Optimization of High Cell density cultivations of Schizosacchraromyces pombe for the production of surface active proteins
Hydrophobins assemble spontaneously at hydrophilic/hydrophobic boundary layers. This way hydrophobic surfaces turn hydrophilic. Due to that feature hydrophobins can be used to alter the surface properties of liquid or solid matters. By the virtue of their biocompatibility these proteins are promising for the application in medical fields, e.g. the modification of the surface characteristics of implant surfaces or the fixation of active ingredients. Prerequisite for the function of the hydrophobin and proteins in general is the correct co- and posttranslational modification in their host cell. Saccharomyces cerevisiae and Schizosaccharomyces pombe, that can be engineered easily, are probably the the most promising candidates for heterologous gene expression. However, these two yeast species differ in both their physiology and metabolism significantly. Recent investigations showed that in some cases proteins can be expressed and excreted more easily in S. pombe, facilitating the down- stream processing.
Objective:
In the project introduced here, two different cloning strategies are realized and the expression- efficiency is evaluated using Flow Cytometry. To measure the expression levels, these the reporter Green Fluorescence Protein (GFP) is used. This way the stability and performance of expression can be evaluated conveniently. Further goal of this project is to develop a cultivation strategy to enhance the the levels of expression and allow an effective production of the hydrophobin HFB I. For fed batch fermentations feed rates, media compositions as well as point in time of the induction should be optimized.
Cooperation partners:
Institute for Genetics, TU-Dresden
Project financing:
Saxonian Ministry for Science and Arts
Contact:
Weber, Bley