Practical Course Dr. Uwe Horn/ Prof Dr. Dirk Hoffmeister

Secondary Metabolites: From Strains to Biotechnological Processes

21 - 25 May 2007, HKI, Biotechnikum

Limited number of  participants :10 * 

*In order to provide each participant a thorough hands on experience we are restring the number of participants to 10. Should there be more interested participants  we would be happy to repeat the course next year.

Wine, sauerkraut or yogurt; since time immemorable the art of fermentation has been known to the man-kind. Metamorphosis of this art to science and then to technology began in the last century with the production of bio-active substances from micro-organisms at the industrial scale.

In the coursework, two applications of fermentation technology would be covered:

1. Periplasmic expression of camelid VHH antibody fragment and antibody purification from the biomass

In contrast to the conventional mammalian antibodies, camelid antibodies bear only single variable heavy domain (VHH) and lack the CH1 domain and the variable light chain. Because of the small size ~15kDa, VHH domains are very useful for recombinant antibody libraries. Specific camelid antibodies have been generated in our department, screened by Phage display technique and engineered for periplasmic expression in E.coli. Herein, E.coli would be cultivated by fed-batch fermentation in a defined glucose mineral salt media followed by downstream procedure of biomass accumulation, cell lysis and antibody purification.

2. Production and downstream processing of secondary metabolite Nourseothricin 

Bioactive nourseothricin along with the resistance confering gene: nourseothricin acetyltransferase, is used as a dominant marker in fungal genetics e.g. Candida albicans¹, Cryptococcus neoformans². Batch fermentation of  Streptomyces noursei would be carried out in complex media. Downstream applications include product recovery, purification using ion exchange chromatography followed by adsorption chromatography and finally recovery as a stable powder.

References:

1. Shen, J., W. Guo, and J.R. Kohler.2005. Infect Immun. 73(2): 1239–1242.
2. 2. McDade, H.C., and G.M. Cox. 2001. Med. Mycol. 39: 151-154