Enrichment of fermentation media and optimization of expression conditions for the production of EAK(16) peptide as fusions with SUMO |
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Authors: | Satakarni Makkapati Koutinas Apostolis A Webb Colin Curtis Robin |
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Institution: | School of Chemical Engineering and Analytical Science, The University of Manchester, PO Box 88, Manchester M601QD, United Kingdom. |
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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. |
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Keywords: | SUMO fusion protein wheat‐based biorefinery |
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