Purification, characterization, and cloning of a bifunctional molybdoenzyme with hydratase and alcohol dehydrogenase activity

A bifunctional hydratase/alcohol dehydrogenase was isolated from the cyclohexanol degrading bacterium Alicycliphilus denitrificans DSMZ 14773. The enzyme catalyzes the addition of water to α,β-unsaturated carbonyl compounds and the subsequent alcohol oxidation. The purified enzyme showed three subunits in SDS gel, and the gene sequence revealed that this enzyme belongs to the molybdopterin binding oxidoreductase family containing molybdopterins, FAD, and iron-sulfur clusters

Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis

For the improved production of vaccines and therapeutic proteins, a detailed understanding of the metabolic dynamics during batch or fed-batch production is requested. To study the new human cell line AGE1.HN, a flexible metabolic flux analysis method was developed that is considering dynamic changes in growth and metabolism during cultivation. This method comprises analysis of formation of cellular components as well as conversion of major substrates and products, spline fitting of dynamic data and flux estimation using metabolite balancing. During batch cultivation of AGE1.HN three distinct phases were observed, an initial one with consumption of pyruvate and high glycolytic activity, a second characterized by a highly efficient metabolism with very little energy spilling waste production and a third with glutamine limitation and decreasing viability. Main events triggering changes in cellular metabolism were depletion of pyruvate and glutamine. Potential targets for the improvement identified from the analysis are (i) reduction of overflow metabolism in the beginning of cultivation, e.g. accomplished by reduction of pyruvate content in the medium and (ii) prolongation of phase 2 with its highly efficient energy metabolism applying e.g. specific feeding strategies. The method presented allows fast and reliable metabolic flux analysis during the development of producer cells and production processes from microtiter plate to large scale reactors with moderate analytical and computational effort. It seems well suited to guide media optimization and genetic engineering of producing cell lines

Studying in times of Corona

In March 2020 the Covid-19 pandemic changed life circumstances drastically for everyone. Exactly one year ago the university had to shift from campus to the internet. This led to huge alterations in how professors lecture and how student university life takes place. After this year, it is time to evaluate the changes and put them into perspective. This means recognizing the benefits of technology to legal education instead of seeing the current situation as a replacement that disappears as soon as a return back to campus is possible

Studying in times of Corona

In March 2020 the Covid-19 pandemic changed life circumstances drastically for everyone. Exactly one year ago the university had to shift from campus to the internet. This led to huge alterations in how professors lecture and how student university life takes place. After this year, it is time to evaluate the changes and put them into perspective. This means recognizing the benefits of technology to legal education instead of seeing the current situation as a replacement that disappears as soon as a return back to campus is possible

Studying in times of Corona

In March 2020 the Covid-19 pandemic changed life circumstances drastically for everyone. Exactly one year ago the university had to shift from campus to the internet. This led to huge alterations in how professors lecture and how student university life takes place. After this year, it is time to evaluate the changes and put them into perspective. This means recognizing the benefits of technology to legal education instead of seeing the current situation as a replacement that disappears as soon as a return back to campus is possible