Utjecaj reakcijskih parametara na imobilizaciju enzima adsorpcijom i unakrsnim vezivanjem

The activity of enzymes after the immobilization by weak interactions such as adsorption or adsorption followed by a cross-linking reaction can change easily not only during further application but during the activity measured via desorption and/or inactivation of enzymes. The changes in activity could be a consequence of the interactions between the enzyme and support. In this work a quick and efficient method is developed that permits studying of the strength and properties of the forces between the enzymes and support by examining the kinetics of desorption and/or inactivation. Applying this new cyclic kinetic desorption method, the effect of reaction parameters on immobilization could also be studied. The efficiency of this method was tested for optimizing the parameters for immobilization of β-glucosidase on Amberlite IRA 900 anion exchange resin by an adsorption followed by a cross-linking with glutaraldehyde to form potential preparation for food industry. The following parameters were chosen: concentration (0.10 M) of the buffer (sodium acetate, pH=5.5) and the ratio of carrier to enzyme (10:1) for the adsorption step, then the time of treatment (1 min) and concentration (0.25, by mass per volume) of glutaraldehyde for the cross-linking by using the kinetic desorption method. The activity of this preparation was 57 μmol/(min·g) in respect of dry resin. It was established that the effect of different parameters on this enzyme immobilization could be characterized by the new cyclic kinetic desorption method in a quick and efficient way; furthermore, it permitted separate testing of the effect of parameters on the adsorption and cross-linking processes.Imobilizacija enzima adsorpcijom ili adsorpcijom s unakrsnim vezivanjem utječe na njegovu aktivnost, koja se može izmjeriti desorpcijom i/ili inaktivacijom enzima. Promjena je aktivnosti vjerojatno posljedica slabih veza između enzima i podloge. Da bi se istražile jakost i snaga tih veza, razvijena je brza i učinkovita metoda kojom je ispitana kinetika desorpcije i/ili inaktivacije, te utjecaj reakcijskih parametara na imobilizaciju. Uspješnost je metode provjerena optimiranjem parametara imobilizacije β-glukozidaze adsorpcijom i unakrsnim vezivanjem s glutaraldehidom na anionskom izmjenjivaču Amberlite IRA 900. Odabrane su ove vrijednosti: koncentracija acetatnog pufera (pH=5,5) od 0,1 M, omjer nosača i enzima od 10:1, vrijeme imobilizacije od 1 min i koncentracija glutaraldehida od 0,25 m/V. Pritom je aktivnost enzima bila 57 µmol/(min·g). Utvrđeno je da se ovom metodom može brzo i učinkovito ispitati utjecaj različitih parametara na imobilizaciju, tj. adsorpciju i unakrsno vezivanje enzima

Magyar Tanítóképző 38 (1925) 7-10

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Magyar Tanítóképző 50 (1937) 7

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Magyar Tanítóképző 50 (1937) 4

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Testing of the Effect of Reaction Parameters on the Enzyme Immobilization by Adsorption and Cross-Linking Processes with Kinetic Desorption Method

The activity of enzymes after the immobilization by weak interactions such as adsorption or adsorption followed by a cross-linking reaction can change easily not only during further application but during the activity measured via desorption and/or inactivation of enzymes. The changes in activity could be a consequence of the interactions between the enzyme and support. In this work a quick and efficient method is developed that permits studying of the strength and properties of the forces between the enzymes and support by examining the kinetics of desorption and/or inactivation. Applying this new cyclic kinetic desorption method, the effect of reaction parameters on immobilization could also be studied. The efficiency of this method was tested for optimizing the parameters for immobilization of β-glucosidase on Amberlite IRA 900 anion exchange resin by an adsorption followed by a cross-linking with glutaraldehyde to form potential preparation for food industry. The following parameters were chosen: concentration (0.10 M) of the buffer (sodium acetate, pH=5.5) and the ratio of carrier to enzyme (10:1) for the adsorption step, then the time of treatment (1 min) and concentration (0.25, by mass per volume) of glutaraldehyde for the cross-linking by using the kinetic desorption method. The activity of this preparation was 57 μmol/(min·g) in respect of dry resin. It was established that the effect of different parameters on this enzyme immobilization could be characterized by the new cyclic kinetic desorption method in a quick and efficient way; furthermore, it permitted separate testing of the effect of parameters on the adsorption and cross-linking processes