TY - RPRT A1 - Siegert, Petra A1 - Bongaerts, Johannes A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Selmer, Thorsten T1 - Abschlussbericht zum Projekt zur Überwachung biotechnologischer Prozesse mittels Diacetyl-/Acetoin-Biosensor und Evaluierung von Acetoin-Reduktasen zur Verwendung in Biotransformationen Y1 - 2022 N1 - Laufzeit: 01.01.2016 – 31.12.2019 (verlängert bis 31.12.2020) Förderkennzeichen: 322-8.03.04.02-FH-Struktur 2016/02 Gefördert durch: Ministerium für Innovation, Wissenschaft und Forschung des Landes Nordrhein-Westfalen CY - Aachen ER - TY - JOUR A1 - Cheenakula, Dheeraja A1 - Hoffstadt, Kevin A1 - Krafft, Simone A1 - Reinecke, Diana A1 - Klose, Holger A1 - Kuperjans, Isabel A1 - Grömping, Markus T1 - Anaerobic digestion of algal–bacterial biomass of an Algal Turf Scrubber system JF - Biomass Conversion and Biorefinery N2 - This study investigated the anaerobic digestion of an algal–bacterial biofilm grown in artificial wastewater in an Algal Turf Scrubber (ATS). The ATS system was located in a greenhouse (50°54′19ʺN, 6°24′55ʺE, Germany) and was exposed to seasonal conditions during the experiment period. The methane (CH4) potential of untreated algal–bacterial biofilm (UAB) and thermally pretreated biofilm (PAB) using different microbial inocula was determined by anaerobic batch fermentation. Methane productivity of UAB differed significantly between microbial inocula of digested wastepaper, a mixture of manure and maize silage, anaerobic sewage sludge, and percolated green waste. UAB using sewage sludge as inoculum showed the highest methane productivity. The share of methane in biogas was dependent on inoculum. Using PAB, a strong positive impact on methane productivity was identified for the digested wastepaper (116.4%) and a mixture of manure and maize silage (107.4%) inocula. By contrast, the methane yield was significantly reduced for the digested anaerobic sewage sludge (50.6%) and percolated green waste (43.5%) inocula. To further evaluate the potential of algal–bacterial biofilm for biogas production in wastewater treatment and biogas plants in a circular bioeconomy, scale-up calculations were conducted. It was found that a 0.116 km2 ATS would be required in an average municipal wastewater treatment plant which can be viewed as problematic in terms of space consumption. However, a substantial amount of energy surplus (4.7–12.5 MWh a−1) can be gained through the addition of algal–bacterial biomass to the anaerobic digester of a municipal wastewater treatment plant. Wastewater treatment and subsequent energy production through algae show dominancy over conventional technologies. KW - Biogas KW - Methane KW - Algal Turf Scrubber KW - Algal–bacterial bioflm KW - Circular bioeconomy Y1 - 2022 U6 - https://doi.org/10.1007/s13399-022-03236-z SN - 2190-6823 N1 - Corresponding author: Dheeraja Cheenakula VL - 13 SP - 15 Seiten PB - Springer CY - Berlin ER - TY - BOOK A1 - Lauth, Jakob T1 - Physikalische Chemie kompakt KW - Physikalische Chemie Y1 - 2022 SN - 978-3-662-64587-1 U6 - https://doi.org/https://doi.org/10.1007/978-3-662-64588-8 PB - Springer Spektrum CY - Berlin ER - TY - JOUR A1 - Falkenberg, Fabian A1 - Rahba, Jade A1 - Fischer, David A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Biochemical characterization of a novel oxidatively stable, halotolerant, and high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T JF - FEBS Open Bio N2 - Halophilic and halotolerant microorganisms represent a promising source of salt-tolerant enzymes suitable for various biotechnological applications where high salt concentrations would otherwise limit enzymatic activity. Considering the current growing enzyme market and the need for more efficient and new biocatalysts, the present study aimed at the characterization of a high-alkaline subtilisin from Alkalihalobacillus okhensis Kh10-101T. The protease gene was cloned and expressed in Bacillus subtilis DB104. The recombinant protease SPAO with 269 amino acids belongs to the subfamily of high-alkaline subtilisins. The biochemical characteristics of purified SPAO were analyzed in comparison with subtilisin Carlsberg, Savinase, and BPN'. SPAO, a monomer with a molecular mass of 27.1 kDa, was active over a wide range of pH 6.0–12.0 and temperature 20–80 °C, optimally at pH 9.0–9.5 and 55 °C. The protease is highly oxidatively stable to hydrogen peroxide and retained 58% of residual activity when incubated at 10 °C with 5% (v/v) H2O2 for 1 h while stimulated at 1% (v/v) H2O2. Furthermore, SPAO was very stable and active at NaCl concentrations up to 5.0 m. This study demonstrates the potential of SPAO for biotechnological applications in the future. KW - Alkalihalobacillus okhensis KW - detergent protease KW - halotolerant protease KW - high-alkaline subtilisin KW - oxidative stable protease Y1 - 2022 U6 - https://doi.org/10.1002/2211-5463.13457 SN - 2211-5463 N1 - Corresponding author: Petra Siegert VL - 12 IS - 10 SP - 1729 EP - 1746 PB - Wiley CY - Hoboken, NJ ER - TY - JOUR A1 - Monakhova, Yulia A1 - Diehl, Bernd W.K. T1 - Multinuclear NMR screening of pharmaceuticals using standardization by 2H integral of a deuterated solvent JF - Journal of Pharmaceutical and Biomedical Analysis N2 - NMR standardization approach that uses the 2H integral of deuterated solvent for quantitative multinuclear analysis of pharmaceuticals is described. As a proof of principle, the existing NMR procedure for the analysis of heparin products according to US Pharmacopeia monograph is extended to the determination of Na+ and Cl- content in this matrix. Quantification is performed based on the ratio of a 23Na (35Cl) NMR integral and 2H NMR signal of deuterated solvent, D2O, acquired using the specific spectrometer hardware. As an alternative, the possibility of 133Cs standardization using the addition of Cs2CO3 stock solution is shown. Validation characteristics (linearity, repeatability, sensitivity) are evaluated. A holistic NMR profiling of heparin products can now also be used for the quantitative determination of inorganic compounds in a single analytical run using a single sample. In general, the new standardization methodology provides an appealing alternative for the NMR screening of inorganic and organic components in pharmaceutical products. KW - NMR spectroscopy KW - Inorganic ions KW - Heparin KW - Standardization Y1 - 2022 SN - 0731-7085 U6 - https://doi.org/10.1016/j.jpba.2021.114530 VL - 209 IS - Article number: 114530 PB - Elsevier ER - TY - JOUR A1 - Welden, Melanie A1 - Poghossian, Arshak A1 - Vahidpour, Farnoosh A1 - Wendlandt, Tim A1 - Keusgen, Michael A1 - Wege, Christina A1 - Schöning, Michael Josef T1 - Towards multi-analyte detection with field-effect capacitors modified with tobacco mosaic virus bioparticles as enzyme nanocarriers JF - Biosensors N2 - Utilizing an appropriate enzyme immobilization strategy is crucial for designing enzyme-based biosensors. Plant virus-like particles represent ideal nanoscaffolds for an extremely dense and precise immobilization of enzymes, due to their regular shape, high surface-to-volume ratio and high density of surface binding sites. In the present work, tobacco mosaic virus (TMV) particles were applied for the co-immobilization of penicillinase and urease onto the gate surface of a field-effect electrolyte-insulator-semiconductor capacitor (EISCAP) with a p-Si-SiO₂-Ta₂O₅ layer structure for the sequential detection of penicillin and urea. The TMV-assisted bi-enzyme EISCAP biosensor exhibited a high urea and penicillin sensitivity of 54 and 85 mV/dec, respectively, in the concentration range of 0.1–3 mM. For comparison, the characteristics of single-enzyme EISCAP biosensors modified with TMV particles immobilized with either penicillinase or urease were also investigated. The surface morphology of the TMV-modified Ta₂O₅-gate was analyzed by scanning electron microscopy. Additionally, the bi-enzyme EISCAP was applied to mimic an XOR (Exclusive OR) enzyme logic gate. KW - urease KW - enzyme-logic gate KW - bi-enzyme biosensor KW - capacitive field-effect sensor KW - tobacco mosaic virus (TMV) KW - penicillinase Y1 - 2022 U6 - https://doi.org/10.3390/bios12010043 SN - 2079-6374 N1 - This article belongs to the Special Issue "Biosensors: 10th Anniversary Feature Papers" VL - 12 IS - 1 PB - MDPI CY - Basel ER - TY - JOUR A1 - Falkenberg, Fabian A1 - Bott, Michael A1 - Bongaerts, Johannes A1 - Siegert, Petra T1 - Phylogenetic survey of the subtilase family and a data-mining-based search for new subtilisins from Bacillaceae JF - Frontiers in Microbiology N2 - The subtilase family (S8), a member of the clan SB of serine proteases are ubiquitous in all kingdoms of life and fulfil different physiological functions. Subtilases are divided in several groups and especially subtilisins are of interest as they are used in various industrial sectors. Therefore, we searched for new subtilisin sequences of the family Bacillaceae using a data mining approach. The obtained 1,400 sequences were phylogenetically classified in the context of the subtilase family. This required an updated comprehensive overview of the different groups within this family. To fill this gap, we conducted a phylogenetic survey of the S8 family with characterised holotypes derived from the MEROPS database. The analysis revealed the presence of eight previously uncharacterised groups and 13 subgroups within the S8 family. The sequences that emerged from the data mining with the set filter parameters were mainly assigned to the subtilisin subgroups of true subtilisins, high-alkaline subtilisins, and phylogenetically intermediate subtilisins and represent an excellent source for new subtilisin candidates. Y1 - 2022 U6 - https://doi.org/10.3389/fmicb.2022.1017978 SN - 1664-302X VL - 2022 IS - 13 PB - Frontiers CY - Lausanne ER - TY - JOUR A1 - Welden, Melanie A1 - Severins, Robin A1 - Poghossian, Arshak A1 - Wege, Christina A1 - Bongaerts, Johannes A1 - Siegert, Petra A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Detection of acetoin and diacetyl by a tobacco mosaic virus-assisted field-effect biosensor JF - Chemosensors N2 - Acetoin and diacetyl have a major impact on the flavor of alcoholic beverages such as wine or beer. Therefore, their measurement is important during the fermentation process. Until now, gas chromatographic techniques have typically been applied; however, these require expensive laboratory equipment and trained staff, and do not allow for online monitoring. In this work, a capacitive electrolyte–insulator–semiconductor sensor modified with tobacco mosaic virus (TMV) particles as enzyme nanocarriers for the detection of acetoin and diacetyl is presented. The enzyme acetoin reductase from Alkalihalobacillus clausii DSM 8716ᵀ is immobilized via biotin–streptavidin affinity, binding to the surface of the TMV particles. The TMV-assisted biosensor is electrochemically characterized by means of leakage–current, capacitance–voltage, and constant capacitance measurements. In this paper, the novel biosensor is studied regarding its sensitivity and long-term stability in buffer solution. Moreover, the TMV-assisted capacitive field-effect sensor is applied for the detection of diacetyl for the first time. The measurement of acetoin and diacetyl with the same sensor setup is demonstrated. Finally, the successive detection of acetoin and diacetyl in buffer and in diluted beer is studied by tuning the sensitivity of the biosensor using the pH value of the measurement solution. Y1 - 2022 U6 - https://doi.org/10.3390/chemosensors10060218 SN - 2227-9040 N1 - This article belongs to the Special Issue "Nanostructured Devices for Biochemical Sensing" VL - 10 IS - 6 PB - MDPI CY - Basel ER - TY - JOUR A1 - Ojovan, Michael I. A1 - Steinmetz, Hans-Jürgen T1 - Approaches to Disposal of Nuclear Waste JF - Energies N2 - We present a concise mini overview on the approaches to the disposal of nuclear waste currently used or deployed. The disposal of nuclear waste is the end point of nuclear waste management (NWM) activities and is the emplacement of waste in an appropriate facility without the intention to retrieve it. The IAEA has developed an internationally accepted classification scheme based on the end points of NWM, which is used as guidance. Retention times needed for safe isolation of waste radionuclides are estimated based on the radiotoxicity of nuclear waste. Disposal facilities usually rely on a multi-barrier defence system to isolate the waste from the biosphere, which comprises the natural geological barrier and the engineered barrier system. Disposal facilities could be of a trench type, vaults, tunnels, shafts, boreholes, or mined repositories. A graded approach relates the depth of the disposal facilities’ location with the level of hazard. Disposal practices demonstrate the reliability of nuclear waste disposal with minimal expected impacts on the environment and humans. KW - borehole disposal KW - geological disposal KW - disposal facility KW - retention time KW - nuclear waste Y1 - 2022 U6 - https://doi.org/10.3390/en15207804 SN - 1996-1073 N1 - This article belongs to the Special Issue "Treatment of Radioactive Waste and Sustainability Energy" VL - 15 IS - 20 PB - MDPI CY - Basel ER - TY - GEN A1 - Braun, Lena A1 - Krafft, Simone A1 - Tippkötter, Nils T1 - Combined supercritical carbon dioxide extraction and chromatography of the algae fatty linoleic and linolenic acid T2 - Chemie Ingenieur Technik N2 - A method for the integrated extraction and separation of fatty acids from algae using supercritical CO2 is presented. Desmodesmus obliquus and Chlorella sorokiniana were used as algae. First, a method for chromatographic separation of fatty acids of different degrees of saturation was established and optimized. Then, an integrated method for supercritical extraction was developed for both algal species. It was also verified whether prior cell disruption was beneficial for extraction. In developing the method for chromatographic separation, statistical experimental design was used to determine the optimal parameter settings. The methanol content in the mobile phase proved to be the most important parameter for successful separation of the three unsaturated fatty acids oleic acid, linoleic acid, and linolenic acid. Supercritical extraction with dried algae showed that about four times more fatty acids can be extracted from C. sorokiniana relative to the dry mass used. Y1 - 2022 U6 - https://doi.org/10.1002/cite.202255308 SN - 0009-286X SN - 1522-2640 (eISSN) N1 - ProcessNet and DECHEMA‐BioTechNet Jahrestagungen 2022 together with 13th ESBES Symposium 2022, 12. - 15. September 2022, Eurogress Aachen VL - 94 IS - 9 SP - 1304 PB - Wiley-VCH CY - Weinheim ER -