Fermentación alcohólica por Saccharomyces cerevisiae y cuantificación de flavonoides del zumo de Citrus x clementina (naranja)

Objetivo: elaborar una bebida por fermentación alcohólica y la cuantificación de flavonoides del zumo de Citrus x clementina (naranja). Metodología: se utilizó el método de fermentación alcohólica por levadura de la variedad Saccharomyces cerevisiae, se fermento el jugo de naranja con una densidad de 1,050 glcm3 por 5 semanas y se cuantificó los flavonoides de la bebida alcohólica por el método de cromatografía HPLC. Resultados: después de las 5 semanas se analizó que la bebida por fermentación alcohólica tuvo un 11 % de alcohol y flavonoides de hesperidina 13,9 mgl100 ml y naringenina 6,3 mg/100 ml en su concentración.

SUMMARY Aim: to elaborate a drink by alcoholic fermentation and the quantification of flavonoids in Citrus x clementine (orange) juice. Methodology: the method of alcoholic fermentation by yeast of the Saccharomyces cerevisiae variety was used, the orange juice was fermented with a density of 1.050 glcm3 for 5 weeks and the flavonoids of the alcoholic beverage were quantified by the HPLC chromatography method. Results: after 5 weeks it was analyzed that the drink by alcoholic fermentation had 11 % alcohol and hesperidin flavonoids 13.9 mgl100 ml and 6.3 mg/100 ml naringenin in its concentration.

Objetivo: elaborar uma bebida por fermentação alcoólica e quantificação de flavonóides no suco Citrus x clementina (laranja). Metodologia: foi utilizado o método de fermentação alcoólica por levedura da variedade Saccharomyces cerevisiae, o suco de laranja foi fermentado com densidade de 1,050 glcm3 por 5 semanas e os flavonóides da bebida alcoólica foram quantificados pelo método de cromatografía HPLC. Resultados: após 5 semanas foi analisado que a bebida por fermentação alcoólica continha álcool a 11 % e flavonóides de hesperidina 13,9 mgl100 ml e 6,3 mg/100 ml naringenina em sua concentração.

Lactobacillus plantarum as a malolactic starter culture in winemaking: a new (old) player?

Malolactic fermentation (MLF) is a process in winemaking responsible for the conversion of L-malic acid to L-lactic acid and CO2, which reduces the total acidity, improves the biological stability, and modifies the aroma profile of wine. MLF takes place during or after alcoholic fermentation and is carried out by one or more species of lactic acid bacteria (LAB), which are either present in grapes and cellars or inoculated with malolactic starters during the winemaking process. Although the main bacterium among LAB used in commercial starter cultures for MLF has traditionally been Oenococcus oeni, in the last decade, Lactobacillus plantarum has also been reported as a malolactic starter, and many works have shown that this species can survive and even grow under harsh conditions of wine (i.e., high ethanol content and low pH values). Furthermore, it has been proved that some strains of L. plantarum are able to conduct MLF just as efficiently as O. oeni. In addition, L. plantarum exhibits a more diverse enzymatic profile than O. oeni, which could play an important role in the modification of the wine aroma profile. This enzymatic diversity allows obtaining several starter cultures composed of different L. plantarum biotypes, which could result in distinctive wines. In this context, this review focuses on showing the relevance of L. plantarum as a MLF starter culture in winemaking.

Cassava Pulp Enzymatic Hydrolysate as a Promising Feedstock for Ethanol Production

ABSTRACT The aim of this study was to produce bioethanol from enzymatic hydrolysates of cassava pulp, a by-product of cassava flour manufacturing, using an alcohol-tolerant Saccharomyces cerevisiae strain. First, the best operational condition of the starch hydrolysis process was determined through a complete factorial design (24), with triplicates at the central point. The independent variables evaluated were: the concentrations of α-amylase (Termamyl 2X) and glucoamylase (AMG 300L) and both liquefaction and saccharification times. The most favorable hydrolysis condition in the assay was achieved using 0.517 mL AMG.g starch-1 and 0.270 mL Termamyl.g starch-1, with liquefaction and saccharification times of 1 and 2 h, respectively. The broth obtained at this hydrolysis condition contained a high glucose concentration (160 g.L-1). Once the best reaction conditions were determined, fermentation tests were carried out in a 3 L bioreactor, in a batch system, at 30 °C, 100 rpm and pH 5.5, using 3 g.L-1 (dry biomass) of yeast as inoculum. After 24 h of fermentation, an ethanol concentration of 68 g.L-1 was obtained, with 0.48 ethanol yield and 2.83 g.L-1.h-1 productivity. These results indicate the potential use of cassava pulp, a by-product of cassava flour industries in Brazil, as a raw material for bioethanol production.

Efecto del anegamiento en la actividad de la enzima alcohol deshidrogenasa en raíces de maracuyá amarillo Passiflora Edulis var. Flavicarpa / Effect of waterlogging on the alcohol dehydrogenase activity in yellow passion fruit roots Passiflora Edulis var. Flavicarpa

Bajo entornos naturales, las plantas de maracuyá amarillo frecuentemente se enfrentan a condiciones de anegamiento, un factor limitante para la producción generalizada de su cultivo, especialmente en tierras bajas inundables. El presente estudio se encargó de identificar la activación de la enzima alcohol deshidrogenasa (ADH) durante la inundación, como un posible mecanismo de sobrevivencia de plántulas de maracuyá amarillo. Se evaluó la actividad de la enzima ADH durante 0, 1, 3, 7, 9 y 14 días de tratamiento con inundación y sin inundación, en raíces de plántulas de maracuyá amarillo de tres meses y medio de germinadas. En las raíces de plántulas de maracuyá amarillo en condiciones de inundación, la actividad de la enzima ADH presentó un aumento significativo respecto a las plántulas en condiciones normales de riego (sin inundación). Esto sugiere que la actividad de la enzima alcohol deshidrogenasa, implicada en el metabolismo anaeróbico, es un posible mecanismo de supervivencia al anegamiento de plántulas de maracuyá amarillo en periodos cortos de inundación.

Under natural environments, yellow passion fruit plants often face flooded conditions as a limiting factor for the widespread production of the crop, especially in flood-prone lowlands. This study was carried out to identify the activation of the enzyme alcohol dehydrogenase (ADH) during the flood, as a possible mechanism for the survival of yellow passion fruit seedlings. The activity of the ADH enzyme was assessed on days 0, 1, 3, 7, 9 and 14, with and without flooding, on rooted yellow passion fruit seedlings germinated during three and a half months. In the roots of the yellow passion fruit seedlings in flood conditions, the activity of the ADH enzyme showed a significant increase compared to seedlings under normal irrigation (no flooding). These results suggest that the activity of the alcohol dehydrogenase enzyme involved in anaerobic metabolism, is a possible mechanism for survival of yellow passion fruit seedlings waterlogged in short periods of flooding.

Características fermentativas e bromatológicas de silagens de cana-de-açúcar com taboa / Fermentative and chemical characteristics of sugarcane silages with "taboa"

Objetivou-se avaliar o potencial da taboa (Typha domingensis) como ingrediente, com diferentes níveis de inclusão em silagem de cana-de-açúcar (Saccharum officinalis), através de estudo dos parâmetros fermentativos. Utilizou-se delineamento experimental inteiramente casualizado com quatro tratamentos e seis repetições. Após 90 dias, com a abertura dos silos, determinaram-se os teores de carboidratos solúveis (CHO-Sol), etanol, ácidos orgânicos (lático, acético, propiônico, butírico), proteína bruta (PB), matéria mineral (MM), nutrientes digestíveis totais (NDT), digestibilidade in vitro a gás da matéria orgânica (DIVMO), pH e poder tampão (PT); estimou-se a perda total de MS, perdas por efluente e gases, densidade, e a recuperação da MS. Os níveis crescentes de taboa propiciaram diminuição (P<0,05) da densidade nas silagens e aumento dos teores de PB, MM, NDT e DIVMO; muito embora não tenha se observado efeito (P>0,05) sobre as perdas e recuperação da MS. As concentrações de etanol foram influenciadas (P<0,05) pelo incremento de taboa, quando se observou redução de 76,92%, com 40% de taboa em relação à silagem exclusiva de cana. A inclusão de taboa aumenta o conteúdo de proteína e minerais, bem como NDT e DIVMO, reduzindo significativamente a produção de etanol da silagem de cana-de-açúcar. A inclusão de taboa na ensilagem de cana-de-açúcar melhorou substancialmente as características fermentativas e o valor nutricional da massa ensilada.

This study aimed to evaluate the potential of Typha domingensis ("taboa") as an ingredient with different inclusion levels in sugar cane (Saccharum officinalis) silage, by the study of the fermentative parameters. The experiment was carried out as a completely randomized design with four treatments and six replicates. After 90 days, of the silos' opening , the levels of water soluble carbohydrates, ethanol, organic acids (lactic, acetic, propionic, butyric acid), crude protein (CP), mineral matter (MM), total digestible nutrients (TDN), in vitro organic matter digestibility by gas (IVDOM), pH and buffer capacity; and, the losses by gas, effluent and total of dry mater, density and dry matter recovery were estimated. Increasing levels of "taboa" reduced (P<0.05) silage density and increased the contents of CP, MM, TDN e IVDOM; although it has not been observed effect (P>0.05) on the loss and recovery of DM. Ethanol concentrations were influenced (P<0.05) by increment of "taboa" where reduction of 76.92% was observed, with 40% of "taboa" relative sugarcane silages exclusively. The inclusion of "taboa" increases the protein content and minerals, as well as TDN and IVDOM, also significantly reduces ethanol production of sugarcane silages. The inclusion of taboa in the ensiling of sugar cane substantially improved fermentation characteristics and nutritional value of the silage mass.

Características fermentativas na ensilagem de cana-de-açúcar com maniçoba / Fermentative characteristics in sugarcane silage with maniçoba

A utilização da silagem de cana-de-açúcar apresenta benefícios que têm despertado interesse de produtores e pesquisadores. No entanto, algumas limitações, principalmente relacionadas ao seu padrão de fermentação, têm constituído grande desafio para utilização desta forrageira. Com objetivo de avaliar a inclusão de maniçoba sobre as características fermentativas na silagem de cana-de-açúcar, foi elaborado experimento em delineamento inteiramente casualizado com quatro tratamentos (níveis de inclusão: 0, 20, 30 e 40%) e seis repetições. Após 90 dias do fechamento dos silos, foram determinadas as perdas na forma de gases, perda total e recuperação de matéria seca e produção de efluentes. Os silos então foram abertos e obtido o extrato aquoso das silagens, foram determinados os teores dos ácidos orgânicos (acético, butírico, lático e propiônico), etanol, N-NH3, NT e valores de pH. À medida que se aumentou o nível de inclusão de maniçoba, foi observada redução de 67% nas perdas por gases e de 23,3% na produção de efluente, enquanto a concentração de etanol foi reduzida de 25,6 a 2,7%. As máximas concentrações de ácido lático (2,4%), acético (2,2%) e propiônico (0,03%) foram encontradas em silagens contendo 40% de maniçoba, enquanto que, para o ácido butírico, decresceu linearmente. Em todas as silagens, o pH encontrado mostrou-se adequado ao processo de ensilagem (variação de 3,4 a 3,7). O maior nível de inclusão de maniçoba em silagens de cana-de-açúcar melhora a fermentação das silagens e reduz as perdas fermentativas.

The use of silage sugar cane has a benefit that has aroused the interest of producers and researchers. However, some limitations mainly related to its fermentation pattern, has become the challenge for forage use. To evaluate the inclusion of maniçoba on silage fermentation characteristics of sugar cane, it was prepared an experiment in a completely randomized design with four treatments (inclusion levels: 0, 20, 30 and 40% ) and six replications. After 90 days of the silos closing o were resoluted losses in the form of gases, total loss and recovery of dry matter production and effluent. The silos were opened and then obtained the aqueous extract of the silages, determined levels of organic acids (acetic, butyric, lactic and propionic), ethanol, NH3-N, NT and pH values. As the level of inclusion of maniçoba was increased, it was observed a 67% reduction in the losses of gases and 23.3% in the effluent production, while the ethanol concentration was reduced from 25.6 to 2.7%. The maximum concentration of lactic acid (2.4%), acetic acid (2.2%) and propionic (0.03%) were found in silages containing 40% of maniçoba, while for butyric acid it decreased linearly. In all silages the pH found was adequate to silage (range 3.4 to 3.7). The highest level of maniçoba inclusion in sugar cane silages improves the fermentation of the silage and reduces fermentation losses.

Características fermentativas de silagens de cana-de-açúcar com erva-sal / Fermentation characteristics of sugarcane silages with saltbush

Objetivou-se com o presente estudo avaliar diferentes níveis (0, 20, 40 e 60%) de inclusão de erva-sal na dinâmica fermentativa da silagem de cana-de-açúcar. Utilizou-se delineamento experimental inteiramente casualizado com quatro tratamentos e seis repetições. Após 90 dias, com a abertura dos silos, determinou-se os teores de carboidratos solúveis (CHO-Sol), etanol, ácidos orgânicos (lático, acético, propiônico, butírico), proteína bruta (PB), matéria mineral (MM), matéria seca (MS), fibra em detergente neutro livre de cinzas (FDNc), pH e poder tampão (PT); estimou-se a perda total de MS, perdas por efluente e gases, densidade e a recuperação da MS. Os níveis crescentes de erva-sal propiciaram diminuição das perdas totais de MS e por efluente, aumento de seu teor e recuperação desta. As silagens apresentaram pH adequado à conservação (≤3,70) e concentração de minerais acrescidos. As concentrações de etanol foram influenciadas (P<0,05) pelo incremento de erva-sal, em que se observou redução de 51,84%, com 40% de erva-sal na silagem de cana. A adição de erva-sal reduz as perdas de MS e a produção de etanol e, melhora a qualidade fermentativa da silagem de cana-de-açúcar.

The objective of this study was to evaluate different inclusion levels (0, 20, 40 and 60%) of old man saltbush in the fermentation dynamics of sugarcane silages. The experiment was carried out as a completely randomized design with four treatments and six replicates. After 90 days, with the opening of the silos, the levels of water soluble carbohydrates, ethanol, organic acids (lactic, acetic, propionic, butyric acid), crude protein (CP), mineral matter (MM), dry matter (DM), neutral detergent fiber ash free (NDFa), pH and buffer capacity were determined; and, the losses by gas, effluent and total of dry mater, density and dry matter recovery were estimated. Increasing levels of saltbush propitiated reduction in losses: by effluent and total dry matter, it also increased the concentration and recovery. The silages showed pH suitable for conservation (≤3.70) and mineral concentration increased. The ethanol concentrations were influenced (P<0.05) by the increase of saltbush, where reduction of 51.84% was observed, with 40% saltbush in sugarcane silage. The addition of saltbush reduces dry matter losses and ethanol production and, improves the quality of sugarcane silage.

Population dynamics of mixed cultures of yeast and lactic acid bacteria in cider conditions

The objective of this work was to study the malolactic bioconversion in low acidity cider, according Brazilian conditions. The apple must was inoculated with Saccharomyces cerevisiae or S. cerevisiae with Oenococcus oeni. The control contained the indigenous microorganisms. Fermentation assays were carried out with clarified apple must from the Gala variety. At the beginning of fermentation, there was a fast growth of the non-Saccharomyces yeast population. Competitive inhibition occurred in all the assays, either with inoculated or indigenous populations of the yeast. The lactic acid bacteria count was ca. 1.41·10²CFU/mL at the beginning and 10(6)CFU/mL after yeast cells autolysis. The lactic bacteria O. oeni reached the highest population (10(7)CFU/mL) when added to the apple must after the decline of the yeast. The malic acid was totally consumed during the alcoholic fermentation period (80.0 to 95.5 %) and lactic acid was still synthesized during the 35 days of malolactic fermentation. These results could be important in order to achieve a high quality brut, or sec cider obtained from the dessert apple must.

Evaluation of stress tolerance and fermentative behavior of indigenous Saccharomyces cerevisiae

Sixty six indigenous Saccharomyces cerevisiae strains were evaluated in stressful conditions (temperature, osmolarity, sulphite and ethanol tolerance) and also ability to flocculate. Eighteen strains showed tolerant characteristics to these stressful conditions, growing at 42 ºC, in 0.04% sulphite, 1 mol L-1 NaCl and 12% ethanol. No flocculent characteristics were observed. These strains were evaluated according to their fermentative performance in sugar cane juice. The conversion factors of substrates into ethanol (Yp/s), glycerol (Yg/s) and acetic acid (Yac/s), were calculated. The highest values of Yp/s in sugar cane juice fermentation were obtained by four strains, one isolated from fruit (0.46) and the others from sugar cane (0.45, 0.44 and 0.43). These values were higher than the value obtained using traditional yeast (0.38) currently employed in the Brazilian bioethanol industry. The parameters Yg/s and Yac/s were low for all strains. The UFLA FW221 presented the higher values for parameter related to bioethanol production. Thus, it was tested in co-culture with Lactobacillus fermentum. Besides this, a 20-L vessel for five consecutive batches of fermentation was performed. This strain was genetically stable and remained viable during all batches, producing high amounts of ethanol. The UFLA FW221 isolated from fruit was suitable to produce bioethanol in sugar cane juice. Therefore, the study of the biodiversity of yeasts from different environmental can reveal strains with desired characteristics to industrial applications.

Produção e caracterização físico-química de fermentado de umbu / Production and physicochemical characterization of fermented umbu

O objetivo deste trabalho foi desenvolver um fermentado de umbu visando a agregar valor a esse fruto e contribuir para a melhoria de renda das famílias do semiárido nordestino. A polpa utilizada nos experimentos foi submetida a análises físico-químicas e microbiológicas. Para a produção da bebida, a polpa foi diluída em água, sendo necessário realizar uma chaptalização com sacarose até atingir 20,5°Brix. A levedura vínica comercial utilizada foi Saccharomyces cerevisiae. A fermentação foi conduzida a 18°C durante 18 dias e posteriormente a bebida foi submetida a uma estabilização com auxílio de agentes de sedimentação, gelatina e bentonita, por 14 dias a 1°C. Após a estabilização, o fermentado de umbu foi filtrado em filtro prensa. O fermentado de umbu obtido apresentou teor alcoolico de 11,20°GL. A bebida foi analisada quanto às suas características físico-químicas e todos os parâmetros estavam em conformidade com a legislação vigente.

The objective of this study was to develop a fermented umbu drink aiming at adding value to this fruit and contribute for improving the families income in Brazil northeast semiarid region. The pulp used in the experiments was subjected to physico-chemical and microbiological analysis. For the production of liquor, the pulp was diluted with water, and needed a sugaring with sucrose up to 20.5°Brix. A commercial Saccharomyces cerevisiae wine yeast was used. The fermentation was conducted at 18°C for 18 days and subsequently underwent a drink with the aid of stabilizing agents of sedimentation, gelatin and bentonite, for 14 days at 1°C. After stabilizing the fermented umbu drink was subjected to filtered in filter press. The fermented alcoholic umbu drink showed an alcohol contents of 11.20°GL. The drink was analyzed for their physicochemical characteristics and all parameters were in accordance with current legislation.

INFLUENCIA DE LA AIREACIÓN EN LA ACTIVIDAD FERMENTATIVA DE Kloeckera apiculata DURANTE LA FERMENTACIÓN DE JUGO DE MANZANA / Influence of Aeration in the Fermentative Activity of Kloeckera apiculata during Fermentation of Apple Juice

Se ha estudiado la influencia de la aireación en la actividad fermentativa de Kloeckera apiculata RIVE 9-2-1 con el objetivo de evaluar la producción de metabolitos de la fermentación. Para ello, la cepa se cultivó en frascos Erlenmeyer conteniendo jugo de manzana esterilizado y sin aroma, y los compuestos químicos producidos durante la fermentación en cultivo agitado (200 min-1) y estático (sin agitación) se determinaron. Los resultados mostraron que la agitación del medio de cultivo incrementa la producción de alcoholes superiores (hasta 591,0 mg/L) comparado al cultivo estático, mientras que por el contrario, la producción de ácido acético, etil acetato y glicerol (260,0 ± 11,0 mg/L, 196,0 ± 10,0 mg/L y 2,6 ± 0,2 g/L) resultaron ser mayores que en cultivo agitado (222,0 ± 8,0 mg/L, 96,0 ± 4,5 mg/L y 1,8 ± 0,2 g/L) respectivamente. Cultivos bacth realizados en biorreactor con un flujo de aire de 25 L/h reportaron una tasa de crecimiento (µ) de 0,17 h-1, producción de etanol (12,5 ± 2,0 g/L) y otros compuestos típicamente producidos durante la fermentación alcohólica. La concentración de oxígeno disuelto en el medio de fermentación afecta su metabolismo así, cantidades insuficientes de oxígeno provocaría un metabolismo respirofermentativo. Los mejores resultados en términos de calidad organoléptica de la bebida fermentada en lo referente al aroma, sabor y olor se obtuvieron en la fermentación en cultivo estático. El control de la aireación del medio de fermentación puede ser usado para controlar la síntesis de compuestos químicos de impacto sensorial en la producción de bebidas fermentadas.

The influence of aireation on the fermentative activity of Kloeckera apiculata RIVE 9-2-1 was studied in order to evaluate the production of metabolites of the fermentation. To achieve this, the strain was cultured in Erlenmeyer flasks containing sterilized and aroma removed apple juice, and the chemical compounds produced during fermentation in shaken (200 min-1) and static (without agitation) cultivation were determined. The results showed that the agitation of the culture medium increases production of higher alcohols (till 591.0 mg/L) compared to static cultivation, whereas on the contrary, the production of acetic acid, ethyl acetate and glycerol (260.0 ± 11.0 mg/L, 196.0 ± 10.0 mg/L y 2.6±0.2 g/L) were higher compared to shaken cultivation (222.0 ± 8.0 mg/L, 96.0 ± 4.5 mg/L and 1.8 ± 0.2 g/L) respectively. Batch cultivations carried out in bioreactor with air flux of 25 l/h reported a growth rate (µ) of 0.17 h-1, production of ethanol (12.5 ± 2.0 g/L) and other compounds typically produced during alcoholic fermentation. The concentration of dissolved oxygen in the fermentation medium affects its metabolism thus; insufficient amounts of oxygen would provoke a respirofermentative metabolism. The best results in terms of organoleptic quality of the fermented beverage regarding to aroma, taste and flavour was obtained when fermented in static cultivation. The control of aeration during fermentation can be used to control the synthesis of chemical compounds of sensory impact in the production of fermented beverages.

Actividad fermentativa de Hanseniaspora uvarum y su importancia en la producción de bebidas fermentadas / Fermentative activity of Hanseniaspora uvarum and its importance in production of fermented beverages

Se estudió la actividad fermentativa de Hanseniaspora uvarum RIVE 6-2-2 con el objetivo de evaluar su importancia en los procesos de producción de bebidas fermentadas. La cepa se cultivó en frascos Erlenmeyer conteniendo jugo de manzana esterilizado y sin aroma, y se determinaron los compuestos químicos de importancia sensorial producidos durante la fermentación en cultivo agitado (200 min-1) y estático (sin agitación). Los resultados mostraron que la cepa fue capaz de producir etanol hasta 4,02±0,1v/v% en cultivo estático a 28 °C. La agitación del medio de cultivo incrementó la producción de alcoholes superiores (hasta 488,2 mg/L) y ácido acético (468,0±10,2 mg/L) comparado al cultivo estático, mientras que por el contrario, la producción de etil acetato y glicerol (189,0±12,0 mg/L y 3,2±0,3 g/L) resultó ser mayor que en cultivo agitado (142,0±8,0 mg/L y 2,3±0,25 g/L) respectivamente. Cultivos bacth realizados adicionalmente reportaron una tasa de crecimiento (μ) de 0,05 h-1 y producción de pequeñas cantidades de compuestos típicamente encontrados en la fermentación alcohólica. Los mejores resultados, en términos de calidad organoléptica (aroma, sabor y olor), se obtuvieron en la fermentación en cultivo estático. El control de la aireación del medio de fermentación es una herramienta importante para controlar la síntesis de compuestos de importancia sensorial en la producción de bebidas fermentadas.

The fermentation activity of Hanseniaspora uvarum RIVE 6-2-2 was studied with the purpose of evaluating its importance in the production process of fermented beverages. The strain was cultured in Erlenmeyer flasks, which contained sterilized and odorless apple juice, and the chemical compounds of sensorial importance produced during fermentation in shaken (200 min-1), and static (without shaking) cultures at 28 ºC were determined. The results showed that the strains were capable of producing ethanol up to 4.20±0.1v/v% in static cultures at 28 ºC. Shaking of the culture medium increased the superior alcohol production (up to 488.2 mg/L) and acetic acid (468.0± 10.2 mg/L), when compared with the static culture; on the other hand, the production of ethyl acetate and glycerol (189.0±12.0 mg/L and 3.2±0.3 g/L) was higher in static than in shaken cultures (142.0±8.0 mg/L and 2.3±0.25 g/L), respectively. Additional batch cultures reported a growth rate (µ) of 0.05 h-1 and production of small amounts of compounds typically found in alcoholic fermentation. The best results, in terms of organoleptic qualities (aroma, taste and smell), were found in the static culture fermentation. The aeration control of the fermentation medium is an important tool for controlling the synthesis of sensorial importance compounds in the production of fermented beverages.

Identification of Saccharomyces cerevisiae strains for alcoholic fermentation by discriminant factorial analysis on electronic nose signals

An electronic nose (E-nose) coupled to gas chromatography was tested to monitor alcoholic fermentation by Saccharomyces cerevisiae ICV-K1 and Saccharomyces cerevisiae T306, two strains well-known for their use in oenology. The biomass and ethanol concentrations and conductance changes were measured during cultivations and allowed to observe the standard growth phases for both yeast strains. The two strains were characterized by a very similar tendency in biomass or ethanol production during the fermentation. E-nose was able to establish a kinetic of the production of aroma compounds production and which was then easy to associate with the fermentation phases. Principal Component Analysis (PCA) showed that the data collected by E-nose during the fermentation mainly contained cultivation course information. Discriminant factorial analysis (DFA) was able to clearly identify differences between the two strains using the four main principal components of PCA as input data. Nevertheless, the electronic nose responses being mainly influenced by cultivation course, a specific data treatment limiting the time influence on data was carried out and permitted to achieve an overall performance of 83.5 percent.

Cultivo contínuo de Arthrospira (Spirulina) platensis em fotobiorreator tubular utilizando uréia como fonte de nitrogênio e CO2 puro ou proveniente de fermentação alcoólica / Continuous cultivation of Arthrospira (Spirulina) platensis in tubular photobioreactor using urea as nitrogen source and CO2 from cylinder or produced by alcoholic fermentation

A aplicabilidade do processo de produção de microrganismos fotossintetizantes depende da obtenção de altas concentrações de biomassa e para isso seria interessante o emprego de fotobiorreatores tubulares. Eles permitem redução da área de cultivo e menor perda de CO2 e nitrogênio amoniacal por volatilização. Em uma primeira etapa deste trabalho, Arthrospira platensis foi cultivada por processo contínuo, avaliando-se diferentes valores de vazão específica de alimentação (D = 0,2 a 1,0 dia-1) e diferentes intensidades luminosas (I = 60 e 120 µmol fótons.m-2.s-1). Verificou-se que 120 µmol fótons.m-2.s-1 associada a D igual a 0,2 dia-1 resultou em maior valor de concentração celular em regime permanente (XP = 2446 ± 74 mg.L-1.d-1), mas o mesmo I associado a maior valor de D (0,6 dia-1) levou ao melhor valor de produtividade em células (PX = 938,73 mg.L-1.d-1). Foi possível a obtenção do regime permanente em quase todos os ensaios, o que indica que o cultivo contínuo de A. platensis em fotobiorreator tubular, usando uréia como fonte de nitrogênio, pode levar a resultados satisfatórios. Considerando a preocupação em relação à substituição de combustíveis fósseis por biocombustíveis, é iminente o crescente aumento da produção de etanol ainda nos próximos anos, e esse trabalho propõe o uso do CO2 liberado pela fermentação alcoólica na produção de microrganismos fotossintetizantes como A. platensis. Para isso, em uma segunda etapa, A. platensis foi cultivada por processo contínuo, com I igual a 120 µmol fótons.m-2.s-1, empregando uréia e CO2 proveniente de fermentação alcoólica para manutenção de pH e reposição da fonte de carbono. O uso desse CO2, sem tratamento prévio, associado a D igual a 0,6 dia-1 e concentração de uréia de 3,2 mM no meio de alimentação, permitiu a obtenção de PX igual a 839 ± 25 mg.L-1.d-1, o que está próximo de 938 ± 30mg.L-1.d-1, obtido com CO2 puro de cilindro. Estes resultados mostram que o uso de CO2 de fermentação alcoólica, associado a...

Appropriately designed tubular photobioreactors seem to be suitable for photosynthetic biomass production. It can reduce the cultivation area and provide lower loss of CO2 and ammoniacal nitrogen by volatilization. In a first step of this study, Arthrospira platensis was cultivated by continuous process, testing different values of dilution rate (D = 0.2 to 1.0 d-1) and light intensities (I = 60 and 120 µmol photons.m-2.s-1). The results of these runs showed that the maximum steady-state cell concentration (XS = 2446 ± 74 mg.L-1.d-1) was achieved at 120 µmol photons.m-2.s-1 and D of 0.2 d-1, but the same light intensity associated to higher dilution rate (0.6 d-1) provided the highest cell productivity (PX = 938 ± 30 mg.L-1.d-1), a value appreciably higher than that reported in other studies. Besides, steady-state conditions were achieved in most of the runs indicating that A. platensis continuous cultivation in the tubular photobioreactor, using urea as nitrogen source, can be performed effectively, thus appearing an interesting alternative for the large scale fixation of carbon dioxide to mitigate the green house effect. Taking into account the concern about the substitution of fossil fuel with biofuels, its evident that the ethanol production is going to increase even more in the next years, and this study propose the use of the CO2 released by the alcoholic fermentation for the production of photosynthetic microorganism such as A. platensis. For this purpose, in a second step, cultivations of A. platensis were carried out with 120 µmol photons.m-2.s-1 by continuous process, using urea and CO2 from Alcoholic fermentation for pH maintenance and carbon source replacement. The use of this CO2, without any treatment, associated with a D of 0.6 d-1 and feed urea concentration of 3.2 mM provide us a PX of 839 ± 25 mg.L-1.d-1, which is slightly lower than 938 ±30 mg.L-1.d-1, obtained with pure CO2 from cylinder. Our results showed that the use of CO2 from...

Cultivo descontínuo alimentado de Arthrospira (Spirulina) platensis em reator tubular utilizando uréia como fonte de nitrogênio e CO2 puro ou proveniente de fermentação alcoólica / Arthrospira (Spirulina) platensis fed batch cultivation using urea as nitrogen source and CO2 pure or from alcoholic fermentation in tubular photobioreactor

A cianobactéria fotossintetizante Arthrospira platensis é conhecida por apresentar em sua biomassa altos teores protéicos (50-70%), presença do ácido graxo essencial γ-linolênico e diversas outras substâncias importantes para a alimentação humana e animal. Esses micro-organismos são capazes de converter o CO2 em biomassa de grande potencial na área de alimentos. Durante a fermentação alcoólica, a produção de CO2 é da mesma ordem de grandeza da produção de etanol e, considerando a crescente demanda interna e externa por esse combustível, seria importante que houvesse um processo que fixasse esse CO2, transformando-o em um produto que poderia ser útil para a nossa população. Adicionalmente, o uso de uma fonte de nitrogênio de baixo custo (uréia) em reatores tubulares contribuiria para a redução do custo de produção da A. platensis. O objetivo desse trabalho foi avaliar os parâmetros cinéticos e bioenergéticos, bem como a composição centesimal da A. platensis cultivadas em biorreator tubular, alimentados com CO2 proveniente de fermentação alcoólica ou com utilização de CO2 puro, para o controle do pH, sob diferentes intensidades luminosas (I) e fontes de nitrogênio (N) adicionadas por meio de processo descontínuo alimentado. Os resultados mostraram que maiores valores de I proporcionaram maiores valores de concentração celular máxima (Xm) e produtividade em células (Px), mas não influenciaram nos valores do fator de conversão de nitrogênio em células (YX/N). As diferentes fontes de CO2 não influenciaram nos valores de Xm, Px, YX/N. O uso da uréia aumentou os valores de YX/N em relação aos cultivos com NO3-. Na composição centesimal, pode-se observar que I influenciou nos teores de clorofila, proteínas e lipídios, mas não influenciou nos teores de cinzas e carboidratos na biomassa final. Em relação aos parâmetros bioenergéticos dos cultivos com CO2 puro, observou-se que os maiores valores de dissipação de energia de Gibbs foram obtidos em tempos mais curtos...

Photosynthetic cyanobacterium A. platensis contains in its biomass high protein content (50-70%), γ-linolenic acid and many other substances important for health human. These microorganisms are capable of converting CO2 into biomass of great potential in the food industry. During the alcoholic fermentation, the production of CO2 has the same magnitude of the production of ethanol. Considering the increasingly global demand for this fuel, a process to fix this CO2 is of utmost importance. Furthermore, the use of low cost nitrogen source (urea) in tubular reactors would contribute to reducing the production cost of A. platensis. Therefore, the purpose of this work was to evaluate the kinetic and bioenergetics parameters, as well as the chemical composition of biomass obtained in A. platensis cultures in tubular bioreactor using CO2 from alcoholic fermentation or pure CO2 to control the pH under different light intensity (I) and nitrogen sources (N). The results showed that higher I induced higher maximum cell concentration (Xm) and cell productivity (Px) values, but it did not exert any influence on the nitrogen-cell conversion factor (YX/N). Urea increased the YX/N values compared to use of NO3-. In the centesimal composition of biomass, it can be observed that I influenced the chlorophyll, protein, and lipid contents, but not influenced the carbohydrate and ash contents. For bioenergetics parameters, it was observed that the highest Gibbs energy dissipation values for cell growth and maintenance were obtained in shorter time at 120-240 mol photons m-2 s-1 in both nitrogen sources, while the moles of photons absorbed by the system to produce one C-mol biomass was higher in cultures with NO3-. The highest values of the molar production of O2 and consumption of H+ were obtained at the highest I values, using NO3-. The estimated percentages of the energy absorbed by the cell showed that, compared with nitrate, the use of urea as nitrogen source allowed the...

Fermentación alcohólica del lactosuero por Kluyveromyces marxianus y solventes orgánicos como extractantes / Lactoserum alcoholic fermentation by Kluyveromyces marxianus and organic solvents as extractants

Las fermentaciones alcohólicas son realizadas a bajas concentraciones de azúcares, para evitar la inhibición por productos o substratos. La fermentación alcohólica extractiva es utilizada en procesos limitados por la concentración de etanol final. Se estudió la fermentación alcohólica del lactosuero evitando la inhibición del Kluyveromyces marxianus, con la adición de solventes de extracción (ácido oleico, hexadecano, butil laurato y aceite de soja) como extractantes del etanol. Se evaluó la biocompatibilidad, cinética de extracción de los solventes, consumo de sustrato, producción de etanol, crecimiento celular (sin solventes y con solventes seleccionados como mejores extractantes y biocompatibles) y rendimientos globales. El ácido oleico y el aceite de soja fueron los mejores solventes de extracción. La velocidad específica de crecimiento se incrementó con el uso de solventes, siendo mayor con el uso de aceite de soja. La lactosa consumida fue de 97 g/L para la fermentación extractiva con ácido oleico, 81,2 g/L con aceite de soja, y 64 g/L para la fermentación convencional. Se obtuvieron 53 y 44 g/L de etanol con ácido oleico y aceite de soja respectivamente, y 35 g/L de etanol con la fermentación convencional. Queda demostrado que la fermentación alcohólica usando acido oleico puede ser usada para reducir la concentración de etanol en el medio y evitar la inhibición de la levadura en el proceso, lo que conlleva a la posibilidad de usar un substrato con altas concentraciones de azúcares, produciendo un incremento en la productividad del sistema.

Alcoholic fermentations are done under low sugar concentrations to avoid product or substrate inhibition. Extractive alcoholic fermentation is used in processes limited by the final ethanol concentration. Lactoserum alcoholic fermentation avoiding the inhibition of Kluyveromyces marxianus by the addition of extraction solvents (oleic acid, hexadecane, butyl laureate and soy oil) as ethanol extractants was studied. Biocompatibility, extraction kinetic of the solvents, substrate use, ethanol production, cell growth (without solvents and with solvents selected as the best extractants and biocompatible), and global yield were evaluated. Oleic acid and soy oil were the best extraction solvents. Specific growth speed was increased by the use of solvents; the greatest increase was obtained with soy oil. Lactose consumption was 97 g/L for oleic acid extractive fermentation, 81.2 g/L for soy oil, and 64 g/L for conventional fermentation. The ethanol yield was 53 and 44 g/L with oleic acid and soy oil fermentation respectively, and 35 g/L with conventional fermentation. This study demonstrates that oleic acid alcoholic fermentation can be used to reduce the ethanol concentration in the medium and to avoid the inhibition of the yeasts during the process, which entails the possibility of using substrates with high sugar concentrations, obtaining an increase in the productivity of the system.

Potencial amilolítico do grão de milho maltado no processo de sacarificação do mesmo cereal / Amylolytic potential of maize malt in the process of sacarification of the same cereal

Objetivou-se avaliar o potencial de sacarificação do malte de milho para produção de álcool deste cereal. Para tal, foi realizada germinação do grão de milho a 20 ºC por 5 dias. O potencial amilolítico do malte de milho foi otimizado por meio de um PlanejamentoFatorial Completo 22 com 3 pontos centrais, sendo as variáveis de estudo a temperatura de incubação e concentração de substrato [S]. A sacarificação foi realizada por meio de um Planejamento de Mistura com três pontos centrais, sendo as variáveis de estudo a concentração de malte e concentração de substrato, durante 6 horas. A fermentação foi conduzida com a melhor condição de sacarificação, em reator de 5 L e volume útil de 2 L. O pH foi fixado em 4,5-5. O Brix foi acertado para o valor de 14º. O mosto hidrolisado foi esterilizado e inoculado com 5,0 g.L-1 de Saccharomyces cerevisiae proveniente de fermento comercial liofilizado. O sistema foi submetido a 2 horas de aeração inicial (170 LO2.h -1) e mantido na temperatura de 30 °C. Durante o processo de fermentação, foram realizadas determinações da concentração de células (g.L-1), açúcares redutores (mg.mL-1) e álcool (g.L-1). As variáveis temperatura de incubação e concentração de substrato foram significativos (p< 0,05) na condição de estudo de 20 % de substrato e incubação a 70 °C. Os experimentos centrais com 50 % de malte apresentaram os melhores parâmetros cinéticos com taxa volumétrica de formação de produto em etanol de 2,81 getanol.L-1.h-1 que corresponde a 8,0 ºGL e conversão de substrato em células 0,127 gcélula.gglicose-1.

The aim of this research was the production of maize malt and the evaluation of its use for alcohol production. The maize grain was allowed to germinate at 20°C for five days. The amylolitic potential of malt was optimized through a Complete Factorial Planning 22 with three central points, where the study variables were the temperature of incubation and the substrate concentration [S]. The sacarification was performed through a Planning of Mixture with three central points, where the study variables were the malt concentration and [S] during six 6 hours. The fermentation was carried out using the best sacarification condition, in a five-liter reactor and a two-liter working volume. The pH was maintained at 4.5 – 5.0. The Brix was corrected to 14º. The sterile hydrolyzedproduct was inoculated with 5.0 g.L-1 Saccharomyces cerevisiae. The system was submitted to two hours of initial aeration (170 LO2.h-1) and kept at 30°C. During the fermentation process, analyses were made to determine the cell concentrations (g.L-1), reducingsugars (mg.mL-1), alcohol (g.L-1) and °Brix. The temperature of incubation and substrate concentration were significant factors (p<0.05) in the determination of the amylolytic potential under the conditions of study utilizing 20% of substrate and incubation at 70°C. The central experiments with 50% malt presented the best kinetic parameters with a volumetric rate of ethanol formation of 2.81 gethanol.L-1.h-1, which correspond to 8.0 ºGL, and a substrate conversion in cells of 0.127gcells.gglucose -1.

STABILITY OF WINE YEAST DURING LONG-TERM USE OF STARTER CULTURES / 微生物学通报

The stability and physiological activity of the wine yeast during long-term tank to tank culture, customary enological practice, were studied under the production conditions. The results indicted that, during about one month of tank to tank culture, the yeast cells could remain the fermentative activity and the fine characters selected of the initiative yeast. On the other hand, it was also observed that, yeast cells'alcoholic fermentation of tank to tank culture might be more active than that of the initiative yeast, perhaps due to the selection by grape musts.