Simulation of Large-Scale Production of a Soluble Recombinant Protein Expressed in Escherichia coli Using an Intein-Mediated Purification System
| Autor: | Sarah W. Harcum, Shaorong Chong, Shamik S. Sharma |
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| Rok vydání: | 2005 |
| Předmět: |
Reducing agent
Recombinant Fusion Proteins Chitin Bioengineering Biology medicine.disease_cause Applied Microbiology and Biotechnology Biochemistry Inteins law.invention Industrial Microbiology chemistry.chemical_compound Affinity chromatography law Escherichia coli medicine Computer Simulation Tromethamine Molecular Biology Temperature General Medicine Hydrogen-Ion Concentration Fusion protein Molecular Weight carbohydrates (lipids) Dithiothreitol Solubility chemistry Recombinant DNA Target protein Intein Software Biotechnology |
| Zdroj: | Applied Biochemistry and Biotechnology. 126:093-118 |
| ISSN: | 0273-2289 |
| DOI: | 10.1385/abab:126:2:093 |
| Popis: | Inteins are self-cleavable proteins that under reducing conditions can be cleaved from a recombinant target protein. Industrially, an intein-based system could potentially reduce production costs of recombinant proteins by facilitating a highly selective affinity purification using an inexpensive substrate such as chitin. In this study, SuperPro Designer was used to simulate the large-scale recovery of a soluble recombinant protein expressed in Escherichia coli using an intein-mediated purification process based on the commercially available IMPACT system. The intein process was also compared with a conventional process simulated by SuperPro. The intein purification process initially simulated was significantly more expensive than the conventional process, primarily owing to the properties of the chitin resin and high reducing-agent (dithiothreitol [DTT]) raw material cost. The intein process was sensitive to the chitin resin binding capacity, cleavage efficiency of the intein fusion protein, the size of the target protein relative to the intein tag, and DTT costs. An optimized intein purification process considerably reduced costs by simulating an improved chitin resin and alternative reducing agents. Thus, to realize the full potential of intein purification processes, research is needed to improve the properties of chitin resin and to find alternative, inexpensive raw materials. |
| Databáze: | OpenAIRE |
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