Preparation of highly purified eluates of acid protease from native solution Aspergillus oryzae by sorption-chromatographic and membrane methods
Abstract
This paper presents the results of preparation of highly purified acid protease eluates from the native solution of Aspergillus oryzae strain 55. The analysis of the component composition of the native solution of the Aspergillus oryzae was carried out by the elution gel chromatography method. In addition, the molecular weight of the acid protease of Aspergillus oryzae was determined. The study of the native solution by the method of electrophoresis confirmed the obtained data. Isolation and purification of acid protease from ballast impurities was performed by sorption on various cation exchangers and by ultrafiltration. Optimal conditions for sorption of acid protease from the native solution of Aspergillus oryzae on cation exchangers KU-23, С-106, KB-2 were chosen, and equilibrium parameters of sorption of acid protease on these sorbents were studied. Sorption isotherms were constructed and the distribution coefficient Kd was calculated. Isotherms of protease sorption on carboxyl cation exchangers (KB-2 and C-106) are characterized by the presence of a maximum. Sorption of the protease on the sulfate cationite KU-23 corresponds to the Langmuir model. The maximum selectivity and sorption capacity were obtained on the KU-23 and C-106 sorbents, the values are comparable and are m=5800 U/g, Kd=36.2 сm3/g for KU-23 and m=5600 U/g, Kd=35.0 сm3/g for
C-106.
A kinetic-dynamic analysis of the sorption of acid protease on ionites C-106 and KU-23 with speed of process 0.4-0.9 ml/min was carried out. The maximum yield of the desired product was obtained on a KU-23 sulfate cationite at a process rate of ω=0.8 сm3/ min. It is shown that the maximum yield of the target
product is achieved on the sulfocationite KU-23, and is 66% at the sorption stage and 76% at the desorptionstage. Purification of acid protease eluate using ultra - and diafiltration can increase the specific activity by 4.5 times.
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