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Purification and properties of a β-galactosidase with potential application as a digestive supplement

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Abstract

Functional-based screening of crude β-galactosidase activities from 42 yeast strains resulted in the selection of a single enzyme of potential interest as a digestive supplement. β-Galactosidase produced by Kluyveromyces marxianus DSM5418 was purified to homogeneity by a combination of gel filtration, ion-exchange, and hydroxylapatite chromatographies. The denatured (123 kDa) and native molecular masses (251 kDa) suggest that the enzyme is a homodimer. The optimum pH and temperature of the purified enzyme were 6.8 and 37°C, respectively. The unpurified β-galactosidase in particular displayed a high level of stability when exposed to simulated intestinal conditions in vitro for 4 h. Matrix-assisted laser desorption ionization mass sectrometry analysis revealed that the enzyme's trypsin-generated peptide mass fingerprint shares several peptide fragment hits with β-galactosidases from Kluyveromyces lactis. This confirms the enzyme's identity and indicates that significant sequence homology exists between these enzymes.

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Authors and Affiliations

  1. Department of Chemical and Environmental Sciences and MSSI, University of Limerick, Limerick City, Ireland

    S. O'Connell & G. Walsh

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  1. S. O'Connell
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  2. G. Walsh
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Correspondence to G. Walsh.

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O'Connell, S., Walsh, G. Purification and properties of a β-galactosidase with potential application as a digestive supplement. Appl Biochem Biotechnol 141, 1–13 (2007). https://doi.org/10.1007/s12010-007-9206-4

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  • DOI: https://doi.org/10.1007/s12010-007-9206-4

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