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The acid tolerant and cold-active β-galactosidase from Lactococcus lactis strain is an attractive biocatalyst for lactose hydrolysis

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Abstract

The gene encoding the β-galactosidase from the dairy Lactococcus lactis IL1403 strain was cloned, sequenced and overexpressed in Escherichia coli. The purified enzyme has a tetrameric arrangement composed of four identical 120 kDa subunits. Biochemical characterization showed that it is optimally active within a wide range of temperatures from 15 to 55 °C and of pH from 6.0 to 7.5. For its maximal activity this enzyme requires only 0.8 mM Fe2+ and 1.6 mM Mg2+. Purified protein displayed a high catalytic efficiency of 102 s−1 mM−1 for lactose. The enzyme stability was increased by immobilization mainly at low pH (from 4.0 to 5.5) and high temperatures (55 and 60 °C). The bioconversion of lactose using the L. lactis β-galactosidase allows the production of lactose with a high bioconversion rate (98 %) within a wide range of pH and temperature.

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Acknowledgments

The authors would like to express their gratefulness to members of the Laboratory of Microbiological Genetics (INRA/Jouy-en-Josas) for their generous help and support. We thank Dr Michel Juy for his kind advices and Magali Dejob for her help. Our acknowledgements are also addressed to Helen Nyampinga for her help with English.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Laboratory for Biocrystallography and Structural Biology of Therapeutic Targets, Bases Moléculaires des Systèmes Infectieux-UMR 5086, CNRS/Université de Lyon 1, Institut de Biologie et Chimie des Protéines-FR3302, 7 Passage du Vercors, 69367, Lyon Cedex 07, France

    Violette Vincent, Nushin Aghajari, Noémie Pollet, Anaïs Boisson & Richard Haser

  2. INRA, UMR 1319 Micalis, 78350, Jouy-en-Josas, France

    Samira Boudebbouze, Emmanuelle Maguin & Moez Rhimi

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  1. Violette Vincent
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  2. Nushin Aghajari
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  3. Noémie Pollet
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  4. Anaïs Boisson
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  5. Samira Boudebbouze
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  6. Richard Haser
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  7. Emmanuelle Maguin
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  8. Moez Rhimi
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Correspondence to Moez Rhimi.

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Vincent, V., Aghajari, N., Pollet, N. et al. The acid tolerant and cold-active β-galactosidase from Lactococcus lactis strain is an attractive biocatalyst for lactose hydrolysis. Antonie van Leeuwenhoek 103, 701–712 (2013). https://doi.org/10.1007/s10482-012-9852-6

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