Biomedical subjects

C Herwig

Publications and source records attributed to C Herwig.

3 recordsLinked to original sources

PubMed2003

Know-how and know-why in biochemical engineering.

This contribution analyzes the position of biochemical engineering in general and bioprocess engineering particularly in the force fields between fundamental science and applications, and between academia and industry. By using culture technology as an example, it can be shown that bioprocess engineering has moved slowly but steadily from an empirical art concerned with mainly know-how to a science elucidating the know-why of culture behavior. Highly powerful monitoring tools enable biochemical engineers to understand and explain quantitatively the activity of cellular culture on a metabolic basis. Among these monitoring tools are not just semi-online analyses of culture broth by HPLC, GC and FIA, but, increasingly, also noninvasive methods such as midrange IR, Raman and capacitance spectroscopy, as well as online calorimetry. The detailed and quantitative insight into the metabolome and the fluxome that bioprocess engineers are establishing offers an unprecedented opportunity for building bridges between molecular biology and engineering biosciences. Thus, one of the major tasks of biochemical engineering sciences is not developing new know-how for industrial applications, but elucidating the know-why in biochemical engineering by conducting research on the underlying scientific fundamentals.

Biochemistry↗

PubMed2001

On-line stoichiometry and identification of metabolic state under dynamic process conditions.

A method for the on-line calculation of conversion rates and yield coefficients under dynamic process conditions was developed. The method is based on cumulated mass balances using a moving average method. Elemental balances were used to test the measured cumulated quantities for gross errors and inappropriate stoichiometry definition followed by data reconciliation and estimation of non-measured conversion rates, using a bioprocess set-up including multiple on-line analysis techniques. The quantitative potential of the proposed method is demonstrated by executing transient experiments in aerobic cultures of Saccharomyces cerevisiae on glucose. Rates and yield coefficients could be consistently quantified in shift-up, shift-down, and accelerostat experiments. The method shows the capability to describe quantitatively transient changes in metabolism including uncoupling of catabolism and anabolism, also for the case when multiple components of metabolism are not measured. The validity of the experiment can be evaluated on-line. Additionally, the method detects with high sensitivity inappropriate stoichiometry definition, such as a change in state of metabolism. It was shown that concentration values can be misleading for the identification of the metabolic state. In contrast, the proposed method provides a clear picture of the metabolic state and new physiological regulations could be revealed. Hence, the novelty of the proposed method is the on-line availability of consistent stoichiometric coefficients allowing a significant speed up in strain characterization and bioprocess development using minimal knowledge of the metabolism. Additionally, it opens up the use of transient experiments for physiological studies.

Bioreactors↗

PubMed2001

Quantitative analysis of the regulation scheme of invertase expression in Saccharomyces cerevisiae.

In Saccharomyces cerevisiae, the expression of invertase, which is the hydrolyzing enzyme of sucrose, is controlled by the presence of monosaccharides, such as glucose and fructose, and referred to as carbon catabolite repression. To date, efforts have been made to identify the mechanism by which cells sense extracellular monosaccharide concentrations and trigger the genes involved in the repression pathway. The aim of the present work was to quantitatively investigate the cellular regulation of invertase expression in the wild-type strain S. cerevisiae CEN.PK113-7D during batch growth containing mixed sugar substrates under different initial conditions. Because of the high frequency and accurate online analysis of multiple components, a tight control of invertase expression could be observed, and threshold concentrations of the monosaccharides for derepression could be determined to 0.5 gl(-1) for glucose and 2 gl(-1) for fructose. Also, the existence of a hitherto undescribed regulatory state, in which cells regulate invertase expression very precisely and operate over long periods at monosaccharide concentrations lower than the above thresholds, could be demonstrated. All experimental observations could be summarized in a formulation of the cellular regulation scheme of invertase expression. A simple kinetic model could show that the regulation scheme explains the observed behavior very well. Additionally, the model was able to explain consequences of the regulation on the global metabolism.

Bioreactors↗