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High yield expression of Lipase A from Candida antarctica in the methylotrophic yeast Pichia pastoris and its purification and characterisation

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Abstract

The current investigation focuses on shedding further light on the characteristics of lipase A from Candida antarctica (CalA), which has attracted growing attention in its suitability for industrial applications. CalA was functionally expressed in the methylotrophic yeast Pichia pastoris, purified and characterised. A classical fed-batch process and a semi-continuous process were developed and tested with regard to their yield capacity. Lipase concentrations of 0.88 and 0.55 g l−1 were obtained using the fed-batch and semi-continuous processes, respectively. The semi-continuous process reaches a total activity of 10,233,000 U and so surpasses the fed-batch process reaching 7,530,000 U. The purified enzyme showed highest activity between 50 and 70 °C at pH 7.0 and a preference for short-chain triglycerides (C4-C8). Significantly reduced activity was observed in the presence of hydrophilic esters.

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Acknowledgement

We thank Nestec Ltd. for its financial support to this work.

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Authors and Affiliations

  1. Institute of Technical Biochemistry, University of Stuttgart, Allmandring 31, 70569, Stuttgart, Germany

    Jan Pfeffer, Sven Richter, Jens Nieveler, Rolf D. Schmid & Monika Rusnak

  2. Nestlé Research Center Lausanne, Nestec Ltd., Vers chez les blanc, P.O. Box 44, 1000, Lausanne 26, Switzerland

    Carl-Erik Hansen & Rachid Bel Rhlid

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  1. Jan Pfeffer
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  2. Sven Richter
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  3. Jens Nieveler
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  4. Carl-Erik Hansen
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  6. Rolf D. Schmid
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  7. Monika Rusnak
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Correspondence to Monika Rusnak.

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Pfeffer, J., Richter, S., Nieveler, J. et al. High yield expression of Lipase A from Candida antarctica in the methylotrophic yeast Pichia pastoris and its purification and characterisation. Appl Microbiol Biotechnol 72, 931–938 (2006). https://doi.org/10.1007/s00253-006-0400-z

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  • DOI: https://doi.org/10.1007/s00253-006-0400-z

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