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

Authors :
Carvalho CT
Oliveira Júnior SD
Lima WBB
Medeiros FGM
Leitão ALOS
Dantas JMM
Santos ESD
Macêdo GR
Sousa Júnior FC
Source :
Anais da Academia Brasileira de Ciencias [An Acad Bras Cienc] 2022 Jan 17; Vol. 94 (1), pp. e20200752. Date of Electronic Publication: 2022 Jan 17 (Print Publication: 2022).
Publication Year :
2022

Abstract

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.

Details

Language :
English
ISSN :
1678-2690
Volume :
94
Issue :
1
Database :
MEDLINE
Journal :
Anais da Academia Brasileira de Ciencias
Publication Type :
Academic Journal
Accession number :
35043852
Full Text :
https://doi.org/10.1590/0001-3765202220200752