Sequential statistical methods were used to maximise carotenoid production by a strain of Rhodotorula mucilaginosa, isolated from the Brazilian ecosystem. Initially, a factorial 2(5-1) experimental design was used, and the variables were pH and the levels of glucose, yeast extract, MgSO4.7H2O and KH2PO4. The nitrogen source (yeast extract) was the most important variable in enhancing carotenoid production; MgSO4.7H2O and KH2PO4 had a negative influence. The initial pH had no significant effect on carotenoid and cell productions. We further investigated the effects of glucose and yeast extract effects, using a second-order central composite design (CCD) to optimise carotenoid production, which was adequately approximated with a full quadratic equation obtained from a two-factor-2-level design. The analysis of quadratic surfaces showed that after 5 days of cultivation at 25ºC, the maximum carotenoid concentration (745 µg l-1) was obtained with 15 g l-1 of yeast extract and 20 g l-1 of glucose. The maximum carotenoid production (152 µg g-1) was obtained with 5 g l-1 yeast extract and 10 g l-1 glucose. Carotenoid formation was more sensitive to changes in yeast extract concentration than to changes in glucose concentration. Maximum cell production was achieved with 15-17 g l-1 of yeast extract and 15-20 g l-1 of glucose.

Propôs-se neste trabalho, otimizar a produção de xilose a partir do bagaço de cana-de-açúcar. O processo de hidrólise foi realizado para avaliar o efeito de temperatura e concentração de ácido sulfúrico na produção de xilose, em 18, 30 e 50 min de hidrólise. Um planejamento completo com pontos centrais foi usado para se estudar duas variáveis do processo, temperatura (111,5; 114,0; 120,0; 126,0 e 128,5 ºC) e concentração de ácido sulfúrico (0,20; 0,70; 1,90; 3,10 e 3,60% v v-1). O ácido sulfúrico teve maior influência no processo de hidrólise do que a temperatura. Nas concentrações de ácido sulfúrico acima de 3,10% (v v-1), o aumento da temperatura teve pouca influência no rendimento da produção de xilose. A melhor condição para obtenção da xilose (266,73 mg g-1 de bagaço seco) foi a 18 min de reação, 126 ºC e 3,10% (v v-1) de concentração de ácido sulfúrico, representando 96,0% do máximo teórico.

The aim of this study was to optimize the production of xylose from sugarcane bagasse. The hydrolysis process was carried out to evaluate the effect of temperature and sulphuric acid concentration on the xylose production at 18, 30, and 50 min of hydrolysis. A Central Composite Rotatable Design (CCRD) was used to study two process variables, temperature (111.5; 114.0; 120.0; 126.0 and 128.5 ºC) and sulphuric acid concentration (0.20; 0.70; 1.90; 3.10 and 3.60% v v-1). Sulphuric acid had a greater influence on the hydrolysis process than temperature. At concentrations of sulphuric acid higher than 3.10% (v v-1), raising temperature had little influence on the yield of xylose production. The best condition to obtain xylose (266.73 mg g-1 of dry bagasse) was at 18 min of reaction, at 126 ºC and 3.10% (v v-1) of sulphuric acid, representing 96.0% of the theoretical maximum.

Este trabalho teve como objetivo selecionar e identificar leveduras encontradas no Brasil capazes de produzir carotenóides. As leveduras pigmentadas foram isoladas de amostras de solos, folhas, frutos, flores e um alimento processado. As amostras foram colocadas em frascos de erlenmeyer, contendo meio de Extrato de Malte e Levedura (YM), e incubadas a 30ºC. Após 48 horas, as amostras foram inoculadas em placas de petri contendo meio YM ágar e incubadas a 30ºC por 120 horas. As colônias, que apresentaram coloração entre amarelo e vermelho, foram transferidas para os tubos de culturas, contendo meio YM ágar e incubadas a 30ºC por 72 horas. Das 242 amostras, somente cinco delas apresentaram coloração intensa entre amarelo e vermelho. Estas colônias de leveduras foram reisoladas, em placas de petri contendo YM ágar e, posteriormente, transferidas para tubos de ensaios contendo GPYM ágar. A identificação das leveduras, baseada nas características morfológicas, de reprodução, além dos testes fisiológicos e bioquímicos, classificou quatro linhagens como Rhodotorula mucilaginosa e uma como Rhodotorula graminis. Os principais pigmentos extraídos destas linhagens foram identificados através da análise de cromatografia de alta eficiência como beta-caroteno e toruleno. As linhagens de leveduras mostraram potencial como microrganismos promissores para a produção comercial de carotenóides por fermentação.

The objective of the present study was to select and identify yeasts from Brazil capable of producing carotenoids. Pigmented yeasts were isolated from soil, leaves, fruits, flowers and a processed product. The samples were incubated at 30ºC in Erlenmeyer flasks, containing YM broth. After 48 hours, they were inoculated in Petri dishes with YM agar, and incubated at 30ºC during 120 hours. The yeast colonies, which presented yellow to red coloration, were transferred to culture tubes containing YM agar, and incubated at 30ºC for 72 hours. Out of 242 samples, only five had yellow to red color at high intensity. These highly pigmented yeasts were re-isolated in Petri dishes with YM agar and then transferred to tubes with GPYM agar. Identification through morphological and reproduction characteristics, along with physiological and biochemical tests, classified four strains as R. mucilaginosa and one strain as R. graminis. The main carotenoids extracted from them were identified through HPLC analysis as beta-carotene and torulene. The strains showed potential as promising microorganisms for the commercial production of carotenoids.

Um método simples e rápido para a preparação da amostra e quantificação simultânea de açúcares e polióis em sorvetes "diet"/"light" por CLAE-IR foi investigado. Os açúcares frutose, glicose, lactose, maltose, sacarose, juntamente com glicerol e sorbitol foram separados. Diferentes variáveis foram testadas na preparação da amostra e nas condições cromatográficas para a separação dos componentes. A melhor condição para a separação dos analitos da matriz foi obtida através de duas extrações consecutivas com água : etanol (1:8 v/v, seguida de 1:4 v/v). Para a separação cromatográfica em uma coluna CLC-NH2 , a fase móvel foi composta de acetonitrila: água (77,5:22,5 v/v) na vazão de 1mL/min, a 30 ºC. O tempo de corrida foi menor do que 20 min. Amostras de sorvetes "diet"/"light" de 3 diferentes marcas, com sabores de morango, chocolate, flocos e baunilha, recolhidas em Campinas-SP, apresentaram valores de açúcares totais entre 9-15%. Somente os sorvetes com sabor morango apresentaram frutose (0,1-0,5%), e somente aqueles sorvetes com sabor de flocos mostraram a presença de sacarose em maiores quantidades. Os maiores valores de açúcares foram encontrados para lactose, independente da marca ou sabor testados (3,5-10%). As quantidades de sorbitol variaram de 3-4%. A mesma análise foi realizada com sorvete normal (controle) que apresentou valor de açúcares totais de aproximadamente 29%, sendo que destes 16,8%, foi sacarose. Não foram detectados polióis. Na amostra de sorvete controle foi realizado teste de recuperação de sorbitol (99,3%). Os valores encontrados foram comparados com os valores permitidos pela legislação.

A simple fast method for sample preparation and the simultaneous quantification of sugars and polyols in diet/light ice creams, using HPLC-RI, was investigated. The sugars fructose, glucose, lactose, maltose and sucrose, together with glycerol and sorbitol, were separated. Different variables were tested both for sample preparation and for the chromatographic conditions used to separate the components. The best conditions to separate these components from the matrix were by way of two extractions with water:ethanol (1:8 v/v followed by 1:4 v/v). The chromatographic separation using an CLC-NH2 column with a mobile phase consisting of acetonitrile:water (77.5:22.5 v/v) with flow rate of 1mL/min, at 30ºC. The run time was less than 20 min. Samples of three different brands of strawberry, chocolate, chocolate chip and vanilla ice cream, all collected in Campinas-S.P., presented values for total sugars between 9 and 15%. Only the strawberry flavored ice creams showed the presence of fructose (0.1-0.5%) and only the chocolate chip ice creams showed the presence of sucrose in higher quantities. The sugar found in greatest amounts was lactose (3.5-10%), whatever the brand or flavor. Sorbitol was present in amounts of 3-4%. The same analyses was applied to non-light ice cream, which presented value for total sugars of approximately 29%, of which 16.8% was sucrose, polyols not being detected. Sorbitol recovery was tested in one of these samples(99.3%). The values encountered were compared with the values legally permitted.