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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.
This paper describes the potential for developing a digital twin of society- a dynamic model that can be used to observe, analyze, and predict the evolution of various societal aspects. Such a digital twin can help governmental agencies and policy makers in interpreting trends, understanding challenges, and making decisions regarding investments or policies necessary to support societal development and ensure future prosperity. The paper reviews related work regarding the digital twin paradigm and its applications. The paper presents a motivating case study- an analysis of opportunities and challenges faced by the German federal employment agency, Bundesagentur f¨ur Arbeit (BA), proposes solutions using digital twins, and describes initial proofs of concept for such solutions.
Dieses Lehr- und Fachbuch vermittelt anschaulich die Grundlagen der HF-Technik, gibt konkrete Beschreibungen für den Entwurf von linearen Komponenten aus Bauteilen wie auch Leitungen für High-Speed- und HF-Schaltungen. Dem Leser wird vermittelt, wie Bauteile modelliert und Schaltungen synthetisiert und optimiert werden. Mit Hilfe frei verfügbarer Simulationssoftware können GHz-Schaltungen selbst entwickelt werden. Viele Übungsbeispiele ermöglichen die Eigenkontrolle des Wissensstandes. Weiterhin werden komplexe nichtlineare Komponenten wie Hochfrequenzmischer, Oszillatoren und Synthesegeneratoren in ihrer Funktionalität dargestellt. Die neuen Mixed-Mode-Streuparameter sowie deren Leitungs- und Schaltungstechnik für Anwendungen der schnellen Digital- und der modernen HF-Technik sind ausführlich beschrieben. Es wird auf Systeme für folgende Bereiche eingegangen: Streuparametermesstechnik, verschiedene Funktechniken, UHF-RFID und Lokalisierung- und Ortung. Dem Leser wird somit ermöglicht, komplexe GHz-Schaltungen insbesondere mit Halbleiter-, SMD- und LTCC-Schaltungen zu entwickeln.
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.
This study describes the development of a new combined polysaccharide-matrix-based technology for the immobilization of Lactobacillus rhamnosus GG (LGG) bacteria in biofilm form. The new composition allows for delivering the bacteria to the digestive tract in a manner that improves their robustness compared with planktonic cells and released biofilm cells. Granules consisting of a polysaccharide matrix with probiotic biofilms (PMPB) with high cell density (>9 log CFU/g) were obtained by immobilization in the optimized nutrient medium. Successful probiotic loading was confirmed by fluorescence microscopy and scanning electron microscopy. The developed prebiotic polysaccharide matrix significantly enhanced LGG viability under acidic (pH 2.0) and bile salt (0.3%) stress conditions. Enzymatic extract of feces, mimicking colon fluid in terms of cellulase activity, was used to evaluate the intestinal release of probiotics. PMPB granules showed the ability to gradually release a large number of viable LGG cells in the model colon fluid. In vivo, the oral administration of PMPB granules in rats resulted in the successful release of probiotics in the colon environment. The biofilm-forming incubation method of immobilization on a complex polysaccharide matrix tested in this study has shown high efficacy and promising potential for the development of innovative biotechnologies.
Damit Sie als Moderator effektiv und professionell moderieren können, sollten Sie die entsprechenden Methoden kennen.
Mit den richtigen Methoden können Sie Diskussionen leiten, Konflikte lösen, die Teilnehmer motivieren und dafür sorgen, dass die Ziele der Veranstaltung erreicht werden. Außerdem helfen sie Ihnen, eine positive Atmosphäre zu schaffen und das Interesse der Teilnehmer zu halten.
In diesem zweiten Beitrag der mehrteiligen Serie lernen Sie die grundsätzlichen Methoden kennen, um erfolgreiche Teamsitzungen, Arbeitsgruppentreffen, Kick-offs und Meetings durchzuführen.
Die Erbringung einer Leistung in der irrigen Vorstellung einer Bestellung (Zu BGH, MDR 2023, 1439)
(2023)
Manufacturing companies across multiple industries face an increasingly dynamic and unpredictable environment. This development can be seen on both the market and supply side. To respond to these challenges, manufacturing companies must implement smart manufacturing systems and become more flexible and agile. The flexibility in operational planning regarding the scheduling and sequencing of customer orders needs to be increased and new structures must be implemented in manufacturing systems’ fundamental design as they constitute much of the operational flexibility available. To this end, smart and more flexible solutions for production planning and control (PPC) are developed. However, scheduling or sequencing is often only considered isolated in a predefined stable environment. Moreover, their orientation on the fundamental logic of the existing IT solutions and their applicability in a dynamic environment is limited. This paper presents a conceptual model for a task-based description logic that can be applied to factory planning, technology planning, and operational control. By using service-oriented architectures, the goal is to generate smart manufacturing systems. The logic is designed to allow for easy and automated maintenance. It is compatible with the existing resource and process allocation logic across operational and strategic factory and production planning.