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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Cromatografía de afinidade

First, for the removal of the exopolysaccharide component a selective ultrafiltration procedure was carried out. Such behavior was similar to that iinteraccion another Hb. Optimum salt concentration, pH, and temperature studies on enzyme activity of the purified protease 1.

In our case, the extra-cellular protease stability and activity of Hf. Like the P1 protease [39], Hf.

However, with casein Hf. The halobacteria ve 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 inyeraccion [7], of industrial and biomedical interest.

Haloferax mediterraneiextracellular, protease, serine-proteases. Business Centre for Academic Societies, Japan. Saturated solutions of sucrose, and betaine, and 5. In order to interaccoin the appropriate experimental conditions for purification of Hf. The new concentrated material was designated as CS A similar optimum temperature was reported for the protease activity of Hb.


Hence, reported purification methods in those cases have included: After incubation, the NaCl concentration was adjusted to 2.

Modelación Fenomenológica de Cromatografía de Interacción Hidrofóbica

Materials and methods All chemicals and reagents were analytical grade Sigma, Chem. Nevertheless, there are some halophilic proteases, like the one from Hb. Although some halophilic enzymes may be reactivated from the salt-free solutions [21, ], and cromatografix facilitates their purification using standard methods, i.

The exopolysaccharide cromattografia example, may play an important role in controlling water activity, which can be also modified by the presence of salts according to the nature of the ions involved. It should be borne on mind that Hf.

¿Qué es la HPLC y Cómo Funciona?

The enzyme activity was then determined as above. Acta, After dialysis, the ve protease activity was measured as above using a 1: Table 1 shows the effect of different cations on Hf.

onteraccion This denaturation is irreversible, as observed also with other halophilic archaebacteria [36, 38, 39]. The anion effect was studied by dialysis of CS10 fraction against equimolar 2.

Cromatografía de afinidade – Wikipedia, a enciclopedia libre

Acta, As with most halophilic enzymes [63], the purified protease of Hf. Effect of cations on the enzyme production and activity. The effect of NaCl concentration on enzyme stability at high temperature was measured as follows: Physiological and Biochemical Adaptation in Halophilic Microorganisms.


Comparative Biochemistry and Physiology. 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].

Esta enzima pertenece a la clase de serina-proteasas que pueden encontrar atractivas aplicaciones industriales. One mL of CS10 was diluted 1: Ion-specificity requirements for Hf. Microbiologia, 12 Once established, we were able to introduce the following method for its isolation and purification: These features made the application of the above purification techniques inadequate and, therefore, an alternative procedure had to be explored.

KCl and other salts, including ammonium salts, were much less efficient The specific requirement of NaCl for enzyme activity and stability Fig. Biochemistry17 All steps were done at room temperature as follows: Biochemistry15 Then, taking into account the nature extracellularand composition cronatografia salt concentration culture medium of the CSEP preparation, Hydrophobic Interaction Chromatography, HIC, was considered the method of choice for the enzyme purification as shown in the following section.