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Research paper thumbnail of Mapping Cellular Microenvironments: Proximity Labeling and Complexome Profiling (Seventh Symposium of the Göttingen Proteomics Forum)

Cells

Mass spectrometry-based proteomics methods are finding increasing use in structural biology resea... more Mass spectrometry-based proteomics methods are finding increasing use in structural biology research. Beyond simple interaction networks, information about stable protein-protein complexes or spatially proximal proteins helps to elucidate the biological functions of proteins in a wider cellular context. To shed light on new developments in this field, the Göttingen Proteomics Forum organized a one-day symposium focused on complexome profiling and proximity labeling, two emerging technologies that are gaining significant attention in biomolecular research. The symposium was held in Göttingen, Germany on 23 May, 2019, as part of a series of regular symposia organized by the Göttingen Proteomics Forum.

Research paper thumbnail of Signal peptide replacement resulted in recombinant homologous expression of laccase Lcc8 in Coprinopsis cinerea

AMB Express

Although the model agaricomycete Coprinopsis cinerea possess 17 different laccase genes, up to no... more Although the model agaricomycete Coprinopsis cinerea possess 17 different laccase genes, up to now only four C. cinerea laccases have been purified and characterized to some degree. By exchanging the nucleotide sequence of the deduced signal peptide of Lcc8 it was possible to homologously express lcc8 in C. cinerea under control of the Agaricus bisporus gdpII promoter and the C. cinerea lcc1 terminator. The purified Lcc8 showed two bands in the SDS-PAGE with a molecular weight of 64 kDa and 77 kDa, respectively. The IEF determined pI values of 3.3 and 3.4 for both bands. The optimal pH for oxidation of the substrates ABTS, 2,6-dimethoxyphenol, guaiacol and syringaldazine was pH 4.0, pH 5.0, pH 4.5 and pH 5.0, respectively. Best pH for enzyme storage was pH 8.0. The optimal temperature for oxidation of ABTS was 63 °C, while Lcc8 showed activity of at least 50% over 300 min at 50 °C. The comparable high stability of Lcc8 at alkaline pH and higher temperatures can be of interest for biotechnical applications.

Research paper thumbnail of Integrative omics - from data to biology

Expert review of proteomics, Jun 1, 2018

Multi-omic approaches are promising a broader view on cellular processes and a deeper understandi... more Multi-omic approaches are promising a broader view on cellular processes and a deeper understanding of biological systems. with strongly improved high-throughput methods the amounts of data generated have become huge, and their handling challenging. Area Covered: New bioinformatic tools and pipelines for the integration of data from different omics disciplines continue to emerge, and will support scientists to reliably interpret data in the context of biological processes. comprehensive data integration strategies will fundamentally improve systems biology and systems medicine. to present recent developments of integrative omics, the göttingen proteomics forum (gpf) organized its 6th symposium on the 23rd of november 2017, as part of a series of regular gpf symposia. more than 140 scientists attended the event that highlighted the challenges and opportunities but also the caveats of integrating data from different omics disciplines. Expert commentary: The continuous exponential grow...

Research paper thumbnail of Process for the decontamination of oxygen containing gases, especially of waste gases

Research paper thumbnail of Wissenschaftler entschlüsseln Genom des Hausschwamms

Research paper thumbnail of Process for the treatment of oxygenated exhaust gases

Research paper thumbnail of A method for synthesis of14C-labelled styrene

J Label Compound Radiopharm, 1993

Research paper thumbnail of Anfälligkeit von lebendem und totem Holz der Küstentanne gegenüber Pilzen

Research paper thumbnail of Polyamines in leaves and roots of Populus maximowiczii grown in differing levels of potassium and phosphorus

Research paper thumbnail of Cell wall-associated redox enzymes in white rot fungi

Research paper thumbnail of Changes in plasmodial pigments of Physarum polycephalum in relation to the age of the culture

Canadian Journal of Microbiology, 1987

Plasmodia of the myxomycete Physarum polycephalum were grown in the dark on a semidefined medium ... more Plasmodia of the myxomycete Physarum polycephalum were grown in the dark on a semidefined medium and pigments were assayed from cultures of different ages. The absorption of the crude pigment extracts, measured at the maximum, increased with an increase in the growth rate of the organisms, but with a time lag of about 2 days when compared to increases in the plasmodial protein content. Anaylses of pigments were performed with a newly elaborated HPLC method, and eight pigments, monitored at 382 m, were obtained from the different aged plasmodia. Throughout the culture, quantitative changes in the pigments were found which could be described as periodical. Individual pigments varied in their quantitative changes. In prolonged culture the amount of two pigments significantly increased, in comparison with the other colored compounds, and attained their maximum values after plasmodial growth ceased.

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

Enzyme and Microbial Technology, Nov 1, 2007

This study presents a combined method to analyze extracellular fungal laccases using a new anti-l... more 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 II 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.

Research paper thumbnail of Expression of laccase gene Icc1 in Coprinopsis cinerea under control of various basidiomycetous promoters

Applied Microbiology and Biotechnology, 2006

Research paper thumbnail of Degradation of polychlorinated biphenyls by white‐rot fungi pleurotus ostreatus and trametes versicolor in a solid state system

Http Dx Doi Org 10 1080 02772249309357947, Sep 19, 2008

Research paper thumbnail of Biotechnological research on basidiomycete fungi

Research paper thumbnail of Method for the treatment of straw as a substrate for the cultivation of mushrooms

Research paper thumbnail of Overexpression of laccases in Coprinop¬sis cinerea

Research paper thumbnail of Bioremediation

Books in Soils, Plants, and the Environment, 1997

Research paper thumbnail of Oxidation of Polycyclic Aromatic Hydrocarbons (PAH) by Laccase of Trametes Versicolor

Laccase of Trametes versicolor was able to oxidize in vitro most of the 14 polycyclic aromatic hy... more Laccase of Trametes versicolor was able to oxidize in vitro most of the 14 polycyclic aromatic hydrocarbons (PAH) tested. Acenaphthylene was removed by 37% followed by anthracene and benzo[a]pyrene which were oxidized by 18 and 19%, respectively. Lower but significant oxidation of about 10% was found for eight additional PAH: acenaphthene, fluoranthene, pyrene, benzo[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, and perylene. Naphthalene, fluorene, and phenanthrene were recovered unchanged after incubation for 72 h with laccase. Addition of 1-hydroxybenzotriazole (HBT) to the reaction mixture increased oxidation of PAH: acenaphthylene, acenaphthene, fluorene, anthracene, benzo[a]pyrene, and perylene were almost completely removed from the reaction mixture.

Research paper thumbnail of Early drought-induced changes to the needle proteome of Norway spruce

Tree Physiology, 2007

To elucidate early drought responses in needles of Norway spruce (Picea abies (Karst.) L.), we su... more To elucidate early drought responses in needles of Norway spruce (Picea abies (Karst.) L.), we subjected 1-year-old seedlings to gradual desiccation for 6 weeks. Four weeks of drought treatment caused a small but significant decrease in photosystem II quantum yield of light-adapted needles (Φ a) compared with that of well-watered controls. Six weeks of drought treatment reduced Φ a and the photosystem II quantum yield of dark-adapted needles (Φ) by 50 and 8%, respectively, and reduced shoot water potential by 0.7 MPa, but had no measurable effect on needle relative water content. After two weeks of drought treatment, and before there was a discernible effect of drought on Φ or a statistically significant effect on shoot water potential, needles were analyzed for changes in protein composition. Five out of several hundred detected proteins in needles of drought-treated plants showed consistent changes compared with control leaves. The proteins were identified by LC-MS/MS as components of the oxygen-evolving complex (oxygen evolving enhancer protein 2), ribulose-1,5-bisphosphate carboxylase/oxgenase large subunit, and one protein of unknown function, whose mRNA was found in a previous screen of wound-and methyl-jasmonateinduced bark proteins.

Research paper thumbnail of Mapping Cellular Microenvironments: Proximity Labeling and Complexome Profiling (Seventh Symposium of the Göttingen Proteomics Forum)

Cells

Mass spectrometry-based proteomics methods are finding increasing use in structural biology resea... more Mass spectrometry-based proteomics methods are finding increasing use in structural biology research. Beyond simple interaction networks, information about stable protein-protein complexes or spatially proximal proteins helps to elucidate the biological functions of proteins in a wider cellular context. To shed light on new developments in this field, the Göttingen Proteomics Forum organized a one-day symposium focused on complexome profiling and proximity labeling, two emerging technologies that are gaining significant attention in biomolecular research. The symposium was held in Göttingen, Germany on 23 May, 2019, as part of a series of regular symposia organized by the Göttingen Proteomics Forum.

Research paper thumbnail of Signal peptide replacement resulted in recombinant homologous expression of laccase Lcc8 in Coprinopsis cinerea

AMB Express

Although the model agaricomycete Coprinopsis cinerea possess 17 different laccase genes, up to no... more Although the model agaricomycete Coprinopsis cinerea possess 17 different laccase genes, up to now only four C. cinerea laccases have been purified and characterized to some degree. By exchanging the nucleotide sequence of the deduced signal peptide of Lcc8 it was possible to homologously express lcc8 in C. cinerea under control of the Agaricus bisporus gdpII promoter and the C. cinerea lcc1 terminator. The purified Lcc8 showed two bands in the SDS-PAGE with a molecular weight of 64 kDa and 77 kDa, respectively. The IEF determined pI values of 3.3 and 3.4 for both bands. The optimal pH for oxidation of the substrates ABTS, 2,6-dimethoxyphenol, guaiacol and syringaldazine was pH 4.0, pH 5.0, pH 4.5 and pH 5.0, respectively. Best pH for enzyme storage was pH 8.0. The optimal temperature for oxidation of ABTS was 63 °C, while Lcc8 showed activity of at least 50% over 300 min at 50 °C. The comparable high stability of Lcc8 at alkaline pH and higher temperatures can be of interest for biotechnical applications.

Research paper thumbnail of Integrative omics - from data to biology

Expert review of proteomics, Jun 1, 2018

Multi-omic approaches are promising a broader view on cellular processes and a deeper understandi... more Multi-omic approaches are promising a broader view on cellular processes and a deeper understanding of biological systems. with strongly improved high-throughput methods the amounts of data generated have become huge, and their handling challenging. Area Covered: New bioinformatic tools and pipelines for the integration of data from different omics disciplines continue to emerge, and will support scientists to reliably interpret data in the context of biological processes. comprehensive data integration strategies will fundamentally improve systems biology and systems medicine. to present recent developments of integrative omics, the göttingen proteomics forum (gpf) organized its 6th symposium on the 23rd of november 2017, as part of a series of regular gpf symposia. more than 140 scientists attended the event that highlighted the challenges and opportunities but also the caveats of integrating data from different omics disciplines. Expert commentary: The continuous exponential grow...

Research paper thumbnail of Process for the decontamination of oxygen containing gases, especially of waste gases

Research paper thumbnail of Wissenschaftler entschlüsseln Genom des Hausschwamms

Research paper thumbnail of Process for the treatment of oxygenated exhaust gases

Research paper thumbnail of A method for synthesis of14C-labelled styrene

J Label Compound Radiopharm, 1993

Research paper thumbnail of Anfälligkeit von lebendem und totem Holz der Küstentanne gegenüber Pilzen

Research paper thumbnail of Polyamines in leaves and roots of Populus maximowiczii grown in differing levels of potassium and phosphorus

Research paper thumbnail of Cell wall-associated redox enzymes in white rot fungi

Research paper thumbnail of Changes in plasmodial pigments of Physarum polycephalum in relation to the age of the culture

Canadian Journal of Microbiology, 1987

Plasmodia of the myxomycete Physarum polycephalum were grown in the dark on a semidefined medium ... more Plasmodia of the myxomycete Physarum polycephalum were grown in the dark on a semidefined medium and pigments were assayed from cultures of different ages. The absorption of the crude pigment extracts, measured at the maximum, increased with an increase in the growth rate of the organisms, but with a time lag of about 2 days when compared to increases in the plasmodial protein content. Anaylses of pigments were performed with a newly elaborated HPLC method, and eight pigments, monitored at 382 m, were obtained from the different aged plasmodia. Throughout the culture, quantitative changes in the pigments were found which could be described as periodical. Individual pigments varied in their quantitative changes. In prolonged culture the amount of two pigments significantly increased, in comparison with the other colored compounds, and attained their maximum values after plasmodial growth ceased.

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

Enzyme and Microbial Technology, Nov 1, 2007

This study presents a combined method to analyze extracellular fungal laccases using a new anti-l... more 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 II 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.

Research paper thumbnail of Expression of laccase gene Icc1 in Coprinopsis cinerea under control of various basidiomycetous promoters

Applied Microbiology and Biotechnology, 2006

Research paper thumbnail of Degradation of polychlorinated biphenyls by white‐rot fungi pleurotus ostreatus and trametes versicolor in a solid state system

Http Dx Doi Org 10 1080 02772249309357947, Sep 19, 2008

Research paper thumbnail of Biotechnological research on basidiomycete fungi

Research paper thumbnail of Method for the treatment of straw as a substrate for the cultivation of mushrooms

Research paper thumbnail of Overexpression of laccases in Coprinop¬sis cinerea

Research paper thumbnail of Bioremediation

Books in Soils, Plants, and the Environment, 1997

Research paper thumbnail of Oxidation of Polycyclic Aromatic Hydrocarbons (PAH) by Laccase of Trametes Versicolor

Laccase of Trametes versicolor was able to oxidize in vitro most of the 14 polycyclic aromatic hy... more Laccase of Trametes versicolor was able to oxidize in vitro most of the 14 polycyclic aromatic hydrocarbons (PAH) tested. Acenaphthylene was removed by 37% followed by anthracene and benzo[a]pyrene which were oxidized by 18 and 19%, respectively. Lower but significant oxidation of about 10% was found for eight additional PAH: acenaphthene, fluoranthene, pyrene, benzo[a]anthracene, chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, and perylene. Naphthalene, fluorene, and phenanthrene were recovered unchanged after incubation for 72 h with laccase. Addition of 1-hydroxybenzotriazole (HBT) to the reaction mixture increased oxidation of PAH: acenaphthylene, acenaphthene, fluorene, anthracene, benzo[a]pyrene, and perylene were almost completely removed from the reaction mixture.

Research paper thumbnail of Early drought-induced changes to the needle proteome of Norway spruce

Tree Physiology, 2007

To elucidate early drought responses in needles of Norway spruce (Picea abies (Karst.) L.), we su... more To elucidate early drought responses in needles of Norway spruce (Picea abies (Karst.) L.), we subjected 1-year-old seedlings to gradual desiccation for 6 weeks. Four weeks of drought treatment caused a small but significant decrease in photosystem II quantum yield of light-adapted needles (Φ a) compared with that of well-watered controls. Six weeks of drought treatment reduced Φ a and the photosystem II quantum yield of dark-adapted needles (Φ) by 50 and 8%, respectively, and reduced shoot water potential by 0.7 MPa, but had no measurable effect on needle relative water content. After two weeks of drought treatment, and before there was a discernible effect of drought on Φ or a statistically significant effect on shoot water potential, needles were analyzed for changes in protein composition. Five out of several hundred detected proteins in needles of drought-treated plants showed consistent changes compared with control leaves. The proteins were identified by LC-MS/MS as components of the oxygen-evolving complex (oxygen evolving enhancer protein 2), ribulose-1,5-bisphosphate carboxylase/oxgenase large subunit, and one protein of unknown function, whose mRNA was found in a previous screen of wound-and methyl-jasmonateinduced bark proteins.