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Recovery of β-galactosidase produced by Kluyveromyces lactis by ion-exchange chromatography: Influence of pH and ionic strength parameters.

Catherine Teixeira de CarvalhoSérgio Dantas de Oliveira JúniorWildson Bernardino de Brito LimaFábio Gonçalves Macêdo de MedeirosAna Laura Oliveira de Sá LeitãoJúlia Maria de MedeirosEveraldo Silvino Dos SantosGorete Ribeiro de MacedoFrancisco Canindé de Sousa Júnior
Published in: Anais da Academia Brasileira de Ciencias (2022)
The use of β-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 β-galactosidase (β-gal) by ion-exchange chromatography in a fixed-bed column. Batch adsorption tests were performed using four types of adsorbents. The β-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 β-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 β-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 β-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 β-gal of K. lactis with potential industrial applications.
Keyphrases
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