TY - JOUR A1 - Silva, Bibiana A1 - Oliveira Costa, Ana Carolina A1 - Tchewonpi, Sorel Sagu A1 - Bönick, Josephine A1 - Huschek, Gerd A1 - Gonzaga, Luciano Valdemiro A1 - Fett, Roseane A1 - Baldermann, Susanne A1 - Rawel, Harshadrai Manilal T1 - Comparative quantification and differentiation of bracatinga (Mimosa scabrella Bentham) honeydew honey proteins using targeted peptide markers identified by high-resolution mass spectrometry JF - Food research international N2 - Honey traceability is an important topic, especially for honeydew honeys, due to the increased incidence of adulteration. This study aimed to establish specific markers to quantify proteins in honey. A proteomics strategy to identify marker peptides from bracatinga honeydew honey was therefore developed. The proteomics approach was based on initial untargeted identification of honey proteins and peptides by LC-ESI-Triple-TOF-MS/MS, which identified the major royal jelly proteins (MRJP) presence. Afterwards, the peptides were selected by the in silico digestion. The marker peptides were quantified by the developed targeted LC-QqQ-MS/MS method, which provided good linearity and specificity, besides recoveries between 92 and 100% to quantify peptides from bracatinga honeydew honey. The uniqueness and high response in mass spectrometry were backed by further complementary protein analysis (SDS-PAGE). The selected marker peptides EALPHVPIFDR (MRJP 1), ILGANVK (MRJP 2), TFVTIER (MRJP 3), QNIDVVAR (MRJP 4), FINNDYNFNEVNFR (MRJP 5) and LLQPYPDWSWTK (MRJP 7), quantified by LC-QqQ-MS/MS, highlighted that the content of QNIDVVAR from MRJP 4 could be used to differentiate bracatinga honeydew honey from floral honeys (p < 0.05) as a potential marker for its authentication. Finally, principal components analysis highlighted the QNIDVVAR content as a good descriptor of the analyzed bracatinga honeydew honey samples. KW - Honeydew honey KW - Major royal jelly proteins KW - Marker peptides KW - High-resolution mass spectrometry KW - Principal component analysis Y1 - 2020 U6 - https://doi.org/10.1016/j.foodres.2020.109991 SN - 0963-9969 SN - 1873-7145 VL - 141 PB - Elsevier CY - New York, NY [u.a.] ER - TY - JOUR A1 - Borremans, An A1 - Bußler, Sara A1 - Sagu Tchewonpi, Sorel A1 - Rawel, Harshadrai Manilal A1 - Schlüter, Oliver K. A1 - Leen, Van Campenhout T1 - Effect of blanching plus fermentation on selected functional properties of mealworm (Tenebrio molitor) powders JF - Foods : open access journal N2 - The aim of this study was to determine the effect of blanching followed by fermentation of mealworms (Tenebrio molitor) with commercial meat starter cultures on the functional properties of powders produced from the larvae. Full fat and defatted powder samples were prepared from non-fermented and fermented mealworm pastes. Then the crude protein, crude fat, and dry matter contents, pH, bulk density, colour, water and oil binding capacity, foaming capacity and stability, emulsion capacity and stability, protein solubility, quantity of free amino groups, and protein composition of the powders were evaluated. Regardless of the starter culture used, the blanching plus fermentation process reduced the crude and soluble protein contents of the full fat powders and in general impaired their water and oil binding, foaming, and emulsifying properties. Defatting of the powders improved most functional properties studied. The o-phthaldialdehyde assay revealed that the amount of free amino groups was higher in the fermented powders while sodium dodecyl sulfate polyacrylamide gel electrophoresis demonstrated that the soluble proteins of the fermented powders were composed of molecules of lower molecular mass compared to non-fermented powders. As molecular sizes of the soluble proteins decreased, it was clear that the protein structure was also modified by the fermentation process, which in turn led to changes in functional properties. In general, it was concluded that fermentation of mealworms with blanching as a pre-treatment does not contribute to the functional properties studied in this work. Nevertheless, the results confirmed that the properties of non-fermented powders are comparable to other food protein sources. KW - mealworm KW - fermentation KW - functional properties KW - insect proteins KW - SDS-PAGE Y1 - 2020 U6 - https://doi.org/10.3390/foods9070917 SN - 2304-8158 VL - 9 IS - 7 PB - MDPI CY - Basel ER - TY - JOUR A1 - Wetzel, Alexandra Nicole A1 - Scholtka, Bettina A1 - Schumacher, Fabian A1 - Rawel, Harshadrai Manilal A1 - Geisendörfer, Birte A1 - Kleuser, Burkhard T1 - Epigenetic DNA methylation of EBI3 modulates human interleukin-35 formation via NFkB signaling BT - a promising therapeutic option in ulcerative colitis JF - International journal of molecular sciences N2 - Ulcerative colitis (UC), a severe chronic disease with unclear etiology that is associated with increased risk for colorectal cancer, is accompanied by dysregulation of cytokines. Epstein-Barr virus-induced gene 3 (EBI3) encodes a subunit in the unique heterodimeric IL-12 cytokine family of either pro- or anti-inflammatory function. After having recently demonstrated that upregulation of EBI3 by histone acetylation alleviates disease symptoms in a dextran sulfate sodium (DSS)-treated mouse model of chronic colitis, we now aimed to examine a possible further epigenetic regulation of EBI3 by DNA methylation under inflammatory conditions. Treatment with the DNA methyltransferase inhibitor (DNMTi) decitabine (DAC) and TNF alpha led to synergistic upregulation of EBI3 in human colon epithelial cells (HCEC). Use of different signaling pathway inhibitors indicated NF kappa B signaling was necessary and proportional to the synergistic EBI3 induction. MALDI-TOF/MS and HPLC-ESIMS/MS analysis of DAC/TNF alpha-treated HCEC identified IL-12p35 as the most probable binding partner to form a functional protein. EBI3/IL-12p35 heterodimers (IL-35) induce their own gene upregulation, something that was indeed observed in HCEC cultured with media from previously DAC/TNF alpha-treated HCEC. These results suggest that under inflammatory and demethylating conditions the upregulation of EBI3 results in the formation of anti-inflammatory IL-35, which might be considered as a therapeutic target in colitis. KW - decitabine KW - DNMT inhibitor KW - EBI3 KW - inhibitory cytokines KW - interleukin-35 KW - TNF alpha KW - Ulcerative colitis Y1 - 2021 U6 - https://doi.org/10.3390/ijms22105329 SN - 1422-0067 VL - 22 IS - 10 PB - MDPI CY - Basel ER - TY - JOUR A1 - Bußler, Sara A1 - Rawel, Harshadrai Manilal A1 - Schlüter, Oliver K. T1 - Impact of plasma processed air (PPA) on phenolic model systems BT - suggested mechanisms and relevance for food applications JF - Innovative food science & emerging technologies : the official journal of the European Federation of Food Science and Technology N2 - Cold plasma is considered to be a novel, non-thermal, chemical-free and eco-friendly disinfection and surface modification technology. Plasma treatment of air to generate the so called plasma processed air (PPA) induces the formation of reactive oxygen (ROS) and nitrogen species (RNS). As a result, PPA has a different chemical composition compared to untreated air and suits therefore as an alternative method for microbial disinfection. However, depending on the product properties of the food matrix and its composition, a number of plasmainduced reactions also need to be taken into consideration. This necessitates also the elucidation and understanding of the basic interactions of plasma species with bioactive compounds. The intention here is to avoid the degradation of these valuable substances and to prevent other undesirable effects in future food related applications. In the present study, the effects of PPA treatment on selected antioxidants such as pyrocatechol and derivatives of hydroxycinnimic acid were investigated in model systems to specify possible reactions induced. Antioxidant capacity, pH value, UV-Vis spectroscopy, RP-HPLC and LC-MS analysis were applied to identify reaction products providing information on possible changes induced in food matrices by PPA treatment. Exposure to PPA caused a perceptible color change towards yellow-brown accompanied by a strong reduction of the pH and the formation of insoluble sediments in the model solutions. The accumulation of nitrate, nitrite, but not of hydrogen peroxide was shown. LC-MS analysis demonstrated the formation of plasma-modified derivatives in all tested systems. The main reactions in liquid model solutions exposed to PPA were attributed to oxidation, nitration and polymerization of the phenolic compounds. KW - cold atmospheric pressure plasma KW - reactive oxygen and nitrogen species KW - food safety KW - antioxidative phenolic ingredients KW - phenol oxidation KW - phenol nitration KW - plasma process indicators Y1 - 2020 U6 - https://doi.org/10.1016/j.ifset.2020.102432 SN - 1466-8564 SN - 1878-5522 VL - 64 PB - Elsevier CY - Oxford ER - TY - GEN A1 - Sagu Tchewonpi, Sorel A1 - Huschek, Gerd A1 - Waldbach Braga, Tess A1 - Rackiewicz, Michal A1 - Homann, Thomas A1 - Rawel, Harshadrai Manilal T1 - Design of Experiment (DoE) for Optimization of HPLC Conditions for the Simultaneous Fractionation of Seven α-Amylase/Trypsin Inhibitors from Wheat (Triticum aestivum L.) T2 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe N2 - Wheat alpha-amylase/trypsin inhibitors remain a subject of interest considering the latest findings showing their implication in wheat-related non-celiac sensitivity (NCWS). Understanding their functions in such a disorder is still unclear and for further study, the need for pure ATI molecules is one of the limiting problems. In this work, a simplified approach based on the successive fractionation of ATI extracts by reverse phase and ion exchange chromatography was developed. ATIs were first extracted from wheat flour using a combination of Tris buffer and chloroform/methanol methods. The separation of the extracts on a C18 column generated two main fractions of interest F1 and F2. The response surface methodology with the Doehlert design allowed optimizing the operating parameters of the strong anion exchange chromatography. Finally, the seven major wheat ATIs namely P01083, P17314, P16850, P01085, P16851, P16159, and P83207 were recovered with purity levels (according to the targeted LC-MS/MS analysis) of 98.2 ± 0.7; 98.1 ± 0.8; 97.9 ± 0.5; 95.1 ± 0.8; 98.3 ± 0.4; 96.9 ± 0.5, and 96.2 ± 0.4%, respectively. MALDI-TOF-MS analysis revealed single peaks in each of the pure fractions and the mass analysis yielded deviations of 0.4, 1.9, 0.1, 0.2, 0.2, 0.9, and 0.1% between the theoretical and the determined masses of P01083, P17314, P16850, P01085, P16851, P16159, and P83207, respectively. Overall, the study allowed establishing an efficient purification process of the most important wheat ATIs. This paves the way for further in-depth investigation of the ATIs to gain more knowledge related to their involvement in NCWS disease and to allow the absolute quantification in wheat samples. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 1260 KW - wheat KW - α-amylase/trypsin inhibitors KW - fractionation KW - purification KW - reversed-phase chromatography KW - ion-exchange chromatography KW - design of experiment KW - LC–MS/MS KW - MALDI-TOF-MS Y1 - 2022 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-559282 SN - 1866-8372 SP - 1 EP - 18 PB - Universitätsverlag Potsdam CY - Potsdam ER - TY - JOUR A1 - Sagu Tchewonpi, Sorel A1 - Huschek, Gerd A1 - Waldbach Braga, Tess A1 - Rackiewicz, Michal A1 - Homann, Thomas A1 - Rawel, Harshadrai Manilal T1 - Design of Experiment (DoE) for Optimization of HPLC Conditions for the Simultaneous Fractionation of Seven α-Amylase/Trypsin Inhibitors from Wheat (Triticum aestivum L.) JF - Processes : open access journal N2 - Wheat alpha-amylase/trypsin inhibitors remain a subject of interest considering the latest findings showing their implication in wheat-related non-celiac sensitivity (NCWS). Understanding their functions in such a disorder is still unclear and for further study, the need for pure ATI molecules is one of the limiting problems. In this work, a simplified approach based on the successive fractionation of ATI extracts by reverse phase and ion exchange chromatography was developed. ATIs were first extracted from wheat flour using a combination of Tris buffer and chloroform/methanol methods. The separation of the extracts on a C18 column generated two main fractions of interest F1 and F2. The response surface methodology with the Doehlert design allowed optimizing the operating parameters of the strong anion exchange chromatography. Finally, the seven major wheat ATIs namely P01083, P17314, P16850, P01085, P16851, P16159, and P83207 were recovered with purity levels (according to the targeted LC-MS/MS analysis) of 98.2 ± 0.7; 98.1 ± 0.8; 97.9 ± 0.5; 95.1 ± 0.8; 98.3 ± 0.4; 96.9 ± 0.5, and 96.2 ± 0.4%, respectively. MALDI-TOF-MS analysis revealed single peaks in each of the pure fractions and the mass analysis yielded deviations of 0.4, 1.9, 0.1, 0.2, 0.2, 0.9, and 0.1% between the theoretical and the determined masses of P01083, P17314, P16850, P01085, P16851, P16159, and P83207, respectively. Overall, the study allowed establishing an efficient purification process of the most important wheat ATIs. This paves the way for further in-depth investigation of the ATIs to gain more knowledge related to their involvement in NCWS disease and to allow the absolute quantification in wheat samples. KW - wheat KW - α-amylase/trypsin inhibitors KW - fractionation KW - purification KW - reversed-phase chromatography KW - ion-exchange chromatography KW - design of experiment KW - LC–MS/MS KW - MALDI-TOF-MS Y1 - 2022 U6 - https://doi.org/10.3390/pr10020259 SN - 2227-9717 VL - 10 SP - 1 EP - 18 PB - MDPI CY - Basel, Schweiz ET - 2 ER - TY - JOUR A1 - Figueroa Campos, Gustavo Adolfo A1 - G. K. T. Kruizenga, Johannes A1 - Sagu Tchewonpi, Sorel A1 - Schwarz, Steffen A1 - Homann, Thomas A1 - Taubert, Andreas A1 - Rawel, Harshadrai Manilal T1 - Effect of the post-harvest processing on protein modification in green coffee beans by phenolic compounds JF - Foods : open access journal N2 - The protein fraction, important for coffee cup quality, is modified during post-harvest treatment prior to roasting. Proteins may interact with phenolic compounds, which constitute the major metabolites of coffee, where the processing affects these interactions. This allows the hypothesis that the proteins are denatured and modified via enzymatic and/or redox activation steps. The present study was initiated to encompass changes in the protein fraction. The investigations were limited to major storage protein of green coffee beans. Fourteen Coffea arabica samples from various processing methods and countries were used. Different extraction protocols were compared to maintain the status quo of the protein modification. The extracts contained about 4–8 µg of chlorogenic acid derivatives per mg of extracted protein. High-resolution chromatography with multiple reaction monitoring was used to detect lysine modifications in the coffee protein. Marker peptides were allocated for the storage protein of the coffee beans. Among these, the modified peptides K.FFLANGPQQGGK.E and R.LGGK.T of the α-chain and R.ITTVNSQK.I and K.VFDDEVK.Q of β-chain were detected. Results showed a significant increase (p < 0.05) of modified peptides from wet processed green beans as compared to the dry ones. The present study contributes to a better understanding of the influence of the different processing methods on protein quality and its role in the scope of coffee cup quality and aroma. View Full-Text KW - Arabica coffee KW - coffee processing KW - protein modification KW - bound phenolic compounds KW - peptide biomarkers KW - LC-MS/MS Y1 - 2022 U6 - https://doi.org/10.3390/foods11020159 SN - 2304-8158 VL - 11 PB - MDPI CY - Basel, Schweiz ET - 2 ER - TY - JOUR A1 - Bönick, Josephine A1 - Huschek, Gerd A1 - Rawel, Harshadrai Manilal T1 - Determination of wheat, rye and spelt authenticity in bread by targeted peptide biomarkers JF - Journal of Food Composition and Analysis N2 - Adulteration of food and mislabeled products in global market is a major financial and reputational risk for food manufacturers and trade companies. Consequently, there is a necessity to develop analytical methods to meet these issues. An analytical strategy to check the authenticity of wheat, spelt and rye addition in bread products was developed based on database research, in silico digestion confirming peptide specificity and finally quantification by liquid chromatography-tandem mass spectrometry analysis. Peptide markers for wheat (SQQQISQQPQQLPQQQQIPQQPQQF; QQHQIPQQPQQFPQQQQF and QPHQPQQPYPQQ), spelt (ASIVVGIGGQ; SQQPGQIIPQQPQQPSPL) and rye (LPQSHKQHVGQGAL; AQVQGIIQPQQL and QQFPQQPQQSFPQQPQQPVPQQPL) were identified, verified by protein Basic Local Alignment Search Tool and database research and used for quantification in bread. The specific use of multi-reaction monitoring transitions of selected peptides permitted the identification of closely related species wheat and spelt. Other cereal species (emmer, einkorn, barley, maize, rye and oat) were also checked. The target peptides were quantified at different levels using own reference baked products (bread) after in-solution chymotryptic digestion. Sensitivity of the identification was 0.5-1% using flour-based (0-25%) matrix calibration and the analytical recovery in bread was 80-125%. The analytical strategy described here supplies an emerging, independent and flexible tool in controlling the labeling of bread. KW - Food analysis KW - Food composition KW - Food safety KW - Plant authentication KW - Wheat KW - Spelt KW - Rye KW - LC-MS-MS KW - Quantification of peptides KW - Food labeling Y1 - 2017 U6 - https://doi.org/10.1016/j.jfca.2017.01.019 SN - 0889-1575 SN - 1096-0481 VL - 58 SP - 82 EP - 91 PB - Elsevier CY - San Diego ER - TY - JOUR A1 - Huschek, Gerd A1 - Bönick, Josephine A1 - Merkel, Dietrich A1 - Huschek, Doreen A1 - Rawel, Harshadrai Manilal T1 - Authentication of leguminous-based products by targeted biomarkers using high resolution time of flight mass spectrometry JF - LWT - food science and technology : an official journal of the Swiss Society of Food Science and Technology (SGLWT/SOSSTA) and the International Union of Food Science and Technology (IUFoST) N2 - A growing number of health-conscious individuals supplements their diet with protein-rich plant-based products to reduce their meat consumption. Analytical methods are needed to authenticate these new vegetarian products not only for the correct labelling of ingredients according to European legislation but also to discourage food fraud. This paper presents new biomarkers for a targeted proteomics LC-MS/MS work-flow that can simultaneously prove the presence/absence of garden pea, a protein-rich legume, meat and honey and quantify their content in processed vegan food. We show a novel rapid strategy to identify biomarkers for species authentication and the steps for the multi-parameter LC-MS/MS method validation and quantification. A high resolution triple time of flight mass spectrometer (HRMS) with SWATH Acquisition was used for the rapid discovery of all measurable trypsin-digested proteins in the individual ingredients. From these proteins, species-selective biomarkers were identified with BLAST and Skyline. Vicilin and convicilin (UniProt: D3VND9, Q9M3X6) allow pea authentication with regard to other legume species. Myostatin (UniProt: 018831) is a single biomarker for all meat types. For honey, we identified three selective proteins (UniProt: C6K481, C6K482, Q3L6329). The final LC-MS/MS method can identity and quantify these markers simultaneously. Quantification occurs via external matrix calibration. KW - Vegan KW - Food authentication KW - Legume KW - Honey KW - Meat peptide biomarker KW - MS quantification of leguminous additives KW - Food labelling Y1 - 2018 U6 - https://doi.org/10.1016/j.lwt.2017.12.034 SN - 0023-6438 SN - 1096-1127 VL - 90 SP - 164 EP - 171 PB - Elsevier CY - Amsterdam ER - TY - GEN A1 - Figueroa Campos, Gustavo A. A1 - Perez, Jeffrey Paulo H. A1 - Block, Inga A1 - Tchewonpi Sagu, Sorel A1 - Saravia Celis, Pedro A1 - Taubert, Andreas A1 - Rawel, Harshadrai Manilal T1 - Preparation of activated carbons from spent coffee and coffee parchment and assessment of their adsorbent efficiency T2 - Postprints der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe N2 - The valorization of coffee wastes through modification to activated carbon has been considered as a low-cost adsorbent with prospective to compete with commercial carbons. So far, very few studies have referred to the valorization of coffee parchment into activated carbon. Moreover, low-cost and efficient activation methods need to be more investigated. The aim of this work was to prepare activated carbon from spent coffee grounds and parchment, and to assess their adsorption performance. The co-calcination processing with calcium carbonate was used to prepare the activated carbons, and their adsorption capacity for organic acids, phenolic compounds and proteins was evaluated. Both spent coffee grounds and parchment showed yields after the calcination and washing treatments of around 9.0%. The adsorption of lactic acid was found to be optimal at pH 2. The maximum adsorption capacity of lactic acid with standard commercial granular activated carbon was 73.78 mg/g, while the values of 32.33 and 14.73 mg/g were registered for the parchment and spent coffee grounds activated carbons, respectively. The Langmuir isotherm showed that lactic acid was adsorbed as a monolayer and distributed homogeneously on the surface. Around 50% of total phenols and protein content from coffee wastewater were adsorbed after treatment with the prepared activated carbons, while 44, 43, and up to 84% of hydrophobic compounds were removed using parchment, spent coffee grounds and commercial activated carbon, respectively; the adsorption efficiencies of hydrophilic compounds ranged between 13 and 48%. Finally, these results illustrate the potential valorization of coffee by-products parchment and spent coffee grounds into activated carbon and their use as low-cost adsorbent for the removal of organic compounds from aqueous solutions. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 1158 KW - coffee by-products KW - spent coffee grounds KW - parchment KW - valorization KW - calcination KW - activated carbon KW - organic compounds adsorption Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-521914 SN - 1866-8372 IS - 8 ER -