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Gewählte Publikation:

Kittl, R; Mueangtoom, K; Gonaus, C; Khazaneh, ST; Sygmund, C; Haltrich, D; Ludwig, R.
(2012): A chloride tolerant laccase from the plant pathogen ascomycete Botrytis aclada expressed at high levels in Pichia pastoris.
J Biotechnol. 2012; 157(2):304-314 FullText FullText_BOKU

Abstract:
Fungal laccases from basidiomycetous fungi are thoroughly investigated in respect of catalytic mechanism and industrial applications, but the number of reported and well characterized ascomycetous laccases is much smaller although they exhibit interesting catalytic properties. We report on a highly chloride tolerant laccase produced by the plant pathogen ascomycete Botrytis aclada, which was recombinantly expressed in Pichia pastoris with an extremely high yield and purified to homogeneity. In a fed-batch fermentation, 495 mg L-1 of laccase was measured in the medium, which is the highest concentration obtained for a laccase by a yeast expression system. The recombinant B. aclada laccase has a typical molecular mass 01 61,565 Da for the amino acid chain. The pl is approximately 2.4, a very low value for a laccase. Glycosyl residues attached to the recombinant protein make up for approximately 27% of the total protein mass. B. aclada laccase exhibits very low Km values and high substrate turnover numbers for phenolic and non-phenolic substrates at acidic and near neutral pH. The enzymexxxs stability increases in the presence of chloride ions and, even more important, its substrate turnover is only weakly inhibited by chloride ions (I-50 = 1.4 M), which is in sharp contrast to most other described laccases. This high chloride tolerance is mandatory for some applications such as implantable biofuel cells and laccase catalyzed reactions, which suffer from the presence of chloride ions. The high expression yield permits fast and easy production for further basic and applied research. (C) 2011 Elsevier B.V. All rights reserved.
Autor/innen der BOKU Wien:
Gonaus Christoph
Haltrich Dietmar
Kittl Roman
Ludwig Roland
Sygmund Christoph
BOKU Gendermonitor:


Find related publications in this database (Keywords)
Ascomycete
Botrytis aclada
Chloride inhibition
Heterologous expression
Laccase
Pichia pastoris


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