Enrichment of fermentation media and optimization of expression conditions for the production of EAK16 peptide as fusions with SUMO
✍ Scribed by Makkapati Satakarni; Apostolis A. Koutinas; Colin Webb; Robin Curtis
- Publisher
- John Wiley and Sons
- Year
- 2009
- Tongue
- English
- Weight
- 271 KB
- Volume
- 102
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
Abstract
EAK~16~ (AEAEAKAKAEAKAEAK) belongs to a novel class of self‐assembling peptides, which is being investigated in research and industry. SUMO belongs to the ubiquitin class of proteins and is a promising fusion partner currently in use. In this study, EAK~16~ peptide fusions with hexa‐histidine tagged SUMO have been constructed using Escherichia coli based pET expression vector. Intracellular expression of the SUMO–EAK~16~ fusion using LB media has been optimized. Low‐cost complex media (fungal autolysates, wheat and gluten hydrolysates) produced via a novel wheat‐based biorefinery have been used as alternative fermentation media to LB. Shake flask cultures using either enriched LB or complex wheat‐derived media containing 2 g/L of glucose resulted in intracellular SUMO–EAK~16~ fusion protein production of approximately 250 mg/L fermentation volume which corresponded to 30–35% of the total bacterial protein expressed being the fusion protein. Fusion protein productivities up to five times higher were achieved when using a bioreactor. Biotechnol. Bioeng. 2009; 102: 725–735. © 2008 Wiley Periodicals, Inc.