Characterization of the growth behavior of Leishmania tarentolae: a new expression system for recombinant proteins.
Biotechnological production of recombinant proteins for human therapy requires a cultivation of the host organism in a nutrient medium free of animal substances. Therefore, various nutrient media for the new expression system Leishmania tarentolae were developed and examined according to their cultivation conditions as static suspension culture and agitated culture. Investigations resulted in the development of a serum-free but hemin containing medium, based on yeast extract and buffer salts. Here we report that a high and stable specific growth rate of 0.103 h(-1) and a maximal cell density of 1 x 10(9) cells ml(-1) is obtained in an alternative medium, the YE-medium. For the newly developed medium, the successful expression of enhanced green fluorescent protein and the adaptation of the cultivation from the agitated culture to the bioreactor could be shown. Furthermore, an analytical method for detection of the essential, organic iron source hemin was established. The consumption of hemin was monitored because hemin is a potentially important process parameter for bioprocess control. With knowledge of these results, an improved expression system is available as an alternative to commonly used cell cultures for the production of recombinant proteins.
Fritsche
Claudia
C
Department of Medical Engineering and Biotechnology, University of Applied Sciences Jena, Jena, Germany. claudia.fritsche@fh-jena.de
Sitz
Mandy
M
Weiland
Norman
N
Breitling
Reinhard
R
Pohl
Hans-Dieter
HD
D000818:Animals; D019149:Bioreactors; D016895:Culture Media, Serum-Free; D049452:Green Fluorescent Proteins / Q000096:biosynthesis*; D006427:Hemin / Q000378:metabolism; D007891:Leishmania / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis*
D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D006427:Hemin