alexa Purification and Characterization of β-galactosidase from Kluveromyces lactis Isolated from a Yoghurt Waste Site
E- ISSN: 2320 - 3528
P- ISSN: 2347 - 2286

Research & Reviews: Journal of Microbiology and Biotechnology
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Research Article

Purification and Characterization of β-galactosidase from Kluveromyces lactis Isolated from a Yoghurt Waste Site

Tolulope T Oluwaniyi1*, Bridget O Omafuvbe2, Femi K. Agboola3

1Department of Microbiology, Faculty of Science Ekiti State University Ado- Ekiti, Nigeria

2Department of Microbiology, Faculty of Science, Obafemi Awolowo University, Ile-Ife, Nigeria

3Department of Biochemistry, Faculty of Science, Obafemi Awolowo University, Ile Ife, Nigeria

*Corresponding Author:
Tolulope T. Oluwaniyi
Department of Microbiology, 2 Department of Biochemistry, Obafemi Awolowo University, Ile Ife 220005, Nigeria
Tel: +234 803 7040414
E-mail: [email protected]

Received date: 26/06/2015; Accepted date: 25/02/2016; Published date: 04/03/2016



Some properties of β-galactosidase extracted and purified from Kluyveromyces lactis isolated from a yoghurt waste site were investigated. The crude β-galactosidase isolated from Kluyveromyces lactis was purified by ammonium sulphate precipitation and DEAE-Sephadex ion exchange chromatography. Specific activity of the partially purified enzyme was 107.50 μmole/min/mg of protein and a 7% yield. The molecular weight of the enzyme was 199 kDa as determined by gel filtration on Bio-gel P-200. The Michealis-Menten constant (Km) and maximum velocity (Vmax) value obtained was 3.56 ± 0.62 mM and 0.61 ± 0.08 μmole/min/ml for the enzyme with ortho-nitrophenyl-β-D-galactopyranoside (oNPG) substrate while the Km and Vmax was 18.01 ± 0.23 mM and 0.15 ± 0.04 mmol/20 min/L with lactose substrate. The optimum pH of purified enzyme was 7.5 and 6.5 with oNPG and lactose substrate respectively. The optimum temperature was 40°C with both substrates. This β-galactosidase may have applications in the conversion of whey a by-product of dairy industries into useful products.


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