Institut für Lebensmittelwissenschaft und Biotechnologie

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Die Erweiterung des genetischen Codes als molekulare Prozesskontrolle
(2025) Hiller, Eric; Hermann, Alexander; Lilge, Lars
Bioproduction of bioactive compounds on a large-scale poses challenges that require molecular strain engineering. By integrating the principle of genetic code expansion (GCE), the biosynthesis of the surfactin biosurfactant has been controlled in the production host Bacillus subtilis depending on the supply of a defined non-canonical amino acid. The regulation of this molecular process control has been transferred to high-cell-density fed-batch bioreactor procedures, allowing an initial upscaling.
Chemical compositions and oxidative stabilities of cold‐pressed walnut oils (Juglans regia L.): effects of chemical refining, water degumming, and molecular distillation
(2024) Liu, Longfei; Cai, Hongling; Zhang, Youfeng; Jin, Qingzhe; Wang, Xingguo; Jin, Jun
Walnut oils are of important academic and economic value, and are becoming one of the most important woody oils. Accurate and moderate refining techniques are required to produce high‐quality walnut oils. In this work, walnut oils obtained from cold processing were refined in three typical techniques, mainly chemical refining, water degumming, and molecular distillation. Physicochemical properties (acid value and peroxide value [POV]), minor components (tocopherol, polyphenols, and phytosterol), oxidative stability indices, and volatile compounds were analyzed to find out the appropriate refining method for the cold‐pressed walnut oils. Quality indices of all the refined oils from the three different refining methods met the requirements of the national standard, of which the POV of chemically refined oil (0.241 g/100 g) was higher than crude oil (0.058 g/100 g). Water degumming was most suitable for retaining of bioactive compounds, for example, the tocopherol was 259.40 mg/kg, the polyphenols was 44.54 mg GAE/kg, and the phytosterol was 987.32 mg/kg, but oxidation stability of the obtained oil (3.09 h) was lower than that of molecular distilled oil (4.18 h). Initial physicochemical properties especially the POV had a significant impact on oxidation stability. There is a trade‐off between the retention of nutrients and extending shelf life, indicating appropriate refining techniques should be developed; that is, water degumming is suggested to be involved in producing high‐quality cold‐pressed walnut oils.
Influence of low oxygen concentrations on color stability of modified atmosphere packaged beef
(2026) Krell, Johannes; Aeckerle, Luis; Poveda-Arteaga, Alejandro; Weiss, Jochen; Terjung, Nino; Gibis, Monika
The influence of low oxygen concentrations on the development of color and the myoglobin redox states over storage time was analyzed, to determine whether there are conditions that increase discoloration. Beef slices were packaged in atmospheres containing nitrogen gas and 0 %, 0.5 %, 1 %, 1.5 %, 3 %, and 5 % of oxygen. The samples were stored at 2 °C for 14 days. During storage, color, reflectance and oxygen concentration were measured optically through the packaging. The color difference ΔE2000 and the relative oxymyoglobin (OMb), deoxymyoglobin (DMb), and metmyoglobin (MMb) levels were calculated. After 14 days, the oxygen concentrations changed to 0.09 % (0 %), 0.36 % (0.5 %), 0.92 % (1 %), 1.28 % (1.5 %) 2.55 % (3 %), and 4.29 % (5 %). Regarding MMb formation, the 0 % samples (ΔMMb0–14d 11.1 %) were significantly (p < 0.05) more stable compared to the other samples, which showed an increase of MMb formation with rising oxygen concentration after 14 days. The other samples reached a ΔMMb0–14d increase of 21.1 % (0.5 %), 26.7 % (1 %), 30.0 % (1.5 %), 31.1 % (3 %), and 34.4 % (5 %). The color stability showed significantly (p < 0.05) increasing ΔE values of 2.49 (0 %), 3.39 (0.5 %), 4.66 (1 %), 5.14 (1.5 %), 6.03 (3 %), and 7.34 (5 %) with rising oxygen contents. These findings suggest that to ensure the color stability of beef with minimal MMb formation, it is important to completely exclude oxygen from the packages, since the destabilizing effect of oxygen already started at 0.5 %. The non-invasive measurement of the oxygen concentration and the reflectance data over 14 days gave new insights into the discoloration process of beef stored in low-oxygen atmospheres.
Toward food-grade production of the Bacteroides helcogenes protein-glutamine glutaminase with an optimized Bacillus subtilis strain
(2026) Senger, Jana; Keutgen, Mario; Roth, Nicole; Seitl, Ines; Fischer, Lutz; Senger, Jana; Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstr 25, 70599, Stuttgart, Germany; Keutgen, Mario; Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstr 25, 70599, Stuttgart, Germany; Roth, Nicole; Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstr 25, 70599, Stuttgart, Germany; Seitl, Ines; Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstr 25, 70599, Stuttgart, Germany; Fischer, Lutz; Institute of Food Science and Biotechnology, Department of Biotechnology and Enzyme Science, University of Hohenheim, Garbenstr 25, 70599, Stuttgart, Germany
Protein-glutamine glutaminases (PGs; EC 3.5.1.44) have gained attention in the food industry due to their application in plant protein products. The recently discovered PG from Bacteroides helcogenes (PGB) has especially been shown to provide promising characteristics for improving the techno-functional properties of plant proteins. A prerequisite for food enzymes, such as the PG, is their production with an expression host that meets food safety and yield requirements. The antibiotic-free and secretory production of the PGB was targeted in this study using the undomesticated Bacillus subtilis 007. The CRISPR/Cas9-mediated approach enabled specific genomic PGB integrations, while simultaneously deleting unwanted B. subtilis traits. Firstly, the PGB expression cassette was integrated into the sigF gene, leading to an asporogenic strain and extracellular activity of 4.1 µkat/Lculture in bioreactor cultivations. However, excessive foaming hampered the production process tremendously. Consequently, a second PGB copy was integrated into the sfp locus, which is involved in the production of lipopeptides, such as surfactin. As a result, the PGB activity was increased to 5.4 µkat/Lculture, and foaming during cultivation was reduced significantly. The introduction of a third PGB copy for preventing cell motility did not increase production; however, the integration into the well-established amyE locus improved the PGB yield during reactor cultivations. A final extracellular activity of 9.5 µkat/Lculture was reached. The multiple genomic integrations of the PGB gene enabled the efficient PGB secretion in an optimized B. subtilis host without the need for antibiotics.Key points• Site-specific PGB integration enabled by genome sequencing of B. subtilis 007.• Antibiotic-free and secretory PGB production with an optimized B. subtilis host.• Increased PGB production reaching 9.5 µkat/Lculture.
Potential of homopolysaccharide-producing starter cultures in the fermentation of coconut yoghurt alternatives enriched with pea protein isolate
(2026) Libberecht, Sophie; Ristevska, Mia; Gibis, Monika; Loeffler, Myriam; Libberecht, Sophie; Meat Technology & Science of Protein-Rich Foods (MTSP), Department of Microbial and Molecular Systems, Leuven Food Science and Nutrition Research Centre, KU Leuven Campus Ghent, 9000 Ghent, Belgium; Ristevska, Mia; Meat Technology & Science of Protein-Rich Foods (MTSP), Department of Microbial and Molecular Systems, Leuven Food Science and Nutrition Research Centre, KU Leuven Campus Ghent, 9000 Ghent, Belgium; Gibis, Monika; Department of Food Material Science, University of Hohenheim, 70593 Stuttgart, Germany;; Loeffler, Myriam; Meat Technology & Science of Protein-Rich Foods (MTSP), Department of Microbial and Molecular Systems, Leuven Food Science and Nutrition Research Centre, KU Leuven Campus Ghent, 9000 Ghent, Belgium; Martínez-Culebras, Pedro Vicente;; Roig, Patricia;; Martínez-Culebras, Pedro Vicente; Roig, Patricia
This study investigates the use of a homopolysaccharide (HoPS)-producing Latilactobacillus sakei strain for the production of protein-enriched plant-based yoghurt analogues based on coconut milk. Formulations varied in added sucrose (2.5% or 5.0% w / w ), pea protein isolate (PPI; 0–5.0% w / w ), and tapioca starch (0%, 1.5% w / w ), and were fermented with either a HoPS-producing strain ( L. sakei 1.411), or a non-exopolysaccharide (EPS)-producing control strain ( L. sakei 1.2037) with very similar acidification kinetics. Microbial growth and pH were monitored, HoPS content was determined via HPLC, and both firmness and syneresis were assessed during 5 days of storage at 4 °C. EPS yields were significantly higher ( p < 0.05) in samples with 5.0% w / w added sucrose compared to those with 2.5% w / w . Fermentation with L. sakei 1.411 generally resulted in firmer gels ( p < 0.05) and reduced syneresis ( p < 0.05) compared to L. sakei 1.2307 and the enhanced viscosity (sample thickness) was also observed in a sensory analysis. Samples containing starch and 5.0% w / w PPI showed the highest firmness-related values. These findings demonstrate the potential of in situ HoPS production to improve the texture and stability of protein-enriched coconut-based yoghurt analogues. It highlights the importance of matrix formulation, strain selection and process control, which all contribute to the final product quality.
Model-based process design for surfactin production with Bacillus subtilis
(2025) Hiller, Eric; Off, Manuel; Dittmann, Holger; Perino, Elvio Henrique Benatto; Lilge, Lars; Hausmann, Rudolf; Hiller, Eric; Department of Bioprocess Engineering, Institute of Food Science and Biotechnology, University of Hohenheim, Stuttgart, Germany; Off, Manuel; Department of Bioprocess Engineering, Institute of Food Science and Biotechnology, University of Hohenheim, Stuttgart, Germany; Dittmann, Holger; Department of Bioprocess Engineering, Institute of Food Science and Biotechnology, University of Hohenheim, Stuttgart, Germany; Perino, Elvio Henrique Benatto; Department of Bioprocess Engineering, Institute of Food Science and Biotechnology, University of Hohenheim, Stuttgart, Germany; Lilge, Lars; Department of Bioprocess Engineering, Institute of Food Science and Biotechnology, University of Hohenheim, Stuttgart, Germany; Hausmann, Rudolf; Department of Bioprocess Engineering, Institute of Food Science and Biotechnology, University of Hohenheim, Stuttgart, Germany
Bacillus subtilis is one of the most important production organisms in industrial biotechnology. However, there is still limited knowledge about the kinetics of fed-batch processes in bioreactors, as well as a lack of biological performance indicators, such as production yields, particularly regarding their variation over time. Understanding these kinetics and changes is crucial for optimizing the productivity in fed-batch processes. Fed-batch bioreactor cultures of Bacillus subtilis BMV9 in high cell density processes for surfactin production have been characterized with a kinetic model composed of first-order ordinary differential equations, describing the time course of biomass, substrate, surfactin and acetate. This model contributes to understanding critical restrictions and the knowledge gained was used to design and implement a model-based process. The model integrates biomass growth based on Monod kinetics, substrate consumption, surfactin synthesis and formation of the by-product acetate. After the model was parameterized for B. subtilis BMV9 using 12 different fed-batch bioreactor experiments, the kinetic model was able to accurately describe biomass accumulation, substrate consumption, product formation rates and, to some extent, the overflow metabolism involving acetate. Based on this, the kinetic model was used for a process design, in which the batch was omitted, which led to a product titre of 46.33 g/L and a space–time-yield of 2.11 g/(L*h) was achieved. The kinetic model developed in this study enables the description of the time course of biomass growth, substrate consumption and product formation and thus significantly improves process understanding. The computation of process parameters, which are not analytically accessible at any time, could be realized. A sensitivity analysis identified the maximum specific growth rate, substrate-related maintenance and the maximum acetate formation rate as key parameters influencing model outputs.
Intestinal dysbiosis associated with non-nutritive sweeteners intake: an effect without a cause?
(2025) Marongiu, Luigi; Brzozowska, Ewa; Hetjens, Svetlana; Hoelzle, Ludwig E.; Venturelli, Sascha; Brzozowska, Ewa; Laboratory of Medical Microbiology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland; Hetjens, Svetlana; Department of Medical Statistics, Biomathematics and Information Processing, University Clinic Mannheim, Mannheim, Germany; Hoelzle, Ludwig E.; Department of Food Microbiology and Hygiene, Institute of Food Science and Biotechnology, University of Hohenheim, Stuttgart, Germany; Venturelli, Sascha; Department of Nutritional Biochemistry, University of Hohenheim, Stuttgart, Germany
Non-nutritive sweeteners (NNS) are present in various commercial articles, from foodstuffs to oral hygiene products. Despite their alleged safety, mounting evidence indicates that NNS intake is associated with an alteration of intestinal bacterial populations (dysbiosis) in animals and humans. Since NNS are commercialized based on the assumption that they are not metabolized by human cells and negligible effect on bacterial, the insurgence of dysbiosis associated with NNS intake remains unexplained. The current review aims to assess the effect of selected NNS (acesulfame potassium, advantame, aspartame, neotame, saccharin, stevia, and sucralose) on the human intestinal microbiota. Findings from this review suggests that NNS intake is linked not only to alterations in human physiology but also to modifications of bacterial biochemistry, including the hindrance of quorum sensing pathways, in a species-specific manner. Moreover, there were suggestions that NNS could also affect the biology of phages, namely by binding to the active sites of proteins involved in the infection process and altering the induction rate of prophages. The studies gathered in the present review provide a framework for understanding how NNS might be connected to dysbiosis, both directly through alterations in bacterial biochemistry and indirectly through impaired phage activity.
Enabling adaptive food monitoring through sampling rate adaptation for efficient, reliable critical event detection
(2025) Jox, Dana; Schweizer, Pia; Henrichs, Elia; Krupitzer, Christian; Jox, Dana; Department of Food Informatics and Computational Science Hub, University of Hohenheim, 70599 Stuttgart, Germany; Schweizer, Pia; Department of Food Informatics and Computational Science Hub, University of Hohenheim, 70599 Stuttgart, Germany; Niu, Jianwei; Department of Food Informatics and Computational Science Hub, University of Hohenheim, 70599 Stuttgart, Germany; Niu, Jianwei
Monitoring systems are essential in many fields, such as food production, storage, and supply, to collect information about applications or their environments to enable decision-making. However, these systems generate massive amounts of data that require substantial processing. To improve data analysis efficiency and reduce data collectors’ energy demand, adaptive monitoring is a promising approach to reduce the gathered data while ensuring the monitoring of critical events. Adaptive monitoring is a system’s ability to adjust its monitoring activity during runtime in response to internal and external changes. This work investigates the application of adaptive monitoring—especially, the adaptation of the sensor sampling rate—in dynamic and unstable environments. This work evaluates 11 distinct approaches, based on threshold determination, statistical analysis techniques, and optimization methods, encompassing 33 customized implementations, regarding their data reduction extent and identification of critical events. Furthermore, analyses of Shannon’s entropy and the oscillation behavior allow for estimating the efficiency of the adaptation algorithms. The results demonstrate the applicability of adaptive monitoring in food storage environments, such as cold storage rooms and transportation containers, but also reveal differences in the approaches’ performance. Generally, some approaches achieve high observation accuracies while significantly reducing the data collected by adapting efficiently.
Toward food-grade production of the Glutamicibacter halophytocola diamine oxidase using Komagataella phaffii
(2025) Bechtel, Anna; Kettner, Lucas; Hessenberger, Jan; Vlassakakis, Kenny; Fischer, Lutz; Bechtel, Anna; Department of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany; Kettner, Lucas; Department of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany; Hessenberger, Jan; Department of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany; Vlassakakis, Kenny; Department of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany; Fischer, Lutz; Department of Biotechnology and Enzyme Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstr. 25, 70599, Stuttgart, Germany
The diamine oxidase from Glutamicibacter halophytocola (DAO-GH) was recombinantly produced in K. phaffii using the constitutive glyceraldehyde-3-phosphate dehydrogenase promoter for methanol-free production. Firstly, K. phaffii clones were generated for intracellular and secretory DAO-GH production that still possessed antibiotic resistance due to the cloning procedure. For intracellular production, a maximum intracellular DAO activity of 15,404 nkat/Lculture was achieved in fed-batch bioreactor cultivations, while for secretory production, the highest extracellular DAO activity of 6,078 nkat/Lculture was achieved using the αMF signal peptide without its EAEA sequence. The intracellularly produced DAO-GH was partially purified in several purification steps with a yield of 80%, a purification factor of about 10 and specific DAO activity of 16.7 nkat/mgprotein. The secretory DAO-GH production resulted in a specific DAO activity of 15.4 nkat/mgprotein already in the cell-free culture supernatant at the end of cultivation without further purification steps. The food industry aims to avoid the use of antimicrobial resistance in enzyme production, therefore, a new cassette plasmid with self-excisable antibiotic resistance markers was constructed for secretory DAO-GH production. The antibiotic-resistance-free K. phaffii clone generated with this plasmid achieved a maximum extracellular DAO activity of 4,770 nkat/Lculture in a fed-batch bioreactor cultivation. The DAO-GH obtained in this cultivation was spray-dried, resulting in a storable powder with 23 nkat/gpowder DAO activity and a water activity value of 0.12. This study demonstrated the secretion of recombinant DAO in a microbial host such as K. phaffii for the first time and provides a strategy for generating antibiotic-resistance-free K. phaffii clones.
Secretion of the cytoplasmic and high molecular weight β-galactosidase of Paenibacillus wynnii with Bacillus subtilis
(2024) Senger, Jana; Seitl, Ines; Pross, Eva; Fischer, Lutz
Background: The gram-positive bacterium Bacillus subtilis is widely used for industrial enzyme production. Its ability to secrete a wide range of enzymes into the extracellular medium especially facilitates downstream processing since cell disruption is avoided. Although various heterologous enzymes have been successfully secreted with B. subtilis, the secretion of cytoplasmic enzymes with high molecular weight is challenging. Only a few studies report on the secretion of cytoplasmic enzymes with a molecular weight > 100 kDa. Results: In this study, the cytoplasmic and 120 kDa β-galactosidase of Paenibacillus wynnii (β-gal-Pw) was expressed and secreted with B. subtilis SCK6. Different strategies were focused on to identify the best secretion conditions. Tailormade codon-optimization of the β-gal-Pw gene led to an increase in extracellular β-gal-Pw production. Consequently, the optimized gene was used to test four signal peptides and two promoters in different combinations. Differences in extracellular β-gal-Pw activity between the recombinant B. subtilis strains were observed with the successful secretion being highly dependent on the specific combination of promoter and signal peptide used. Interestingly, signal peptides of both the general secretory- and the twin-arginine translocation pathway mediated secretion. The highest extracellular activity of 55.2 ± 6 µkat/Lculture was reached when secretion was mediated by the PhoD signal peptide and expression was controlled by the PAprE promoter. Production of extracellular β-gal-Pw was further enhanced 1.4-fold in a bioreactor cultivation to 77.5 ± 10 µkat/Lculture with secretion efficiencies of more than 80%. Conclusion: For the first time, the β-gal-Pw was efficiently secreted with B. subtilis SCK6, demonstrating the potential of this strain for secretory production of cytoplasmic, high molecular weight enzymes.
Interactions between lipid oxidation and anthocyanins from black carrots in ω-3 fatty acid-rich flaxseed oil-in-water emulsions
(2024) Klinger, Evelyn; Salminen, Hanna; Bause, Karola; Weiss, Jochen
The application of anthocyanins as red colorants in lipid-containing foods such as oil-in-water emulsions is challenging due to their ability to act as antioxidants and their instability under various environmental conditions. In this study, we investigated the kinetics of black carrot anthocyanin degradation and the subsequent color loss influenced by lipid oxidation reactions in 1% ( w/w ) flaxseed oil-in-water emulsions stabilized by 0.1% ( w/w ) sodium dodecyl sulfate (SDS) at pH 2 upon storage at 35 °C for ten days under light and in the dark. Oxidatively stable Miglyol oil-in-water emulsions and SDS solutions were used as reference. The results showed simultaneous formation of lipid hydroperoxides and degradation of anthocyanins. The addition of anthocyanins decreased lipid hydroperoxide formation, confirming the antioxidant activity of anthocyanins through radical scavenging mechanism. The kinetic modelling of lipid oxidation and anthocyanin degradation are particularly important for estimating color stability in colored emulsion-based food systems such as dairy or non-dairy milk or yoghurt drinks.
Transforming agricultural waste from mediterranean fruits into renewable materials and products with a circular and digital approach
(2025) Castagna, Antonella; Aboudia, Aouatif; Guendouz, Amine; Scieuzo, Carmen; Falabella, Patrizia; Matthes, Julia; Schmid, Markus; Drissner, David; Allais, Florent; Chadni, Morad; Cravotto, Christian; Senge, Julia; Krupitzer, Christian; Canesi, Ilaria; Spinelli, Daniele; Drira, Fadoua; Ben Hlima, Hajer; Abdelkafi, Slim; Konstantinou, Ioannis; Albanis, Triantafyllos; Yfanti, Paraskevi; Lekka, Marilena E.; Lazzeri, Andrea; Aliotta, Laura; Gigante, Vito; Coltelli, Maria-Beatrice; Hilliou, Loic
The Mediterranean area is one of the major global producers of agricultural food. However, along the entire supply chain—from farming to food distribution and consumption—food waste represents a significant fraction. Additionally, plant waste residues generated during the cultivation of specific fruits and vegetables must also be considered. This heterogeneous biomass is a valuable source of bioactive compounds and materials that can be transformed into high-performance functional products. By analyzing technical and scientific literature, this review identifies extraction, composite production, and bioconversion as the main strategies for valorizing agricultural by-products and waste. The advantages of these approaches as well as efficiency gains through digitalization are discussed, along with their potential applications in the Mediterranean region to support new research activities and bioeconomic initiatives. Moreover, the review highlights the challenges and disadvantages associated with waste valorization, providing a critical comparison of different studies to offer a comprehensive perspective on the topic. The objective of this review is to evaluate the potential of agricultural waste valorization, identifying effective strategies while also considering their limitations, to contribute to the development of sustainable and innovative solutions in Mediterranean bioeconomy.
Synthesis of biologically active Shiga toxins in cell-free systems
(2024) Ramm, Franziska; Kaser, Danny; König, Irina; Fellendorf, Juliane; Wenzel, Dana; Zemella, Anne; Papatheodorou, Panagiotis; Barth, Holger; Schmidt, Herbert
Shiga toxins (Stx) produced by pathogenic bacteria can cause mild to severe diseases in humans. Thus, the analysis of such toxins is of utmost importance. As an AB5 toxin, Stx consist of a catalytic A-subunit acting as a ribosome-inactivating protein (RIP) and a B-pentamer binding domain. In this study we synthesized the subunits and holotoxins from Stx and Stx2a using different cell-free systems, namely an E. coli- and CHO-based cell-free protein synthesis (CFPS) system. The functional activity of the protein toxins was analyzed in two ways. First, activity of the A-subunits was assessed using an in vitro protein inhibition assay. StxA produced in an E. coli cell-free system showed significant RIP activity at concentrations of 0.02 nM, whereas toxins synthesized in a CHO cell-free system revealed significant activity at concentrations of 0.2 nM. Cell-free synthesized StxA2a was compared to StxA2a expressed in E. coli cells. Cell-based StxA2a had to be added at concentrations of 20 to 200 nM to yield a significant RIP activity. Furthermore, holotoxin analysis on cultured HeLa cells using an O-propargyl-puromycin assay showed significant protein translation reduction at concentrations of 10 nM and 5 nM for cell-free synthesized toxins derived from E. coli and CHO systems, respectively. Overall, these results show that Stx can be synthesized using different cell-free systems while remaining functionally active. In addition, we were able to use CFPS to assess the activity of different Stx variants which can further be used for RIPs in general.
Hydration-induced reduction strategies of acrylamide in bakery products
(2025) Świacka, Jagoda; Jekle, Mario
Bread is a product made from cereals and/or cereal-based ingredients, which are combined with water or other water-containing liquids. Also, additional ingredients such as oilseeds, legumes, potato products, dairy products, spices, and vegetables may be incorporated into the dough. Products containing these ingredients can be referred to as special bakery products. However, the use of these additional ingredients has been raising food-safety concerns, particularly because of the possibly elevated levels of acrylamide detected in the final products. Acrylamide is a process contaminant, considered undesirable due to its potential carcinogenicity and is currently subject to various European food safety regulations. However, the acrylamide mitigation strategies defined so far for standard bakery products do not appear to be sufficiently effective for special bakery products. This is presumably due to the high levels of precursor molecules and the varying quality of the special ingredients. As a result, there are large variations in the levels of precursors, namely reducing sugars and asparagine in the dough, which consequently lead to unpredictably elevated acrylamide contamination. To address these problems and to gain a deeper understanding of acrylamide formation in special bakery products, further investigation is essential. Therefore, this thesis aimed to identify specific acrylamide precursors, to analyze acrylamide levels, and to assess the effectiveness of hydration-based approaches as mitigation measures for special bakery products. Mitigation was to be achieved either by eliminating the precursors or by adjusting processing conditions. After a theoretical introduction in Chapter I, a systematic investigation of two special ingredients, with a focus on their specific acrylamide precursors, was presented in Chapter II and Chapter III. Particular attention was drawn to potato flakes, known for their high asparagine content, and carrot strips, awaited to be high in reducing sugars. The effects of these ingredients in combination with varying water additions as hydration approach, were investigated using a model dough recipe. The model dough consisted of wheat flour type 550, water, NaCl, yeast and selected special ingredients. After baking, both bread crust and crumb were analyzed for acrylamide and in parallel, baking qualities of these bakery products were evaluated. Considering acrylamide contamination, increased water addition showed highly raw material-specific effects. In high-asparagine doughs, higher dough water addition tended even to increase acrylamide formation, presumably due to enhanced mobility of precursors. Interestingly, for the carrot-based doughs, the polyphenols emerged as a potential factor significantly reducing acrylamide formation at higher water additions, highlighting their relevance for future mitigation strategies. Due to the significant differences in the qualities of available carrot strips, shown in Chapter III, Chapter IV described the effects of origin, processing and drying methods on different quality features of dried carrot strips. For that, three carrot varieties were purchased and processed. Processing steps included blanching and drying methods, such as hot-air drying, vacuum drying, or freeze-drying. The impact of each processing method on quality of carrot strips, their acrylamide-forming potential, and the resulting acrylamide levels in model bakery products was analyzed. Blanching in particular, significantly reduced the acrylamide formation potential of the carrot strips. Freeze-drying consistently resulted in the highest concentrations of sugars, free asparagine, and β‑carotene for all carrot varieties, whereas the retainment of these compounds was inconsistent with other drying methods. Asparagine levels during storage were shown to fluctuate, with an initial increase and a following decrease, underlining the importance of timed processing to warrant constant product quality. Overall, the potential of carrot strips for acrylamide formation depended mostly on their variety and storage conditions, which determined primarily their precursor profile. To investigate a variety-unspecific acrylamide mitigation strategy, soaking was evaluated in Chapter V as an approach for products with dried carrot strips. Additionally, ultrasound-assisted rehydration was investigated as a method to enhance the diffusion of carrot matrix constituents. Regular soaking was compared with ultrasound-assisted rehydration (ultrasonic bath or sonotrode) regarding rehydration rate and leaching of sugars from the carrot strips. The kinetics of rehydration and diffusion were modelled using the ExpAssoc and the Peleg equations. Rehydrated carrot strips were then implemented in model breads and acrylamide concentration in the crust along with baking qualities were analyzed. Overall, longer rehydration led to higher water retention in the carrot strips and increased leaching of sugars and polyphenols. Both ultrasound-assisted rehydration and regular soaking led to reduced acrylamide levels in model breads up to 75%. Summarizing, this thesis systematically examined the addition of potato flakes and carrot strips to bakery products, identifying their critical acrylamide precursors, expected acrylamide levels, and characteristic changes in bread quality. It proved that rehydration is an effective acrylamide mitigation strategy for special bakery products, primarily by promoting precursor leaching and influencing reaction conditions. Overall, this thesis provides guidance on the target raw material qualities and presents (easy-to-apply) mitigation approaches for various manufacturers.
On the structural analysis and optimal input design for joint state and parameter estimation
(2025) Lepsien, Arthur; Kügler, Philipp; Schaum, Alexander
This paper addresses the problem of joint state and parameter estimation for nonlinear affine-input systems with positive parameters including the design of a closed-loop optimal input adaptation to increase an identifiability measure for the system. The identifiability itself is considered in the context of structural observability of the system dynamics based on structural analysis of the system including the unknown parameters as additional states. In particular, the network graph-based interpretation of structural observability is employed at this point. This analysis motivates to include time derivatives of the measurements as additional system outputs to enhance the structural observability properties. For this purpose robust exact differentiation is considered, relying on the super-twisting algorithm to obtain finite time convergent estimates of these signals. Using the extended measurement signal, a continuous-discrete Extended Kalman filter is proposed that ensures strictly positive estimates for the parameters. Based on the estimates of states and parameters the input signal is determined using a moving horizon optimal predictive control that evaluates the condition number of the Fisher information matrix, thus maximizing the information content of the measurements with respect to the parameters. The proposed scheme extends and combines different previously discussed approaches from the literature and is evaluated by means of a thermal process example in simulation and experiment, showing high potential for similar system identification problems.
Sustainability certification in the digital era: a qualitative analysis of sustainability reports across product categories
(2026) Boller, Meta Leonie; Grabinger, Toni; Zurwehme, Annikka; Krupitzer, Christian
The food industry faces growing pressure to align sustainability goals with the rapid advance of digital technologies. This study explores how digitalization supports and transforms sustainability certification practices across diverse food product categories. Novelty lies in examining the interplay between digital technologies and sustainability reporting intensity, offering insights into how firms communicate and operationalize sustainability commitments. A qualitative content analysis was conducted on the sustainability reports of 21 companies using MAXQDA software, assessing the frequency and depth of reporting on sustainability focus topics and digital tools applied. The study includes multiple food product categories, such as coffee and tea, chocolate, grain products, dairy products, beverages, meat and fish, and tropical fruits. Results show that larger firms and those with higher turnover report more frequently and comprehensively on sustainability and digitalization. Three clusters of companies were identified, each reflecting shared sustainability challenges rather than specific product categories. The findings suggest that the relevance of digital technologies is more closely linked to addressing these cross-cutting sustainability challenges, and that high-value product segments provide the most promising starting points for integrating digital technologies into certification processes.
Monitoring a coffee roasting process based on near‐infrared and raman spectroscopy coupled with chemometrics
(2025) Munyendo, Leah; Schuster, Katharina; Armbruster, Wolfgang; Babor, Majharulislam; Njoroge, Daniel; Zhang, Yanyan; von Wrochem, Almut; Schaum, Alexander; Hitzmann, Bernd
Roasting is a fundamental step in coffee processing, where complex reactions form chemical compounds related to the coffee flavor and its health‐beneficial effects. These reactions occur on various time scales depending on the roasting conditions. To monitor the process and ensure reproducibility, the study proposes simple and fast techniques based on spectroscopy. This work uses analytical tools based on near‐infrared (NIR) and Raman spectroscopy to monitor the coffee roasting process by predicting chemical changes in coffee beans during roasting. Green coffee beans of Robusta and Arabica species were roasted at 240°C for different roasting times. The spectra of the samples were taken using the spectrometers and modeled by the k‐nearest neighbor regression (KNR), partial least squares regression (PLSR), and multiple linear regression (MLR) to predict concentrations from the spectral data sets. For NIR spectra, all the models provided satisfactory results for the prediction of chlorogenic acid, trigonelline, and DPPH radical scavenging activity with low relative root mean square error of prediction (pRMSEP < 9.649%) and high coefficient of determination ( R 2 > 0.915). The predictions for ABTS radical scavenging activity were reasonably good. On the contrary, the models poorly predicted the caffeine and total phenolic content (TPC). Similarly, all the models based on the Raman spectra provided good prediction accuracies for monitoring the dynamics of chlorogenic acid, trigonelline, and DPPH radical scavenging activity (pRMSEP < 7.849% and R 2 > 0.944). The results for ABTS radical scavenging activity, caffeine, and TPC were similar to those of NIR spectra. These findings demonstrate the potential of Raman and NIR spectroscopy methods in tracking chemical changes in coffee during roasting. By doing so, it may be possible to control the quality of coffee in terms of its aroma, flavor, and roast level.
From spectra to traits: advancements in predicting wheat characteristics
(2025) Nagel-Held, Johannes Henrich; Hitzmann, Bernd
This thesis is driven by two primary objectives: To evaluate the potential for improving quality assessment throughout the wheat value chain and the potential for replacing protein content as the primary determinant of price and quality with spectroscopic methods. An early, fast, and well-founded decision on wheat quality leads to improved processing properties and product quality of wheat products. Given that these attributes are often more closely tied to protein quality or other components than protein quantity, a direct and precise quantification is highly desirable. The noteworthy advantage of protein content lies in its prediction through near-infrared spectroscopy (NIRS). To augment the information available from near-infrared spectra, which are inherently limited in their insights into molecular compounds within samples, NIRS was supplemented with Raman and fluorescence spectroscopy. The thesis`s foundation comprises four distinct sample sets, encompassing both common wheat and spelt. In total, a staggering 4,237 samples of whole grain, whole grain flour, and extracted flour were analyzed to predict a wide array of 100 diverse quality parameters, spanning from measures of protein quality to dough rheological properties, baking behavior, physical and chemical characteristics and agronomic traits. To enable accurate predictions several spectra pre-processing and regression techniques were applied. Among the spectrometers and algorithms tested, no clear recommendation can be given. In this work, NIRS was found to perform well over different levels of sample preparation, while Raman and fluorescence spectroscopy performed better on flour. A glimpse into the findings is best exemplified by considering loaf volume, a measure of end-product quality. Prediction errors (RMSECV) ranged between 29 and 43 mL/100 g of flour, accompanied by R² values spanning 0.66 to 0.78. One major factor influencing the improvable prediction accuracy is the inherent measurement error associated with the baking trial. However, when this measurement error is known, a straightforward solution emerges: By measuring a few samples repeatedly, the prediction error can be corrected from 43 mL/100 g of flour to 28 mL/100 g, aligning with the magnitude of the measurement error itself. Simultaneously, the corrected R² value improves from 0.66 to 0.86. It has been shown that measurement error correction works for other quality characteristics, such as water absorption or baking loss, and may be applied to other parameters. The robustness of the models was evaluated by successfully predicting complex traits of unknown cultivars. However, the models proved less effective in predicting across separate locations and years. Similar robustness results were found for protein content, water absorption and grain yield. Since protein content is a well-established and predictable parameter, robust models for complex traits are also feasible. Other well predictable parameters are protein content (R² = 0.97-0.98, RMSECV = 0.16-0.20 %), wet gluten content (R² = 0.89-0.98, RMSECV = 0.80-0.84 %), water absorption (R² = 0.68-0.83, RMSECV = 0.9-1.2 mL/100 g) and grain yield (R² = 0.73-0.85, RMSECV = 5.9-6.8 dt/ha). Examples for non-predictable parameters are starch properties measured with a rapid visco analyzer or enzyme activity. And for some parameters no clear conclusion could be drawn, such as SDS sedimentation volume, plant height, thousand kernel weight, extensograph or farinograph parameters. The successful prediction of intricate traits can be further refined by reducing measurement errors of the reference values and using better suited spectrometers. The robustness can be increased by expanding the sample set by including data from a broader spectrum of locations and years.Implementing this proposed methodology holds the potential to instigate positive changes within the wheat supply chain, with implications for product quality, environment, and economic aspects.
Storable cheese curd—effect of milk homogenization as a pre-treatment and freezing and extrusion of cheese curd on production of pasta filata style cheese
(2024) Schmidt, Florian; Graf, Britta; Hinrichs, Jörg
This study investigates the production of pasta filata style cheese from a storable, frozen intermediate material. Homogenization (2–16 MPa, single-staged) of milk (fat/protein = 0.9) was used as a tool to decrease fat globule size and consequently fat losses. Plasticization was achieved by using a single-screw extruder set up with double-jacketed hot water cycle. Non-frozen and frozen cheese curd as well as the extruded pasta filata style cheese pre-treated with different homogenization pressure was analyzed regarding the thermo-rheological properties. Fat and protein gain/loss during extrusion was evaluated by analyzing fat in dry matter (FDM) and protein in dry matter (PDM) before and after extrusion. Homogenization of cheese milk leads to a reduction of tan δ for thereof produced raw cheese curd material as well as the extruded products. Freezing and extrusion counteract the reduction of tan δ . A homogenization pressure of 8 MPa is sufficient to prevent fat losses during extrusion while still maintaining plasticization of the product for fresh and frozen material, respectively. The FDM after extrusion is 0.8% higher for fresh material and 4.9% higher for frozen material, which means that the fat concentrates during extrusion due to water loss. Moreover, there is no loss of PDM for all samples, regardless of the homogenization pressure. A combination of homogenization pressure, freezing, and extrusion leads to a plasticizable product without losses of fat and protein. Hence, frozen cheese curd can be used as a storable intermediate.
Comparative assessment of ethanol production from six typical German waste baked products
(2024) Almuhammad, Mervat; Kölling, Ralf; Einfalt, Daniel
This study investigates the potential for bioethanol production of six types of typical German leftover baked products: bread rolls, pretzel rolls, fine rye bread, white bread, pastry, and cream cakes. The experimental setup consisted of two experiments—one as a control and another with the addition of diammonium phosphate (DAP) to the mash. In terms of monosaccharide concentration at 30% dry matter (DM), white bread mash exhibited the highest level at 251.5 g/L, while cream cakes mash had the lowest at 186 g/L. The highest ethanol production occurred after 96 h of fermentation with rye bread, yielding 78.4 g/L. In contrast, despite having the highest monosaccharide levels, white bread produced only 21.5 g/L of ethanol after 96 h. The addition of DAP accelerated monosaccharide consumption in all baked products, with cream cakes completing the process in just 24 h. Bread rolls, pretzel rolls, pastry, and white bread fermentations finished within 72 h. Ethanol yields significantly increased in three DAP samples, with pretzel rolls yielding the highest ethanol concentration at 98.5 g/L, followed by white bread with 90.6 g/L, and bread rolls with 87.7 g/L. DAP had a substantial impact on all samples, reducing fermentation time and/or increasing ethanol yield. This effect was particularly pronounced with white bread, where it improved conversion efficiency from 17 to 72%, resulting in 90.6 g/L of ethanol. These results demonstrate that waste baked products hold substantial potential for bioethanol production, and this potential can be further enhanced through the addition of DAP.

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