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Molecular and enzymatic characterisation of extra- and intracellular laccases from the acidophilic ascomycete Hortaea acidophila

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Abstract

The pigmented ascomycete Hortaea acidophila is able to grow at a pH as low as 0.6 and produces laccases that are involved in melanin synthesis. We now present data on an extracellular and an intracellular laccase which exhibit a high stability at low pH. Furthermore, the optimum for enzyme acitivity is extraordinarily low with pH 1.5 for the intracellular laccase with 2,6-dimethoxyphenol (DMOP) as substrate. Two complete laccase gene sequences of H. acidophila were amplified by inverse polymerase chain reaction (PCR). Whereas the deduced protein laccase I contains an predicted N-terminal signal sequence for protein export, laccase II does not and thus may represent the intracellular laccase. The acidophilic character of both laccases seems to be reflected in their primary structure.

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Abbreviations

ABTS:

2,2′-azino-bis(3-ethyl-2,3-dihydro-1,3-benzthiazole-6-sulphonat

BCIP:

5-Brom-4-chlor-3-indolylhydrogenphos- phat

cbs:

copper binding sequence/site

DHN:

dihydroxynaphthalene

DMOP:

dimethoxyphenol

MRE:

metal responsive element

NBT:

Nitroblue tetrazolium chloride

PCR:

poylmerase chain reaction

pI:

isoelectric point

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Acknowledgements

Our thanks are due to Dr. Hans Peter Call (Übach-Palenberg) for provision of the laccase-specific antibody.

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Authors and Affiliations

  1. Institut für Zelluläre und Molekulare Botanik, Rheinische Friedrich-Wilhelms-Universität, Kirschallee 1, D-53115, Bonn, Germany

    Larissa Tetsch, Jutta Bend & Udo Hölker

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  1. Larissa Tetsch
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  2. Jutta Bend
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  3. Udo Hölker
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Correspondence to Udo Hölker.

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Tetsch, L., Bend, J. & Hölker, U. Molecular and enzymatic characterisation of extra- and intracellular laccases from the acidophilic ascomycete Hortaea acidophila . Antonie Van Leeuwenhoek 90, 183–194 (2006). https://doi.org/10.1007/s10482-006-9064-z

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  • DOI: https://doi.org/10.1007/s10482-006-9064-z

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