@article{FalkenbergKohnBottetal.2023, author = {Falkenberg, Fabian and Kohn, Sophie and Bott, Michael and Bongaerts, Johannes and Siegert, Petra}, title = {Biochemical characterisation of a novel broad pH spectrum subtilisin from Fictibacillus arsenicus DSM 15822ᵀ}, series = {FEBS Open Bio}, volume = {13}, journal = {FEBS Open Bio}, number = {11}, publisher = {Wiley}, address = {Hoboken, NJ}, issn = {2211-5463}, doi = {10.1002/2211-5463.13701}, pages = {2035 -- 2046}, year = {2023}, abstract = {Subtilisins from microbial sources, especially from the Bacillaceae family, are of particular interest for biotechnological applications and serve the currently growing enzyme market as efficient and novel biocatalysts. Biotechnological applications include use in detergents, cosmetics, leather processing, wastewater treatment and pharmaceuticals. To identify a possible candidate for the enzyme market, here we cloned the gene of the subtilisin SPFA from Fictibacillus arsenicus DSM 15822ᵀ (obtained through a data mining-based search) and expressed it in Bacillus subtilis DB104. After production and purification, the protease showed a molecular mass of 27.57 kDa and a pI of 5.8. SPFA displayed hydrolytic activity at a temperature optimum of 80 °C and a very broad pH optimum between 8.5 and 11.5, with high activity up to pH 12.5. SPFA displayed no NaCl dependence but a high NaCl tolerance, with decreasing activity up to concentrations of 5 m NaCl. The stability enhanced with increasing NaCl concentration. Based on its substrate preference for 10 synthetic peptide 4-nitroanilide substrates with three or four amino acids and its phylogenetic classification, SPFA can be assigned to the subgroup of true subtilisins. Moreover, SPFA exhibited high tolerance to 5\% (w/v) SDS and 5\% H₂O₂ (v/v). The biochemical properties of SPFA, especially its tolerance of remarkably high pH, SDS and H₂O₂, suggest it has potential for biotechnological applications.}, language = {en} } @article{CheenakulaPaulsenOttetal.2023, author = {Cheenakula, Dheeraja and Paulsen, Svea and Ott, Fabian and Gr{\"o}mping, Markus}, title = {Operational window of a deammonifying sludge for mainstream application in a municipal wastewater treatment plant}, series = {Water and Environment Journal}, volume = {38}, journal = {Water and Environment Journal}, number = {1}, publisher = {Wiley}, address = {Chichester}, issn = {1747-6593}, doi = {10.1111/wej.12898}, pages = {59 -- 70}, year = {2023}, abstract = {The present work aimed to study the mainstream feasibility of the deammonifying sludge of side stream of municipal wastewater treatment plant (MWWTP) in Kaster, Germany. For this purpose, the deammonifying sludge available at the side stream was investigated for nitrogen (N) removal with respect to the operational factors temperature (15-30°C), pH value (6.0-8.0) and chemical oxygen demand (COD)/N ratio (≤1.5-6.0). The highest and lowest N-removal rates of 0.13 and 0.045 kg/(m³ d) are achieved at 30 and 15°C, respectively. Different conditions of pH and COD/N ratios in the SBRs of Partial nitritation/anammox (PN/A) significantly influenced both the metabolic processes and associated N-removal rates. The scientific insights gained from the current work signifies the possibility of mainstream PN/A at WWTPs. The current study forms a solid basis of operational window for the upcoming semi-technical trails to be conducted prior to the full-scale mainstream PN/A at WWTP Kaster and WWTPs globally.}, language = {en} } @article{HoffstadtCheenakulaNikolauszetal.2023, author = {Hoffstadt, Kevin and Cheenakula, Dheeraja and Nikolausz, Marcell and Krafft, Simone and Harms, Hauke and Kuperjans, Isabel}, title = {Design and construction of a new reactor for flexible biomethanation of hydrogen}, series = {Fermentation}, volume = {9}, journal = {Fermentation}, number = {8}, publisher = {MDPI}, address = {Basel}, issn = {2311-5637}, doi = {10.3390/fermentation9080774}, pages = {1 -- 16}, year = {2023}, abstract = {The increasing share of renewable electricity in the grid drives the need for sufficient storage capacity. Especially for seasonal storage, power-to-gas can be a promising approach. Biologically produced methane from hydrogen produced from surplus electricity can be used to substitute natural gas in the existing infrastructure. Current reactor types are not or are poorly optimized for flexible methanation. Therefore, this work proposes a new reactor type with a plug flow reactor (PFR) design. Simulations in COMSOL Multiphysics ® showed promising properties for operation in laminar flow. An experiment was conducted to support the simulation results and to determine the gas fraction of the novel reactor, which was measured to be 29\%. Based on these simulations and experimental results, the reactor was constructed as a 14 m long, 50 mm diameter tube with a meandering orientation. Data processing was established, and a step experiment was performed. In addition, a kLa of 1 h-1 was determined. The results revealed that the experimental outcomes of the type of flow and gas fractions are in line with the theoretical simulation. The new design shows promising properties for flexible methanation and will be tested.}, language = {en} } @article{GrajewskiKleefeld2023, author = {Grajewski, Matthias and Kleefeld, Andreas}, title = {Detecting and approximating decision boundaries in low-dimensional spaces}, series = {Numerical Algorithms}, volume = {93}, journal = {Numerical Algorithms}, number = {4}, publisher = {Springer Science+Business Media}, address = {Dordrecht}, issn = {1572-9265}, pages = {35 Seiten}, year = {2023}, abstract = {A method for detecting and approximating fault lines or surfaces, respectively, or decision curves in two and three dimensions with guaranteed accuracy is presented. Reformulated as a classification problem, our method starts from a set of scattered points along with the corresponding classification algorithm to construct a representation of a decision curve by points with prescribed maximal distance to the true decision curve. Hereby, our algorithm ensures that the representing point set covers the decision curve in its entire extent and features local refinement based on the geometric properties of the decision curve. We demonstrate applications of our method to problems related to the detection of faults, to multi-criteria decision aid and, in combination with Kirsch's factorization method, to solving an inverse acoustic scattering problem. In all applications we considered in this work, our method requires significantly less pointwise classifications than previously employed algorithms.}, language = {en} } @article{LuftBremenLuft2023, author = {Luft, Angela and Bremen, Sebastian and Luft, Nils}, title = {A cost/benefit and flexibility evaluation framework for additive technologies in strategic factory planning}, series = {Processes}, volume = {11}, journal = {Processes}, number = {7}, publisher = {MDPI}, address = {Basel}, issn = {2227-9717}, doi = {10.3390/pr11071968}, pages = {Artikel 1968}, year = {2023}, abstract = {There is a growing demand for more flexibility in manufacturing to counter the volatility and unpredictability of the markets and provide more individualization for customers. However, the design and implementation of flexibility within manufacturing systems are costly and only economically viable if applicable to actual demand fluctuations. To this end, companies are considering additive manufacturing (AM) to make production more flexible. This paper develops a conceptual model for the impact quantification of AM on volume and mix flexibility within production systems in the early stages of the factory-planning process. Together with the model, an application guideline is presented to help planners with the flexibility quantification and the factory design process. Following the development of the model and guideline, a case study is presented to indicate the potential impact additive technologies can have on manufacturing flexibility Within the case study, various scenarios with different production system configurations and production programs are analyzed, and the impact of the additive technologies on volume and mix flexibility is calculated. This work will allow factory planners to determine the potential impacts of AM on manufacturing flexibility in an early planning stage and design their production systems accordingly.}, language = {en} } @article{JanusAchtsnichtTempeletal.2023, author = {Janus, Kevin Alexander and Achtsnicht, Stefan and Tempel, Laura and Drinic, Aleksaner and Kopp, Alexander and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Influence of fibroin membrane composition and curing parameters on the performance of a biodegradable enzymatic biosensor manufactured from Silicon-Free Carbon}, series = {Physica status solidi : pss. A, Applications and materials science}, volume = {220}, journal = {Physica status solidi : pss. A, Applications and materials science}, number = {22}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6300 (Print)}, doi = {10.1002/pssa.202300081}, pages = {10 Seiten}, year = {2023}, abstract = {Herein, fibroin, polylactide (PLA), and carbon are investigated for their suitability as biocompatible and biodegradable materials for amperometric biosensors. For this purpose, screen-printed carbon electrodes on the biodegradable substrates fibroin and PLA are modified with a glucose oxidase membrane and then encapsulated with the biocompatible material Ecoflex. The influence of different curing parameters of the carbon electrodes on the resulting biosensor characteristics is studied. The morphology of the electrodes is investigated by scanning electron microscopy, and the biosensor performance is examined by amperometric measurements of glucose (0.5-10 mM) in phosphate buffer solution, pH 7.4, at an applied potential of 1.2 V versus a Ag/AgCl reference electrode. Instead of Ecoflex, fibroin, PLA, and wound adhesive are tested as alternative encapsulation compounds: a series of swelling tests with different fibroin compositions, PLA, and Ecoflex has been performed before characterizing the most promising candidates by chronoamperometry. Therefore, the carbon electrodes are completely covered with the particular encapsulation material. Chronoamperometric measurements with H2O2 concentrations between 0.5 and 10 mM enable studying the leakage current behavior.}, language = {en} } @article{FalkenbergVossBottetal.2023, author = {Falkenberg, Fabian and Voß, Leonie and Bott, Michael and Bongaerts, Johannes and Siegert, Petra}, title = {New robust subtilisins from halotolerant and halophilic Bacillaceae}, series = {Applied Microbiology and Biotechnology}, volume = {107}, journal = {Applied Microbiology and Biotechnology}, publisher = {Springer Nature}, address = {Berlin}, issn = {1432-0614}, doi = {10.1007/s00253-023-12553-w}, pages = {3939 -- 3954}, year = {2023}, abstract = {The aim of the present study was the characterisation of three true subtilisins and one phylogenetically intermediate subtilisin from halotolerant and halophilic microorganisms. Considering the currently growing enzyme market for efficient and novel biocatalysts, data mining is a promising source for novel, as yet uncharacterised enzymes, especially from halophilic or halotolerant Bacillaceae, which offer great potential to meet industrial needs. Both halophilic bacteria Pontibacillus marinus DSM 16465ᵀ and Alkalibacillus haloalkaliphilus DSM 5271ᵀ and both halotolerant bacteria Metabacillus indicus DSM 16189 and Litchfieldia alkalitelluris DSM 16976ᵀ served as a source for the four new subtilisins SPPM, SPAH, SPMI and SPLA. The protease genes were cloned and expressed in Bacillus subtilis DB104. Purification to apparent homogeneity was achieved by ethanol precipitation, desalting and ion-exchange chromatography. Enzyme activity could be observed between pH 5.0-12.0 with an optimum for SPPM, SPMI and SPLA around pH 9.0 and for SPAH at pH 10.0. The optimal temperature for SPMI and SPLA was 70 °C and for SPPM and SPAH 55 °C and 50 °C, respectively. All proteases showed high stability towards 5\% (w/v) SDS and were active even at NaCl concentrations of 5 M. The four proteases demonstrate potential for future biotechnological applications.}, language = {en} } @article{MoehrenBergmannJanseretal.2023, author = {M{\"o}hren, Felix and Bergmann, Ole and Janser, Frank and Braun, Carsten}, title = {On the influence of elasticity on propeller performance: a parametric study}, series = {CEAS Aeronautical Journal}, volume = {14}, journal = {CEAS Aeronautical Journal}, publisher = {Springer Nature}, address = {Berlin}, issn = {1869-5590 (Online)}, doi = {10.1007/s13272-023-00649-y}, pages = {311 -- 323}, year = {2023}, abstract = {The aerodynamic performance of propellers strongly depends on their geometry and, consequently, on aeroelastic deformations. Knowledge of the extent of the impact is crucial for overall aircraft performance. An integrated simulation environment for steady aeroelastic propeller simulations is presented. The simulation environment is applied to determine the impact of elastic deformations on the aerodynamic propeller performance. The aerodynamic module includes a blade element momentum approach to calculate aerodynamic loads. The structural module is based on finite beam elements, according to Timoshenko theory, including moderate deflections. Several fixed-pitch propellers with thin-walled cross sections made of both isotropic and non-isotropic materials are investigated. The essential parameters are varied: diameter, disc loading, sweep, material, rotational, and flight velocity. The relative change of thrust between rigid and elastic blades quantifies the impact of propeller elasticity. Swept propellers of large diameters or low disc loadings can decrease the thrust significantly. High flight velocities and low material stiffness amplify this tendency. Performance calculations without consideration of propeller elasticity can lead to decreased efficiency. To avoid cost- and time-intense redesigns, propeller elasticity should be considered for swept planforms and low disc loadings.}, language = {en} } @article{BlockMayWetzeletal.2023, author = {Block, Franziska and May, Alexander and Wetzel, Katharina and Adels, Klaudia and Elbers, Gereon and Schulze, Margit and Monakhova, Yulia}, title = {What is the best spectroscopic method for simultaneous analysis of organic acids and (poly)saccharides in biological matrices: Example of Aloe vera extracts?}, series = {Talanta Open}, volume = {7}, journal = {Talanta Open}, number = {Art. No. 100220}, publisher = {Elsevier}, address = {Amsterdam}, issn = {2666-8319}, doi = {10.1016/j.talo.2023.100220}, pages = {1 -- 9}, year = {2023}, abstract = {Several species of (poly)saccharides and organic acids can be found often simultaneously in various biological matrices, e.g., fruits, plant materials, and biological fluids. The analysis of such matrices sometimes represents a challenging task. Using Aloe vera (A. vera) plant materials as an example, the performance of several spectro-scopic methods (80 MHz benchtop NMR, NIR, ATR-FTIR and UV-vis) for the simultaneous analysis of quality parameters of this plant material was compared. The determined parameters include (poly)saccharides such as aloverose, fructose and glucose as well as organic acids (malic, lactic, citric, isocitric, acetic, fumaric, benzoic and sorbic acids). 500 MHz NMR and high-performance liquid chromatography (HPLC) were used as the reference methods. UV-vis data can be used only for identification of added preservatives (benzoic and sorbic acids) and drying agent (maltodextrin) and semiquantitative analysis of malic acid. NIR and MIR spectroscopies combined with multivariate regression can deliver more informative overview of A. vera extracts being able to additionally quantify glucose, aloverose, citric, isocitric, malic, lactic acids and fructose. Low-field NMR measurements can be used for the quantification of aloverose, glucose, malic, lactic, acetic, and benzoic acids. The benchtop NMR method was successfully validated in terms of robustness, stability, precision, reproducibility and limit of detection (LOD) and quantification (LOQ), respectively. All spectroscopic techniques are useful for the screening of (poly)saccharides and organic acids in plant extracts and should be applied according to its availability as well as information and confidence required for the specific analytical goal. Benchtop NMR spectroscopy seems to be the most feasible solution for quality control of A. vera products.}, language = {en} } @article{HaegerWirgesTanzmannetal.2023, author = {Haeger, Gerrit and Wirges, Jessika and Tanzmann, Nicole and Oyen, Sven and Jolmes, Tristan and Jaeger, Karl-Erich and Sch{\"o}rken, Ulrich and Bongaerts, Johannes and Siegert, Petra}, title = {Chaperone assisted recombinant expression of a mycobacterial aminoacylase in Vibrio natriegens and Escherichia coli capable of N-lauroyl-L-amino acid synthesis}, series = {Microbial Cell Factories}, journal = {Microbial Cell Factories}, number = {22}, publisher = {Springer Nature}, issn = {1475-2859}, doi = {10.1186/s12934-023-02079-1}, pages = {Article number: 77 (2023)}, year = {2023}, abstract = {Background Aminoacylases are highly promising enzymes for the green synthesis of acyl-amino acids, potentially replacing the environmentally harmful Schotten-Baumann reaction. Long-chain acyl-amino acids can serve as strong surfactants and emulsifiers, with application in cosmetic industries. Heterologous expression of these enzymes, however, is often hampered, limiting their use in industrial processes. Results We identified a novel mycobacterial aminoacylase gene from Mycolicibacterium smegmatis MKD 8, cloned and expressed it in Escherichia coli and Vibrio natriegens using the T7 overexpression system. The recombinant enzyme was prone to aggregate as inclusion bodies, and while V. natriegens Vmax™ could produce soluble aminoacylase upon induction with isopropyl β-d-1-thiogalactopyranoside (IPTG), E. coli BL21 (DE3) needed autoinduction with lactose to produce soluble recombinant protein. We successfully conducted a chaperone co-expression study in both organisms to further enhance aminoacylase production and found that overexpression of chaperones GroEL/S enhanced aminoacylase activity in the cell-free extract 1.8-fold in V. natriegens and E. coli. Eventually, E. coli ArcticExpress™ (DE3), which co-expresses cold-adapted chaperonins Cpn60/10 from Oleispira antarctica, cultivated at 12 °C, rendered the most suitable expression system for this aminoacylase and exhibited twice the aminoacylase activity in the cell-free extract compared to E. coli BL21 (DE3) with GroEL/S co-expression at 20 °C. The purified aminoacylase was characterized based on hydrolytic activities, being most stable and active at pH 7.0, with a maximum activity at 70 °C, and stability at 40 °C and pH 7.0 for 5 days. The aminoacylase strongly prefers short-chain acyl-amino acids with smaller, hydrophobic amino acid residues. Several long-chain amino acids were fairly accepted in hydrolysis as well, especially N-lauroyl-L-methionine. To initially evaluate the relevance of this aminoacylase for the synthesis of N-acyl-amino acids, we demonstrated that lauroyl-methionine can be synthesized from lauric acid and methionine in an aqueous system. Conclusion Our results suggest that the recombinant enzyme is well suited for synthesis reactions and will thus be further investigated.}, language = {en} }