Institut für Lebensmittelwissenschaft und Biotechnologie

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Advancing 2D fluorescence online monitoring in microtiter plates by separating scattered light and fluorescence measurement, using a tunable emission monochromator
(2023) Berg, Christoph; Busch, Selma; Alawiyah, Muthia Dewi; Finger, Maurice; Ihling, Nina; Paquet-Durand, Olivier; Hitzmann, Bernd; Büchs, Jochen
Online fluorescence monitoring has become a key technology in modern bioprocess development, as it provides in‐depth process knowledge at comparably low costs. In particular, the technology is widely established for high‐throughput microbioreactor cultivation systems, due to its noninvasive character. For microtiter plates, previously also multi‐wavelength 2D fluorescence monitoring was developed. To overcome an observed limitation of fluorescence sensitivity, this study presents a modified spectroscopic setup, including a tunable emission monochromator. The new optical component enables the separation of the scattered and fluorescent light measurements, which allows for the adjustment of integration times of the charge‐coupled device detector. The resulting increased fluorescence sensitivity positively affected the performance of principal component analysis for spectral data of Escherichia coli batch cultivation experiments with varying sorbitol concentration supplementation. In direct comparison with spectral data recorded at short integration times, more biologically consistent signal dynamics were calculated. Furthermore, during partial least square regression for E. coli cultivation experiments with varying glucose concentrations, improved modeling performance was observed. Especially, for the growth‐uncoupled acetate concentration, a considerable improvement of the root‐mean‐square error from 0.25 to 0.17 g/L was achieved. In conclusion, the modified setup represents another important step in advancing 2D fluorescence monitoring in microtiter plates.
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.
Bacillus subtilis high cell density fermentation using a sporulation-deficient strain for the production of surfactin
(2021) Klausmann, Peter; Hennemann, Katja; Hoffmann, Mareen; Treinen, Chantal; Aschern, Moritz; Lilge, Lars; Morabbi Heravi, Kambiz; Henkel, Marius; Hausmann, Rudolf
Bacillus subtilis 3NA is a strain capable of reaching high cell densities. A surfactin producing sfp+ variant of this strain, named JABs32, was utilized in fed-batch cultivation processes. Both a glucose and an ammonia solution were fed to set a steady growth rate μ of 0.1 h-1. In this process, a cell dry weight of up to 88 g L-1 was reached after 38 h of cultivation, and surfactin titers of up to 26.5 g L-1 were detected in this high cell density fermentation process, achieving a YP/X value of 0.23 g g-1 as well as a qP/X of 0.007 g g-1 h-1. In sum, a 21-fold increase in surfactin titer was obtained compared with cultivations in shake flasks. In contrast to fed-batch operations using Bacillus subtilis JABs24, an sfp+ variant derived from B. subtilis 168, JABs32, reached an up to fourfold increase in surfactin titers using the same fed-batch protocol. Additionally, a two-stage feed process was established utilizing strain JABs32. Using an optimized mineral salt medium in this high cell density fermentation approach, after 31 h of cultivation, surfactin titers of 23.7 g L-1 were reached with a biomass concentration of 41.3 g L-1, thus achieving an enhanced YP/X value of 0.57 g g-1 as well as a qP/X of 0.018 g g-1 h-1. The mutation of spo0A locus and an elongation of AbrB in the strain utilized in combination with a high cell density fed-batch process represents a promising new route for future enhancements on surfactin production.
Barley-sorghum craft beer production with Saccharomyces cerevisiae, Torulaspora delbrueckii and Metschnikowia pulcherrima yeast strains
(2020) Einfalt, Daniel
The use of different yeast strains contributes to obtain insights into beer products with diverse sensory characteristics. In this study, three yeast species of different genera were selected to evaluate their fermentation performance and sensory profile for barley-sorghum beer production. Baley-sorghum wort was produced with 12.5°P and fermented with Saccharomyces cerevisiae, Torulaspora delbrueckii and Metschnikowia pulcherrima yeast strains. Differences were observed in terms of fermentation time and ability to ferment maltose. S. cerevisiae attenuated initial maltose concentration within 72 h, while M. pulcherrima and T. delbrueckii performed fermentation within 120 and 192 h, respectively. Both yeast strains simultaneously produced 11% and 23% lower ethanol concentrations, compared to S. cerevisiae with 37.9 g/L. Wort fermented with T. delbrueckii showed residual maltose concentration of 19.7 ± 4.1 g/L, resulting in significantly enhanced beer sweetness. S. cerevisiae produced significantly increased levels of higher alcohols, and obtained the highest scores for the sensory attribute body perception. Beer produced with T. delbrueckii contained significantly lower fermentative 2,3-butanediol and 2-methyl-1-butanol volatiles; this beer also showed reduced body perception. Beer conditioned with T. delbrueckii was significantly preferred over M. pulcherrima. Besides S. cerevisiae with high fermentative power, T. delbrueckii and M. pulcherrima were found to have reduced maltose fermenting abilities and provide significantly different sensory attributes to barley-sorghum beers.
Characterization of Bacillus velezensis UTB96, demonstrating improved lipopeptide production compared to the strain B. velezensis FZB42
(2022) Vahidinasab, Maliheh; Adiek, Isabel; Hosseini, Behnoush; Akintayo, Stephen Olusanmi; Abrishamchi, Bahar; Pfannstiel, Jens; Henkel, Marius; Lilge, Lars; Vögele, Ralf ; Hausmann, Rudolf
Bacillus strains can produce various lipopeptides, known for their antifungal properties. This makes them attractive metabolites for applications in agriculture. Therefore, identification of productive wild-type strains is essential for the development of biopesticides. Bacillus velezensis FZB42 is a well-established strain for biocontrol of plant pathogens in agriculture. Here, we characterized an alternative strain, B. velezensis UTB96, that can produce higher amounts of all three major lipopeptide families, namely surfactin, fengycin, and iturin. UTB96 produces iturin A. Furthermore, UTB96 showed superior antifungal activity towards the soybean fungal pathogen Diaporthe longicolla compared to FZB42. Moreover, the additional provision of different amino acids for lipopeptide production in UTB96 was investigated. Lysine and alanine had stimulatory effects on the production of all three lipopeptide families, while supplementation of leucine, valine and isoleucine decreased the lipopeptide bioproduction. Using a 45-litre bioreactor system for upscaling in batch culture, lipopeptide titers of about 140 mg/L surfactin, 620 mg/L iturin A, and 45 mg/L fengycin were achieved. In conclusion, it becomes clear that B. velezensis UTB96 is a promising strain for further research application in the field of agricultural biological controls of fungal diseases.
Characterization of the aroma profile of food smoke at controllable pyrolysis temperatures
(2023) Rigling, Marina; Höckmeier, Laura; Leible, Malte; Herrmann, Kurt; Gibis, Monika; Weiss, Jochen; Zhang, Yanyan
Smoking is used to give food its typical aroma and to obtain the desired techno-functional properties of the product. To gain a deeper knowledge of the whole process of food smoking, a controllable smoking process was developed, and the influence of wood pyrolysis temperature (150–900 °C) on the volatile compounds in the smoking chamber atmosphere was investigated. The aroma profile of smoke was decoded by headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS). Subsequently, the correlations in the most important substance classes, as well as in individual target components, were investigated by the Pearson test. Phenols and lactones showed an increase over the entire applied temperature range (rT = 0.94 and rT = 0.90), whereas furans and carbonyls showed no strict temperature dependence (rT < 0.6). Investigations on single aroma compounds showed that not all compounds of one substance class showed the same behavior, e.g., guaiacol showed no significant increase over the applied pyrolysis temperature, whereas syringol and hydoxyacetone showed a plateau after 450 °C, and phenol and cyclotene increased linear over the applied temperature range. These findings will help to better understand the production of aroma-active compounds during smoke generation in order to meet consumers preferences.
Correction: Schubert et al. Microencapsulation of bacteriophages for the delivery to and modulation of the human gut microbiota through milk and cereal products. Appl. Sci. 2022, 12, 6299
(2023) Schubert, Christina; Fischer, Sabina; Dorsch, Kathrin; Teßmer, Lutz; Hinrichs, Jörg; Atamer, Zeynep
Editorial: Flavor chemistry of food: Mechanism, interaction, new advances
(2023) Huang, Mingquan; Fan, Gang; Zhang, Yanyan
Effect of refrigerated storage on some physicochemical characteristics of a teff‐based fermented beverage and the viability of the fermenting Lactiplantibacillus plantarum and Lacticaseibacillus rhamnosus used
(2022) Alemneh, Sendeku Takele; Emire, Shimelis Admassu; Jekle, Mario; Hitzmann, Bernd
Probiotic beverages made from cereals become interesting in the recent food industries. In this contribution, a fermented teff-based probiotic beverage was produced using the whole grain teff flour and co-culture strains of Lactiplantibacillus plantarum (LPA6) and Lacticaseibacillus rhamnosus GG (LCGG). Then, the effect of 25 days of refrigerated storage on cell viability (LPA6 and LCGG), and contents of sugars, organic acids, and titratable acidity (TA), as well as pH values were examined. Furthermore, pathogenic microorganisms, hygiene indicators, and sensory tests of the beverage were analyzed. Presumptive cell counts of LPA6 and LCGG were observed to decrease throughout refrigerated storage. Glucose, lactic acid, maltose, and acetic acid contents were significantly (p < 0.05) increased over storage time. Also, pH reduction and TA increment were observed in storage time. Examined pathogenic microorganisms and hygiene indicators were not detected in the beverage. Sensory analysis of the beverage after 10 days of refrigerated storage was accepted by the panelists. Novelty Impact Statement Throughout refrigerated storage of teff-based probiotic beverage sugars and organic acids were produced. Sensory attributes of the newly produced teff-based probiotic beverage were accepted by the panelist after 10 days of refrigerated storage. The pH of the teff-based probiotic beverage became more acidic throughout 25 days of refrigerated storage.
Effect of storage temperature on volatile marker compounds in cured loins fermented with Staphylococcus carnosus by brine injection
(2020) Bosse, Ramona; Wirth, Melanie; Weiss, Jochen; Gibis, Monika
In this study, the influence of low (5 °C), intermediate (15 °C) and high (25 °C) storage temperatures on the profile of volatile compounds of North European cured loins fermented with Staphylococcus carnosus strains was investigated. In this context, proteolytic activity, bacterial growth, key volatile compounds and sensory attributes were studied. In conclusion, storage temperature significantly affected the volatile marker compounds. A multiple regression indicated significant effects of seven volatile compounds (acetophenone, benzaldehyde, butanone, 3-methylbutanal, 1-octen-3-ol, nonanal and pentanone) on the overall odor (R2 = 95.9%) and overall flavor (R2 = 81.1%). The sum of the marker volatiles aldehydes, ketones and alcohol increased with rising temperatures and the highest amounts of the odor active 3-methylbutanal up to 155 and 166 ng/g meat were detected in high temperature-stored loins. Moreover, the addition of S. carnosus strain LTH 3838 showed maximum effect at 5 °C-storage temperature in comparison to the control.
Effects of fingerroot (Boesenbergia pandurata) oil on microflora as an antimicrobial agent and on the formation of heterocyclic amines in fried meatballs
(2024) Soikam, Panida; Rachtanapun, Chitsiri; Suriyarak, Sarisa; Weiss, Jochen; Gibis, Monika
This study aimed to determine the antibacterial activity of the essential oil of fingerroot (Boesenbergia pandurata) (EOF) as a natural preservative in ground meat and its effect on the formation of heterocyclic amines (HAs) in pan-fried meatballs. EOF was applied either by adding it to ground pork or marinating pork in it before grinding. In addition, the antibacterial activity of EOF was tested. Aerobic mesophilic total viable count (TVC), lactic acid bacteria (LAB), and Enterobacteriaceae bacteria were monitored. The results show that EOF exhibited strong antibacterial activity when added at concentrations of 1.0 and 2.5 wt%. Antimicrobial activity against TVC, LAB, and especially Enterobacteriaceae bacteria was observed at all EOF concentrations (0.25, 0.5, 1.0, and 2.5 wt%). A 2.5% concentration of EOF applied by marinating trimmings can extend the shelf-life of ground pork to 18 days, while 2.5% EOF applied via addition can extend the shelf-life to 15 days, compared with 3 days for the control sample. After frying the meatballs, the inhibitory effect on the formation of heterocyclic amines was only significant for MeIQx with the highest addition of EOF (2.5 wt%). Significant increases in the concentrations of all other HAs were determined by adding EOF (2.5 wt%).
Effects of Ugali maize flour fortification with chia seeds (Salvia hispanica L.) on its physico-chemical properties and consumer acceptability
(2024) Chemutai, Susan; Mburu, Monica; Njoroge, Daniel; Zettel, Viktoria
The study investigated the effect of incorporating whole chia seeds (WCS) and defatted chia seed flour (DCF) into whole maize meal for ugali preparation. Both were incorporated at substitution levels of 3%, 6%, and 9% separately, and the resulting treatments subjected to laboratory analysis. In addition, ugali samples were prepared from all the resulting flour formulations and subjected to consumer acceptability assessment. Incorporation of both DCF and WCS resulted in increased water absorption capacity (ranging from 0.78 to 0.98 g/mL), swelling index (ranging from 0.15 to 3.25 mL/g), and swelling capacity (ranging from 2.46 to 5.74 g/g). WCS decreased the bulk density and oil absorption capacity. DCF, however, resulted in an increase in bulk density and oil absorption capacity. Both DCF and WCS lowered the lightness (L*) of the products. Proximate composition ranged from 4.78 to 7.46% for crude fat, 7.22% to 9.16% for crude protein, and 1.74 to 4.27% for crude fiber. The obtained results show the potential of chia seeds as a good fortificant of maize flour since it resulted in nutritionally superior products (crude ash, crude protein, crude fat, and energy value) when compared to control. The freshly prepared ugali samples were generally acceptable to the panelists up to 9% WCS and 6% DCF substitution levels.
Evaluation of an external foam column for in situ product removal in aerated surfactin production processes
(2023) Treinen, Chantal; Claassen, Linda; Hoffmann, Mareen; Lilge, Lars; Henkel, Marius; Hausmann, Rudolf
In Bacillus fermentation processes, severe foam formation may occur in aerated bioreactor systems caused by surface-active lipopeptides. Although they represent interesting compounds for industrial biotechnology, their property of foaming excessively during aeration may pose challenges for bioproduction. One option to turn this obstacle into an advantage is to apply foam fractionation and thus realize in situ product removal as an initial downstream step. Here we present and evaluate a method for integrated foam fractionation. A special feature of this setup is the external foam column that operates separately in terms of, e.g., aeration rates from the bioreactor system and allows recycling of cells and media. This provides additional control points in contrast to an internal foam column or a foam trap. To demonstrate the applicability of this method, the foam column was exemplarily operated during an aerated batch process using the surfactin-producing Bacillus subtilis strain JABs24. It was also investigated how the presence of lipopeptides and bacterial cells affected functionality. As expected, the major foam formation resulted in fermentation difficulties during aerated processes, partially resulting in reactor overflow. However, an overall robust performance of the foam fractionation could be demonstrated. A maximum surfactin concentration of 7.7 g/L in the foamate and enrichments of up to 4 were achieved. It was further observed that high lipopeptide enrichments were associated with low sampling flow rates of the foamate. This relation could be influenced by changing the operating parameters of the foam column. With the methodology presented here, an enrichment of biosurfactants with simultaneous retention of the production cells was possible. Since both process aeration and foam fractionation can be individually controlled and designed, this method offers the prospect of being transferred beyond aerated batch processes.
Evaluation of an oxygen‐dependent self‐inducible surfactin synthesis in B. subtilis by substitution of native promoter PsrfA by anaerobically active PnarG and PnasD
(2021) Hoffmann, Mareen; Braig, Alina; Fernandez Cano Luna, Diana Stephanie; Rief, Katharina; Becker, Philipp; Treinen, Chantal; Klausmann, Peter; Morabbi Heravi, Kambiz; Henkel, Marius; Lilge, Lars; Hausmann, Rudolf
A novel approach targeting self-inducible surfactin synthesis under oxygen-limited conditions is presented. Because both the nitrate (NarGHI) and nitrite (NasDE) reductase are highly expressed during anaerobic growth of B. subtilis, the native promoter PsrfA of the surfactin operon in strain B. subtilis JABs24 was replaced by promoters PnarG and PnasD to induce surfactin synthesis anaerobically. Shake flask cultivations with varying oxygen availabilities indicated no significant differences in native PsrfA expression. As hypothesized, activity of PnarG and PnasD increased with lower oxygen levels and surfactin was not produced by PsrfA::PnarG as well as PsrfA::PnasD mutant strains under conditions with highest oxygen availability. PnarG showed expressions similar to PsrfA at lowest oxygen availability, while maximum value of PnasD was more than 5.5-fold higher. Although the promoter exchange PsrfA::PnarG resulted in a decreased surfactin titer at lowest oxygen availability, the strain carrying PsrfA::PnasD reached a 1.4-fold increased surfactin concentration with 696 mg/L and revealed an exceptional high overall YP/X of 1.007 g/g. This value also surpassed the YP/X of the reference strain JABs24 at highest and moderate oxygen availability. Bioreactor cultivations illustrated that significant cell lysis occurred when the process of “anaerobization” was performed too fast. However, processes with a constantly low agitation and aeration rate showed promising potential for process improvement, especially by employing the strain carrying PsrfA::PnasD promoter exchange. Additionally, replacement of other native promoters by nitrite reductase promoter PnasD represents a promising tool for anaerobic-inducible bioprocesses in Bacillus.
Exploration of surfactin production by newly isolated Bacillus and Lysinibacillus strains from food‐related sources
(2022) Akintayo, Stephen Olusanmi; Treinen, Chantal; Vahidinasab, Maliheh; Pfannstiel, Jens; Bertsche, Ute; Fadahunsi, I.; Oellig, Claudia; Granvogl, Michael; Henkel, Marius; Lilge, Lars; Hausmann, Rudolf
As a lipopeptide (LP), surfactin exhibits properties, such as emulsifying and dispersing ability, which are useful in food industry. Discovery of new LP‐producing strains from food sources is an important step towards possible application of surfactin in foods. A total of 211 spore‐forming, Gram‐positive, and catalase‐positive bacterial strains were isolated from fermented African locust beans (iru) and palm oil mill effluents in a screening process and examined for their ability to produce surfactin. This was achieved by a combination of methods, which included microbiological and molecular classification of strains, along with chemical analysis of surfactin production. Altogether, 29 isolates, positive for oil spreading and emulsification assays, were further identified with 16S rDNA analysis. The strains belonged to nine species including less commonly reported strains of Lysinibacillus, Bacillus flexus, B. tequilensis, and B. aryabhattai. The surfactin production was quantitatively and qualitatively analysed by high‐performance thin‐layer chromatography and liquid chromatography‐mass spectrometry (LC–MS). Confirmation of surfactin by MS was achieved in all the 29 strains. Highest surfactin production capability was found in B. subtilis IRB2‐A1 with a titre of 1444·1 mg L−1.
Formation of key aroma compounds during 30 weeks of ripening in gouda-type cheese produced from pasteurized and raw milk
(2024) Duensing, Philipp W.; Hinrichs, Jörg; Schieberle, Peter
Gouda-type cheeses were produced on a pilot-scale from raw milk (RM-G) and pasteurized milk (PM-G). Sixteen key aroma compounds previously characterized by the sensomics approach were quantitated in the unripened cheeses and at five different ripening stages (4, 7, 11, 19, and 30 weeks) by means of stable isotope dilution assays. Different trends were observed in the formation of the key aroma compounds. Short-chain free fatty acids and ethyl butanoate as well as ethyl hexanoate continuously increased during ripening but to a greater extent in RM-G. Branched-chain fatty acids such as 3-methylbutanoic acid were also continuously formed and reached a 60-fold concentration after 30 weeks, in particular in PM-G. 3-Methylbutanal and butane-2,3-dione reached a maximum concentration after 7 weeks and decreased with longer ripening. Lactones were high in the unripened cheeses and increased only slightly during ripening. Recent results have shown that free amino acids were released during ripening. The aroma compounds 3-methylbutanal, 3-methyl-1-butanol, and 3-methylbutanoic acid are suggested to be formed by microbial enzymes degrading the amino acid l-leucine following the Ehrlich pathway. To gain insight into the quantitative formation of each of the three aroma compounds, the conversion of the labeled precursors (13C6)-l-leucine and (2H3)-2-keto-4-methylpentanoic acid into the isotopically labeled aroma compounds was studied. By applying the CAMOLA approach (defined mixture of labeled and unlabeled precursor), l-leucine was confirmed as the only precursor of the three aroma compounds in the cheese with the preferential formation of 3-methylbutanoic acid.
Four-dimensional (4D) printing of dynamic foods - definitions, considerations, and current scientific status
(2023) Fahmy, Ahmed Raouf; Derossi, Antonio; Jekle, Mario
Since its conception, the application of 3D printing in the structuring of food materials has been focused on the processing of novel material formulations and customized textures for innovative food applications, such as personalized nutrition and full sensory design. The continuous evolution of the used methods, approaches, and materials has created a solid foundation for technology to process dynamic food structures. Four-dimensional food printing is an extension of 3D printing where food structures are designed and printed to perform time-dependent changes activated by internal or external stimuli. In 4D food printing, structures are engineered through material tailoring and custom designs to achieve a transformation from one configuration to another. Different engineered 4D behaviors include stimulated color change, shape morphing, and biological growth. As 4D food printing is considered an emerging application, imperatively, this article proposes new considerations and definitions in 4D food printing. Moreover, this article presents an overview of 4D food printing within the current scientific progress, status, and approaches.
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.
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.
Homogenization improves foaming properties of insoluble pea proteins
(2022) Moll, Pascal; Salminen, Hanna; Griesshaber, Elena; Schmitt, Christophe; Weiss, Jochen
Foams are essential in many food applications and require surface-active ingredients such as proteins for formation and stabilization. We investigated the influence of high-pressure homogenization on foaming properties of insoluble pea protein dispersions (5% w/w) at pH 3 and 5. Unhomogenized insoluble pea protein dispersions did not foam at either pH 3 or 5, as they consisted of large insoluble pea protein aggregates with limited surface activity. At pH 3, the homogenized pea protein dispersions generated foams due to higher protein solubility and surface activity through disruption of large protein aggregates into smaller particles. The foam stability decreased with increasing homogenization pressure and number of cycles due to a reduction in continuous phase viscosity. At pH 5, the insoluble pea proteins foamed when the homogenization resulted in formation of aggregates made of smaller protein entities, which was the case for homogenization ≥ 100 MPa and three cycles. In general, the foam capacity (amount of formed foam) was higher at pH 3 due to improved protein solubility and surface activity that facilitated incorporation of air, while the foam stability (resistance against foam collapse) was better at pH 5 because of the presence of larger protein aggregates that formed thicker and more viscous films around the air bubbles benefitting retention of gas bubbles. Overall, homogenization improved the foaming properties of insoluble pea proteins at pH 3 and 5.

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