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An Acad Bras Cienc ; 94(1): e20200752, 2022.
Article in English | MEDLINE | ID: mdl-35043852

ABSTRACT

The use of ß-galactosidase in food products has been a major focus of the industry. Therefore, the development of efficient and inexpensive methodologies to purify it is essential. Thus, this study aimed to recover the enzyme ß-galactosidase (ß-gal) by ion-exchange chromatography in a fixed-bed column. Batch adsorption tests were performed using four types of adsorbents. The ß-gal adsorption capacity in batch mode using Streamline DEAE resin presented the best performance, with a retention capacity of 18.77 ± 0.14 U/g at pH 6.0. A 22 experimental design was applied to optimize the ß-gal recovery using an AKTA Start system, evaluating the ionic strength and the pH as process parameters. The results showed that ionic strength exerted a greater influence on fold purification (FP). The ß-gal fraction in elution using 0.1-0.4 M of NaCl showed a yield of 51.65 ± 0.17% and FP of 2.00 ± 0.43. Electrophoresis confirmed the ß-gal recovery, where an evident band with a molecular weight between 60 and 120 kDa was observed. These results point to the recovery of a stable ß-gal of K. lactis with potential industrial applications.


Subject(s)
beta-Galactosidase , Chromatography, Ion Exchange , Hydrogen-Ion Concentration , Kluyveromyces , Osmolar Concentration
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