Detection, quantification and identification of fungal extracellular laccases using polyclonal antibody and mass spectrometry - Research portal of Justus Liebig University Giessen:

Journal article

Detection, quantification and identification of fungal extracellular laccases using polyclonal antibody and mass spectrometry

Authors list: Kellner, H; Jehmlich, N; Benndorf, D; Hoffmann, R; Rühl, M; Hoegger, PJ; Majcherczyk, A; Kües, U; von Bergen, M; Buscot, F

Publication year: 2007

Pages: 694-701

Journal: Enzyme and Microbial Technology

Volume number: 41

Issue number: 6-7

ISSN: 0141-0229

DOI Link: https://doi.org/10.1016/j.enzmictec.2007.06.002

Publisher: Elsevier

Abstract:
This study presents a combined method to analyze extracellular fungal laccases using a new anti-laccase antibody together with the identification of tryptic laccase peptides by mass spectrometry (nanoLC-ESI-MS/MS). The polyclonal anti-laccase antibody LccCbr2 was raised against peptides designed from the copper binding region 11 of fungal laccases using in silico data obtained from GenBank database. As a consequence, detection requires denaturation of the enzymes due to the stable conformation of the copper binding region II. The specificity of the antibody was shown with denatured laccase Lcc1 of Coprinopsis cinerea and laccase of Hypholoma fasciculare. LccCbr2 detected amounts as low as 5 ng of highly purified laccase, indicating a possible use of the antibody for quantification of laccase proteins. Denatured extracellular laccases from culture supernatants of the basidiomycetes C. cinerea, H. fasciculare, Lentinula edodes, Mycena sp., Piriformospora indica, Pleurotus cornucopiae, Pleurotus ostreatus, Pycnoporus cinnabarinus, Trametes versicolor and furthermore the ascomycete Verpa conica were detected with apparent molecular masses between 60 and 70 kDa by LccCbr2. The identity of extracellular laccases from C cinerea, H. fasciculare, P ostreatus, P. cinnabarinus and T versicolor were verified by tryptic peptides using nanoLC-ESI-MS/MS.

Authors/Editors

- Uni.-Prof. Dr. Martin Georg Wilhelm Rühl

Citation Styles

Harvard Citation style: Kellner, H., Jehmlich, N., Benndorf, D., Hoffmann, R., Rühl, M., Hoegger, P., et al. (2007) Detection, quantification and identification of fungal extracellular laccases using polyclonal antibody and mass spectrometry, Enzyme and Microbial Technology, 41(6-7), pp. 694-701. https://doi.org/10.1016/j.enzmictec.2007.06.002

APA Citation style: Kellner, H., Jehmlich, N., Benndorf, D., Hoffmann, R., Rühl, M., Hoegger, P., Majcherczyk, A., Kües, U., von Bergen, M., & Buscot, F. (2007). Detection, quantification and identification of fungal extracellular laccases using polyclonal antibody and mass spectrometry. Enzyme and Microbial Technology. 41(6-7), 694-701. https://doi.org/10.1016/j.enzmictec.2007.06.002