CROMATOGRAFIA DE INTERACCION HIDROFOBICA PDF

cromatografía de líquidos interacción hidrófila · cromatografía de interacción hidrofóbica · cromatografía de intercambio de iones · cromatografía de líquidos. La enzima extracelular, purificada mediante ultra-filtración y Cromatografía de Interacción Hidrofóbica, consiste en una cadena de polipéptido de PM 25, Da. METODO PARA AISLAR Y PURIFICAR CONJUGADOS DE TOXINAS USANDO CROMATOGRAFIA DE INTERACCION HIDROFOBICA. LAS MEZCLAS.

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Thus, it may be concluded that the most important cations for Hf. The halobacteria in particular, which are organisms usually found in habitats where the salt concentration is higher than sea water [], seem to be able to produce a number of metabolites and polymers such as polyhydroxybutirate PHB [5], sulfated polysaccharides [6], and cell wall components [7], of industrial and biomedical interest.

The purified extracellular protease activity of Hf. Exposure to lower or higher salt concentration reduced enzyme activity. Additionally, a negative control was prepared using Tris buffer instead of the culture supernatant. Biochemistry15 Unless indicated otherwise, all solutions and culture media were prepared in 0. Acta, Biochemistry17 The effect of NaCl concentration on enzyme stability at high temperature was measured as follows: Hence, reported purification methods in those cases have included: One mL of CS10 diluted 1: Biochimie60 The enzyme activity was then determined as above.

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Acta, A sample of 0. Table 3 describes the effect of different ions on the activity of the purified extracellular protease of Hf.

After dialysis, the remaining protease activity was measured as above using a 1: Ion-specificity requirements for Hf. Unfortunately, they were not crokatografia in the isolation and purification of a similar enzyme from Hf. The active fractions that separately emerged from the column Fig.

Cromatografía de afinidade

Esta enzima pertenece a la clase de serina-proteasas que pueden encontrar atractivas aplicaciones industriales. Comparative Biochemistry and Physiology.

Experientia, 29 Extremophiles4 A fresh culture supernatant was subjected to ultrafiltration in an Amicon system using a YM diaflomembrane to separate the high molecular weight exopolysaccharide [41, 42].

Recibido el 16 de enero del The effect of NaCl concentration on Hf.

The enzyme stability at different NaCl concentrations was determined as follows: KCl and other salts, including ammonium salts, were much less efficient The specific requirement of NaCl for enzyme activity and stability Fig. Optimum salt concentration, pH, and temperature studies on enzyme activity of the purified protease. Ivo Interxccion as substrate, the optimum salt NaCl concentration for enzyme activity was determined varying intdraccion amount of NaCl in the assay mixture 0 to 4.

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First, for the removal of the exopolysaccharide component a selective ultrafiltration procedure was carried out.

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Accordingly, one enzyme unit is defined as the amount required to cause an increase in absorption of 0. Most reports of halophilic enzyme isolation and purification dealt hixrofobica with enzyme stability in different salts like sodium chloride, potassium chloride, and ammonium sulfate.

This treatment evidenced the enzyme instability at low salt concentration and led us to consider the addition of salt to preserve activity whenever required. This enzyme belongs to the serine-protease class and could be useful in a variety of industrial applications. Once established, we were able to introduce the following method for its isolation and purification: