Evaluation of different expression systems for the heterologous expression of pyranose 2-oxidase from Trametes multicolor in E. coli

The heterologous production of the industrially relevant fungal enzyme pyranose 2-oxidase in the prokaryotic host E. coli was investigated using 3 different expression systems, i.e. the well-studied T7 RNA polymerase based pET21d+, the L-arabinose inducible pBAD and the pCOLD system. Preliminary experiments were done in shaking flasks at 25°C and optimized induction conditions to compare the productivity levels of the different expression systems. The pET21d+ and the pCOLD system gave 29 U/L·h and 14 U/L·h of active pyranose 2-oxidase, respectively, whereas the pBAD system only produced 6 U/L·h. Process conditions for batch fermentations were optimized for the pET21d+ and the pCOLD systems in order to reduce the formation of inactive inclusion bodies. The highest productivity rate with the pET21d+ expression system in batch fermentations was determined at 25°C with 32 U/L·h. The pCOLD system showed the highest productivity rate (19 U/L·h) at 25°C and induction from the start of the cultivation. Using the pCOLD system in a fed batch fermentation at 25°C with a specific growth rate of μ = 0.15 h-1resulted in the highest productivity rate of active pyranose oxidase with 206 U/L·h

An Empirical Survey on Co-simulation: Promising Standards, Challenges and Research Needs

Co-simulation is a promising approach for the modelling and simulation of complex systems, that makes use of mature simulation tools in the respective domains. It has been applied in wildly different domains, oftentimes without a comprehensive study of the impact to the simulation results. As a consequence, over the recent years, researchers have set out to understand the essential challenges arising from the application of this technique. This paper complements the existing surveys in that the social and empirical aspects were addressed. More than 50 experts participated in a two-stage Delphi study to determine current challenges, research needs and promising standards and tools. Furthermore, an analysis of the strengths, weakness, opportunities and threats of co-simulation utilizing the analytic hierarchy process resulting in a SWOT-AHP analysis is presented. The empirical results of this study show that experts consider the FMI standard to be the most promising standard for continuous time, discrete event and hybrid co-simulation. The results of the SWOT-AHP analysis indicate that factors related to strengths and opportunities predominate

Analysis of the cell surface layer ultrastructure of the oral pathogen Tannerella forsythia

The Gram-negative oral pathogen Tannerella forsythia is decorated with a 2D crystalline surface (S-) layer, with two different S-layer glycoprotein species being present. Prompted by the predicted virulence potential of the S-layer, this study focused on the analysis of the arrangement of the individual S-layer glycoproteins by a combination of microscopic, genetic, and biochemical analyses. The two S-layer genes are transcribed into mRNA and expressed into protein in equal amounts. The S-layer was investigated on intact bacterial cells by transmission electron microscopy, by immune fluorescence microscopy, and by atomic force microscopy. The analyses of wild-type cells revealed a distinct square S-layer lattice with an overall lattice constant of 10.1 ± 0.7 nm. In contrast, a blurred lattice with a lattice constant of 9.0 nm was found on S-layer single-mutant cells. This together with in vitro self-assembly studies using purified (glyco)protein species indicated their increased structural flexibility after self-assembly and/or impaired self-assembly capability. In conjunction with TEM analyses of thin-sectioned cells, this study demonstrates the unusual case that two S-layer glycoproteins are co-assembled into a single S-layer. Additionally, flagella and pilus-like structures were observed on T. forsythia cells, which might impact the pathogenicity of this bacterium

On-treatment measurements of circulating tumor DNA during FOLFOX therapy in patients with colorectal cancer.

We addressed a significant unknown feature of circulating tumor DNA (ctDNA), i.e., how ctDNA levels change during chemotherapy, by serially monitoring ctDNA in patients with colorectal cancer during the 48-h application of FOLFOX. Surprisingly, we did not observe a spike in ctDNA as a sign of a responsive tumor, but instead ctDNA levels initially decreased and remained low in patients with stable disease or partial response. Our observations reveal further insights into cell destruction during chemotherapy with important implications for the management of patients

District Heating Systems: An Analysis of Strengths, Weaknesses, Opportunities, and Threats of the 4GDH

Fourth-generation district heating networks (4GDH) can play a special role in the efficient and climate-friendly use of energy. In this study, we have examined the strengths, weaknesses, opportunities, and threats (SWOT) of this innovative technology. Using a combination of qualitative and quantitative research methods, we assessed the SWOT-factors in terms of their importance. Among the factors that were weighted with the highest relative importance were the ability of 4GDH to serve as a label bundling and stimulating considerations with respect to the further development of district heating systems and the increased value creation within the national economy through the inclusion of local, renewable energy sources. Moreover, the interviewed experts agreed that regulatory frameworks in the context of 4GDH have to be further developed

Modeling and simulation of large-scale systems: A systematic comparison of modeling paradigms

A trend across most areas where simulation-driven development is used is the ever in- creasing size and complexity of the systems under consideration, pushing established methods of modeling and simulation towards their limits. This paper complements existing surveys on large-scale modeling and simulation of physical systems by conducting expert surveys. We conducted a two-stage empirical survey in order to investigate research needs, current challenges as well as promising modeling and simulation paradigms. Furthermore, we applied the analytic hierarchy process method to prioritise the strengths and weak- ness of different modeling paradigms. The results of this study show that experts consider acausal modeling techniques to be suitable for modeling large scale systems, while causal techniques are considered less suitable.Validerad;2019;Nivå 2;2019-09-18 (johcin)</p