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Enrichment of fermentation media and optimization of expression conditions for the production of EAK 16 peptide as fusions with SUMO
Author(s) -
Satakarni Makkapati,
Koutinas Apostolis A.,
Webb Colin,
Curtis Robin
Publication year - 2008
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.22114
Subject(s) - biorefinery , fermentation , fusion protein , hydrolysate , escherichia coli , biochemistry , histidine , intracellular , food science , bioreactor , chemistry , biology , recombinant dna , microbiology and biotechnology , amino acid , botany , hydrolysis , gene , biofuel
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.