Cell permeabilization of Kluyveromyces and Saccharomyces species to obtain potential biocatalysts for lactose hydrolysis

Yeast's beta-galactosidase is an intracellular enzyme, through which it is possible to determine in vivo its activity as a biocatalyst in the lactose hydrolysis. Permeabilization process was used for transforming the microorganisms cells into biocatalysts with an enhanced enzyme activity. The potential application of this enzyme technology in industrial process depends mainly on the enzyme activity. Beta-galactosidase enzyme that hydrolyzes lactose, for instance, is largely dependent on the reaction time and its stability under different physical conditions, such as pH, temperature and enzyme concentration. The objective of this study was to optimize the cellular permeabilization process of Kluyveromyces marxianusCCT 3172 and Saccharomyces fragilisCCT 7586 cultured in cheese whey for lactose hydrolysis. Box-Behnken design was carried out for cell permeabilization with three independent variables, ethanol concentration, permeabilization time and temperature. The best permeability conditions for K. marxianusCCT 3172 were 27% (v v-1) ethanol, 3 min at 20ºC, with specific enzymatic activity of 0.98 U mg-1.For S. fragilisCCT 7586, a specific enzymatic activity of 1.31 U mg-1was achieved using 45% (v v-1) of ethanol, 17 min. of reaction under 17ºC. Thus, it was concluded that cellular permeabilization with ethanolis an efficient process to determine beta-galactosidase activity.(AU)

What do patents tell us about microalgae in agriculture?

Microalgae have been used widely as a biological source for several industries, such as biofuel, pharmaceutical and food. Recently, the agricultural industry has also began using microalgae as an alternative source for sustainable products to replace agrochemicals. Due to the lack of scientific articles in this research area, the objective of this study was to search for applications of microalgae and to characterize its use in agriculture using the patent documents available in three patent databases, World Intellectual Property Organization (WIPO), European Patent Office (EPO) and Brazilian Institute of Industrial Property (INPI). The search was carried out using the keyword "microalgae" and applying the filter for International Patent Classification (IPC) code "A01N" which corresponds to patents related to agriculture and cultivation of microalgae. Our patent database search returned 669 documents and 132 patents were selected for the study based on their abstracts. The first patent was registered in 1982 and described the use of microalgae Chlorella extract as a plant growth promoter. After that, no patent was registered for 15 years. From 2005 to 2014, only seven patents were found. However, the scenario changed from 2015 when the number of patents increased mainly in the United States, China and Europe. The patent analysis showed several applications for microalgae in the agricultural sector, such as plant growth promotion, biofertilization, plant disease control, weed management, and post-harvest quality. This review confirmed the increasing interest in microalgae-derived products in agriculture and the value of using patent documents to assess innovative areas.

Permeabilization of Saccharomyces fragilis IZ 275 cells with ethanol to obtain a biocatalyst with lactose hydrolysis capacity / Permeabilização de células de Saccharomyces fragilis IZ 275 com etanol para obtenção de biocatalisador com capacidade de hidrólise de lactose

The permeabilization was used to transform microorganisms in cell biocatalysts with high enzymatic activity. The Saccharomyces fragilis IZ 275 yeast cells were permeabilized with ethanol, as permeabilizing agent. To optimize the permeabilization conditions were used the design of Box-Behnken 15 trials (3 central points). The independent variables and their levels were ethanol (29, 32 and 35%), temperature (15, 20 and 25°C) and time (15, 20 and 25 min). The answer (Y) function has beta-galactosidase activity (U mg-1). The optimum conditions for obtaining a high enzymatic activity were observed in 35% ethanol concentration, temperature 15ºC and 20 min. treatment time. The maximum activity of the enzyme beta-galactosidase obtained was 10.59 U mg-1. The permeabilization of the S. fragilis IZ 275 cells was efficient.

A permeabilização foi usada para transformar células de microrganismos em biocatalisadores com alta atividade enzimática. As células de levedura de Saccharomyces fragilis IZ 275 foram permeabilizadas com etanol, como agente permeabilizante. Para otimizar as condições de permeabilização foi utilizado o delineamento de Box-Behnken com 15 ensaios (3 repetições no ponto central) . As variáveis independentes e seus níveis foram etanol (29, 32 e 35%), temperatura (15, 20 e 25ºC) e tempo (15, 20 e 25 min.). A função resposta (Y) foi atividade de beta-galactosidase (U mg-1). As condições ótimas para a obtenção de uma alta atividade enzimática foram observadas em 35% de concentração de etanol, temperatura de 15°C e tempo de tratamento de 20 minutos. A máxima atividade da enzima beta-galactosidase obtida foi de 10.59 U mg-1. A permeabilização das células de S. fragilis IZ 275 foi eficiente.

Permeabilization of Saccharomyces fragilis IZ 275 cells with ethanol to obtain a biocatalyst with lactose hydrolysis capacity / Permeabilização de células de Saccharomyces fragilis IZ 275 com etanol para obtenção de biocatalisador com capacidade de hidrólise de lactose

The permeabilization was used to transform microorganisms in cell biocatalysts with high enzymatic activity. The Saccharomyces fragilis IZ 275 yeast cells were permeabilized with ethanol, as permeabilizing agent. To optimize the permeabilization conditions were used the design of Box-Behnken 15 trials (3 central points). The independent variables and their levels were ethanol (29, 32 and 35%), temperature (15, 20 and 25°C) and time (15, 20 and 25 min). The answer (Y) function has betagalactosidase activity (U mg-1). The optimum conditions for obtaining a high enzymatic activity were observed in 35% ethanol concentration, temperature 15ºC and 20 min. treatment time. The maximum activity of the enzyme beta-galactosidase obtained was 10.59 U mg-1. The permeabilization of the S. fragilis IZ 275 cells was efficient.(AU)

A permeabilização foi usada para transformar células de microrganismos em biocatalisadores com alta atividade enzimática. As células de levedura de Saccharomyces fragilis IZ 275 foram permeabilizadas com etanol, como agente permeabilizante. Para otimizar as condições de permeabilização foi utilizado o delineamento de Box-Behnken com 15 ensaios (3 repetições no ponto central). As variáveis independentes e seus níveis foram etanol (29, 32 e 35%), temperatura (15, 20 e 25ºC) e tempo (15, 20 e 25 min.). A função resposta (Y) foi atividade de beta-galactosidase (U mg-1). As condições ótimas para a obtenção de uma alta atividade enzimática foram observadas em 35% de concentração de etanol, temperatura de 15°C e tempo de tratamento de 20 minutos. A máxima atividade da enzima beta-galactosidase obtida foi de 10.59 U mg-1. A permeabilização das células de S. fragilis IZ 275 foi eficiente.(AU)

Dynamics of ethanol production from deproteinized whey by Kluyveromyces marxianus: An analysis about buffering capacity,thermal and nitrogen tolerance

The production of value-added products could be a valuable option for cheese wastewater management. However, this kind of study cannot just focus alone on getting the final product. This also necessitates studies on the dynamics of bioprocesses. With these as background, the present investigation aimed at evaluating the buffering capacity of deproteinized whey and effect of temperature and nitrogen source on ethanol yields from it. The batch fermentation conditions used to evaluate ethanol production were temperatures 30, 35, 40°C and pH 4.5, 5.0, 5.5, 6.0. To study the influence of nitrogen source on ethanol yield, a design matrix was applied using yeast extract and (NH4)2SO4.The final pH was analyzed to evaluate the buffering capacity. The results showed that the Kluyveromyces marxianus was thermotolerance to produce ethanol at 35 and 40°C, which was not observed at 30°C. Results also showed that the deproteinization procedure did not affect the buffering capacity of cheese whey. Finally, higher ethanol production was obtained using yeast extract (3% v/v). These results could be important for developing low-cost method for industrial production of ethanol from deproteinized whey.