Institut für Lebensmittelwissenschaft und Biotechnologie

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Bioprocess strategies for efficient microbial conversion of acetate as alternative biotechnological carbon source derived from lignocellulose streams
(2024) Kiefer, Dirk; Hausmann, Rudolf
In near future, humanity will be faced with the global challenges from climate change, fossil fuel depletion to food shortage. Our current economic system will thus need to make a transition into a sustainable bioeconomy which uses key technologies like biotechnology for conversion of renewable feedstocks. The majority of all biotechnological carbon sources is generated from food plants. In context to the global population growth coming along with food insecurity, competing use of these food plants for human and animal nutrition creates a substrate dilemma for future biotechnology. Future biorefineries will thus need to realize a paradigm shift to carbon sources generated from non-competing or waste resources. As such, especially lignocellulosic biomass is regarded as the central feedstock for a bio-based industry. Among the different substrate streams obtained by lignocellulose refining strategies, the C2 carboxylic acid acetic acid (from now on referred to as acetate) is one of the most common occurring components. In contrast to its high economic role as building block in the chemical industry, acetate shows low relevance as biotechnological carbon source yet. This is mainly related to its inhibitory characteristic, the reason for why acetate has been used in food preservation for thousands of years. However, most industrial platform organisms are also capable to grow on acetate as sole carbon source. It also shows several attractive characteristics for bioprocessing which is why research studies on microbial acetate conversion have significantly increased in the past years. Together with the potential routes to generate non-fossil bio-acetate in large quantities from non-food competing resources like lignocellulose and biomass-derived/waste C1 gas streams, acetate might become a next-generation platform substrate for a future bio-industry. In order to make acetate-based bioprocesses competitive to those using conventional sugar-based carbon sources, adapted process strategies for efficient acetate conversion need to be established. The present thesis is aimed to address efficient bioprocess strategies for biotechnological conversion of lignocellulosic acetate into microbial biomass and growth-coupled products under fed-batch culture conditions. The experimental studies presented in this work were divided into two parts: In Part I, the potential of acetate as carbon source for high cell density cultivation of industrial platform bacterium Corynebacterium glutamicum was evaluated. Preliminary growth studies with different initial acetate concentrations revealed a high natural acetate tolerance for wild-type ATCC 13032 with growth at high acetate levels up to 60 g/L. In addition, it was shown that maximum growth rates (μmax) of 0.47 h-1 for acetate concentrations below 10 g/L are competitive to that on D-glucose as common carbon source. By implementation of an online feeding control which enables automated supply of pure acetic acid (HAc) via pH control, high cell density cultivation on lignocellulosic acetate was demonstrated for the first time in a 42 L stirred-tank bioreactor. Optimization of the molar carbon-to-nitrogen feeding ratio resulted in a highly efficient bioprocess yielding cell densities up to 80.2 g/L CDW after 28.9 h with biomass yields (YXIS) of 0.35 g/g and space-time yields (STY) of 66.6 g/L·d. Part II focused on the potential of acetate as carbon source for integrated protein production in C. glutamicum. Batch cultures of genome-reduced strains MB001/MB001(DE3) demonstrated that acetate clearly shows no inhibitory effect on protein production of eYFP as model protein using C. glutamicum as production host. Interestingly, comparative expression studies with C. glutamicum T7 expression system indicated an up to 83 % higher biomass-specific production on acetate compared to D-glucose as carbon source. By transferring the implemented pH-coupled online feeding control for high cell density cultivation of strain MB001(DE3) pMKEX2-eyfp in a 42 L stirred-tank bioreactor, this study showed efficient protein accumulation on lignocellulosic acetate yielding final protein titers of 2.7 g/L after 27 h with product yields (YPIX) of 40 mg/g and volumetric productivities (PV) of 0.10 g/L·h. In conclusion, the presented results demonstrate the high biotechnological potential of acetate as alternative carbon source. In contrast to most other studies before, it is shown that a suitable process strategy based on pH-coupled online feeding control allows efficient microbial acetate conversion under industry-relevant fed-batch culture conditions. This work provides exemplary proof-of-concept bioprocesses for high cell density cultivation on lignocellulosic acetate and its growth-coupled conversion into protein using industrial platform organism C. glutamicum. Therefore, the presented thesis contributes to the development of efficient concepts for microbial conversion of next-generation platform substrates like lignocellulosic acetate.
Characterization of aroma and sensorial variation of basil and ginger during short-time drying process
(2024) Liang Jiaqi; Zhang, Yanyan
Industries are increasingly interested in aromatic herbs and spices, such as basil and ginger, which play a crucial role in daily life as flavoring agents in foods, beverages, and pharmaceuticals. Basil, an annual herb from the mint family (Lamiaceae), is renowned for its wonderful “royal” fragrance and numerous health benefits. Ginger, one of the most widely consumed dietary spices worldwide, has seen a surge in interest due to its significant healthpromoting properties. In 2023, global market sales for basil leaves and ginger reached 50 million euros and 4.2 billion euros, respectively. As may other herbs and spices, basil and ginger are typically used in dried form to extend shelf life and facilitate transportation. However, the drying process significantly alter the aroma from fresh samples, reduce the quality of the final products and generate numerous by-products. This study aims to identify the off-odor compound(s) produced during basil drying process using molecular sensory techniques and to develop effective strategies to inhibit their formation. Additionally, ginger peel and ginger fiber by-products generated during the industrial and domestic use, as well as the spray drying process, were investigated and reintroduced into the procedure to preserve the aroma compounds of ginger and promote environmental sustainability. In the first part of the study, a method using stir bar sorptive extraction combined with gas chromatography-mass spectrometry-olfactory (SBSE-GC-MS-O) was established to analyse the aroma changes of basil and ginger during short-term spray drying. This approach was designed to maximize desorption efficiency for various odorants with diverse physicochemical properties under different desorption conditions through four types of mathematical modelling. Among these models, the Random Forest model demonstrated the highest performance and minimal errors, with an R value of 0.910 after validation using a dataset of six new compounds. In addition, the model determined that cryo-focusing temperature was the most important factor, followed by molecular weight, log P, boiling point, desorption temperature, desorption time, and helium flow. This algorithm can be further utilized to predict the optimal parameters for maximizing desorption efficiency in aroma analysis of basil and ginger by SBSE-GC-MS-O. Subsequently, to investigate the formation of the hay-like off-odor that lowers consumer acceptability in dried basil products, the responsible compound was identified in thawed, airdried, and spray-dried basil samples using a trained human panel (n = 10) and gas chromatography–mass spectrometry–olfactometry. 3-Methylnonane-2,4-dione (3-MND) was found to be the odorant contributing to the hay-like off-odor in all basil samples. In order to reduce this odorant, the effects of light, oxygen, and temperature on the formation of 3-MND during the processes of thawing, spray drying, and air drying were studied based on a potential pathway involving 3-MND precursors. The results revealed that controlling light exposure, employing nitrogen protective environment, and maintaining low temperatures are critical processing parameters for minimizing the generation of the hay-like compound 3-MND, thereby meeting consumer demands for high-quality dried basil products. Regarding the processing of ginger, the drying process generates a significant amount of ginger peel as an industrial by-product. To facilitate environmental sustainability and establish a reference for its potential applications, the effect of ginger peel on aroma, sensory profiles, and nutrition-related physicochemical properties was investigated. The total concentration of aroma compounds in unpeeled ginger was 1.3 times higher than that in peeled ginger, according to SBSE-GC-MS-O analysis. Sensory evaluation data indicated that unpeeled ginger had significantly enhanced citrus-like and fresh impressions compared to peeled ginger, which is associated with the higher odor activity values of odorants such as β-myrcene (pungent, citruslike), geranial (citrus-like), citronellal (citrus-like, sourish), and linalool (floral, fresh). Additionally, the total polyphenol content in unpeeled ginger was 7.96 mg/100 g higher than that in peeled ginger. Unpeeled ginger not only demonstrates more intense aromatic and sensory qualities but also offers superior nutritional and environmental benefits, making it a promising option for future global spice use and reducing ginger side streams. In the spray drying process of ginger, ginger fibre is also typically regarded as industrial byproduct, as ginger juice is often used as a feed material. To achieve sustainable development and produce a clean-labeled product, a split-stream spray-drying process was developed to address sugar-rich feed solutions by reintroducing ginger fiber in their natural composition as a carrier material, rather than treating them as a by-product. The characterizations of both feed materials were then compared. The method was optimized for aroma retention by adjusting the inlet and outlet temperatures to 220 ℃ and 80 ℃, respectively. Aroma decoding results using SBSE-GC-MS-O demonstrated that reintegrating ginger fiber significantly increased the concentration of eight key odorants, including hexanal, linalool, neral, borneol, geranial, citronellol, nonanoic acid, and α-bisabolol, in comparison to the concentration observed in ginger juice. This presents a promising solution for maximizing the utilization of ginger in spray drying, enhancing the aroma profiles of feed material, and addressing sustainability considerations in the food industry. Overall, the presented work comprehensively explores the challenges and potential solutions associated with the drying process of basil and ginger, highlighting the impact of various processing conditions on aroma and sensory properties. By identifying key off-odor compounds and optimizing techniques to reduce their formation, as well as investigating aroma profiles and sustainable approaches for utilizing by-products such as ginger peel and ginger fiber, this research offers valuable insights into improving both the aromatic quality and environmental sustainability of dried herb and spice products.
The influence of growth rate-controlling feeding strategy on the surfactin production in Bacillus subtilis bioreactor processes
(2024) Hiller, Eric; Off, Manuel; Hermann, Alexander; Vahidinasab, Maliheh; Benatto Perino, Elvio Henrique; Lilge, Lars; Hausmann, Rudolf
Background The production of surfactin, an extracellular accumulating lipopeptide produced by various Bacillus species, is a well-known representative of microbial biosurfactant. However, only limited information is available on the correlation between the growth rate of the production strain, such as B. subtilis BMV9, and surfactin production. To understand the correlation between biomass formation over time and surfactin production, the availability of glucose as carbon source was considered as main point. In fed-batch bioreactor processes, the B. subtilis BMV9 was used, a strain well-suited for high cell density fermentation. By adjusting the exponential feeding rates, the growth rate of the surfactin-producing strain, was controlled. Results Using different growth rates in the range of 0.075 and 0.4 h-1, highest surfactin titres of 36 g/L were reached at 0.25 h-1 with production yields YP/S of 0.21 g/g and YP/X of 0.7 g/g, while growth rates lower than 0.2 h-1 resulted in insufficient and slowed biomass formation as well as surfactin production (YP/S of 0.11 g/g and YP/X of 0.47 g/g for 0.075 h-1). In contrast, feeding rates higher than 0.25 h-1 led to a stimulation of overflow metabolism, resulting in increased acetate formation of up to 3 g/L and an accumulation of glucose due to insufficient conversion, leading to production yields YP/S of 0.15 g/g and YP/X of 0.46 g/g for 0.4 h-1. Conclusions Overall, the parameter of adjusting exponential feeding rates have an important impact on the B. subtilis productivity in terms of surfactin production in fed-batch bioreactor processes. A growth rate of 0.25 h-1 allowed the highest surfactin production yield, while the total conversion of substrate to biomass remained constant at the different growth rates.
Development of rapid analytical methods for coffee quality assessment: Spectroscopy and chemometrics approach
(2024) Munyendo, Leah Masakhwe; Hitzmann, Bernd; Zhang, Yanyan
The assessment of coffee quality is based on the physical characteristics (bean quality), chemical constituents, and cup quality. Different factors, including altitude, genetics, management conditions, presence of adulterants, roasting, geographical origin, processing methods, and storage, affect the coffee quality. To meet the consumers' expectations regarding quality, the development of fast, new, and advanced analytical techniques for assessing the factors affecting coffee quality is a central aspect. Therefore, this research aimed to develop spectroscopic techniques complemented with chemometrics for evaluating the factors affecting coffee quality. The first specific objective was to investigate the ability of a deep autoencoder neural network to detect adulterants in roasted Arabica coffee and to determine a coffee’s geographical origin using near‐infrared (NIR) spectroscopy. Arabica coffee was adulterated with Robusta coffee or chicory at adulteration levels ranging from 2.5 % to 30 % in increments of 2.5 % at light, medium, and dark roast levels. Based on the results, all the samples adulterated with chicory were detectable by the autoencoder at all roast levels. For Robusta-adulterated samples, the detection was possible at adulteration levels above 7.5 % at medium and dark roasts. One can attribute the observations to potential differences in the chemical composition among the samples. Additionally, it was possible to differentiate coffee samples from different geographical origins. As a continuation of the first objective, the potential of NIR spectroscopy to quantify Robusta coffee or chicory in roasted Arabica coffee using different regression models constructed from the linear discriminant analysis (LDA) or principal component analysis (PCA) features was investigated. In addition, two classification methods (k-nearest neighbor regression (KNR) and LDA) were used. The regression models derived from LDA-extracted features exhibited better accuracies than those derived from PCA-extracted features. The two feature extraction methods exhibit differences in their working principle. PCA focuses on identifying the direction of maximum variance regardless of the adulteration levels. In contrast, LDA identifies the feature subspace that optimizes the separability of the classes (adulteration levels) and minimizes the variance within the class. Therefore, LDA extracted the features better than PCA, explaining the better performance of the regression models constructed from its features. The models provided satisfactory results with the coefficient of determination (R2) values above 0.92 for both the adulterants, indicating their efficiency in quantifying Robusta coffee or chicory in roasted Arabica coffee. For the classification methods, the LDA model performed better than KNR. Another focus of this doctoral research was to develop analytical tools based on Raman and NIR spectroscopy for real-time monitoring of 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 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, and 29 minutes. Four process runs were performed for each coffee species. The spectra of the ground samples were taken using the two spectrometers and modeled by the KNR, partial least squares regression (PLSR), and multiple linear regression (MLR). All the models based on the NIR spectra 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 < 9.469 %) and high R2 (> 0.916) values. Similarly, all the models based on the Raman spectra provided acceptable prediction accuracies for monitoring the dynamics of chlorogenic acid, trigonelline, and DPPH radical scavenging activity (pRMSEP < 7.849 % and R2> 0.944). In conclusion, this research proposes different approaches that would allow valuable decisions regarding coffee quality to be made quickly and efficiently. The study suggests using NIR spectroscopy to determine a coffee’s geographical origin and detect and quantify adulterants in roasted coffee. The findings reveal that the method could be a promising tool for routine coffee quality control applications in the coffee industry and other legal sectors. The study also proposes using different spectroscopic methods (NIR and Raman) to monitor a coffee roasting process. One can consider the presented approaches as essential steps toward optimizing the roasting process at an industrial scale as they permit instantaneously taking significant process decisions.
Bioprospecting for novel lipopeptide-producing strains for potential application in food and agriculture
(2024) Akintayo, Stephen Olusanmi; Hausmann, Rudolf
The need for sustainable alternatives to chemical products has been a huge topic in recent years and has put a demand on researchers and biotechnological companies to come up with bio-based alternatives to several chemical products. In line with this, interest in biosurfactants as alternatives to chemical surfactants is on the rise. Biosurfactants produced by microorganisms have great potential for application in detergents, personal care products, and pharmaceuticals, as well as in environmental, food processing, and agricultural applications. There are a few types of biosurfactants, including lipopeptides, which are primarily produced by Bacillus species and exhibit antimicrobial properties in addition to the well-known surface activity, surface tension reduction, and emulsifying ability of biosurfactants. Like other biosurfactants, lipopeptides have found more use in environmental applications such as bioremediation and microbial enhanced oil recovery (MEOR), while their use in agriculture and food industries remains limited due to concerns that may be related to acceptability, compatibility, and low yield by wild-type strains. To overcome these challenges, this thesis sought to find novel wild-type lipopeptide-producing strains from food-related sources that could be presumably safe for use in agriculture and food applications. To achieve this goal, a screening approach that combined several methods was adopted to identify potential high-yield wild-type, and possibly novel lipopeptide-producing strains. The ability of selected strains as promising biocontrol agents in agriculture was also evaluated. In Publication 1, potential lipopeptide-producing strains were isolated from food-related sources and screened for lipopeptide production. The screening approach combined microbiological and molecular identification of strains, with screening methods based on biosurfactant properties, as well as chemical analysis of surfactin production. Strains with promising lipopeptide-production potential belonging to three genera of Bacillus, Lysinibacillus and Priestia were identified. These strains included several exotic species that were either previously unknown or minimally studied with respect to LP production. Multiple strains that produced more than 150 mg L-1 surfactin, including a B. subtilis strain with a yield of about 1.5 g L−1 were discovered. In Publication 2, two promising LP-producing B. velezensis strains ES1-02 and EFSO2-04 were evaluated for their biocontrol potential and compared with commercial biocontrol strains B. velezensis QST713 and FZB42. The isolated strains demonstrated biocontrol ability comparable to QST713 against Diaporthe spp., which are notorious fungal pathogens of soybeans and other economically important crops. Co-incubation of strain ES1-02 with the phytopathogen D. longicolla induced a 10-fold increase in surfactin production. The broader molecular response of B. velezensis to plant pathogens investigated through an associated global proteome analysis showed the adaptation and response mechanisms of B. velezensis to plant pathogens. In general, B. velezensis seemed to adopt LP- modulation, physiological adaptation, and increased abundance of antimicrobial compounds as antagonistic and adaptation strategies for interaction with the phytopathogen D. longicolla. In Publication 3, genomic techniques were used in the discovery and description of a novel lipopeptides-producing species of the genus Lysinibacillus for which the name Lysinibacillus irui sp. nov. was proposed. This Gram-positive, motile, aerobic, rod-shaped, endospore-forming strain designated IRB4-01T was isolated from fermented African locust beans (Iru) and as such was named after Iru. A comprehensive chemotaxonomic analysis of the strain showed that the cell wall peptidoglycan type is A4α (Lys–Asp), and MK-7 is the major respiratory quinone. Detailed information about the polar lipids and major cellular fatty acids was also obtained. The G+C content of the genomic DNA was 37.4 mol%. Surfactin production by this novel strain was described in Publication 1 of this work.
Optimization of no-wait flowshop scheduling problem in bakery production with modified PSO, NEH and SA
(2021) Babor, Majharulislam; Senge, Julia; Rosell, Cristina M.; Rodrigo, Dolores; Hitzmann, Bernd
In bakery production, to perform a processing task there might be multiple alternative machines that have the same functionalities. Finding an efficient production schedule is challenging due to the significant nondeterministic polynomial time (NP)-hardness of the problem when the number of products, processing tasks, and alternative machines are higher. In addition, many tasks are performed manually as small and medium-size bakeries are not fully automated. Therefore, along with machines, the integration of employees in production planning is essential. This paper presents a hybrid no-wait flowshop scheduling model (NWFSSM) comprising the constraints of common practice in bakeries. The schedule of an existing production line is simulated to examine the model and is optimized by performing particle swarm optimization (PSO), modified particle swarm optimization (MPSO), simulated annealing (SA), and Nawaz-Enscore-Ham (NEH) algorithms. The computational results reveal that the performance of PSO is significantly influenced by the weight distribution of exploration and exploitation in a run time. Due to the modification to the acceleration parameter, MPSO outperforms PSO, SA, and NEH in respect to effectively finding an optimized schedule. The best solution to the real case problem obtained by MPSO shows a reduction of the total idle time (TIDT) of the machines by 12% and makespan by 30%. The result of the optimized schedule indicates that for small- and medium-sized bakery industries, the application of the hybrid NWFSSM along with nature-inspired optimization algorithms can be a powerful tool to make the production system efficient.
Influence of muscle type and microstructure on iridescence in cooked, cured pork meat products
(2021) Ruedt, Chiara; Gibis, Monika; Weiss, Jochen
Microstructural factors associated with surface iridescence in cooked, cured pork products were investigated. Meat iridescence is a commonly observed physical phenomenon in raw meat and meat products that consist of intact mus- cle tissue. Since the purchase decision of consumers is mainly driven by the first impression of meat color and appearance, products showing colorful iri- descence may be rejected. Four different muscles (RF: M. rectus femoris, BF: M. biceps femoris, ST: M. semitendinosus, and LD: M. longissimus thoracis et lum- borum) were brine-injected, cooked, sliced, and iridescence was evaluated by digital image analysis and sensory analysis. Sarcomere lengths, fiber diameters, and surface microstructure were analyzed in iridescent and noniridescent sec- tions. Highest iridescence extent by image analysis was found in LD (37.3 ± 16.4%), and highest overall iridescence score (extent and intensity, 6.11 ± 1.78) was observed in BF. Sarcomere lengths did not differ significantly between iridescent (1.05 ± 0.09 µm LD) and noniridescent areas (1.08 ± 0.94 µm LD) within mus- cles (p > 0.05). Iridescent sections showed smooth and ordered surface structures with cross-sectioned myofibers, whereas in noniridescent sections, surfaces were more unstructured and myofibers obliquely cut. The results of the study indicate that the sarcomere length and fiber diameters may thus be only of minor impor- tance for the explanation of meat iridescence in cooked meat products and are rather related to multiple scattering and absorption effects on smaller structural entities such as the myofilament lattice or larger entities such as fiber bundles. Practical application: Iridescence can be a problem for the meat industry due to consumers concerns about green-iridescent colors in meat. The underlying mechanisms and structures have not yet been fully clarified, and thus no prac- tical solutions to eliminate iridescence have been found so far. This research presents new insights into the structural attributes that are interrelated with meat iridescence and shows that iridescence is rather influenced by cutting angle of muscle fibers and surface homogeneity than by muscle fiber diameters or sar- comere lengths. This should be considered by the industry when seeking for ways to reduce the potential problem of iridescence
Micro-scale shear kneading: Gluten network development under multiple stress-relaxation steps and evaluation via multiwave rheology
(2022) Vidal, Leonhard Maria; Braun, Andre; Jekle, Mario; Becker, Thomas
To evaluate the kneading process of wheat flour dough, the state of the art is a subsequent and static measuring step on kneaded dough samples. In this study, an in-line measurement setup was set up in a rheometer based on previously validated shear kneading processes. With this approach, the challenge of sample transfer between the kneader and a measurement device was overcome. With the developed approach, an analysis of the dynamic development of the dough is possible. Through consecutive stress–relaxation steps with increasing deformation, a kneading setup in a conventional rheometer is implemented. Fitting of the shear stress curve with a linearization approach, as well as fitting of the relaxation modulus after each kneading step, is a new way to evaluate the matrix development. Subsequently, multiwave rheology is used to validate the kneading process in-line. The shear kneading setup was capable of producing an optimally developed dough matrix close to the reference kneading time of 150 ± 7.9 s (n = 3). The linearization approach as well as the power-law fit of the relaxation modulus revealed gluten network development comparable to the reference dough. With this approach, a deeper insight into gluten network development and crosslinking processes during wheat flour dough kneading is given.
Generic chemometric models for metabolite concentration prediction based on Raman spectra
(2022) Yousefi-Darani, Abdolrahim; Paquet-Durand, Olivier; von Wrochem, Almut; Classen, Jens; Tränkle, Jens; Mertens, Mario; Snelders, Jeroen; Chotteau, Veronique; Mäkinen, Meeri; Handl, Alina; Kadisch, Marvin; Lang, Dietmar; Dumas, Patrick; Hitzmann, Bernd
Chemometric models for on-line process monitoring have become well established in pharmaceutical bioprocesses. The main drawback is the required calibration effort and the inflexibility regarding system or process changes. So, a recalibration is necessary whenever the process or the setup changes even slightly. With a large and diverse Raman dataset, however, it was possible to generate generic partial least squares regression models to reliably predict the concentrations of important metabolic compounds, such as glucose-, lactate-, and glutamine-indifferent CHO cell cultivations. The data for calibration were collected from various cell cultures from different sites in different companies using different Raman spectrophotometers. In testing, the developed “generic” models were capable of predicting the concentrations of said compounds from a dilution series in FMX-8 mod medium, as well as from an independent CHO cell culture. These spectra were taken with a completely different setup and with different Raman spectrometers, demonstrating the model flexibility. The prediction errors for the tests were mostly in an acceptable range (<10% relative error). This demonstrates that, under the right circumstances and by choosing the calibration data carefully, it is possible to create generic and reliable chemometric models that are transferrable from one process to another without recalibration.
Application of two-dimensional fluorescence spectroscopy for the on-line monitoring of teff-based substrate fermentation inoculated with certain probiotic bacteria
(2022) Alemneh, Sendeku Takele; Emire, Shimelis Admassu; Jekle, Mario; Paquet-Durand, Olivier; von Wrochem, Almut; Hitzmann, Bernd
There is increasing demand for cereal-based probiotic fermented beverages as an alternative to dairy-based products due to their limitations. However, analyzing and monitoring the fermentation process is usually time consuming, costly, and labor intensive. This research therefore aims to apply two-dimensional (2D)-fluorescence spectroscopy coupled with partial least-squares regression (PLSR) and artificial neural networks (ANN) for the on-line quantitative analysis of cell growth and concentrations of lactic acid and glucose during the fermentation of a teff-based substrate. This substrate was inoculated with mixed strains of Lactiplantibacillus plantarum A6 (LPA6) and Lacticaseibacillus rhamnosus GG (LCGG). The fermentation was performed under two different conditions: condition 1 (7 g/100 mL substrate inoculated with 6 log cfu/mL) and condition 2 (4 g/100 mL substrate inoculated with 6 log cfu/mL). For the prediction of LPA6 and LCGG cell growth, the relative root mean square error of prediction (pRMSEP) was measured between 2.5 and 4.5%. The highest pRMSEP (4.5%) was observed for the prediction of LPA6 cell growth under condition 2 using ANN, but the lowest pRMSEP (2.5%) was observed for the prediction of LCGG cell growth under condition 1 with ANN. A slightly more accurate prediction was found with ANN under condition 1. However, under condition 2, a superior prediction was observed with PLSR as compared to ANN. Moreover, for the prediction of lactic acid concentration, the observed values of pRMSEP were 7.6 and 7.7% using PLSR and ANN, respectively. The highest error rates of 13 and 14% were observed for the prediction of glucose concentration using PLSR and ANN, respectively. Most of the predicted values had a coefficient of determination (R2) of more than 0.85. In conclusion, a 2D-fluorescence spectroscopy combined with PLSR and ANN can be used to accurately monitor LPA6 and LCGG cell counts and lactic acid concentration in the fermentation process of a teff-based substrate. The prediction of glucose concentration, however, showed a rather high error rate.
A robust fermentation process for natural chocolate-like flavor production with Mycetinis scorodonius
(2022) Rigling, Marina; Heger, Fabienne; Graule, Maria; Liu, Zhibin; Zhang, Chen; Ni, Li; Zhang, Yanyan
Submerged fermentation of green tea with the basidiomycete Mycetinis scorodonius resulted in a pleasant chocolate-like and malty aroma, which could be a promising chocolate flavor alternative to current synthetic aroma mixtures in demand of consumer preferences towards healthy natural and ‘clean label’ ingredients. To understand the sensorial molecular base on the chocolate-like aroma formation, key aroma compounds of the fermented green tea were elucidated using a direct immersion stir bar sorptive extraction combined with gas chromatography–mass spectrometry–olfactometry (DI-SBSE-GC-MS-O) followed by semi-quantification with internal standard. Fifteen key aroma compounds were determined, the most important of which were dihydroactinidiolide (odor activity value OAV 345), isovaleraldehyde (OAV 79), and coumarin (OAV 24), which were also confirmed by a recombination study. Furthermore, effects of the fermentation parameters (medium volume, light protection, agitation rate, pH, temperature, and aeration) on the aroma profile were investigated in a lab-scale bioreactor at batch fermentation. Variation of the fermentation parameters resulted in similar sensory perception of the broth, where up-scaling in volume evoked longer growth cycles and aeration significantly boosted the concentrations yet added a green note to the overall flavor impression. All findings prove the robustness of the established fermentation process with M. scorodonius for natural chocolate-like flavor production.
Surfactin shows relatively low antimicrobial activity against Bacillus subtilis and other bacterial model organisms in the absence of synergistic metabolites
(2022) Lilge, Lars; Ersig, Nadine; Hubel, Philipp; Aschern, Moritz; Pillai, Evelina; Klausmann, Peter; Pfannstiel, Jens; Henkel, Marius; Morabbi Heravi, Kambiz; Hausmann, Rudolf
Surfactin is described as a powerful biosurfactant and is natively produced by Bacillus subtilis in notable quantities. Among other industrially relevant characteristics, antimicrobial properties have been attributed to surfactin-producing Bacillus isolates. To investigate this property, stress approaches were carried out with biotechnologically established strains of Corynebacterium glutamicum, Bacillus subtilis, Escherichia coli and Pseudomonas putida with the highest possible amounts of surfactin. Contrary to the popular opinion, the highest growth-reducing effects were detectable in B. subtilis and E. coli after surfactin treatment of 100 g/L with 35 and 33%, respectively, while P. putida showed no growth-specific response. In contrast, other antimicrobial biosurfactants, like rhamnolipids and sophorolipids, showed significantly stronger effects on bacterial growth. Since the addition of high amounts of surfactin in defined mineral salt medium reduced the cell growth of B. subtilis by about 40%, the initial stress response at the protein level was analyzed by mass spectrometry, showing induction of stress proteins under control of alternative sigma factors σB and σW as well as the activation of LiaRS two-component system. Overall, although surfactin is associated with antimicrobial properties, relatively low growth-reducing effects could be demonstrated after the surfactin addition, challenging the general claim of the antimicrobial properties of surfactin.
Orotic acid production by Yarrowia lipolytica under conditions of limited pyrimidine
(2021) Swietalski, Paul; Hetzel, Frank; Klaiber, Iris; Pross, Eva; Seitl, Ines; Fischer, Lutz
Orotic acid (OA) is an intermediate of the pyrimidine biosynthesis with high industrial relevance due to its use as precursor for production of biochemical pyrimidines or its use as carrier molecule in drug formulations. It can be produced by fermentation of microorganisms with engineered pyrimidine metabolism. In this study, we surprisingly discovered the yeast Yarrowia lipolytica as a powerful producer of OA. The overproduction of OA in the Y. lipolytica strain PO1f was found to be caused by the deletion of the URA3 gene which prevents the irreversible decarboxylation of OA to uridine monophosphate. It was shown that the lack of orotidine‐5′‐phosphate decarboxylase was the reason for the accumulation of OA inside the cell since a rescue mutant of the URA3 deletion in Y. lipolytica PO1f completely prevented the OA secretion into the medium. In addition, pyrimidine limitation in the cell massively enhanced the OA accumulation followed by secretion due to intense overflow metabolism during bioreactor cultivations. Accordingly, supplementation of the medium with 200 mg/L uracil drastically decreased the OA overproduction by 91%. OA productivity was further enhanced in fed‐batch cultivation with glucose and ammonium sulfate feed to a maximal yield of 9.62 ± 0.21 g/L. Y. lipolytica is one of three OA overproducing yeasts described in the literature so far, and in this study, the highest productivity was shown. This work demonstrates the potential of Y. lipolytica as a possible production organism for OA and provides a basis for further metabolic pathway engineering to optimize OA productivity.
High molecular weight λ-carrageenan improves the color stability of phycocyanin by associative interactions
(2022) Buecker, Stephan; Grossmann, Lutz; Loeffler, Myriam; Leeb, Elena; Weiss, Jochen
Phycocyanin is a protein-chromophore structure present in Arthrospira platensis commonly used as a blue-colorant in food. Color losses of phycocyanin can be reduced by electrostatic complexation with λ-carrageenan. The aim of this study was to investigate the effect of molecular weight (MW) of λ-carrageenan on the color stabilization of electrostatic complexes formed with phycocyanin and λ-carrageenan. Samples were heated to 70 or 90°C at pH 3.0 and stored at 25°C for 14 days. The MW of λ-carrageenan was reduced by ultrasound treatments for 15, 30, 60, and 90 min. Prolonged ultrasonication had a pronounced effect on the Mw, which decreased from 2,341 to 228 kDa (0–90 min). Complexes prepared with low MW λ-carrageenan showed greater color changes compared to complexes prepared with high MW λ-carrageenan. The MW had no visible effect on color stability on day 0, but green/yellow shifts were observed during storage and after heating to 70°C. Medium MW showed less color stabilization effects compared to low MW when heated to 70°C. Moreover, for solutions prepared with ultrasonicated λ-carrageenan, significant hue shifts toward green/yellow, and precipitation were observed after a heat treatment at 90°C. In addition, the sizes of the complexes were significantly reduced (646–102 nm) by using ultrasonicated λ-carrageenan, except for the lowest MW λ-carrageenan when heated to 90°C. Overall, these findings demonstrated that decreasing the MW of λC had adverse effects on the color stability of PC:λC complexes heated to 70 and 90°C.
Influence of processing steps on structural, functional, and quality properties of beef hamburgers
(2022) Berger, Lisa M.; Witte, Franziska; Terjung, Nino; Weiss, Jochen; Gibis, Monika
In hamburger manufacturing, meat is subjected to four main processing steps (pre-grinding, mixing, grinding, and forming), whereby muscle fibers are disintegrated. In this study, the influence of these process steps was characterized by structural (amount of non-intact cells (ANIC), CLS-Microscopy), functional (drip loss) and qualitative (soluble protein content, lactate dehydrogenase (LDH) activity, myoglobin content (Mb)) parameters of the meat. Therefore, meat samples were analyzed after each process step. Histological analyses revealed an increased ANIC with progressive processing. Thereby, the first and second grinding steps caused the strongest increases (factors 2.43 and 2.69). Comparable results were found in the relative LDH activity (factor 2.20 and 1.62) and the Mb concentration (factor 2.24 and 1.33) of the extracted meat solution. The findings suggest that the disintegration of the meat structure increases with progressive processing, causing more vulnerable structures which result in increased leakage of intramuscular substances. Further, the type of stress acting on the meat determines the extent of the changes. The presented findings enable manufacturers to precisely adjust their process towards more gentle production parameters and thus, to meet the legal regulations.
Jahresbericht 2012 / Institut für Lebensmittelwissenschaft und Biotechnologie
(2013)
Jahresbericht 2011 / Institut für Lebensmittelwissenschaft und Biotechnologie
(2012)
Consumption of antioxidant-rich “Cerrado” cashew pseudofruit affects hepatic gene expression in obese C57BL/6J high fat-fed mice
(2022) Egea, Mariana Buranelo; Pierce, Gavin; Park, Si-Hong; Lee, Sang-In; Heger, Fabienne; Shay, Neil
The pseudofruit of A. othonianum Rizzini, “Cerrado” cashew pulp, has been described as rich in flavonoids, phenolic compounds, and vitamin C. The objective of this work was to evaluate the beneficial health effects seen with the addition of “Cerrado” cashew pulp (CP) to an obesogenic high fat diet provided to C57BL/6J male mice. In week 9, the HF-fed group had a significantly higher baseline glucose concentration than the LF- or HF+CP-fed groups. In RNAseq analysis, 4669 of 5520 genes were found to be differentially expressed. Among the genes most upregulated with the ingestion of the CP compared to HF were Ph1da1, SLc6a9, Clec4f, and Ica1 which are related to glucose homeostasis; Mt2 that may be involved steroid biosynthetic process; and Ciart which has a role in the regulation of circadian rhythm. Although “Cerrado” CP intake did not cause changes in the food intake or body weight of fed mice with HF diet, carbohydrate metabolism appeared to be improved based on the observed changes in gene expression.
A robust one-step recombineering system for enterohemorrhagic escherichia coli
(2022) Peng, Lang; Dumevi, Rexford Mawunyo; Chitto, Marco; Haarmann, Nadja; Berger, Petya; Koudelka, Gerald; Schmidt, Herbert; Mellmann, Alexander; Dobrindt, Ulrich; Berger, Michael
Enterohemorrhagic Escherichia coli (EHEC) can cause severe diarrheic in humans. To improve therapy options, a better understanding of EHEC pathogenicity is essential. The genetic manipulation of EHEC with classical one-step methods, such as the transient overexpression of the phage lambda (λ) Red functions, is not very efficient. Here, we provide a robust and reliable method for increasing recombineering efficiency in EHEC based on the transient coexpression of recX together with gam, beta, and exo. We demonstrate that the genetic manipulation is 3–4 times more efficient in EHEC O157:H7 EDL933 Δstx1/2 with our method when compared to the overexpression of the λ Red functions alone. Both recombineering systems demonstrated similar efficiencies in Escherichia coli K-12 MG1655. Coexpression of recX did not enhance the Gam-mediated inhibition of sparfloxacin-mediated SOS response. Therefore, the additional inhibition of the RecFOR pathway rather than a stronger inhibition of the RecBCD pathway of SOS response induction might have resulted in the increased recombineering efficiency by indirectly blocking phage induction. Even though additional experiments are required to unravel the precise mechanistic details of the improved recombineering efficiency, we recommend the use of our method for the robust genetic manipulation of EHEC and other prophage-carrying E. coli isolates.
CortexVR: Immersive analysis and training of cognitive executive functions of soccer players using virtual reality and machine learning
(2022) Krupitzer, Christian; Naber, Jens; Stauffert, Jan-Philipp; Mayer, Jan; Spielmann, Jan; Ehmann, Paul; Boci, Noel; Bürkle, Maurice; Ho, André; Komorek, Clemens; Heinickel, Felix; Kounev, Samuel; Becker, Christian; Latoschik, Marc Erich
Goal: This paper presents an immersive Virtual Reality (VR) system to analyze and train Executive Functions (EFs) of soccer players. EFs are important cognitive functions for athletes. They are a relevant quality that distinguishes amateurs from professionals. Method: The system is based on immersive technology, hence, the user interacts naturally and experiences a training session in a virtual world. The proposed system has a modular design supporting the extension of various so-called game modes. Game modes combine selected game mechanics with specific simulation content to target particular training aspects. The system architecture decouples selection/parameterization and analysis of training sessions via a coaching app from an Unity3D-based VR simulation core. Monitoring of user performance and progress is recorded by a database that sends the necessary feedback to the coaching app for analysis. Results: The system is tested for VR-critical performance criteria to reveal the usefulness of a new interaction paradigm in the cognitive training and analysis of EFs. Subjective ratings for overall usability show that the design as VR application enhances the user experience compared to a traditional desktop app; whereas the new, unfamiliar interaction paradigm does not negatively impact the effort for using the application. Conclusion: The system can provide immersive training of EF in a fully virtual environment, eliminating potential distraction. It further provides an easy-to-use analyzes tool to compare user but also an automatic, adaptive training mode.

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