Abstract:The identification of yeasts isolated from spoiled Jubileu peach puree using the API 20C AUX method and a commercial yeast as witness were studied. Subsequently, the yeast's growth potential using two batch culture treatments were performed to evaluate number of colonies (N), reducing sugar concentration (RS), free-invertase (FI), and culture-invertase activity (CI). Stock cultures were maintained on potato dextrose agar (PDA) slants at 4 °C and pH 5 for later use for batch-culture (150 rpm) at 30°C for 24 h, … Show more
“…To ensure the experiment's accuracy, selection of 3-5 generations of cells of well-grown is for the experiment. 5: Strictly complying with aseptic operation to avoid contamination (Ferreira et al, 2016).…”
This study amied to explore the culture method of biliary tract fibroblasts from Diannan small-ear pigs, and lay the foundation for further exploration on biliary tract benign stenosis. The biliary tract injury model was constructed via adopting Diannan smallear pigs, and the tissue adherence method was adopted. After trying different medium and fetal bovine serum concentrations, the culture method of mature Diannan small-ear pig biliary tract fibroblast was explored. Additionally, vimentin and desmin was examined and identified as fibroblasts. The biliary tract fibroblasts that can be passaged were available to be cultured via tissue adherence method. Biliary tract fibroblast, a sort of cell with proliferation and multidirectional differentiation potential, is difficult to be cultured and passed on, and its successful culture lays a good foundation for further exploration on drug intervention.
“…To ensure the experiment's accuracy, selection of 3-5 generations of cells of well-grown is for the experiment. 5: Strictly complying with aseptic operation to avoid contamination (Ferreira et al, 2016).…”
This study amied to explore the culture method of biliary tract fibroblasts from Diannan small-ear pigs, and lay the foundation for further exploration on biliary tract benign stenosis. The biliary tract injury model was constructed via adopting Diannan smallear pigs, and the tissue adherence method was adopted. After trying different medium and fetal bovine serum concentrations, the culture method of mature Diannan small-ear pig biliary tract fibroblast was explored. Additionally, vimentin and desmin was examined and identified as fibroblasts. The biliary tract fibroblasts that can be passaged were available to be cultured via tissue adherence method. Biliary tract fibroblast, a sort of cell with proliferation and multidirectional differentiation potential, is difficult to be cultured and passed on, and its successful culture lays a good foundation for further exploration on drug intervention.
“…O microrganismo usado neste experimento foi isolado a partir de resíduo de pêssego e identificado como C. guilliermondii pelo sistema API 20C AUX, com 95% de exatidão e perfil numérico 6776277 (FERREIRA et al, 2016). Os extratos enzimáticos foram obtidos usando o método de extração com bicarbonato de sódio segundo Ferreira et al (2018) e a purificação destes foi feita através de precipitação das proteínas com acetona seguido de diálise.…”
Section: Metodologiaunclassified
“…Outra levedura, a Candida utilis, também apresenta duas isoformas (GUIMARÃES et al, 2007). Em purê de pêssego e seus resíduos do processamento foram identificados e quantificados vários micro-organismos destacando-se a levedura S. cerevisiae (LOPEZ et al 2014) com potencial para produção de invertase (FERREIRA et al 2016). O objetivo desse trabalho foi estudar o efeito do pH, da temperatura e concentração de sacarose na atividade da invertase de C. guilliermondii (ICg) extraída de resíduo de pêssego e recuperada por precipitação com acetona bem como identificar as isoenzimas por eletroforese.…”
RESUMO -O objetivo deste trabalho foi verificar o efeito do pH, da temperatura e da concentração de sacarose na atividade da invertase de Candida guilliermondii (ICg) extraída de resíduos de pêssego e identificada pelo sistema API 20C AUX. Os extratos enzimáticos foram obtidos com solução de bicarbonato de sódio, purificados por precipitação com acetona e as isoenzimas foram detectadas por eletroforese em gel de poliacrilamida a 6% usando BSA como marcador de proteína. A atividade enzimática e o teor de proteínas foram determinados com os métodos do ácido 3,5-DNS e Lowry respectivamente. A recuperação da invertase da ICg foi de 56% com atividade ótima a pH 5 a 50°C e na eletroforese foram detectadas três frações enzimáticas. A cinética enzimática demonstrou que a invertase da ICg teve comportamento michaeliano sendo mais específica a 50°C do que a 25°C.
Resumen -La extracción fue realizada a través de un proceso de autolisis, donde las enzimas degradan las estructuras celulares y liberan el contenido citoplasmático al medio extracelular. El proceso de autolisis fue realizado en shaker con diferentes velocidades de agitación (50 -350 rpm) y concentraciones de bicarbonato de sodio (50 -350 mM NaHCO 3 ) a 40 °C por 24 horas, usando un diseño experimental compuesto central adaptado para dos variables y cinco niveles, así como una metodología de superficie de r e s p u e s t a (MSR) y análisis canónico para definir las condiciones óptimas de extracción de invertasa de S. cerevisiae aislada de puré de durazno (ISc). En la condición optima de extracción, se estudió el efecto de la liofilización y purificación en la actividad de la invertasa, usando el método colorimétrico del ácido 3,5-dinitrosalicílico (DNS) a 490 nm para acompañar su actividad expresada en unidades. Una unidad (U) es definida como 1 mg glucosa.min -1 . El valor de proteínas fue cuantificado por el método Lowry y la invertasa de levadura comercial (ILC) como testigo. La influencia de la concentración de NaHCO 3 y de la velocidad de agitación en la extracción de la enzima invertasa indicó efecto lineal, cuadrático e interactivo. El modelo de forma polinómica de segundo orden explicó el fenómeno de extracción de la invertasa con un R 2 de 0,80, y con actividad significativamente dependiente de ambas variables. La máxima extracción fue observada en el punto central experimental (200 mM NaHCO 3 y 200 rpm) con una actividad de 14,7 U.mg -1 . El punto estacionario es un punto de máxima extracción, el cual difiere en menos de 10% del punto central experimental. De acuerdo a estas características, las óptimas condiciones para extracción de la invertasa de S. cerevisiae aislada de puré de durazno son 200 mM NaHCO 3 como agente de autolisis, 200 rpm de agitación orbital a 40 o C durante 24 horas y usando una biomasa de levaduras liofilizadas. El etanol es más efectivo que la acetona para la recuperación de la actividad específica. Términos para indexación: levadura; autolisis; MSR; análisis canónico.
Optimized extraction and partial purification of S. Cerevisiae invertase from peach pureeAbstract -The extraction was performed using an autolysis process where enzymes degrade cell structures and release the cytoplasmic contents to the extracellular medium. The autolysis process was carried out in shake flasks with different agitation speeds (50 -350 rpm) and the concentration of sodium bicarbonate (50 -350 mM NaHCO 3 ) at 40 ° C for 24 hours, using a central composite design adapted for two variables and five levels and response surface methodology (RSM) and canonical analysis to define the optimal conditions of extraction of invertase S. cerevisiae from peach puree (ISc). In great condition extraction, we studied the effect of the liofilization and purified in the invertase activity using colorimetric method of 3,5-dinitrosalicílico (DNS) at 490 nm to monitor their activity. One unit (U) defined as 1 mg glicose....
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