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Uml2 is a novel CalB-type lipase of Ustilago maydis with phospholipase A activity

Applied Microbiology and Biotechnology volume 98, pages 4963–4973 (2014)Cite this article

Abstract

CalB of Pseudozyma aphidis (formerly named Candida antarctica) is one of the most widely applied enzymes in industrial biocatalysis. Here, we describe a protein with 66 % sequence identity to CalB, designated Ustilago maydis lipase 2 (Uml2), which was identified as the product of gene um01422 of the corn smut fungus U. maydis. Sequence analysis of Uml2 revealed the presence of a typical lipase catalytic triad, Ser-His-Asp with Ser125 located in a Thr-Xaa-Ser-Xaa-Gly pentapeptide. Deletion of the uml2 gene in U. maydis diminished the ability of cells to hydrolyse fatty acids from tributyrin or Tween 20/80 substrates, thus demonstrating that Uml2 functions as a lipase that may contribute to nutrition of this fungal pathogen. Uml2 was heterologously produced in Pichia pastoris and recombinant N-glycosylated Uml2 protein was purified from the culture medium. Purified Uml2 released short- and long-chain fatty acids from p-nitrophenyl esters and Tween 20/80 substrates. Furthermore, phosphatidylcholine substrates containing long-chain saturated or unsaturated fatty acids were effectively hydrolysed. Both esterase and phospholipase A activity of Uml2 depended on the Ser125 catalytic residue. These results indicate that Uml2, in contrast to CalB, exhibits not only esterase and lipase activity but also phospholipase A activity. Thus, by genome mining, we identified a novel CalB-like lipase with different substrate specificities.

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Acknowledgments

This work was partly financed by the Ministry of Innovation, Science and Research of the German state of North-Rhine Westphalia (MIWF) and the European Regional Development Fund (EFRE). We thank the Ministry of Innovation, Science and Research of North Rhine-Westphalia and Heinrich Heine University Düsseldorf for a scholarship within the CLIB2021-Graduate Cluster Industrial Biotechnology.

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  1. Institut für Molekulare Mykologie, Heinrich-Heine-Universität, Universitätsstr. 1/26.12, 40225 Düsseldorf, Germany

    Christoph Buerth, Joachim F. Ernst & Denis Tielker

  2. Institut für Molekulare Enzymtechnologie, Heinrich-Heine-Universität, Düsseldorf, Germany

    Filip Kovacic, Susanne Wilhelm & Karl-Erich Jaeger

  3. Institut für Mikrobiologie, Heinrich-Heine-Universität, Düsseldorf, Germany

    Janpeter Stock, Marius Terfrüchte, Michael Feldbrügge & Kerstin Schipper

  4. Institut für Molekulare Mykologie, Heinrich-Heine-Universität, Universitätsstrasse 1/26.12, 40225, Düsseldorf, Germany

    Joachim F. Ernst

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  1. Christoph Buerth
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Buerth, C., Kovacic, F., Stock, J. et al. Uml2 is a novel CalB-type lipase of Ustilago maydis with phospholipase A activity. Appl Microbiol Biotechnol 98, 4963–4973 (2014). https://doi.org/10.1007/s00253-013-5493-6

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Keywords

  • Uml2
  • CalB
  • Esterase
  • Lipase
  • Phospholipase A
  • Ustilago maydis