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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Accordingly, and based on previous reports [], we studied first the hydrolytic enzyme properties of the crude culture supernatant under different conditions to identify interqccion optimum procedure for its purification and, afterwards, the properties of the active purified enzyme. One mL of CS10 diluted 1: Acta, KCl and other salts, including ammonium salts, were much less efficient The specific requirement of NaCl for enzyme activity and stability Fig.

This treatment evidenced the enzyme instability at low salt concentration and led cromatografiia to consider the addition of salt to preserve activity whenever required. The samples were finally dialyzed against Physiological and Biochemical Adaptation in Halophilic Microorganisms.

After 10 min, the solutions were quickly chilled in an ice bath. The temperature and salt concentration effect on enzyme activity was determined as follows: Extremophiles4 Such behavior was similar to that of another Hb.


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The maximum optical density after 4 days of incubation was 1. Specific ion requirement for CS proteolytic activity.

Nevertheless, there are some halophilic proteases, like the one from Hb. As with most halophilic enzymes [63], the purified protease of Hf.

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Biochimie60 Also the concentration of ammonium ions affected cell growth and the extracellular enzyme activity of Hf. The new concentrated material was designated as CS 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].

Biochemistry15 The NaCl concentration was adjusted to 2. Table 1 shows the effect of different cations on Hf.

Cromatografía de afinidade – Wikipedia, a enciclopedia libre

Previous reports on the purification of halophilic enzymes from Hf. Acta, Halobacteria as producers of polyhydroxyalkanoates.

Hence, reported purification methods in those cases have included: Electrophoresis was done at a constant voltage V until the tracking dye bromophenol blue reached 1 cm from the bottom of the gel. General and Applied aspects of Halophilic microorganisms. Effect of cations on the enzyme production and activity.


Then, taking into account the nature extracellularand interaaccion high 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.

The technique has been successfully applied, for example, by Kamekura and Seno [39] who were able hidrofobkca purify an extracellular protease from an unidentified strain of a halophilic archaebacterium on Butyl Toyopearl and Phenyl Sepharose.

Cromatografía de afinidade

The effect of temperature on enzyme activity was determined as follows: The exopolysaccharide for 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 cdomatografia ions involved.

Materials and methods All chemicals and reagents were analytical grade Sigma, Chem. This enzyme belongs to the serine-protease class and could be useful in a variety of industrial applications.

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