e peso inicial de 1,400Â0,100 kg, distribuÃdos no delineamento em blocos ao acaso, com dois tratamentos e quatorze repetiÃÃes, utilizando mÃtodo de coleta total de fezes. Os tratamentos foram: raÃÃo referÃncia (RR) e raÃÃo teste (RT) composta por 60% da raÃÃo referÃncia e 40% de resÃduo desidratado de cervejaria. O resÃduo desidratado de cervejaria apresentou 90,43% de matÃria seca, 37,94% de proteÃna bruta, 51,72% de fibra em detergente neutro, 22,08% de fibra em detergente Ãcido, 8,06% de extrato etÃreo, 5,30% de matÃria mineral e 3371,24 kcal de energia digestÃvel/kg. Para o ensaio de desempenho foram utilizados 80 coellhos (40 machos e 40 fÃmeas) oriundos do cruzamento das raÃas Nova ZelÃndia Branco x CalifÃrnia, no perÃodo de 40 a 90 dias de idade, distribuÃdos no delineamento em blocos ao acaso em esquema fatorial 5x2, considerando cinco nÃveis de inclusÃo do RDC (0, 7, 14, 21 e 28%) e o sexo (macho e fÃmea), com quatro repetiÃÃes de dois animais. NÃo houve efeito dos tratamentos sobre o desempenho dos coelhos, no entanto, observou-se que o peso de carcaÃa decresceu linearmente enquanto o peso do pÃncreas aumentou, em funÃÃo do maior nÃvel de inclusÃo do RDC. O peso do fÃgado dos animais alimentados com raÃÃes contendo RDC acima de 14% foi menor em relaÃÃo aos animais que receberam raÃÃo sem este ingrediente. NÃo houve efeito dos tratamentos sobre a relaÃÃo carne/osso e caracterÃsticas qualitativas da carne avaliadas. Em funÃÃo do menor valor de custo com alimentaÃÃo bem como dos melhores Ãndices de eficiÃncia econÃmica e de custo, o RDC pode ser incluÃdo em atà 28% em raÃÃes para coelhos em crescimento.

The brewing process is an energy intensive process that uses large quantities of heat and electricity. To produce this energy requires a high, mainly fossil fuel consumption and the cost of this is increasing each year due to rising fuel costs. One of the main by-products from the brewing process is Brewers Spent Grain (BSG), an organic residue with very high moisture content. It is widely available each year and is often given away as cattle feed or disposed of to landfill as waste. Currently these methods of disposal are also costly to the brewing process. The focus of this work was to investigate the energy potential of BSG via pyrolysis, gasification and catalytic steam reforming, in order to produce a tar-free useable fuel gas that can be combusted in a CHP plant to develop heat and electricity. The heat and electricity can either be used on site or exported. The first stage of this work was the drying and pre-treatment of BSG followed by characterisation to determine its basic composition and structure so it can be evaluated for its usefulness as a fuel. A thorough analysis of the characterisation results helps to better understand the thermal behaviour of BSG feedstock so it can be evaluated as a fuel when subjected to thermal conversion processes either by pyrolysis or gasification. The second stage was thermochemical conversion of the feedstock. Gasification of BSG was explored in a fixed bed downdraft gasifier unit. The study investigated whether BSG can be successfully converted by fixed bed downdraft gasification operation and whether it can produce a product gas that can potentially run an engine for heat and power. In addition the pyrolysis of BSG was explored using a novel “Pyroformer” intermediate pyrolysis reactor to investigate the behaviour of BSG under these processing conditions. The physicochemical properties and compositions of the pyrolysis fractions obtained (bio-oil, char and permanent gases) were investigated for their applicability in a combined heat power (CHP) application.

The accumulation of huge amounts of food wastes every year has led to environmental degradation and especially to significant loss of valuable material. Steady consumption of beer results in a steady output of residues. Brewer’s spent grain (BSG) is the most abundant by-product generated in the beer-brewing process. BSG consists of the barley grain husks together with parts of the pericarp and seed coat layer that is obtained as solid residue after the production of beer wort. Despite the wide range of possible applications, its main use is limited to animal feed or simply deposition to landfills. The study of alternative uses of BSG is appropriate, both from the perspective of the brewer who can benefit from the valorisation but also from an environmental outlook. However, a significant understanding gap regarding the variability of this low-cost residue, its influence in the valorisation process and its use in biorefineries exists. This study attempts to characterize two different types of BSGs and the malts by which they are composed in terms of elemental composition, higher heating value, content of volatiles, fixed carbon, humidity, ash yield and to study the kinetic mechanism of decomposition during thermo-oxidative combustion. In terms of methodology, proximate analysis, ultimate analysis and calorific values were performed, as like as thermogravimetric analysis. Thermogravimetric thermograms were determined at different heating rates, using oxygen as oxidative atmosphere. The thermo-chemical reactions were mathematically described through the definition of the main kinetic parameters: activation energy, pre-exponential factor, model of reaction. The so-called kinetic triplet was calculated through the application of iso-conversional methods, Master-Curves and Perez-Maqueda criterion. The results of this study will serve as the baseline for the description of the energy recovery process in a reactor that uses brewer’s spent grain residues as feedstock.

Brewer's spent grain (BSG) is the most abundant by-product of the brewing industry and its main application is limited to low-value cattle feed. Since BSG contains 20 to 25% of proteins, it has the potential to provide a new protein source to the food industry. In this research, an ultrasound-assisted enzymatic extraction was designed to extract protein hydrolysates from BSG. Original BSG and ultrasound pretreated BSG were hydrolyzed under different enzyme (Alcalase) loadings and incubation times. Centrifugation was applied to separate solubilized proteins from insoluble BSG residue. When the enzyme loading increased from 1 to 40 uL /g BSG, the solubilized proteins increased from 34% to 64.8%. The application of ultrasound further increased the solubilized proteins from 64.8% to 69.8%. Solubilized proteins from ultrasound pretreated BSG was significantly higher (p < 0.05) than that from the original BSG. Particle size distribution analysis showed that the application of ultrasound pretreatment reduced the BSG particle size from 331.2 to 215.7 um. Scanning electron microscopy images revealed that the BSG particle surface was partially ruptured by the ultrasound pretreatment. These two phenomena might have contributed to the increased protein separation efficiency with ultrasound pretreatment. The solubility (pH 1.0 to 11.0) of protein hydrolysate increased by the application of ultrasound and the ultrasound did not lead to the change of the amino acid composition of the separated protein hydrolysates. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile, the protein was degraded to peptides which had molecular weights lower than 15 kDa. The color of the separated protein hydrolysates by enzymatic hydrolysis was brighter and lighter than the original BSG. The application of ultrasound did not affect the color of the separated protein hydrolysates. Overall, the ultrasound pretreatment prior to enzymatic hydrolysis enhanced the extraction of proteins from BSG in terms of higher protein separation efficiency, lower enzyme loadings, and reduced incubation time. This study developed a novel and green method to effectively extract value-added protein hydrolysates from the low-value food processing byproducts.
MSLFS

In recent years bioethanol (fuel ethanol derived through fermentation) has become the leading alternative to currently utilised liquid transportation fuels, possessing the benefits of being both sustainable and carbon neutral. If this position is to be maintained, it is clear that the feedstocks and processes used during its production have to be a major focus for the future direction of research, not only within the context of bioethanol but all biofuels in general. In terms of future sustainability, it is important that biofuel production should be derived from waste materials which exhibit limited potential for use in other applications. In this manner, food-to-fuel debates can be circumvented. Lignocellulose represents a readily available biowaste material for biofuel generation. The brewing and distilling industries in particular are uniquely placed to exploit the conversion of lignocellulose to bioethanol, through the utilisation of the millions of tons of spent grains produced annually by distilleries across the globe. Conversion of spent grains (SG) to bioethanol represents one of the most attractive and indeed challenging opportunities for meeting demand for sustainable bioethanol production. However it presents considerable challenges in terms of costs of production and associated energy balances. Enzyme costs are a particularly challenging issue and any technological developments which have potential to increase cellulolysis and decrease cellulase enzyme dosage are of distinct importance. Additionally, current pre-treatment approaches rely heavily of the use of acids during the pre-treatment stage. This generates a variety of compounds which are inhibitory to fermentation (e.g. acetic acid, furfural) and lead to low ethanol yields. Research which can reduce enzyme loading or improve low temperature pre-treatment are of significant importance. The research described here sought to evaluate the use of ultrasound in the pre-treatment and enzymolysis of spent grains. The use of acid in the pre-treatment and enzymolysis of SG was optimised to provide a benchmark for novel hydrolysis techniques. Ultrasound (20 kHz) was assessed as a pre-treatment technique in its own right as well as in combination with various oxidising chemicals such as ozone and hydrogen peroxide. Additionally, high frequency ultrasound (>500 kHz) was investigated in terms of its effect upon the activity of the enzymes involved in SG hydrolysis. A number of yeast species (S.cerevisiae, P.stipitis, K.marxianus, P. tannophilus and C. shehatae) were then evaluated for their ability to ferment the mix of five and six carbon sugars liberated during enzymatic hydrolysis of SG. Ultrasound was found to be effective in enhancing combined ozone and hydrogen peroxide pre-treatment of SG, although not as effective as the use of acid. Additionally, ultrasound was shown to significantly enhance the activity of cellulose, xylanase and ~- glucosidase, with the magnitude of the increase highly dependent on frequency and output power. The research presented here has expanded knowledge in terms of the effect of ultrasound upon lignocellulose as well as the enzymes involved in its digestion.

Thesis (Master of Engineering in Chemical Engineering)--Cape Peninsula University of Technology, 2018.
Brewer’s spent grain (BSG), a solid residue obtained from brewing beer, is gaining attention in the food, cosmetics and pharmaceutical industry due to its use as natural source of colorants, texturisers, functional ingredients and preservatives. It is therefore necessary to develop an economically viable method for the extraction, isolation or enrichment of these compounds. Although literature shows the technical feasibility of extraction of bioactive compounds from BSG at laboratory bench scale, none of the reviewed literature could provide adequate information necessary to determine the economic feasibility of the process at commercial scale. The aim of this study was to investigate the technical and economic viability of a commercial process for the recovery of antioxidant rich polyphenolic compounds from brewers spent grain using organic solvents and/or water. The objectives were to select the best solvent, perform the optimisation and kinetic study, as well as to model and simulate the extraction process with the aim of performing an economic analysis. In selecting the best solvent, maceration and soxhlet extraction were used for the recovery of polyphenolic compounds. Acetone and acetone: water mixtures, ethanol and ethanol: water mixtures as well as pure water were used as solvents. The evaluation of the best solvent was measured by the total phenolic content (TPC), flavonol content, the antioxidant activity using 2.2-diphenyl-1-picrylhydrazyl (DPPH) assay and ferric reducing antioxidant power (FRAP) assay. The study performed optimisation for various operational parameters (time, temperature, solvent to feed ratio and shaking speed) using response surface method. The effect of temperature on the extraction kinetics was also investigated with experiments being carried out at 20°C, 40°C and 80°C. Antioxidant activity was detected in all BSG extracts, but water showed the highest global yield and rates of extraction. The optimum conditions were found at 15 min reaction time, temperature of 40 °C, shaking speed of 185 rpm and solvent to solid ratio of 27.5: 1. So-Macdonald model was a suitable fit for the experimental data with a R2-value range of (0.85 < r2 < 0.995). A processing scenario was proposed as a base case, upon which subsequent scenarios were generated to improve the operation or the economics. SuperPro Designer® (Intelligen, Inc) was used for modelling the proposed process, for simulation and for the economic evaluation. Four alternative schemes from the base case simulation were developed for optimisation of the process. The process was found to be economically feasible and attractive with a return of investment (ROI) of 48.45 % for alternative scheme 4. The results in this thesis highlight the likely economic feasibility of the extraction of polyphenolic compounds from BSG at commercial scale by the maceration method.

Remediation of contaminated land needs to be carried out using methods that are both cost effective and minimise environmental pollution. However, the remediation option currently chosen by practitioners is often based upon limited economic information with the true environmental costs not being considered. This can result in the least sustainable option being chosen. This study has developed a methodology to evaluate the sustainability, in terms of economic and environmental costs, for a range of treatments available for the remediation of diesel contaminated land, including bioremediation (with and without the addition of brewery spent grain), disposal to landfill and thermal treatment. Initial laboratory investigations indicated that the use of brewery spent grain decreased the time taken for the clean-up of soil contaminated with diesel, suggesting that bioremediation augmented by the addition of this organic material was a viable option. A costing model was then developed that included all of the costs associated with the remediation options chosen. This included both direct and indirect costs. The results show that considering the indirect costs of remediation such as costs associated with delayed development the land, make bioremediation in this study an economically feasible option. Finally environmental costs were considered with a focus on the release of carbon dioxide a known greenhouse gas. Respirometry was used to determine the volume of carbon dioxide released during the bioremediation process. This information was then combined with data collected from a range of other sources and the impact of the chosen remediation options on atmospheric greenhouse gas release was evaluated. Other environmental impacts were also determined including land and water pollution. The results indicate that bioremediation with brewery spent grain has one of the lowest environmental costs and showed that emission from pollutants such as NOx, PM1.0, PM2.5, NH3 and SO2 could contribute to the limit values in the area covered by remediation work. The model developed in this study has indicated that the use of bioremediation with and without the use of brewery spent grain is a sustainable remediation option providing both direct and indirect economic costs are included. The results have indicated that, the strategy of using brewery spent grain to augment bioremediation v process promotes the re-use of by-product material, reduces waste and conserve resources. There is a need for the remediation industry to adopt similar models in order that decisions made, as to the remediation option chosen, are based upon accurate costings of their sustainability.

Breweries generate extensive by-products particularly spent grain, slurry and yeast. Much of the brewery spent grain (BSG) from large breweries is processed on a large scale for animal feed because of its high protein and fibre content. Bulk processing is economic and provides a useful income to the large brewery. Grain from small, local breweries is delivered intermittently in smaller volumes. Such use is much less economic and may incur losses if not carefully assessed for transport costs. The main aim of this research is to provide an analysis of using food by- products in agriculture with specific focus on spent grains from beverage production. This aim is investigated using a survey addressed to UK breweries. This survey produced a range of data regarding spent grain production, collection methods, vehicles size, average load, time of loading and unloading, etc. The data allowed the breweries to be grouped into three sizes, small, medium and large according to the scale of production of BSG. In order to achieve the research objectives, three models were applied; namely spent grain costing model (SGCM), Linear regression and Co-product Optimizer Decision Evaluator model (Cattle CODE). In addition, initial experiments were conducted to investigate alternative uses for wet brewery spent grain (WBSG) particularly soil fertilization. The SGCM model revealed that the majority of vehicles used to transport WBSG from three different sizes of brewery (small, medium and large) to farms were between 1 to 6 tonne loading capacity and the average distance from these breweries to farms was 5 miles. Data analyses were conducted by three vehicles classes and they were categorized in terms of loading capacity 1 tonne, 3 tonnes and up to 6 tonnes. This analysis was conducted to determine the average cost of transporting WBSG from breweries to farms. The results indicated that the average transport cost per tonne of WBSG for vehicle with the sizes of 1 tonne, 3 tonne and 6 tonnes were £ 10.11, £5.20 and £3.27 respectively. Linear regression based on SGCM model results was used to investigate the relationship between the distance between the three sizes of brewery and farms, and the average transport cost per tonne loaded of WBSG using three sizes of vehicle (vehicle 1, vehicle 2 and vehicle 3). The results indicated that the estimated average transport cost per tonne loaded of WBSG rises by £1.99, £1.31 and £0.45 per one mile increase in the distance for the three sizes of vehicle respectively. The Cattle CODE model results indicated that with WBSG available at £0 per tonne the collection system had a small but clear impact on the value of the grains with WBSG from medium and large breweries giving higher returns than WBSG from smaller breweries. At a maximal substitution rate to cattle feed of 50% this difference is around £28 per head of cattle. With the profitability of cattle being so low this difference may make a considerable impact on the viability of small scale cattle production. Primary experiments were conducted to investigate alternative uses for wet brewers spent grains WBSG as agents to improve soil fertilization. These experiments were conducted to assess alternative agricultural uses of spent grains and were focused on assessing the effect of grains on plant growth using cabbage, lettuce, radish and leek. The experimental results showed that in most cases the WBSG produces a clear effect on growth at the 40% addition level. This is most evident for cabbage wet weight and dry weight which increased by up to 70% compared to control plants. A similar pattern is suggested for leek wet and dry weight but is clearly not so for radish where the 10% WBSG level has most effect. These differences may allow specific recommendations of use to be developed to allow optimal application of WBSG.

This master‘s thesis deals with the influence of disintegration of brewer's grains for the production of biogas by anaerobic fermentation. The first part introduces us with the issue of biodegradable wastes. There are given their characteristics, ways of handling with them and legislative restrictions associated with them. In other sections there is a detailed description of anaerobic fermentation itself, including characteristics of biogas. Then there is issue of disintegration during fermentation including a list and characteristic of some disintegration methods. The main part of the thesis consists of description of the experiments on which it is observed increment of biogas based on the disintegration of brewer's grains and the evaluation of the individual experiments including the calculation of economic of the disintegration in laboratory conditions.

Mestrado em Engenharia Alimentar - Instituto Superior de Agronomia
Resulting from the filtration of wort, BSG is, among others, a by-product of the brewing industry. With the purpose of transforming this by-product in raw material to use in the baking industry, some of the bioactive components have been quantified, namely minerals, polysaccharides, phenolic components and antioxidant capacity of its components (AOA). The AOA was measured directly on BSG flour by DPPH method and ranged from 0.034 to 0.011 TEAC (M Trolox/100 g flour). When using the FRAP method, values varied between 0,066 and 0,031 TEAC (M Trolox/100g flour). These values were dependent on the concentration of flour and extract in the mixture and are lower than the ones obtained in tomato repiso, oregano and gorse. As for the mineral fraction, 100 gr of flour contains 260 mg of calcium, 255 mg of phosphorus, 186 mg of magnesium, 9 mg of sodium, 7 mg of iron and 4 mg of zinc, these levels are higher when compared to the ones for wheat and rye flours. Using spectrophotometry at 280 nm in concentrated aqueous solution, it was observed that the total phenolic content was of 18,63 mg EAG/g extract and by using the aqueous extraction process it was obtained a yield of 7% lyophilized extract, 80% insoluble solids and 2% polysaccharides. From all the results obtained it can be concluded that BSG flour has some antioxidant activity and high contents of minerals leading to the conclusion that can be incorporated in bakery products as a source to increase bioactive components.

First part of this master´s thesis decribes the basic properties of hydrogen, its utilization in industry and economy. Greater attention is focused on the possibilities of hydrogen produciton. Next part describes anaerobic fermentation and optimum conditions for hydrogen production. The main part of the thesis deals with posibility of hydrogen production in an experimental laboratory unit using brewer´s grains as a primary material. The experiment was aimed to find suitable process conditions for hydrogen production. The quantity of produced gas and his quality depending on the amount of brewer´s grains and frequency of the dosage was observed. Based on the experiment, some recommendations are proposed for further research. An automatic feeding system is the most important of them. It will improve the quality of experimental work. A separate chapter is devoted to the automatic feeding system.

Sustainable development is a growing concern for inhabitants of the planet earth. Consumption of fossil sources keeps up the depletion of nature’s capital and causes environmental impacts. One solution to have a sustainable society is to reduce the dependency on fossil fuels and substitute them by renewable energy sources. Among different types of renewable energy, biofuels have great potential for development and improvement. Though, the production of biofuels is criticized by many experts from the energy efficiency, environmental and economical points of view. Biogas as one type of first generation biofuels is achieved from the wastes and by-products of other industries, and can be used as a transportation fuel in the form of biomethane. The use of by-products may give added value as inputs to the biogas production process, a process which may be called "upcycling." The aim of upcycling is to convert wastes into new materials with higher quality or higher environmental value in order to reduce the consumption of raw materials which results in decreasing of energy usage and environmental impacts The aim of this thesis is to study the possibility of a biogas plant to act as an upcycler of wastes and by-products through anaerobic digestion process by the use of exergy analysis and economic evaluation. An imaginary biogas plant which uses a major by-product of brewing industry, i.e. Brewer’s Spent Grain (BSG), is considered to quantify the added value by biogas production process. The results of the exergy analysis show that the exergy of the input BSG (78,320 MJ) is upgraded into two main products as biomethane (47,430 MJ) and biofertilizer (37,026 MJ) with a total exergy amount of 84,456 MJ. On the other hand, the economic analysis of the studied biogas production process indicates that the biogas plant has the added value for the input material. In the economic analysis, the annual costs and benefits of the biogas production is calculated. The results show that the production of biomethane and biofertilizer from the by-product of brewing industry is profitable. However, the price of input BSG and also the variation price of the biofertilizer in different seasons, have great impact on the economy of a biogas plant. The outcomes from exergy and economic analysis are indicated that the biogas production process is an "upcycling" process which has the added value for the inputs, from both economic and quality points of view. The exergy and economic evaluation may be used as indicators of the sustainable development, but only increasing the exergy and the economic value of a production process alone should not be considered as the sustainability of a system.

O presente trabalho se propôs a avaliar o efeito de uma etapa de tratamento alcalino no bagaço de malte visando à extração de acetato e de compostos fenólicos, e como consequência diminuir a concentração destes no hidrolisado hemicelulósico. Foi ainda objetivo deste trabalho, estudar o efeito do tratamento alcalino na bioconversão das frações celulósica e hemicelulósica do bagaço de malte, por meio de fermentação com as leveduras Kluyveromyces marxianus e Pichia stipitis, respectivamente. Os resultados mostraram que para todas as condições de tratamento alcalino avaliadas, ocorreu solubilização de acetato, compostos fenólicos e furanos, não sendo observada solubilização de açúcares. A capacidade de solubilização destes compostos aumentou com o aumento da concentração de hidróxido de sódio empregada até o tempo de 40 min, sendo observada uma concentração máxima de 0,86 g/L, 13,0 g/L e 0,58 g/L empregando 1%%, 2% e 2% de NaOH, respectivamente. Para todas as condições avaliadas, o tratamento alcalino do bagaço de malte reduziu a concentração de ácido acético no hidrolisado hemicelulósico. A menor concentração (0,026 g/L) foi obtida após 20 minutos de tratamento com 2% de NaOH. Com relação à concentração xilose, o menor teor desta pentose (8,3 g/L) foi obtido após o tratamento do bagaço com 0,25 % de NaOH por 2 minutos e o maior (22,0 g/L) com 0,5% de NaOH por 60 minutos. Visando estabelecer as condições de tratamento alcalino que permitissem uma redução na concentração de ácido acético, concomitantemente ao favorecimento da solubilização de xilose no hidrolisado hemicelulósico e de glicose no hidrolisado celulósico, as condições de temperatura e concentração de NaOH do tratamento alcalino do bagaço de malte foram otimizadas através de um planejamento estatístico. Nas condições otimizadas (concentração de NaOH de 0,63%, temperatura de 67,6°C e tempo de 20 min), o tratamento alcalino do bagaço de malte promoveu uma redução de 90% na concentração de ácido acético, concomitante ao aumento de 4,5% e 15% na concentração de xilose no hidrolisado hemicelulósico e concentração de glicose no hidrolisado celulósico, respectivamente. Os resultados referentes à bioconversão da fração celulósica demonstraram que para ambas as configurações de processo avaliadas (SHF e SSF) a fermentação da celulignina do material tratado (CLT) forneceu maior concentração de etanol, e essa diferença foi mais pronunciada na SSF. Além disso, na SSF, a produtividade volumétrica total em etanol, a qual considera o tempo de sacarificação e fermentação no processo foi aumentada em 2,2 vezes em relação à SHF. Com relação a bioconversão da hemicelulose, os resultados mostraram que tanto para o hidrolisado do bagaço referência não tratado (HHR) quanto para o previamente tratado com álcali (HHT), a concentração máxima de etanol foi obtida após 89 horas de fermentação, porém na fermentação do (HHT) a produção de etanol foi cerca de 3 vezes superior a obtida no (HHR), passando de 5,8 para 17,4 g/L. Com base nos resultados obtidos, pode-se concluir que a etapa de tratamento alcalino do bagaço de malte, provavelmente promoveu mudanças estruturais na celulignina aumentando a liberação de glicose no hidrolisado e ainda foi capaz de remover substâncias inibidoras do metabolismo microbiano, especialmente o ácido acético, favorecendo o processo fermentativo da fração hemicelulósica.
This study aimed to evaluate the effect of alkaline treatment in brewer\'s spent grain on extraction of acetate and phenolic compounds, and consequently to decrease their levels on hemicellulosic hydrolyzate. In addition, the effect of the alkali treatment on the bioconversion of cellulose and hemicellulose hydrolyzates employing Kluyveromyces marxianus and Pichia stipitis, respectively, was also evaluated. The results showed that for all conditions of treatment studied occurred solubilization of acetate, phenolics and furans, but not sugars. The solubilizing capacity of these compounds increased with increasing concentration of sodium hydroxide employed until the time of 40 min, and the maximum concentration observed was 0.86 g / L 13.0 g / L and 0.58 g / L from 1%, 2% NaOH and 2%, respectively. For all conditions evaluated, alkali treatment of brewer\'s spent grain reduced the acetic acid concentration in hemicellulosic hydrolyzate. The lowest concentration (0.026 g / L) was obtained after 20 minutes with 2% NaOH. With respect to xylose concentration, the lower content of this pentose (8.3 g / L) was obtained after the treatment with 0.25% NaOH for 2 minutes and the highest (22.0 g / L) with 0.5 % NaOH for 60 minutes. To establish optimal conditions of temperature and NaOH concentration in the alkaline treatment aiming to decrease the concentration of acetic acid and, at the same time, increase the solubilization of xylose in the hemicellulose hydrolyzate and glucose in the cellulosic hydrolyzed, a statistical design was used. Under optimized conditions (NaOH concentration of 0.63%, a temperature of 67.6 °C and time of 20 min) alkaline treatment of the brewer\'s spent grain promoted a decrease of 90% in acetic acid concentration, concomitant to an increase of 4.5% and 15% on xylose concentration in the hemicellulose hydrolyzate and on glucose concentration in the cellulosic hydrolyzate, respectively. The results concerning the bioconversion of cellulosic fraction showed that for both configurations of the process evaluated (SHF and SSF) the treated material cellulignin (CLT) provided higher ethanol concentration, and this difference was more pronounced in SSF. In addition, in SSF, the total ethanol volumetric productivity, which considers time saccharification and fermentation process was increased by 2.2 times compared to SHF. In relation to the hemicellulose bioconversion, the results showed that for both hydrolyzate employed, i.e untreated (HHR) and previously treated with alkali (HHT), the maximum ethanol concentration was achieved after 89 hours of fermentation, however the ethanol production in HHT was about 3 times higher obtained in HHR (from 5.8 to 17.4 g/L). Based on these results, we conclude that the alkali treatment, probably induced structural changes in the fraction of cellulignin regarding glucose released in the hydrolyzate and still was able to remove substances inhibitory to microbial metabolism, especially acetic acid, favoring the fermentation of hemicellulose fraction.

Mestrado em Engenharia Alimentar - Instituto Superior de Agronomia
The present work aimed to study the incorporation of Draff (about 20% fibre and 25% protein)and Sodium Stearoil Lactilate (SSL) at wheat flour to produce bread supplemented with fibre and protein. An experimental design was adopted based on the Response Surface Methodology with 5 levels and two variables: addition of draff and addition of SSL; given rise to 9 formulations plus two replicates of the central point (10% draff and 0.25% SSL). Farinografic tests were previously made to define the hydration levels of the formulations as well as their mixing times and dough stability. The breadmaking experiments were conducted after a previously designed protocol, producing 4 breads for each formulation. These breads were evaluated after 24h for appearance (supported by photographs and scanning of the bread slices); colour Lch parameters (Minolta colorimeter); volume (rape seed method); aw (rotronic equipment); texture of the crust and crumb (Texturometer Tax-Tplus da Stable Microsystems, UK) and weight loss as well as texture and moisture at 48h and 72h after baking to follow bread aging. It was concluded that it is possible to add up to 10% draff using about 0.15% SSL and produce breads with good properties

O presente estudo teve como objetivo avaliar a produção de etanol pela levedura Pichia stipitis, a partir do hidrolisado hemicelulósico do bagaço de malte (HHBM). Primeiramente estudou-se o efeito da suplementação nutricional do hidrolisado adicionando-se extrato de farelo de arroz (0 a 20% v/v), uréia (0 a 3 g/L) e extrato de levedura (0 a 3 g/L). Os resultados mostraram que o hidrolisado suplementado apenas com extrato de levedura proporcionou os melhores resultados de conversão (YP/S = 0,44 g/g) e produtividade em etanol (QP = 0,33 g/L.h). Em seguida foi avaliado o nível ótimo deste nutriente sobre a bioconversão, sendo confirmada a suplementação do HHBM com 3,0 g/L de extrato de levedura. Após o estabelecimento das condições nutricionais, a levedura foi cultivada em HHBM após o crescimento de 24 horas em meio semissintético, visando aclimatar as células aos inibidores presentes no hidrolisado. Este estudo resultou no aumento das velocidades do processo, sendo 16% sobre a produtividade volumétrica em etanol (QP) e 10% sobre a velocidade de consumo de substrato (QS), quando comparado ao cultivo das células somente em meio semissintético. Na etapa seguinte utilizou-se um planejamento fatorial 22 com face centrada para otimização das condições de pH e concentração inicial de células (X0) na fermentação em HHBM não destoxificado e após a destoxificação com carvão ativado. Foram obtidos modelos para descrever os valores de YP/S e QP na região estudada, sendo que para o HHBM não destoxificado os valores destes parâmetros foram 0,40 g/g e 0,65 g/L.h, respectivamente, em pH 6,4 e X0 = 5,0 g/L (modelo 1). Em HHBM destoxificado, as condições ótimas foram obtidas em pH 6,0 e 1,36 g/L de células (modelo 2), obtendo-se YP/S de 0,40 g/g e QP de 0,46 g/L.h. Nas condições otimizadas foram então realizados ensaios que confirmaram a validade dos modelos 1 e 2, obtendo-se a máxima concentração de etanol (23,4 g/L), YP/S de 0,41 g/g e QP de 0,65 g/L.h em HHBM não destoxificado. Realizouse ensaios para avaliação do efeito do ácido acético sobre a fermentação em meio semissintético por P. stipitis, empregando-se as condições de pH e X0 otimizadas em HHBM não destoxificado (modelo 1) e destoxificado (modelo 2). Este estudo mostrou que nas condições do modelo 1, o ácido acético favoreceu a bioconversão sendo os melhores resultados obtidos na presença deste ácido (YP/S = 0,47 g/g e QP = 1,08 g/L.h). Por outro lado, nas condições do modelo 2, os valores de YP/S foram similares, enquanto que com a adição de ácido acético ao meio de fermentação, o valor de QP foi reduzido em 53%. Na fermentação em biorreator, o emprego das condições otimizadas em frascos (pH 6,4 e 5,0 g/L de células) resultaram em valores de QP 48% inferiores ao obtido em frascos (0,65 para 0,44 g/L.h), entretanto YP/S foi apenas 10% inferior (0,41 para 0,37 g/g). No presente estudo, conclui-se que a suplementação nutricional do HHBM e a otimização das condições de pH e X0 resultaram em valores promissores para os principais parâmetros da fermentação por P. stipitis, ressaltando o potencial deste hidrolisado em processos biotecnológicos para produção de etanol.
This study aimed to evaluate the ethanol production by Pichia stipitis in brewer\'s spent grain hemicellulosic hydrolysate (BSGHH). Initially, the effect of nutritional supplementation was evaluated by adding rice bran extract, urea and yeast extract. The results showed that supplementation only with yeast extract promoted the highest conversion values (YP/S = 0.44 g/L) and ethanol productivity (QP = 0.33 g/L.h). Additional assays showed that the optimal concentration of this nutrient was 3.0 g/L. To acclimate the cells to inhibitors present in BSGHH the yeast was cultivated in hydrolysate after growth for 24 hours in semisynthetic medium. This study resulted in increased of the process rates, 16% of the ethanol productivity (QP) and 10% on the substrate consumption (QS) when compared to growing cells only in the semisynthetic medium. In the second step a 22 full factorial centeredface design was employed to optimized the conditions of pH and initial cells concentration (X0) in fermentation of hydrolysate undetoxified and after detoxification with activated charcoal. Mathematical models that relate the YP/S and QP were obtained. For non-detoxified BSGHH (model 1) the optimal conditions of pH (6.4) and X0 (5.0 g/L) showed values parameters of 0.41 g/g and 0.65 g/L.h, respectively. In detoxified BSGHH (model 2) the optimum conditions of pH (6.0) and X0 (1.36 g/L), resulted in YP/S and QP values of 0.40 g/g and 0.46 g/L.h, respectively. Under these conditions, the the validity of the models were confirmed. The effect of acetic acid on fermentation by P. stipitis in semisynthetic medium, employing optimized conditions of pH and X0 in model 1 and model 2 was evaluated. The results showed that under the conditions of model 1 and in a concentration of 2,9 g/L, the acetic acid favored the bioconversion by P. stipitis, increasing the YP/S (15 %) and QP (66 %). On the other hand, in the conditions of the model 2 the YP/S values were similar, whereas the QP values were reduced by 53% when the acetic acid was added. By using these optimized conditions in bioreactor fermentation it was obtained the ethanol productivity was approximately 48% lower (0.65 to 0.44 g/L.h), however the ethanol production was similar as compared to fermentation flasks. It is possible conclude that the HHBSG requires nutritional supplementation and that the optimized conditions of pH and initial cells concentration can be used as a strategy in order to raising the fermentation parameters.

Pivski trop čini približno 85% od ukupnih sporednih proizvoda proizvodnje piva, i dostupan je po veoma niskim cenama tokom čitave godine. Pivski trop ima veliku perspektivu za primenu u biotehnologiji i proizvodnji visoko vrednih proizvoda. Jedna od veoma ekološki i ekonomski isplativih alternativa je upotreba pivskog tropa u proizvodnji mlečne kiseline, jer se poslednjih par decenija uočava intenzivan rast potražnje za mlečnom kiselinom. Mlečna kiselina je najvažnija hidroksikarbonska kiselina široko rasprostranjena u prirodi, sa velikom primenom u prehrambenoj, farmaceutskoj, tekstilnoj i hemijskoj industriji i industriji prerade kože.Cilj istraživanja ove doktorske disertacije je ispitivanje primene pivskog tropa u proizvodnji mlečne kiseline. Prvo je izvršena optimizacija enzimske hidrolize pivskog tropa u cilju dobijanja što je moguće veće koncentacije redukujućih šećera neophodne za mlečno-kiselu fermentaciju. Hidrolizat pivskog tropa je dobijen enzimskom hidrolizom dodatkom komercijalnih enzima za razgradnju skroba i celuloze. Parametri čiji je uticaj na efikasnost enzimske hidrolize ispitanu su: pH vrednost, temperatura hidrolize i količina dodatih enzima. Nakon što su određeni najbolji uslovi razgranje pivskog tropa, dobijeni postupak hidrolize je primenjen u proizvodnji hidrolizata pivskog tropa koji je korišćen u mlečno-kiselim fermentacijama.Nakon toga je ispitana mlečno-kisela fermentacija sa dva proizvodna mikoorganizma. Kao proizvodni mikroorganizmi u mlečno-kiselim fermentacijama primenjena su dva soja bakterija mlečne kiseline: Lactobacillus fermentum PL-1 i Lactobacillus rhamnosus ATCC 7469. Ispitan je uticaj dodatka različitih koncentracija ekstrakta kvasca (0,5-5,0%) uz korekciju pH vrednosti tokom fermentacije sa dodatkom kalcijum-karbonata. U zavisnosti od udela L-(+)- i D-(-)-mlečne kiseline koje nastaju tokom fermentacije izabran je proizvodni mikroorganizam koji proizvodi više L-(+)-mlečne kiseline.U daljim ispitivanjima je ispitan uticaj korekcije pH pomoću natrijum-hidroksida kao i dodatak različitih koncentracija ekstrakta kvasca (0,5-5,0%) i redukujućih šećera (2,7; 5,4 i 8,1%) u hidrolizatu pivskog tropa na mlečno-kiselu fermentaciju pomoću odabranog soja bakterija mlečne kiseline. Na osnovu dobijenih rezultata izabrana je najbolja koncentracija redukujućih šećera i ekstrakta kvasca koji će se koristiti u daljim istraživanjima.Takođe je ispitana i mogućnost zamene skupog ekstrakta kvasca i glukoze sa obnovljivim sirovinama, kao što su pivski kvasac, džibra i bistra džibra.Ispitan je uticaj dodatka različitih koncentracija pivskog kvasca (0,5-5,0%), džibre (5-20%) i bistre džibre (5-50%) pre fermentacije kao i dodatak bistre džibre u dolivnoj fermentaciji, na mlečno-kiselu fermentaciju hidrolizata pivskog tropa.Ispitan je i dolivni postupak fermentacije hidrolizata pivskog tropa dodatkom glukoze, glukoze i ekstrakta kvasca i sladovine. Takođe je ispitana mogućnost izvođenja više uzastopnih fermentacija sa imobilisanim ćelijama odabranog soja bakterija mlečne kiseline u kalcijum-alginatu.Na osnovu eksperimentalnih rezultata zaključujeno je da je dodatak kalcijum-karbonata imao pozitivan uticaj na proizvodnju mlečne kiseline sa L. fermentum i L. rhamnosus. Sa dodatkom kalcijum-karbonata povećali su se utrošak redukujućih šećera, koncentracija i prinos mlečne kiseline i vijabilnost ćelija L. fermentum i L. rhamnosus. Ekstrakt kvasca i kalcijum-karbonat su imali značajan uticaj na proizvodnju mlečne kiseline sa L. fermentum i L. rhamnosus. U fermentacijama sa L. fermentum najveći prinos ukupne mlečne kiseline (44%) je postignut sa dodatkom 5,0% ekstrakta kvasca i 2,0% kalcijum-karbonata. U fermentacijama sa L. rhamnosus najveći prinos ukupne mlečne kiseline (98%) i L-(+)-mlečne kiseline (96%) je ostvaren u fermentaciji sa dodatkom 2,0% ekstrakta kvasca i 2,0% kalcijum-karbonata. Na osnovu rezultata odlučeno je da se u daljim ispitivanjima mlečno-kisele fermentacije hidrolizata pivskog tropa kao proizvodni mikoorganizam koristi L. rhamnosus.Primenom natrijum-hidroksida za korekciju pH je skratila fermentaciju za 48 sati a ostvareno je i značajno povećanje zapreminske produktivnosti L-(+)-mlečne kiseline (za 200%, povećanje sa 0,21 na 0,63 g/l·h-1). Korekcija pH u svim daljim istraživanjima je vršena sa dodatkom natrijum-hidroksida.U mlečno-kiselim fermentacijama sa različitim početnim koncentracijama redukujućih šećera (2,7; 5,4 i 8,1%) i sa dodatkom različitih koncentracija ekstrakta kvasca (0,5-5,0%), najveći prinos L-(+)-mlečne kiseline i zapreminska produktivnost od 91,29% i 1,69 g/l·h-1, kao i vijabilnost ćelija L. rhamnosus od 9,7·109 CFU/ml ostvareni su u fermentaciji sa početniom koncentracijom redukujućih šećera od 5,4% i dodatkom 5,0% ekstrakta kvasca.Na osnovu ostvarenih rezultata u istraživanjima sa dodatkom džibre i dodacima tokom fermentacije kao i u fermentacijama sa imobilisanim ćelijama je korišćen hidrolizat pivskog tropa sa početnom koncentracijom redukujućih šećera od 5,4%.U mlečno kiseloj fermentaciji sa dodatkom pivskog kvasca najveći prinos L-(+)-mlečne kiseline (89,01%) i zapreminska produktivnost (0,89 g/l·h-1) L-(+)-mlečne kiseline su ostvareni u fermentaciji sa dodatkom 5,0% pivskog kvasca i korekcijom početne koncentracije redukujućih šećera na 5,0%. Na osnovu rezultata utvrđeno je da se može izvršiti delimična ili potpuna zamena ekstrakta kvasca pivskim kvascem uz značajno smanjenje cene podloge za mlečno-kiselu fermentaciju, bez značajnog smanjenja efikasnosti mlečno-kisele fermentacije.U mlečno-kiseloj fermentaciji sa dodatkom džibre i bistre džibre najveć koncetracija, prinos i zapreminska produktivnost L-(+)-mlečne kiseline od 31,03 g/l, 86,15% i 0,93 g/l·h-1, ostvareni su u fermentaciji sa dodatkom 50% bistre džibre. Najviša koncentracija, prinos i zapreminska produktivnost L-(+)-mlečne kiseline ostvareni u dolivnoj fermentaciji sa dodatkom glukoze i bistre džibre tokom mlečno-kisele fermentacije su iznosili su 48,02 g/l, 87,82% i 0,96 g/l·h-1.U fermentacijama sa dodatkom nutritijenata tokom mlečno-kisele fermentacije najveća vrednost koncetracije, prinosa i zapreminske produktivnosti L-(+)-mlečne kiseline od 116,08 g/l, 93,32% i 2,04 g/L·h-1, su ostvarene u fermentaciji sa dodatkom glukoze i ekstrakta kvasca tokom fermentacije. Na osnovu rezultata utvrđeno je da se dolivni postupak fermentacije može koristiti u cilju povećanja efikasnosti mlečno-kisele fermentacije.Izvršena je imobilizacija ćelija L. rhamnosus u kalcijum-alginatu uz izuzetno visoku vijabilnost (1010 CFU/ml). Imobilisane ćelije L. rhamnosus su uspešno korišćene u tri mlečno-kisele fermentacije. Prinos L-(+)-mlečne kiseline i zapreminska produktivnost su u sve tri fermentacije bili izuzetno visoki, pri čemu su najveći prinos L-(+)-mlečne kiseline i zapreminska produktivnost od 95,2% i 1,76 g/l·h-1, ostvareni u drugoj fermentaciji. Upotrebom imobilisanih ćelija L. rhamnosus je osim povećanja prinosa i zapreminske produktivnosti L-(+)-mlečne kiseline skraćena fermentacija za 12 sati u poređenju sa šaržnim fermentacijama.
Brewers spent grain represents (BSG) about 85% of the total by-products from brewing process and is available at low price during the whole year. Due to its chemical composition BSG has great potential use in biotechnology and production of high-value products. One of very eco-friendly and economical alternative uses of BSG is in production of lactic acid (LA), since in the last few decades the demand for the LA has significantly risen, mostly because of development of biodegradable lactic polymers, which are eco-friendly and nontoxic.Lactic acid is the most important hydrocarboxylic acid with an asymmetrical carbon atom, widely distributed in nature, and it has shown great potential in fields of food, pharmaceutical, textile, leather and chemical industries.The aim of this doctoral thesis was to investigate the application of BSG in lactic acid production. First, the optimization of enzymatic hydrolysis of BSG was conducted, with the goal to achieve high reducing sugar concentrations, as much as possible, that are necessary on LA fermentation. BSG hydrolysis was conducted by usage of commercial enzymes for degradation of starch and cellulose. Effect of pH value, temperature and enzyme dosage on BSG hydrolysis efficiency was investigated. After the best conditions for BSG hydrolysis were determined, the optimized procedure for BSG hydrolysis was used for the production of BSG hydrolysate that will be used in LA fermentations.After optimization of BSG hydrolysis, LA fermentation by two LA producing microorganisms was investigated. The strains investigated were two LA bacteria strains: Lactobacillus fermentum PL-1 and Lactobacillus rhamnosus ATCC 7469. The effect of yeast extract (0.5; 1.0; 2.0; 3.0; 4.0, and 5.0%) addition in BSG hydrolysate, with the correction of pH value during LA fermentation by the addition of calcium-carbonate, on LA fermentation was investigated. Based on the results achieved for L-(+)- and D-(-)-LA ratio the LAB strains that produced more L-(+)-LA was chosen for further research.In further research the effect of pH correction (with addition of NaOH), yeast extract (0.5, 1.0, 2.0, 3.0, 4.0, and 5.0%) addition and reducing sugar concentration (2.7; 5.4 and 8.1%) in BSG hydrolysate on LA fermentation was investigated. Based on the results achieved the best yeast extract and reducing sugars concentrations was determined and used in further analysis or research. Also the possible replacement of expensive yeast extract and glucose with cheap alternatives, like brewer`s spent grain and stillage was investigated. The effect of brewer`s spent grain (0.5; 1.0; 2.0; 3.0; 4.0, and 5.0%), whole stillage (5, 10, 15 i 20%) and thin stillage (5, 10, 15, 20, 30, 40, 50%) addition before fermentation as well as thin stillage addition in fed-batch fermentation in BSG hydrolysate on LA fermentation were investigated.Also fed-batch fermentation procedure (addition of glucose, glucose and yeast extract and wort during fermentation) was investigated. The possible application of cells immobilized in Ca-alginate for LA fermentation of BSG hydrolysate was also investigated.Based on the results it was concluded that BSG can be successfully utilized as a raw material in production of LA, after optimization of hydrolysis and addition of nitrogen source.According to the results of chemical composition before and after optimized hydrolysis 78.6% of total cellulose was hydrolyzed.Addition of calcium-carbonate had positive effect on LA production by L. fermentum i L. rhamnosus. With the addition of calcium-carbonate reducing sugar utilization, LA yield and concentration and cell viability (both L. fermentum i L. rhamnosus) increased. Addition of calcium-carbonate and yeast extract had a positive effect on LA fermentation by L. fermentum and L. rhamnosus. In LA fermentation by L. fermentum the highest LA yield (44%) was achieved with addition of 5.0% of yeast extract and 2.0% of calcium-carbonate. In L. rhamnosus fermentations the highest total LA yield (98%) and L-(+)-LA yield (96%) was reached when 2.0% of yeast extract and 2.0% of calcium-carbonate were added.Based on the results achieved it was concluded that BSG hydrolysate, with the addition of yeast extract, is a good fermentation media for LA fermentation with L. rhamnosus, and it was decided that L. rhamnosus will be used in further research of LA fermentation on BSG hydrolysate.Addition of NaOH instead of calcium-carbonate for the pH correction shortened the fermentation time by 48 h and increased the L-(+)-LA volumetric productivity (by 200%, from 0.21 to 0.63 g/L·h-1). Based on this results pH correction in further experiments was done by addition of NaOH.In LA fermentation with different reducing sugar (2.7, 5.4 and 8.1%) and yeast extract concentrations (0.5-5.0%), the highest L-(+)-LA yield and volumetric productivity of 91.29%, and 1.69 g/L·h-1, respectively, as well as L. rhamnosus cell viability (9.67 log CFU/mL), were achieved with the reducing sugar content of 5.4% and yeast extract content of 5.0%.Based on this results in further experiment with the addition of stillage, in fed-batch fermentation and fermentation with immobilized cell BSG hydrolysate with 5.4% of reducing sugars and 5.0% yeast extract was used.In fermentation with the addition of brewer’s spent yeast the highest L-(+)-LA yield (89.01%) and volumetric productivity (0.89 g/L·h-1) were achieved in the fermentation of BSG hydrolysate with 5.0% of reducing sugar and 5.0% of brewer’s yeast. Based on the results achieved it was concluded that yeast extract can be partial or complete replaced by brewer’s spent yeast with significant decrease of media cost, without the decrease in LA fermentation efficiency.In fermentation with the addition of thin stillage the highest L-(+)-LA concentration, yield, and volumetric productivity of 31.03 g/L, 86.15%, and 0.93 g/L·h-1, respectively, was obtained in fermentation with the addition of 50% of thin stillage. The highest L-(+)-LA concentration, yield, and volumetric productivity achieved in fed-batch fermentation with the addition of glucose and thin stillage during fermentation, were 48,02 g/L, 87,82% i 0,96 g/L·h-1.In fed-batch fermentation the highest L-(+)-LA concentration, yield, and volumetric productivity of 116.08 g/L, 93.32%, and, 2.04 g/L h-1, respectively, were achieved in fermentation with glucose and yeast extract addition during fermentation. The results showerd that fed-batch fermentation could be used to increase L-(+)-LA fermentation efficiencyImmobilization of L. rhamnosus cells with high viability (1010 CFU/mL) in Ca-alginate was conducted. Immobilized cells we successfully utilized in three repeated batch fermentation. L-(+)-LA yield and volumetric productivity were very high in all three batch fermentation, with the highest results achieved (95.20% and 1.76 g/L·h-1, respectively) in second fermentation. Application of immobilized L. rhamnosus cells increased L-(+)-LA yield and volumetric productivity and shortened the fermentation time for 12 h in comparison with batch fermentation.

Atualmente observa-se um aumento significativo no consumo de cervejas com baixo teor alcoólico e cervejas sem álcool. Existe um crescente interesse por este tipo de produto tanto por parte das indústrias cervejeiras como por parte dos consumidores. Este aumento é principalmente devido a questões de saúde e a razões de segurança no trabalho e nas estradas. Além disso, há países onde o consumo de álcool é proibido por lei. Tendo em vista que o consumidor está em busca de novos produtos com características mais próximas possíveis de uma cerveja convencional, o presente trabalho visa analisar uma nova abordagem para a produção de cerveja lager com baixo teor alcoólico, reutilizando o bagaço de malte, um subproduto da indústria cervejeira, como substituinte de parte do malte. Foram obtidas cervejas com as seguintes proporções de bagaço/malte: 0:100, 25/75, 50/50 e 75/25. Nas cervejas com 50 e 75% de bagaço de malte, observou-se uma redução no tempo de fermentação quando comparadas com a cerveja puro malte. A atenuação real de fermentação das cervejas produzidas também foi menor devido a menor fermentabilidade dos mostos. O teor alcoólico das cervejas obtidas em escala de bancada apresentou valores entre 0,665 e 1,615 % (v/v), o que as caracterizam como cervejas de baixo teor alcoólico A cerveja com 25% de bagaço recebeu as maiores notas na análise sensorial, tendo boa aceitação entre os provadores, e foi escolhida para a produção em escala piloto. O perfil de fermentação da cerveja produzida em escala piloto foi similar ao perfil de fermentação da cerveja produzida em escala de bancada. Também foi realizada uma análise sensorial, comparando a cerveja produzida em escala piloto com duas cervejas comercias. A utilização do bagaço de malte como substituinte do malte na fabricação de cervejas com baixo teor alcoólico é uma estratégia viável, produzindo uma bebida com aceitação sensorial semelhante ao de uma cerveja de baixo teor alcoólico comercial.
Nowadays there has been a significant increase in the consumption of beer with low alcohol content and non-alcoholic beers. There is a growing interest on this type of product by both brewers industries and consumers. This increase is mainly due to health issues and safety reasons at work and on the roads. In addition, there are countries where the consumption of alcohol is prohibited by law. Given that the consumer is looking for new products with characteristics as similar as possible of conventional beer, this study aims to analyze a new approach for the production of lager beer with low alcohol content, by reusing brewer\'s spent grain, a by-product of brewing industry, as a substitute of part of the malt. Beers were obtained with the following proportions of bagasse/malt: 0/100, 25/75, 50/50 and 75/25. In beers with 50 and 75% of bagasse, it was observed a reduction in the fermentation time compared with all-malt beer. The Real Attenuation Fermentation of Beer produced was also lower due to lower fermentability of wort. The beers\' alcohol content obtained in laboratory scale showed values between 0.665 and 1.615% (v/v), which characterize them as low-alcohol beers. Beer with 25% of bagasse received the highest scores in sensory analysis, having a good acceptance among the tasters, therefore it was chosen for the production on a pilot scale. The beer\'s fermentation profile produced on pilot scale was similar to the beer\'s fermentation profile produced in laboratory scale. Sensory analysis was also performed, comparing the beer produced in pilot scale with two commercial beers. The use of brewer\'s spent grain as a substitute in the manufacture of beers with low alcohol content is a viable strategy, producing a beverage with sensory acceptance similar to a commercial low-alcohol beer.

Zadatak ove teze je da se ispita uticaj zamene dela pšeničnog brašna ekstrudiranim snek proizvodima obogaćenim dodatkom pšenične klice, pivskog tropa i tropa jabuke na svojstva testa (boja, fizička, reološka) kao i na karakteristike čajnog peciva (fizičke, senzorske, nutritivne, trajnost, mikrobiološke). U toku preliminarnih ispitivanja (prva faza eksperimentalnog rada) ispitan je uticaj obezmašćene pšenične klice na kvalitet testa i čajnog peciva u cilju definisanja optimalnog nivoa zamene pšeničnog brašna ekstrudatima kukuruzne krupice sa dodatkom sporednih proizvoda prehrambene industrije i optimalnog sadržaja vlage testa. Pšenično brašno je u količini od 5, 10 i 15 % zamenjeno obezmašćenom pšeničnom klicom. Pored uticaja nivoa zamene pšeničnog brašna pšeničnom klicom, ispitan je i uticaj veličine čestica pšenične klice (<150 μm, 150-1000 μm i 800-2000 μm), kao i vlage testa pri zamesu (20, 22 i 24 %). U toku druge faze eksperimentalnog rada ove doktorske disertacije, pripremljeni su ekstrudati kukuruzne krupice sa dodatkom sporednih proizvoda prehrambene industrije i to: pšenične klice, pivskog tropa i tropa jabuke u udelima 15, 30 i 45 % (odnos kukuruzna krupica:sporedni proizvod bio je 85:15, 70:30 i 55:45). Dobijeni ekstrudati su samleveni i prosejani na sitima u cilju dobijanja tri frakcije sa različitim veličinama čestica (< 250 μm, 250 - 1000 μm i 1000 - 2000 μm). Izvršena je karakterizacija ekstrudata u smislu određivanja hemijskog sastava, raspodele veličine čestica, boje i mikrobiološke analize. Ovako dobijeni ekstrudati su zatim korišćeni u proizvodnji čajnog peciva kao zamena pšeničnog brašna u količinama od 5, 10 i 15 % računato na masu brašna. Za procenu uticaja veličine čestica ekstrudata, udela sporednog proizvoda u ekstrudatu i udela ekstrudata u čajnom pecivu na osobine testa i čajnog peciva takođe je korišćen Box-Behnken eksperimentalni dizajn. Rezultati su pokazali da je moguća proizvodnja funkcionalnog čajnog peciva sa dodatkom ekstrudata kukuruzne krupice obogaćene sporednim proizvodima prehrambene industrije, pri čemu se poboljšava nutritivni profil čajnog peciva, bez negativnih efekata na fizičke i senzorske karakteristike gotovog proizvoda.
The aim of this study is to examine the effect of replacing a portion of wheat flour with extruded snack products enriched with the addition of wheat germ, brewer's spent grain and apple pomace on the rheological and textural properties of dough and on characteristics of cookies (physical, sensory, nutritive, microbiological). During the preliminary analysis (the first phase of the experimental work), the influence of defatted wheat germ particle size, wheat germ content and dough moisture content on the quality of the dough and the cookies was investigated using the Box-Behnken experimental design. During the second phase of the experimental work, the extruded corn meal with the addition of by-products of the food industry (wheat germ, brewer's spent grain and apple pomace) were prepared (the ratio of corn meal: by-product was 85:15, 70:30 and 55:45). The obtained snack products are milled and sieved to obtain 3 fractions with different particle sizes (<250 μm, 250-1000 μm and 1000-2000 μm). Characterization of the extrudates in terms of determining the chemical composition, particle size distribution, color and microbiological analysis was performed. The obtained extrudates were then used in the production of cookies for the replacement of wheat flour in quantities of 5, 10 and 15%. The Box-Behnken experimental design was used to evaluate the influence of the extrudate particle size, the share of the by-product in the extrudate, and the share of extrudates in the cookies, on the dough properties and cookies characteristics.The results showed that the corn snack products enriched with food industry by-products (brewer's spent grain, wheat germ and apple pomace) can be used for production of functional cookies. These additives in an amount up to 15% improving the nutritional profile of the cookies, without adverse effects on the physical and sensory characteristics of the final product.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES
The majority of agroindustrial by-products are rich in low digestible fiber, and the bioconversion process for the production of microbial protein can be a practical and promising alternative to increase the protein content and nutritional value of substrate and food quality, turning these fibers into digestible components for the feeding of non-ruminant animals. Thus, this paper was aimed at protein enrichment of brewery spent grain by Rhizopus oligosporus CCT 4134 by solid state fermentation to be added in diets of juvenile Nile tilapia (Oreochromis niloticus). Solid state fermentation experiments were performed in order to determine the highest protein increment studying variables initial moisture (50, 60 and 70%) and supplemental nitrogen sources (ammonium sulfate, urea and sodium nitrate). To study the addition of fermented brewery spent grain in diets of Nile tilapia, 120 juveniles were used, divided into 24 boxes representing the six levels of addition (0, 2, 4, 6, 8 and 10%) of fermented brewery spent grain and four replications of each treatment. Fish were fed for 67 days between May and August 2015, the period where the water temperature was below the thermal comfort for the species. Solid state fermentation provided protein enrichment of brewery spent grain in about 2 and 4 times the content of crude and soluble protein, respectively, and is considered an alternative to use of industrial by-products to replace traditional ingredients in diets of juvenile Nile tilapia, because with the addition of fermented brewery spent grain, no significant difference in productive performance, hematological and biochemical parameters of juveniles Nile tilapia was found. Thus, the fermentation of brewery spent grain is a good alternative to be used as substrate for the cultivation of Rhizopus oligosporus and microbial protein production, allowing its use as a protein supplement in diets for juvenile Nile tilapia, with addition of up to 10 % without compromising growth performance and hematological parameters of the species.
A maioria dos subprodutos agroindustriais é rica em fibras com baixa digestibilidade, e o processo de bioconversão para produção de proteína microbiana, pode ser uma alternativa prática e promissora para aumentar o teor proteico, o valor nutritivo do substrato e a qualidade da alimentação, transformando estas fibras em componentes digestíveis para alimentação de animais não ruminantes. Diante disso, o presente trabalho teve como objetivo o enriquecimento proteico do bagaço de malte por Rhizopus oligosporus CCT 4134 via fermentação em estado sólido e adição em dietas de juvenis de tilápia do Nilo (Oreochromis niloticus). Na fermentação em estado sólido foram realizados experimentos visando determinar o maior aumento proteico, estudando as variáveis umidade inicial (50, 60 e 70%) e suplementação de fontes de nitrogênio (sulfato de amônio, ureia e nitrato de sódio). Para estudar a adição do bagaço de malte fermentado em dietas de tilápia do Nilo, foram utilizados 120 juvenis, distribuídos em 24 caixas que representa os seis níveis de adição (0, 2, 4, 6, 8 e 10%) do bagaço de malte fermentado e quatro repetições de cada tratamento. Os peixes foram alimentados por 67 dias compreendidos entre os meses de maio a agosto de 2015, período que a temperatura da água estava abaixo do conforto térmico da espécie. A fermentação em estado sólido proporcionou enriquecimento proteico do bagaço de malte em aproximadamente 2 e 4 vezes no conteúdo de proteína bruta e proteína solúvel, respectivamente, sendo considerada uma alternativa para aproveitamento dos subprodutos industriais na substituição de ingredientes tradicionais em dietas de tilápia do Nilo, pois com a adição do bagaço de malte fermentado não observou diferença significativa nos parâmetros de desempenho produtivo e parâmetros hematológicos e bioquímicos dos juvenis de tilápia do Nilo. Com isso, a fermentação do bagaço de malte é uma boa alternativa para seu aproveitamento como substrato para o cultivo de Rhizopus oligosporus e produção de proteína microbiana, permitindo sua utilização como suplemento proteico em dietas para juvenis de tilápia do Nilo, com adição de até 10%, sem comprometer o desempenho produtivo e parâmetros hematológicos da espécie.

碩士
國立中興大學
畜牧學系
85
The aim of this study was to evaluate the effects of brewers grains and bean curd pomance supplementation on the performance of lactating cows and the characteristics of their rumen (by-products accounting for 10％ of TMR, DM basis), so as to create a way to solve the problem of waste and pollution from these by-products. Thirty-two Holstein lactating cows were alloted radomly into four groups for treatment. The treatments were control; brewers grains group; bean curd pomance group and mixed by-products group(containing the same amount of brewers grains and bean curd pomance), respectively. The mixed rations of treatments were formulated to be iso- energetic and iso-nitrogenous according to the nutrient commendation of NRC(1989); and containing 35％ corn silage、 20％ alfalfa hay and 45％ concentrate(DM basis). The experimental period lasted for 8 weeks after a week of acclaimation period. In addition, four dry Holstein cows fitted with rumen cannula were used in a 4×4 Latin square with 10-d periods for collecting rumen liquid samples for analysis. Results showed that cows fed the bean curd pomance diet produced significantly more milk than the others(P<0.05), and cows fed the brewers grains and mixed by-products diet produced significantly more milk than the control group(P<0.05). Both the control and brewers grains groups took in more feed than those in the bean curd pomance and mixed by-products groups(P<0.05). The cows fed the mixed by-products diet produced a significantly lower percentage of milk fat, total solids, milk protein and milk lactose than the others(P<0.05). The milk lactose percentage was significantly more high for cows fed the bean curd pomance diet than those fed the brewers grains diet(P<0.05). Regarding the amount of milk fat, total solids, milk protein and milk lactose produced by cows; Cows fed a bean curd pomance diet produced significantly more than the others(P<0.05). Cows fed the mixed by-products diet produced significantly lower amounts of milk fat, total solids and milk protein than the control and brewers grains groups(P<0.05). This diet did not significantly influence the body weight of the cows or the ruminal pH values(P>0.05).

碩士
國立臺灣大學
生物產業機電工程學研究所
99
Spent grain is the main waste in brewery industry, contributing around 85% of total byproducts. It is mainly used as the animal feed additives. However, other applications should be concerned about since it contains abundant organics. Hydrogen has being considered as an ideal alternative energy for its high heating value and it produce only clean water after combustion, and it could be produced through biodegradation of the organic material. The objective of this study is to find out the optimum operational condition for using the spent grain as substrate. The composition of the spent grain used in this study is: 5.35% cellulose, 34.83% hemi-cellulose, 9.01% lignin and 2.63% ash. The seeding bacteria were obtained from the anaerobic sludge in a dairy farm. Heating pretreatment of 15 minutes for the sludge was conducted to kill those hydrogen utilizing bacteria. Thermophilic fermentation tests were also conducted to tests its effect in inhibiting the methanogen. Dilute acid was added to the spent grain for pretreatment and was heated to boiling for 30 minutes. The pretreated spent grain was then mixed with the centrifuged cold sludge homogeneously and fed into the reactor. Fermentation tests were designed and conducted under conditions of different temperature (37℃, 45℃, 50℃, 55℃ and 60℃) and different pH (5, 6, 6.5 and 7). Experimental results showed that 2% HCl pretreatment had higher hydrogen production than 0.365% HCl. The maximal hydrogen yield of 46 mL H2/g TVS and the hydrogen content of 73.17% were achieved under the optimal operational condition: pH 6 and 37℃. Addition of nutrient broth could shorten the hydrogen-producing time within 24 hrs and effectively decrease the methane production.

Brewers spent grain (BSG) is an abundant waste material from the brewing process, representing approximately 85% of total by-products generated, is rich in cellulose and non-cellulosic polysaccharides and has a strong potential to be recycled. One of the main components of the BSG is protein, which has a potential to be extracted, modified and used as an adhesive for the woodworking adhesive. This work deals with the protein extraction through alkali treatment and its subsequent modification, crosslinking with glyoxal. The crosslinked adhesive was tested through ABES method to find, that ideal content of glyoxal is 20%. Subsequent adjusted standard tests for the lap shear strength and bend strength (MOE and MOR values) were used to compare its values to commercially available PVAc adhesive. The lap shear strength results were 2,7 times lower to PVAc, the bend strength values were, on the other hand, 1,5 times higher. The PVAc was partially replaced (50 and 25%) by the protein adhesive showing that the extracted adhesive could serve as a partial replacement to lower the ecological impact of the PVAc adhesive.

Polyhydroxyalkanoates (PHA) are fully biodegradable polyesters, synthesized by microorganisms from renewable resources, which reveal an alternative to petroleum-based plastics. Its production by mixed microbial cultures, unlike pure cultures, allows the use of industrial by-products in the absence of sterility conditions, potentially reducing process costs. The study described focused on the valorization of a waste from a brewery industry, brewer’s spent grain (BSG), through organic acids production, towards PHA production. PHA production using mixed microbial cultures involves three steps: (1) acidogenic fermentation of BSG for organic acids production, (2) selection of the mixed cultures able to accumulate PHA and (3) PHA production. In order to extract the sugars necessary to produce organic acids (phase 1), pre-treatment methods such as acid hydrolysis and subcritical water have been studied. Acid hydrolysis was selected for further experiments as the hydrolysis yield on sugars reached was higher than using subcritical water (0.152 g total sugars g BSG-1 versus 0.008 and 0.020 g total sugars g BSG-1 at 200 °C and 230 °C, respectively). Afterwards, the pH (5.0, 5.5 and 6.0) and organic loads (10, 20 and 30 gCOD L-1) effect on the acidogenic fermentation of hydrolyzed BSG was studied in batch. It was verified, regardless of the condition, that the most produced acids are acetic and butyric. As pH and organic load affects the fermentation products profile, it is possible to manipulate PHA composition. At last, the acidogenic fermentation of hydrolyzed BSG was studied in a continuous mode, using an expanded granular sludge bed reactor (EGSB). Due to the reduced operating time, it was not possible to reach stability. However, it was observed that sugars were consumed to produce organic acids. This revealed the potential of EGSB to be used for BSG acidogenic fermentation towards PHA production.

碩士
國立中興大學
食品暨應用生物科技學系所
102
Brewery spent grains(BSG) is generated during the production of beer and is considered to be a maximum industrial by-product. In 2010, 1.69 billion hectolitre of beer from barley was produced worldwide (FAOSTAT, 2010). Approximately 15–20 kilogram of BSG is produced per every hectolitre of beer, resulting in annual production of 25.3-33.8 million tonnes of BSG worldwide BSG is rich dietary fiber, that has promote gastrointestinal peristalsis, prevent cardiovascular disease, lower blood cholesterol and stabilize blood sugar. In the diet, dietary fiber can increase satiety and reduce calories intake to achieve weight control. Collect commercial granola bar on the market and evaluate demands, nutrients, each serving and other relevant information as a reference. Investigate the effect of different preparation process for using drying BSG and oatmeal as a raw material to make high-fiber granola bar, and analysis that chemical composition and physical properties. The experiment conditions include the particle size and added percentage of BSG, baking time, baking temperature, calorie calculation and increase the water-soluble dietary fiber, shape and formability of the finished product, mouthful and flavor enhance, those to get the most suitable conditions for making of high-fiber granola bar.After drying BSG of total dietary fiber (TDF) content was 65.82 ± 2.72%. Added 25% BSG for high-fiber granola bar, baking temperatures 160/140 ℃(up/under), 30 minutes were more suitable conditions for color, texture and dietary fiber content. Finally,half butter and half Fibersol-2 formulation that panelists who like it.Each per serving of granola bar is 40 g, 175Kcal and 12.62g content of dietary fiber. Per serving of dietary fiber content can reach recommended daily intake of 1/2.SDF source of the granola bar are oatmeal、Fibersol-2 and Isomatooligosaccharides.SDF content of the granola bar from the original 1.08 g / serving increased to 3.03 g / serving. Not only energy reduced 27% and SDF content increased 2.8 fold.Milling BSG fine also affects the body base structure and mouthful of the granola bar.Added Fibersol-2 that decreased calories and shape of bar which cutting surface had making better, in addition to ,water-soluble dietary fiber (SDF) content was increased. Replaced milk, white egg and sugar powder with glycerol and sorbitol which really upgrade the mouthful and wettability of the granola bar.

Tese doutoramento em Chemical and Biological Engineering
The current project aimed at solving technical problems involved on the implementation of a bioreactor (airlift) for continuous primary beer fermentation. In order to operate in continuous mode, the reactor must be capable of retaining a large amount of immobilized biomass. The first strategy taken towards this achievement was to effectively obtain a yeast carrier from Brewer´s Spent Grains (BSG) by suggesting several different chemical treatments. The ideal yeast carrier was obtained from a 40 min caustic treatment and had about 56% of cellulose content. This material was then used for yeast immobilization in two continuous beer fermentations. In these fermentations, foam fractionation was suggested as a method for continuous skimming non-viable biomass out from the bioreactor. The suggested setup presented good performance (rs = 2.95-3.62 g.L-1.h-1; and rp = 1.33-1.73 g.L-1.h-1) on continuous beer fermentation and a viability difference between yeast subpopulations from foam and remaining yeast in the reactor has been observed suggesting the feasibility of foam fractionation for dead biomass skimming. Yet, it has also been observed that flocculated biomass surrounding the biocatalysts were contributing in a large extent for the total immobilized biomass in the reactor. Thus, an assay testing the feasibility of flocculation as sole way of biomass immobilization in an airlift reactor was carried out. This new and simpler setup has shown much better performance than carriers’ based setups for biomass loading capacity (25 gcell dry wt..L-1), presenting very similar fermentation rates (rs = 3.4 g.L-1.h-1; and rp = 1.54 g.L-1.h-1). Then, two other continuous primary beer fermentations were carried out in order to test the feasibility and quality of beer produced using similar flocculation-based setups. In the first and longer fermentation up to 52 gcell dry wt..L-1 were attained through flocculation. Fermentation rates were higher than any other setup suggested by the literature for continuous beer fermentation (rs = 8.7 g.L-1.h-1; and rp = 3.7 g.L-1.h-1). This experiment was also very relevant as it was possible to correlate many other variables (such as FAN consumption rate influencing higher alcohols and diacetyl formation) contributing for the success of the suggested system. The last experiment was carried out at the laboratories of a large commercial brewery where some important parameters could be better controlled (fast feedback of results by automated beer analyzer, fresh wort always available, etc.). The results were consistent with the ones observed in the last mentioned experiment, replicating its outstanding performance (rs = 9.43 g.L-1.h-1; and rp = 3.75 g.L-1.h-1). Moreover, the beer from flocculated based setup was of excellent quality, confirmed by a professional panel of beer specialists.
Este projeto teve como principal objetivo resolver problemas técnicos associados à implementação de um bioreactor (do tipo airlift) para fermentação primária e contínua de cerveja. Para operar em contínuo, o reator deve ser capaz de reter uma grande quantidade de biomassa imobilizada. A primeira estratégia tomada foi obter suportes para imobilização da levedura a partir de drêche cervejeira através de tratamentos químicos. O suporte ideal foi obtido por um tratamento cáustico de 40 min e tinha aproximadamente 56% de pureza em celulose. Este material foi utilizado na imobilização de leveduras em duas fermentações contínuas. Nestas fermentações, o fracionamento pela espuma foi sugerido como método para escumar a biomassa morta do reactor. A configuração sugerida apresentou bom desempenho (rs = 2,95-3,62 g.L- 1.h-1; and rp = 1,33-1,73 g.L-1.h-1) ao mesmo tempo que foi possível observar uma diferença de viabilidade entre as sub-populações de levedura presentes na espuma e que permaneciam no reactor. Ainda foi possível observar que a biomassa floculada em redor dos biocatalizadores estavam a contribuir em grande parte para biomassa total imobilizada no airlift. Assim, foi realizado um experimento para testar a viabilidade de utilizar a floculação como única forma de imobilização de leveduras no airlift. Esta nova configuração demonstrou não só um desempenho muito superior ao anterior em relação à capacidade de imobilização celular (25 gpeso seco de células..L-1), mas também apresentou uma capacidade fermentativa muito semelhante (rs = 3,4 g.L-1.h-1; and rp = 1,54 g.L-1.h-1). A seguir, duas outras fermentações foram realizadas no intúito de validar a nova configuração e avaliar a qualidade da cerveja produzida. Na primeira e mais longa fermentação, um total de até 52 gpeso seco de células..L-1 foi obtido por floculação e as taxas de fermentação foram superiores todas configurações apresentadas na literatura envolvendo a fermentação contínua de cerveja (rs = 8,7 g.L-1.h-1; and rp = 3,7 g.L-1.h-1). Ainda foi possível correlacionar muitas variáveis (como o consumo de FAN influenciando na formação de alcoois superiores e diacetil) contribuindo para o sucesso desta nova configuração. O último experimento foi realizado nos laboratórios de uma grande indústria cervejeira, aonde foi possível controlar de forma mais rigorosa alguns parâmetros (rápido retorno de resultados através de equipamentos automatizados, mosto fresco sempre disponível, etc.). Os resultados do sistema foram consistentes com o do experimento anterior, constatando-se novamente seu excelente desempenho (rs = 9,43 g.L-1.h-1; and rp = 3,75 g.L-1.h-1). Ainda, a cerveja produzida pelo sistema baseado em floculação foi de excelente qualidade, comprovada por um painel profissional de especialistas cervejeiros.
Fundação para a Ciência e Tecnologia (FCT) - PhD scholarship SFRH / BD / 61777 / 2009.
This thesis was financially supported by a PhD scholarship from Fundação para a Ciência e Tecnologia (Ref. number: SFRH/BD/61777/2009), inserted in the Programa Potencial Humano Quadro de Referência Estratégico Nacional (POPH – QREN) - Tipologia 4.1 - Formação Avançada. The POPH-QREN is co-financed by Fundo Social Europeu (FSE) and by Ministério da Ciência, Tecnologia e Ensino Superior (MCTES). It was also supported by UNICER Bebidas de Portugal, S.A. (S. Mamede de Infesta, Portugal).

碩士
國立屏東科技大學
食品科學系所
101
Brewer’s spent grain (BSG) is the major by-product of the brewing industry. In recent years, many studies have pointed out that the components in BSG such as proteins and fiber are inexpensive and yield high which are suitable for food applications, and can be used to develop various products. So, in this study, different levels (0, 10, 20, 30, 40%) of BSG were added in the extrusion process, and adjusted the proportion of vegetable protein (soybean protein isolate, wheat protein) to ensure that the feed protein content reached 55%, for the development of texturized vegetable protein (TVP) and were further used to investigate the influences on its texturization and physicochemical properties. Results of the study showed that the TVP with less than 20% BSG addition could successful texturization, and the L value, moisture retention, water absorption index and cooking loss of TVP (soybean protein isolate group) would be decreased as the addition level of BSG increased, whereas the cutting force had no significant difference (p > 0.05); The L value, b value, cooking loss and cutting force of TVP (wheat protein group) would be decreased as the addition level of BSG increased. Furthermore, both insoluble dietary fiber and total dietary fiber contents of TVP would be increased as the addition level of BSG increased, TVP (soybean protein isolate group) with 40% BSG had the highest total dietary fiber 36.83% (dry basis), whereas TVP (wheat protein group) with 0% BSG had the lowest total dietary fiber 25.06% (dry basis), the results showed a significant difference (p &;lt; 0.05). The content of soluble dietary fiber would be increased after extrusion, the amount of soluble dietary fiber increased was basically equal to the amount of insoluble dietary fiber decreased. On the whole, adding BSG in the TVP production is possible, and TVP (soybean protein isolate group) with 10% BSG is the best group in this study, not only the total dietary fiber content of the TVP has been increased, but also the similar texturization and physicochemical properties (moisture retention, water absorption index, cooking loss and cutting force) were obtained comparing to those of the control group.

Mestrado em Engenharia Zootécnica - Produção Animal - Instituto Superior de Agronomia / Faculdade de Medicina Veterinária
To compare the effect of nature of fibre in total tract apparent digestibility, in productive parameters and development of the rabbit’s digestive device, four experimental diets with 50% of the neutral detergent fibre (NDF) from different fibre sources were set up: alfafa (AL), not washed brewer’s spent grain (DNL), washed brewer’s spent grain (DL) and beet pulp (BEET). The diets were distributed by 144 rabbits at 21 days old, 3 x 12 cages per diet. From 28 to 70 days old, 1 x 12 cages per diet remained. The source of fibre affected intake, growth and feed conversion ratios. Intake was 117, 96, 95 and 83 g day-1, in AL, DNL, DL and BEET, respectively. Growth was 44, 40, 41 and 35 g day-1, in AL, DNL, DL and BEET, respectively. Finally, feed conversion ratios were 2.70, 2.42, 2.30 and 2.40, in AL, DNL, DL and BEET, respectively. Therefore, there was an inferior value in the final weight of BEET fed rabbits. The source of fibre significantly affected the coefficients of total tract apparent digestibility (CTTAD), calculated at 8 weeks old (0.642, 0.718, 0.711 and 0.723 of DM and 0.648, 0.724, 0.717 and 0.728 of OM, in AL, DNL, DL and BEET, respectively), not affecting CP digestibility. Cellular wall components were more digestible in BEET diet. The nature of fibre significantly affected the weight of carcass, full digestive device, full and empty caeca and pH values of digestive and caecal contents. The liver in alfafa and brewer’s spent grain diets were 30% heavier than in BEET. The intestinal morphology was not affected by the nature of fibre in the last 6 weeks of assay. ----RESUMO---- Com o objectivo de comparar o efeito da natureza da fibra na digestibilidade dos componentes da dieta, nos parâmetros produtivos e no desenvolvimento do aparelho digestivo do coelho, foram elaboradas quatro dietas experimentais com 50% do NDF de fontes de fibra diferentes: luzerna (AL), dreches não lavados (DNL), dreches lavados (DL) e polpa de beterraba (BEET). As dietas foram distribuídas por 144 coelhos aos 21 dias, 3 x 12 gaiolas por regime. Dos 28 aos 70 dias manteve-se 1 x 12 gaiolas por regime. A fonte de fibra afectou o consumo de alimento (117, 96, 95 e 83 g dia-1, para AL, DNL, DL e BEET, respectivamente), o aumento de peso diário (44, 40, 41, 35 g dia-1, para AL, DNL, DL e BEET, respectivamente) e o índice de conversão (2.70, 2.42, 2.30 e 2.40, para AL, DNL, DL e BEET, respectivamente), verificando-se um valor inferior no peso final dos coelhos do regime BEET (1937 g). A fonte de fibra afectou significativamente os coeficientes de utilização digestiva (CUD) calculados às 8 semanas (0.642, 0.718, 0.711 e 0.723 da MS e 0.648, 0.724, 0.717 e 0.728 da MO, para AL, DNL, DL e BEET, respectivamente), não afectando a digestibilidade da PB. Os constituintes da parede celular foram mais digestíveis no regime BEET. A natureza da fibra afectou significativamente o peso da carcaça, do aparelho digestivo cheio e do ceco cheio e vazio. O fígado nos regimes de luzerna e dreches apresentou-se 30% mais pesado do que em BEET. A morfologia intestinal não foi afectada pela natureza da fibra nas últimas 6 semanas de ensaio.

Tese de Doutoramento em Bioengenharia
The demand of prebiotic ingredients, such as xylooligosaccharides (XOS), has been growing over the years as consumers pay more attention to their health and well-being. XOS are considered emergent and competitively priced prebiotics, presenting high potential as food ingredients. As a result, the industry is focused on developing new approaches to improve production efficiency that can meet the increasing demand while reducing costs. Motivated by the industry needs, the main purpose of this thesis was to develop an integrated bioprocess, based on one-step fermentation, for the production of prebiotic XOS, towards the simplification and cost reduction of the process. The one-step fermentation of 13 agro-residues was performed using two Trichoderma species. The most promising results were found for T. reesei using brewers’ spent grain (BSG) as substrate. BSG is an inexpensive and abundant brewery agro-industrial residue that was herein proven interesting for the production of prebiotic arabino-xylooligosaccharides (AXOS). Under optimal conditions (3 days, pH 7.0, 30°C and 20 g/L of BSG), a production yield of 38.3 ± 1.8 mg/g (xylose equivalents/g of BSG) was achieved. AXOS with degree of polymerization (DP) from 2 to 5 were obtained. The onestep fermentation using T. reesei was compared with the application of a commercial xylanase from T. longibrachiatum. The first revealed to be a more attractive and low-cost strategy to hydrolyze sterilized raw BSG (without conventional pretreatment) and produce AXOS. In order to reduce the production time obtained with T. reesei (3 days), the Bacillus subtilis 3610 wild type (wt) was successfully used, for the first time, to produce AXOS through direct fermentation of BSG, reducing the production time to 12 h. Genetic engineering was used to further optimize the microorganism performance, by cloning the T. reesei xylanase gene coupled with a secretion tag into the B. subtilis chromosome (B. subtilis 3610 clone 2). After optimization by Box-Behnken experimental design, AXOS with a DP ranging from 2 to 6 were obtained at 12 h of fermentation with B. subtilis 3610 clone 2, using 20 g/L of BSG, at pH 7.0 and 45°C. The maximum production yield was 54.2 ± 1.1 mg/g (xylose equivalents/g of BSG), representing an increase of 33% comparing to the wt, and 29% comparing to the T. reesei. B. subtilis 3610 clone 2 was also selected for downscale production of XOS by direct fermentation of commercial beechwood xylan. The use of a commercial substrate allowed to better evaluate strategies to overcome common process limitations associated to the use of agro-residues as substrates, namely biomass measurement, mass transfer issues and substrate inhibition. The maximum production yield, 306 ± 4 mg/g (XOS/xylan), was achieved after 8 h of fermentation operating under one-time impulse fed-batch regimen. The optimal conditions found were pH 6.0 and 42.5°C, using 2.5 g/L of initial concentration of xylan increased up to 5.0 g/L at 3 h. A mixture of XOS with a DP ranging from 4 to 6 was obtained, presenting high stability after a static in vitro digestion (98.5 ± 0.2)%. Operating under fed-batch mode allowed to minimize the effect of substrate inhibition, which led to a great increase of the production yield. In vitro studies using human fecal inocula of two healthy donors were performed to evaluate and compare the potential prebiotic effect of commercial lactulose and the XOS herein produced through direct fermentation of beechwood xylan by B. subtilis clone 2. For both substrates, acetate was the main short-chain fatty acid (SCFA) produced during fermentation at 48 h. Lactulose led to the highest production of lactate, while XOS led to the maximum production of butyrate (9.0 ± 0.6 mM for donor 1 samples, and 10.5 ± 0.8 mM for donor 2 samples). The addition of XOS also resulted in the largest production of CO2. The significant increase in the production of SCFAs and CO2, added to the reduction of pH and ammonia concentration suggest that the XOS herein obtained hold potential functional properties for human health. The results gathered in this thesis are very promising, specially from the industrial perspective, providing important insights for the development of new integrated strategies for XOS production from agro-residues.
A procura de ingredientes prebióticos, como os xiloligossacarídeos (XOS), tem crescido ao longo dos anos, à medida que os consumidores prestam mais atenção à sua saúde e bem-estar. Os XOS são considerados prebióticos emergentes e com preços competitivos, apresentando alto potencial como ingredientes alimentares. Consequentemente, a indústria está focada no desenvolvimento de novas abordagens para melhorar a eficiência da produção, para reduzir custos e atender à crescente procura. Motivada pelas necessidades da indústria, o objetivo principal desta tese foi desenvolver um bioprocesso integrado, baseado numa única fermentação para a produção de XOS, visando a simplificação e redução de custos do processo. A fermentação direta de 13 agro-resíduos foi realizada utilizando duas espécies Trichoderma. Os resultados mais promissores foram observados para o T. reesei, usando dreche (BSG) como substrato. A dreche é um resíduo agro-industrial barato e abundante, proveniente da indústria cervejeira que provou ser interessante para a produção de arabino-xilooligossacarídeos (AXOS). Sob condições ótimas (3 dias, pH 7,0, 30°C e 20 g/L de BSG), obteve-se um rendimento de produção de 38,3 ± 1,8 mg/g (equivalentes de xilose/g de BSG). Os AXOS obtidos apresentaram um grau de polimerização (DP) entre 2 e 5. A produção de AXOS através de fermentação com T. reesei foi comparada com a aplicação de uma xilanase comercial de T. longibrachiatum. A fermentação revelou ser uma estratégia mais atrativa e de baixo custo para hidrolisar a dreche esterilizada por autoclave (sem pré-tratamento convencional) e para produzir AXOS. De modo a reduzir o tempo de produção obtido com o T. reesei (3 dias), estudou-se, pela primeira vez, a produção AXOS através de fermentação direta da dreche usando o Bacillus subtilis 3610 wild type (wt), o que levou à redução do tempo de produção para 12 h. Recorrendo a engenharia genética, o desempenho do microrganismo foi otimizado, através da integração no seu cromossoma do gene da xilanase do T. reesei acoplado a um tag de secreção (B. subtilis 3610 clone 2). Após otimização do processo com recurso a um desenho experimental de Box-Behnken, foram produzidos AXOS com DP de 2 a 6 após 12 h de fermentação com o B. subtilis 3610 clone 2, utilizando 20 g/L de BSG, pH 7,0 e 45°C. O rendimento máximo de produção foi de 54,2 ± 1,1 mg/g (equivalentes de xilose/g de BSG), representando um aumento de 33% em relação à wt e 29% em relação ao T. reesei. O B. subtilis 3610 clone 2 foi também selecionado para a produção em menor escala de XOS através da fermentação direta de xilano comercial obtido de madeira de faia. O uso de um substrato comercial permitiu uma melhor avaliação das estratégias necessárias para superar as limitações que são geralmente associadas ao uso de agro-resíduos, nomeadamente, a quantificação da biomassa, os problemas de transferência de massa e a inibição pelo substrato. O rendimento máximo de produção, 306 ± 4 mg/g (XOS/xilano), foi observado após 8 h de fermentação, operando sob regime de fed-batch com um impulso. As condições ótimas encontradas foram pH 6,0 e 42,5°C, utilizando 2,5 g/L de concentração inicial de xilano aumentada para 5,0 g/L às 3 h. Nestas condições obteve-se uma mistura de XOS com um DP entre 4 e 6, que apresentou elevada estabilidade (98,5 ± 0,2%) após ter sido sujeita a um processo de digestão estática in vitro. A produção em modo fed-batch permitiu ainda minimizar o efeito da inibição do substrato, o que levou a um aumento significativo do rendimento de produção. Foram realizados estudos in vitro com inóculos fecais humanos de dois dadores saudáveis para avaliar e comparar o potencial efeito prebiótico da lactulose comercial e da mistura de XOS produzida por fermentação direta de xilano de madeira de faia usando o B. subtilis clone 2. Para ambos os substratos, o acetato foi o principal ácido gordo de cadeia curta (AGCC) produzido após 48 h de fermentação. A lactulose apresentou a produção mais elevada de lactato, enquanto que os XOS apresentaram a produção mais elevada de butirato (9,0 ± 0,6 mM para o doador 1 e 10,5 ± 0,8 mM para o doador 2). A adição de XOS ao meio de cultura também resultou numa produção mais elevada de CO2. O aumento significativo na produção de AGCCs e CO2, bem como a redução do pH e da concentração de amónia, sugere que os XOS produzidos neste trabalho podem apresentar propriedades funcionais importantes para a saúde humana. Os resultados obtidos nesta tese são bastante promissores, especialmente do ponto de vista industrial, apresentando-se informação relevante para o desenvolvimento de novas estratégias integradas para a produção de XOS a partir de agro-resíduos.

Mestrado em Engenharia Zootécnica - Produção Animal - Instituto Superior de Agronomia
In order to study the effect of brewer’s spent grain in digestibility, consistency of faeces, intestinal fermentations and morphology of the intestinal wall, 32 piglets, weaned at 28 days of age were distributed in 4 blocks of 8 animals and 4 experimental diets were provided: a control diet and three other diets enriched with different fiber sources (wheat bran, unwashed brewer spent grain and washed brewer spent grain). To study the effect of brewer spent grain, the control diet was used as the negative control and the wheat bran as the positive control. The values of fecal apparent digestibility of dry matter, protein and mineral fraction with brewer spent grain were higher (P <0.05) then those obtained with wheat bran. The consistency of faeces and intestinal morphology showed better results (P <0.05) with the adding of brewer’s grain, when compared to the control and the bran. We found unexpectedly low values for intestinal fermentations. In conclusion, the fiber of brewer’s spent grain has some beneficial effects, mainly when compared to wheat bran, but there were no clear indications of the existence of a prebiotic effect

Com o aumento da popularidade da cerveja artesanal e o subsequente aumento do número de cervejeiras artesanais e brewpubs em ambientes urbanos, a necessidade de encontrar soluções para a redução de resíduos e a recuperação de valor dos subprodutos é cada vez uma questão mais importante. Caraterísticas chave destes pequenos produtores incluem a sua tendência para utilizar as melhores (e mais caras) matérias-primas disponíveis, ter uma oferta diversificada e estar localizado próximo do cliente final. Isto leva à geração de quantidades elevados de subprodutos (>20 kg por 100 litros de cerveja produzida) com potencial de valorização e opções limitadas para a sua gestão. Assim, as atuais tendências relativamente a preocupações ecológicas, de sustentabilidade e de saúde, podem colocar as cervejeiras artesanais numa posição desfavorável. Este projeto foi realizado com uma cervejeira artesanal e consistiu no acompanhamento do processo de produção de cerveja artesanal, na identificação de resíduos/subprodutos e no desenvolvimento de propostas para a valorização dos mesmos. Representando 85% dos resíduos da indústria cervejeira, a dreche (resíduo do malte) tornou-se o foco deste projeto. Como solução para a valorização deste subproduto/resíduo, pretendeu-se criar um snack para consumir com cerveja, incorporando o máximo de dreche possível, para ser consumido no local de onde provém. O snack desenvolvido incorpora dreche húmida de forma a diminuir os custos de processamento e para que o seu processo de produção seja suficientemente simples para poder ser replicado em qualquer cozinha comercial. O snack criado resulta da mistura de dreche húmida, farinha de trigo e especiarias, tendo um custo potencialmente inferior a 0,55 euros/kg e um teor de dreche acima de 35%. Embora estes snacks possam não ser uma solução para a totalidade da dreche produzida numa cervejeira, podem contribuir significativamente para a redução de desperdícios. Podem também servir como fator de diferenciação de mercado das cervejeiras e brewpubs, ao mesmo tempo que sensibilizam para questões associadas aos resíduos na indústria alimentar e de bebidas.
With the increase in popularity of craft-beer and subsequent increase in the number of small breweries and brewpubs in urban settings, the need for solutions for waste reduction and value recovery from by-products is an issue of growing importance. Key characteristics of these small producers include their tendency to use the best (and more expensive) raw materials available, to have a diverse offer and to be located close to the final customer. This leaves them with large quantities (>20 kg per 100 litres of beer produced) of potentially valuable by-products and limited options to manage them. Thus the current consumer trends regarding ecological concerns, sustainability and healthy lifestyles, might place such small, urban breweries and brewpubs in an unfavourable position. This project counted on the collaboration of a craft beer producer and consisted of a study of the brewing process, the identification and analysis of the brewery’s by products and the development of proposals for their valorisation. Representing 85% of a brewery’s waste, brewers spent grain (BSG) became the focus of this project. As a proposed solution for valorisation of BSG, the intention of this project was to create a snack for pairing with beers that contains the highest level of spent grain possible, intended to be enjoyed in the brewery where they were created. The snack was created using moist spent grain to maintain the processing costs low and maintaining the process simple enough to be easily replicated in any commercial kitchen. The snack is composed of spent grain, flour and herbs and spices, resulting in a potential production cost bellow 0.55 €/kg and a total content of BSG above 35%. Whilst these snacks will likely not be a solution for the totality of BSG produced in a brewery, they may contribute greatly to the reduction of waste. They may also serve as a factor for market differentiation in breweries and brewpubs while raising awareness for waste issues and their potential solutions in the food and beverage industry.