Thesis--Helsingfors. / "Literaturverzeichnis": p. [71]-74.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-06-21T15:01:16Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertação versão final.pdf: 2119371 bytes, checksum: 450a30d2e9d1d1f7c89323a179ac7bea (MD5) / Made available in DSpace on 2016-06-21T15:01:16Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Dissertação versão final.pdf: 2119371 bytes, checksum: 450a30d2e9d1d1f7c89323a179ac7bea (MD5) Previous issue date: 2015-12-09 / CAPEs / A aplicação de processos biotecnológicos para a produção de aromas alimentares tem apresentado um crescimento bastante significativo, já que esses aromas são classificados como aromas naturais. Acetoína (3-hidroxi 2-butanona ou metil carbinol acetil) é uma sustância volátil bastante utilizada em produtos alimentícios, com a finalidade de intensificar o sabor e o aroma dos alimentos, deixando-os mais atrativos aos consumidores. Este trabalho tem como objetivo produzir o bioaroma acetoína por via bioquímica a partir de hidrolisado de melaço utilizando as bactérias do gênero Bacillus isoladas do sedimento de mangue da Lagoa do Araçá (Recife-PE). Foi possível identificar nove espécies desse gênero (B.alvei, B. badius, B. circulans “A”, B. circulans “B”, B. coagulans, B. larvae, B. licheniformis, B. polymyxa, B. popilliae e B.subtilis.) sendo cinco espécies (B. subtilis, B. licheniformis, B. polymyxa, B. alvei e B. coagulans)produtoras do bioaroma acetoína. Foi realizado um planejamento fatorial 23com as variáveis independentes: agitação (0 rpm a 200 rpm), pH (6,5 a 8,5), concentração de melaço (60 g·L-1 a 100 g·L-1) e como variáveis dependentes a conversão de substrato em produto por três bactérias (Bacillus licheniformis, Bacillus alvei e Bacillus polymyxa).Com esse estudo foi possível observar que a agitação foi a principal variável que afetou os três processos de conversão de substrato em acetoína e que a interação do pH com agitação mostrou-se bastante significativo favorecendo essa conversão com os três micro-organimos. Os Bacillus licheniformis foi a bactéria queapresentou maior percentual de YP/S (52,88%) com pH 6,5, agitação de 200 rpm e concentração de melaço de 100 g·L-1. A partir deste resultado foi realizado um estudo cinético em reator batelada de 3L, do tipo tanque agitado, com sistema de aeração. O processo foi acompanhado por determinação de açúcares redutores totais (ART), biomassa (Peso seco), produto (Cromatografia Gasosa). A síntese bioquímica foi interrompida em 144 horas com consumo de 54% dos açúcares. No final do processo a concentração de açúcar residual estabilizou-se em 23,44 g·L-1 e a produção máxima de acetoína foi de 16,11 g·L-1. A produtividade de acetoína foi Pv = 0,11 g·L-1·h. A velocidade específica máxima de crescimento do B. licheniformis foi μmáx = 0,025 h-1. Os coeficientes de conversão de substrato em células e substrato em produto foram respectivamente: YX/S = 0,1 g/g e YP/S = 0,58 g/g. Com esse estudo pode-se concluir que a produção de bioaromas, utilizando hidrolisado de melaço por Bacillus pode ser uma alternativa de produção para gerar produtos com baixo custo e alto valor agregado. / The application of biotechnological processes for producing flavoring aromas has shown a significant growth, since these are classified as natural flavors. Acetoin (3-hidroxi 2- butanone or acetyl methyl carbinol) is a volatile substance widely used in food products, in order to intensify the taste and flavor of food, making it more atractive for consumers. This work aims to biochemically produce acetoin aroma from hydrolyzate of molasses,using the bacterias of bacillus genus isolated of the sediment mangrove of the Araçá lagoon (Recife-PE). It was possible to identify nine species of this genus (B.alvei, B. badius, B. circulans “A”, B. circulans “B”, B. coagulans, B. larvae, B. licheniformis, B. polymyxa, B. popilliaee B.subtilis.), but only five among these (B. subtilis, B. licheniformis, B. polymyxa, B. alvei e B. coagulans) are able to produce the acetoin bioaroma. It’s made a factorial planning 23with the independent variables: agitation (0 rpm to 200 rpm), pH (6.5 to 8.5), molasses concentration (60 g·L-1 a 100 g·L-1) and as independent variables the conversion of substrate in product for three bacteria(Bacillus licheniformis, Bacillus alvei e Bacillus polymyxa).With this study, it was possible to notice that the agitation was the main variable to affect the three processes of conversion of substrate into acetoin, and also that the pH interaction along with the agitation has been proven very significant, contributing to this conversion with the three microorganisms. B.licheniformis was the bacteria that showed the highest percentage of YP/S (52.88%) with pH 6.5, agitation of 200 rpm and molasses concentration of 100 g·L-1. Using these results, a kinetic study was performe batch reactor a three-liter, busy type tank with aeration system.The process was accompanied by determination of total reducing sugars (ART), biomass (dry weight), product (gaseous chromatography). The biochemical synthesis was interrupte dater 144 hours, with consumption of 54% of sugars. At the end of the process, the concentration of residual sugar stabilized in 23.44 g·L-1and the maximum production of acetoin was 16.11 g·L-1. The productivity of acetoin was Pv = 0.11 g·L-1·h. The maximum specific speed of increase of B. licheniformis was μmax = 0.025 h-1. The substrate conversion coefficients in cells and substrate in product respectively were: YX/S = 0.1 g/g and YP/S = 0.58 g/g. With this study, it can be understood that the bio-aroma production using hydrolyzate of molasses by Bacillus licheniformis can be an alternative way to generate products with low cost and high aggregate value.

Thesis (PhD (Chemical Engineering))--Cape Peninsula University of Technology, 2020 / The use of bioproducts in different commercial sectors such as medicine, agriculture, cosmetics, food, and chemical industries motivates the need for easy production and recovery techniques of bioproducts at laboratory and pilot scale. This study aims at the production and recovery of some selected bioproducts using supercritical fluid extraction techniques. Three products are chosen as case studies: these are ethanol, acetoin, and vanillin, since the ease of separation is influenced by the concentration of the product in the broth, these compounds were selected based on their concentration in the fermentation broth, according to literature sources. A standard method was developed in a spectrophotometer for quantifying the targeted product in the broth, while the product recovery studies was carried out using a supercritical fluid extraction pilot plant. Saccharomyces and Bacillus species were chosen for the bioproduction of the selected bioproducts. Experimental design and statistical analysis of results were carried out using response surface methodology (RSM) and artificial neural network (ANN). Studies on each of the selected bioproducts are as justified in the paragraphs below. Bioethanol production has recently become an increasing trend in research, with a focus on increasing its economic viability. Hence, the need to develop a low-cost fermentation medium with minimum redundant nutritional supplements, thereby minimizing the costs associated with nutritional supplements whereby inoculum preparation becomes necessary for ethanol production. Corn steep liquor (CSL) in glucose fermentation by Saccharomyces Type 1 (ST1) strain and Anchor Instant Yeast (AIY), which are low-cost media, are used as replacements for yeast extract (YE). The fermentation process parameters were optimized using artificial neural networks (ANN) and the response surface methodology (RSM). The study shows that for CSL, a maximum average ethanol concentration of 41.92 and 45.16 g/L representing 82% and 88% of the theoretical yield were obtained after 36 h of fermentation in a shake flask for ST1 and AIY respectively. For YE, ethanol concentration equivalent to 86% and 88% of theoretical yield were obtained with ST1 and AIY respectively after 48 h. Although, ANN predicted the responses of ethanol yield better than RSM, optimum conditions for ethanol production were better predicted by RSM. The consumers’ preference for ‘naturally’ produced aromas drives the development of bioproduction of acetoin from glucose with a view to optimize its production. The results revealed that by using a cheap nitrogen source, corn steep liquor, the yield of acetoin was similar to those of yeast and beef extracts. Furthermore, it was shown that by using Box-Behnken design, the optimum parameters such as glucose concentration, corn steep liquor, and inoculum size to maximize the concentration of acetoin produced were 78.40 g/L, 15.00% w/v and 2.70% v/v respectively. The validated concentration of acetoin produced in a triplicate analysis, 10.7 g/L, was 0.06% less than the predicted value. Increasing awareness of consumers of healthy, eco-friendly flavors and fragrances motivates the bioproduction of vanillin. The interactive effects of three variables on vanillin yield were evaluated by response surface methodology (RSM) with Box-Behnken design (BBD) model. The results showed the optimum conditions for the biotransformation of ferulic acid into vanillin can be achieved with maximum overall desirability (D) of 1.0 and a significant (p<0.05) quadratic model with regression coefficient (R2) of 0.995. Corn steep liquor, initial ferulic acid concentration and pH significantly influence the concentration of vanillin in the broth. The results in triplicate experiments confirmed vanillin yield of 386 mg/L after validation, which was in agreement with the prediction of the model. The maximum vanillin yield of 384.40 mg/L was predicted when corn steep liquor, ferulic acid concentration and pH were 7.72 g/L, 2.33 g/L, and 9.34 respectively. Fermentation system in a bioreactor has been proven to be an efficient system for the study of controlled fermentation variables when compared to a shake flask study. The influence of agitation, aeration, time and pH were analysed by Taguchi orthogonal array design for the upscale of acetoin in a bioreactor. The optimized parameters in 1.3L of fermentation vessel were as follows: 300 rpm agitation, 1.5 slpm aeration; 2 days’ fermentation time and 6.5 pH value. Agitation with above 70% was the most contributing factor and other variables were less than 30% in the percentage analysis of variance of each fermentation variables in the batch study of acetoin. A fourfold gain in acetoin titre (42.30 g/L) was obtained with the same substrate concentration in a lab-scale bioreactor on scaling up when compared with the shake flask batch study. The validated acetoin concentration of 41.72 g/L was obtained after a triplicate experiment to confirm the possibility of reproducing acetoin using the optimized conditions. Many separation techniques have been proven to recover value-added products from fermentation broth with a preference for several methods above other and new techniques that are emerging. Supercritical fluids separation using CO2 is one such technique. The feasibility of acetoin concentration and recovery was studied in supercritical CO2 pilot plant with pressure ranges of 100 to 300 bar, CO2 feed rate of 5 to 15 kg/h, at a process temperature of 37 and 80 °C in simulated and fermentation broth, respectively. The validated conditions for the fractionation of acetoin by supercritical fluid extraction (SFE) were determined as follows: extraction pressure, 300 bar; CO2 feed rate, 15 kg/h; extraction temperature 37 °C; and fractionation time of 30 minutes. At these operating conditions, the percentage recovery of acetoin with respect to the feed solution at the raffinate for the simulated and actual ermentation broth was 77.8% (0.20 g/L) and 77% (0.15 g/L) respectively. A two-fold extract increase was obtained after 30 minutes of fractionation. The study provides the technical feasibility and the base case data which are critical to the development and design of processes for production and recovery of acetoin. The lesson gleaned from this study may be extended to develop processes for the production and recovery of other bioproducts (ethanol and vanillin).

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