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Influence of trace elements supplementation on the production of recombinant frutalin by Pichia pastoris KM71H in fed-batch process
Authors:Marcela S O Wanderley  Carla Oliveira  Danyelly Bruneska  Lucília Domingues  José L Lima Filho  José A Teixeira  Solange I Mussatto
Institution:1363. Molecular Prospection and Bioinformatics Group (ProspecMol), Universidade Federal de Pernambuco, 50670-901, Recife, Brazil
2363. Laboratory of Immunopathology Keizo Asami, Universidade Federal de Pernambuco, 50670-901, Recife, Brazil
3363. Centre of Biological Science, Universidade Federal de Pernambuco, 50670-901, Recife, Brazil
4363. Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, 4710-057, Braga, Portugal
Abstract:Frutalin, a galactose-specific lectin used to detect specific tumour markers, is a protein with low expression level in breadfruit. In the present study, fed-batch fermentation in a stirred tank bioreactor was used as a strategy to enhance protein production by a recombinant Pichia pastoris KM71H. By using this process, the production of recombinant frutalin was 4-fold higher than the value obtained in shaker flasks batch assays. Supplementation of the fermentation medium with trace elements (Pichia trace minerals, PTM) was also evaluated in order to stimulate production of the recombinant protein. The addition of PTM to the minimum medium afforded a recombinant protein production of 13.4 mg L?1, which was 2.5-fold higher than that achieved from the culture medium without PTM supplementation. These results are significant as the development of strategies to improve the production of recombinant frutalin may broaden its application in cancer diagnosis.
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