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Max Wellerdiek Dajana Winterhoff Waldemar Reule Jürgen Brandner Marco Oldiges 《Bioprocess and biosystems engineering》2009,32(5):581-592
Representative and valid cytoplasmic concentrations are essential for ensuring the significance of results in the field of
metabolome analysis. One of the most crucial points in this respect is the sampling itself. A rapid and sudden stopping of
the metabolism on a timescale that is much faster than the conversion rates of investigated metabolites is worthwhile. This
can be achieved by applying of cold methanol quenching combined with reproducible, fast, and automated sampling. Unfortunately,
quenching the metabolism by a sharp temperature shift leads to what is known as cold shock or the cell-leakage effect. In
the present work, we applied a microstructure heat exchanger to analyze the cold shock effect using Corynebacterium glutamicum as a model microorganism. Using this apparatus together with a silicon pipe, it was possible to assay the leakage effect
on a timescale starting at 1 s after cooling cell suspension. The high turnover rates not only require a rapid quenching technique,
but also the correct application. Moreover, we succeeded in showing that even when the required appropriate setup of methanol
quenching is not used, the metabolism is not stopped within the required timescale. By applying robust techniques like rapid
sampling in combination with reproducible sample processing, we ensured fast and reliable metabolic inactivation during all
steps. 相似文献
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The decomposition of urea by Nitellopsis obtusa from Characeae was investigated. The intact cells were exposed to the inhibition by two typical urease inhibitors: boric acid and fluoride
ion, used as a criterion to define if urease or UAL-ase is responsible for the ureolytic activity of the algae. It was found
that boric acid and fluoride ion are simple competitive and slow-binding competitive inhibitors of Nitellopsis obtusa enzyme respectively, which is the response characteristic of urease. The inhibition constants equal to 2.3 and 0.1 mM for
boric acid and fluoride ion, when compared to those of jack bean urease, indicate that in the observed kinetic behaviour of
Nitellopsis obtusa urease partake transport processes taking place in the intact cells. 相似文献
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