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Development and validation of a medium for recombinant endo-β-1,4-xylanase production by Kluyveromyces lactis using a statistical experimental design

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Abstract

Kluyveromyces lactis is an excellent host for a high cell density culture, which allows high expression levels of recombinant enzymes. Nutrient composition and culture conditions affect the secretion, production level and stability of the recombinant host. Therefore, it is technologically important to formulate a medium that stimulates high cell density and enhances the desired enzyme production using K. lactis GG799. In this study, six media were initially compared, and a Plackett-Burman experimental design was employed to screen for important components and trace elements. Nitrogen sources such as ammonium sulfate and free amino acid (casamino acid) as well as compounds like MgSO4∙7H2O, Na2SO4, ZnSO4∙6H2O, MnSO4∙4H2O and KH2PO4 affected biomass concentrations (5.67 g/l) and recombinant endo-β-1,4-xylanase (Xyn2) production (49.73 U/ml). Optimum productivity was obtained at shorter incubation times (i.e., 6 h), making the medium suitable for use when seeking efficient production. Expression of recombinant Xyn2 by K. lactis GG799 in the designed medium resulted in satisfactory recombinant Xyn2 volumetric productivity (vp) at 8.29 U/ml/h. When compared to the rich, non-selective YPD medium, the designed medium improved biomass output and recombinant Xyn2 production in K. lactis GG799 by approximately 9 and 22%, respectively.

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Acknowledgement

This project was supported by the Genomics and Molecular Biology Initiatives Programme of the Malaysia Genome Institute, Ministry of Science, Technology and Innovation Malaysia (Project No. 07-05-16-MGI-GMB12).

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Authors and Affiliations

  1. Department of Bioprocess Engineering, Faculty of Chemical Engineering, Universiti Teknologi Malaysia, 81310, Skudai, Johor, Malaysia

    Siti Fatimah Zaharah Mohamad Fuzi, Roshanida A. Rahman & Rosli Md. Illias

  2. Malaysia Genome Institute, Universiti Kebangsaan Malaysia, Bangi, Selangor, Malaysia

    Nor Muhammad Mahadi

  3. Department of Chemical and Process Engineering, Faculty of Engineering and Built Environment, Universiti Kebangsaan Malaysia, 43600, UKM Bangi, Selangor, Malaysia

    Jamaliah Md. Jahim

  4. School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, UKM Bangi, Selangor, Malaysia

    Abdul Munir Abd. Murad

  5. Centre for Gene Analysis and Technology, of Science and Technology, Universiti Kebangsaan Malaysia, 43600, UKM Bangi, Selangor, Malaysia

    Farah Diba Abu Bakar

  6. Department of Chemical Engineering, Faculty of Chemical Engineering, Universiti Teknologi Malaysia, Skudai, Johor, Malaysia

    Mazura Jusoh

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  1. Siti Fatimah Zaharah Mohamad Fuzi
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  2. Nor Muhammad Mahadi
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Correspondence to Rosli Md. Illias.

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Fuzi, S.F.Z.M., Mahadi, N.M., Jahim, J.M. et al. Development and validation of a medium for recombinant endo-β-1,4-xylanase production by Kluyveromyces lactis using a statistical experimental design. Ann Microbiol 62, 283–292 (2012). https://doi.org/10.1007/s13213-011-0258-x

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  • DOI: https://doi.org/10.1007/s13213-011-0258-x

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