@article{AliHomannKreiseletal.2012, author = {Ali, Mostafa and Homann, Thomas and Kreisel, Janka and Khalil, Mahmoud and Puhlmann, Ralf and Kruse, Hans-Peter and Rawel, Harshadrai Manilal}, title = {Characterization and modeling of the interactions between coffee storage proteins and phenolic compounds}, series = {Journal of agricultural and food chemistry : a publication of the American Chemical Society}, volume = {60}, journal = {Journal of agricultural and food chemistry : a publication of the American Chemical Society}, number = {46}, publisher = {American Chemical Society}, address = {Washington}, issn = {0021-8561}, doi = {10.1021/jf303372a}, pages = {11601 -- 11608}, year = {2012}, abstract = {This study addresses the interactions of coffee storage proteins with coffee-specific phenolic compounds. Protein profiles, of Coffea arabica and Coffea canephora (var robusta) were compared. Major Phenolic compounds were extracted and analyzed with appropriate methods. The polyphenol-protein interactions during protein extraction have been addressed by different analytical setups [reversed-phase high-performance liquid chromatography (RP-HPLC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), matrix-assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS), and Trolox equivalent antioxidant capacity (TEAC) assays], with focus directed toward identification of covalent adduct formation. The results indicate that C. arabica proteins are more susceptible to these interactions and the polyphenol oxidase activity seems to be a crucial factor for the formation of these addition products. A tentative allocation of the modification type and site in the protein has been attempted. Thus, the first available in silico modeling of modified coffee proteins is reported. The extent of these modifications may contribute to the structure and function of "coffee melanoidins" and are discussed in the context of coffee flavor formation.}, language = {en} } @article{WeinertWieseRaweletal.2012, author = {Weinert, Christoph H. and Wiese, Stefanie and Rawel, Harshadrai Manilal and Esatbeyoglu, Tuba and Winterhalter, Peter and Homann, Thomas and Kulling, Sabine E.}, title = {Methylation of catechins and procyanidins by rat and human Catechol-O-Methyltransferase metabolite profiling and molecular modeling studies}, series = {Drug metabolism and disposition : the biological fate of chemicals}, volume = {40}, journal = {Drug metabolism and disposition : the biological fate of chemicals}, number = {2}, publisher = {American Society for Pharmacology and Experimental Therapeutics}, address = {Bethesda}, issn = {0090-9556}, doi = {10.1124/dmd.111.041871}, pages = {353 -- 359}, year = {2012}, abstract = {Catechins and procyanidins are major polyphenols in plant-derived foods. Despite intensive studies in recent years, neither their biochemical nor their toxicological properties have been clarified sufficiently. This study aimed to compare the methylation of catechins and procyanidins by the enzyme catechol-O-methyltransferase (COMT) in vitro. We conducted incubations with rat liver cytosol and human placental cytosol including S-adenosyl-L-methionine. The set of substrates comprised the catechins (-)-epicatechin (EC) and (+)catechin (CAT), the procyanidin dimers B1, B2, B3, B4, B5, and B7 as well as procyanidin trimer C1. After extraction, metabolites were analyzed by means of liquid chromatography-electrospray ionizationmass spectrometry and liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry. EC and CAT were converted to two monomethylated metabolites each by human and rat COMT, with the 3'-O-methyl derivatives being consistently the main metabolites. Furthermore, the flavanyl units of procyanidins were methylated consecutively, leading to monomethylated and dimethylated dimeric metabolites as well as monomethylated, dimethylated, and trimethylated C1 metabolites. The methylation status of each flavanyl unit was determined by means of mass spectrometric quinone-methide fragmentation patterns. In addition, molecular modeling studies were performed with the aim to predict the preferred site of methylation and to verify the experimental data. In conclusion, our results indicate that the degree and position of methylation depend clearly on the three-dimensional structure of the entire substrate molecule.}, language = {en} } @article{AliHomannKhaliletal.2013, author = {Ali, Mostafa and Homann, Thomas and Khalil, Mahmoud and Kruse, Hans-Peter and Rawel, Harshadrai Manilal}, title = {Milk whey protein modification by coffee-specific phenolics effect on structural and functional properties}, series = {Journal of agricultural and food chemistry : a publication of the American Chemical Society}, volume = {61}, journal = {Journal of agricultural and food chemistry : a publication of the American Chemical Society}, number = {28}, publisher = {American Chemical Society}, address = {Washington}, issn = {0021-8561}, doi = {10.1021/jf402221m}, pages = {6911 -- 6920}, year = {2013}, abstract = {A suitable vehicle for integration of bioactive plant constituents is proposed. It involves modification of proteins using phenolics and applying these for protection of labile constituents. It dissects the noncovalent and covalent interactions of beta-lactoglobulin with coffee-specific phenolics. Alkaline and polyphenol oxidase modulated covalent reactions were compared. Tryptic digestion combined with MALDI-TOF-MS provided tentative allocation of the modification type and site in the protein, and an in silico modeling of modified beta-lactoglobulin is proposed. The modification delivers proteins with enhanced antioxidative properties. Changed structural properties and differences in solubility, surface hydrophobicity, and emulsification were observed. The polyphenol oxidase modulated reaction provides a modified beta-lactoglobulin with a high antioxidative power, is thermally more stable, requires less energy to unfold, and, when emulsified with lutein esters, exhibits their higher stability against UV light. Thus, adaptation of this modification provides an innovative approach for functionalizing proteins and their uses in the food industry.}, language = {en} } @article{UhrBuchholzHomannetal.2014, author = {Uhr, Linda and Buchholz, Tina and Homann, Thomas and Huschek, Gerd and Rawel, Harshadrai Manilal}, title = {Targeted proteomics-based analysis of technical enzymes from fungal origin in baked products}, series = {Journal of cereal science}, volume = {60}, journal = {Journal of cereal science}, number = {2}, publisher = {Elsevier}, address = {London}, issn = {0733-5210}, doi = {10.1016/j.jcs.2014.04.007}, pages = {440 -- 447}, year = {2014}, abstract = {The application of technical enzymes is a potential tool in modulating the dough and baking quality of cereal products. No endogenous amylases (alpha- and beta-forms) are present in mature wheat grains; they may be synthesized or activated during germination. Hence, microbial alpha-amylases are added to the dough, being resistant to the endogenous alpha-amylase/trypsin inhibitors. Here, we report on the initial identification of two technical enzymes from a commercial sample based on an in-gel tryptic digestion coupled with MALDI-MS analysis. The primary component of the protein fraction with 51.3 kDa was alpha-amylase from Aspergillus species. A second major protein with 24.8 kDa was identified as endo-1,4-xylanase from Thermomyces lanuginosus. In the following experimental work up, a targeted proteomics approach utilizing the combination of specific proteolytic digestion of the added amylase and xylanase in wheat flour, dough or baked products, solid phase extraction of released peptides and their detection using LC-MS/MS was optimized. The targeted (MRM) MS/MS peptide signals showed that the peptide "ALSSALHER" (MW = 983) originating from amylase and "GWNPGLNAR" (MW = 983) from xylanase can be used to identify the corresponding technical enzymes added. Consequently, locally available baked products were tested and found to contain these enzymes as supplementary ingredients. (C) 2014 Elsevier Ltd. All rights reserved.}, language = {en} } @article{GoetzChmielewskiHomannetal.2014, author = {Goetz, Klaus-Peter and Chmielewski, Frank M. and Homann, Thomas and Huschek, Gerd and Matzneller, Philipp and Rawel, Harshadrai Manilal}, title = {Seasonal changes of physiological parameters in sweet cherry (Prunus avium L.) buds}, series = {Scientia horticulturae : an international journal sponsored by the International Society for Horticultural Science}, volume = {172}, journal = {Scientia horticulturae : an international journal sponsored by the International Society for Horticultural Science}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0304-4238}, doi = {10.1016/j.scienta.2014.04.012}, pages = {183 -- 190}, year = {2014}, abstract = {The transition from dormant stage to the beginning of growth was first obvious by markedly changes of the water content. The phase from green tip to tight cluster, with a length of only 4 days, was the period of the most physiological activity in single buds, because of the highest daily accumulation rates of fresh/dry weight, C, N. We assume a concentration dependant regulation of the member of the aspartate family (asparagine, aspartic acid, isoleucine) during dormancy, growth and development in sweet cherry buds. The ABA content showed 2011/12 a clear bimodal pattern which was at lower level similar in 2012/13, but not so strong incisive. In both years, the first peak was probably related to the end of endodormancy. However the ABA-isomer content showed in both seasons a unimodal pattern. The maximum of the ratio of ABA-isomer/ABA indicated the beginning of ontogenetic development which starts 3 and 2 weeks later, respectively. Our results suggest that ABA and the ABA-isomer in the sweet cherry buds regulate differentiated metabolic processes in the dormant stage and during bud growth and development. After replication in the season 2013/14 the estimated dates of release of endodormancy, beginning of ecodormancy and start of ontogenetic development will be used to validate and improve phenological models for the beginning of cherry blossom. (C) 2014 Elsevier B.V. All rights reserved.}, language = {en} } @article{SaguTchewonpiNsoHomannetal.2015, author = {Sagu Tchewonpi, Sorel and Nso, Emmanuel Jong and Homann, Thomas and Kapseu, Cesar and Rawel, Harshadrai Manilal}, title = {Extraction and purification of beta-amylase from stems of Abrus precatorius by three phase partitioning}, series = {Food chemistry}, volume = {183}, journal = {Food chemistry}, publisher = {Elsevier}, address = {Oxford}, issn = {0308-8146}, doi = {10.1016/j.foodchem.2015.03.028}, pages = {144 -- 153}, year = {2015}, abstract = {The stems of Abrus precatorius were used to extract a beta-amylase enriched fraction. A three phase partitioning method and a Doehlert design with 3 variables (ratio of crude extract/t-butanol, the ammonium sulphate saturation and pH) were used. The data was fitted in a second-order polynomial model and the parameters were optimized to enrich beta-amylase. Experimental responses for the modulation were recovery of activity and the purification factor. The optimal conditions were: a ratio of crude extract/t-butanol of 0.87 (v/v), saturation in ammonium sulphate of 49.46\% (w/v) and a pH of 5.2. An activity recovery of 156.2\% and a purification factor of 10.17 were found. The enriched enzyme was identified as a beta-amylase and its molecular weight was 60.1 kDa. K-m and V-max values were 79.37 mg/ml and 5.13 U/ml, respectively and the highest activity was registered at a temperature of 70 degrees C and a pH between 6 and 6.5. A significant stabilization of the beta-amylase was observed up to 65 degrees C. (C) 2015 Elsevier Ltd. All rights reserved.}, language = {en} } @article{ReinkensmeierSteinbrennerHomannetal.2016, author = {Reinkensmeier, Annika and Steinbrenner, Katrin and Homann, Thomas and Bussler, Sara and Rohn, Sascha and Rawel, Harshadrai Manilal}, title = {Monitoring the apple polyphenol oxidase-modulated adduct formation of phenolic and amino compounds}, series = {Food chemistry}, volume = {194}, journal = {Food chemistry}, publisher = {Elsevier}, address = {Oxford}, issn = {0308-8146}, doi = {10.1016/j.foodchem.2015.07.145}, pages = {76 -- 85}, year = {2016}, abstract = {Minimally processed fruit products such as smoothies are increasingly coming into demand. However, they are often combined with dairy ingredients. In this combination, phenolic compounds, polyphenoloxidases, and amino compounds could interact. In this work, a model approach is presented where apple serves as a source for a high polyphenoloxidase activity for modulating the reactions. The polyphenoloxidase activity ranged from 128 to 333 nakt/mL in different apple varieties. From these, 'Braeburn' was found to provide the highest enzymatic activity. The formation and stability of resulting chromogenic conjugates was investigated. The results show that such adducts are not stable and possible degradation mechanisms leading to follow-up products formed are proposed. Finally, apple extracts were used to modify proteins and their functional properties characterized. There were retaining antioxidant properties inherent to phenolic compounds after adduct formation. Consequently, such interactions may also be utilized to improve the textural quality of food products.}, language = {en} } @article{UhrWielandHomannetal.2016, author = {Uhr, Linda and Wieland, Phillis and Homann, Thomas and Huschek, Gerd and Rawel, Harshadrai Manilal}, title = {Identification and LC-MS/MS-based analyses of technical enzymes in wheat flour and baked products}, series = {European food research and technology : official organ of the EuCheMS, Division of Food Chemistry}, volume = {242}, journal = {European food research and technology : official organ of the EuCheMS, Division of Food Chemistry}, publisher = {Springer}, address = {New York}, issn = {1438-2377}, doi = {10.1007/s00217-015-2536-5}, pages = {247 -- 257}, year = {2016}, abstract = {The use of technical enzymes in bakery industry is necessary for a consistent and good quality of baked products. Since the cultivation of cereals leads to low amounts of endogenous enzymes being present, a need of their commercial alternatives is becoming a routine process in order to meet the consumer quality demands. Targeted quantification proteomics-based methods are necessary for their detection to meet the regulatory criteria. Here, we initially report on the identification of Lipase FE-01, a lipase from fungus Thermomyces lanuginosus, as analyzed by SDS-PAGE, in-Gel digestion, and MALDI-TOF-MS. In further experiments, the focus of the study was directed toward an extensive use and optimization of in-solution enzymatic digestion in combination with LC-MS/MS techniques in identification of specific peptide markers and finally in utilization of the latter in delivering reproducible quantification data for several different technical enzymes (alpha-amylases, xylanase, and lipases from microbial origin) in complex matrices such as baked bread and wheat flour. Two digestion protocols (a fast option using thermocycler program and the well-established overnight method) were tested, and both of these can be successfully applied. The application of isotopically labeled analogs of the MRM targeted peptides as internal standards and the addition of an internal protein standard during the extraction/digestion experiment were compared to determine the optimal quantification algorithm of the recovered enzyme concentrations. Thus, a standardized sensitive LC-MS/MS method could be developed to determine technical enzymes as forthcoming ingredients in the prefabricated food formulations in concentrations lower than 10 ppm.}, language = {en} } @article{GoetzChmielewskiGoedekeetal.2017, author = {Goetz, Klaus-Peter and Chmielewski, Frank M. and Goedeke, Kristin and Wolf, Kristine and Jander, Elisabeth and Sievers, Steven and Homann, Thomas and Huschek, Gerd and Rawel, Harshadrai Manilal}, title = {Assessment of amino acids during winter rest and ontogenetic development in sweet cherry buds (Prunus avium. L.)}, series = {Scientia horticulturae : an international journal sponsored by the International Society for Horticultural Science}, volume = {222}, journal = {Scientia horticulturae : an international journal sponsored by the International Society for Horticultural Science}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0304-4238}, doi = {10.1016/j.scienta.2017.05.001}, pages = {102 -- 110}, year = {2017}, abstract = {This study examined changes in sweet cherry buds of 'Summit' cultivar in four seasons (2011/12-2014/15) with respect to the nitrogen (N) content and the profile of eight free amino acids (asparagine (Asn), aspartic acid (Asp), isoleucine (Ile), glutamine (Gln), glutamic acid (Glu), arginine (Arg), alanine (Ala), histidine (His)). The presented results are to our knowledge the first under natural conditions in fruit tree orchards with a high temporal resolution from the dormant stage until cluster development. The N content in the buds from October, during endo- and ecodormancy until the beginning of ontogenetic development was a relatively stable parameter in each of the four seasons. The N accumulation into the buds began after 'swollen bud' and significant differences were visible at 'green tip' with an N content of 3.24, 3.12, 3.08, 2.40 which increased markedly to the mean of 'tight' and 'open cluster' by 3.77\%, 3.78\%, 3.44\% and 3.10\% in 2012-2015, respectively. In the buds, levels of asparagine were higher (up to 44 mg g\&\#8722;1 DW\&\#8722;1) than aspartic acid (up to 2 mg g\&\#8722;1 DW\&\#8722;1) and aspartic acid higher than isoleucine (up to 0.83 mg g\&\#8722;1 DW\&\#8722;1). Levels of glutamine were higher (up to 25 mg g\&\#8722;1 DW\&\#8722;1) than glutamic acid (up to 20 mg g\&\#8722;1 DW\&\#8722;1). The course of the arginine content was higher in 2011/12 compared to 2012/13, 2013/14 and 2014/15 which showed only slight differences. The alanine content in the buds was denoted in the four seasons only by relatively minor changes. The histidine content was higher in 2011/12 and 2012/13 compared to 2013/14 and 2014/15 which showed a comparable pattern. For 6 amino acids (Asn, Asp, Ile, Glu, Arg, Ala), the highest content was observed in 2012/13, the warmest period between swollen bud and open cluster. However in 2014/15, the season with the lowest mean temperature of 8.8 °C, only the content of Gln was the lowest. It was not possible to explain any seasonal differences in the amino acid content by environmental factors (air temperature) on the basis of few seasons. From none of the measured free amino acids could a clear determination of the date of endodormancy release (t1) or the beginning of the ontogenetic development (t1*) be derived. Therefore, these amino acids are no suitable markers to improve phenological models for the beginning of cherry blossom.}, language = {en} } @article{ChmielewskiGoetzHomannetal.2017, author = {Chmielewski, Frank M. and G{\"o}tz, Klaus-Peter and Homann, Thomas and Rawel, Harshadrai Manilal}, title = {Identification of Endodormancy Release for Cherries (Prunus Avium L.) by Abscisic Acid and Sugars}, series = {Journal of Horticulture}, volume = {4}, journal = {Journal of Horticulture}, number = {3}, issn = {2376-0354}, doi = {10.4172/2376-0354.1000210}, pages = {9}, year = {2017}, abstract = {In order to develop reliable and physiologically sound models for the plant development in spring, the date of endodormancy release is always a crucial and mostly unknown model parameter. Until present, classical approaches - such as climate chamber experiments - are used to derive this unknown parameter. In these experiments, progressive plant development or significant changes in bud's fresh weight or water content are measurable markers for dormancy release. This study presents an alternative approach, which is based on four well-known metabolites. For 5 seasons (2011/12-2015/16), the content of abscisic acid (ABA) and sugars such as fructose, sucrose and glucose in sweet cherry flower buds (cultivar 'Summit') were weekly analysed between beginning of October and April. These data allow comparing the annual course of these metabolites with the date of endodormancy release, derived from a classical climate chamber experiment, published in a previous study. Results showed that ABA and sucrose are two important metabolites which can help to identify the date of endodormancy release of sweet cherries. On average, ABA content reached a plateau of 5.65 μg g-1 DW-1 during endodormancy, which was maintained for 3-6 weeks. The significant reduction of the ABA content after this period to 4.41 μg g-1 DW-1 on average during ecodormancy was nearly in agreement with the date of endodormancy release of 'Summit' on 28 November (332 DOY). The annual cycle of sucrose, which has a cryoprotective effect during winter, is well comprehensible and showed a close relationship to the annual course of minimum air temperature after leaf fall(r=-0.90). The nearly constant level of sucrose during ecodormancy (21.0 mg g-1 DW-1, 5 yr. mean) did not only allow deriving the date of endodormancy release but can also be helpful to define the beginning of ontogenetic development.}, language = {en} }