Engineering-enhanced Protein Secretory Expression in Yeast with Application to Insulin
| Autor: | Per Balschmidt, Ivan Diers, Svend Ludvigsen, Anders Robert Sorensen, Thomas Kjeldsen, Niels C. Kaarsholm |
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| Rok vydání: | 2002 |
| Předmět: |
Models
Molecular chemistry.chemical_classification Insulin medicine.medical_treatment Peptide Saccharomyces cerevisiae Cell Biology Biology Protein Engineering Biochemistry Yeast Protein Structure Tertiary Insulin receptor chemistry.chemical_compound chemistry biology.protein medicine Aromatic amino acids Secretion Heterologous expression Binding site Molecular Biology |
| Zdroj: | Journal of Biological Chemistry. 277:18245-18248 |
| ISSN: | 0021-9258 |
| Popis: | Adaptation to efficient heterologous expression is a prerequisite for recombinant proteins to fulfill their clinical and biotechnological potential. We describe a rational strategy to optimize the secretion efficiency in yeast of an insulin precursor by structure-based engineering of the folding stability. The yield of a fast-acting insulin analogue (Asp(B28)) expressed in yeast was enhanced 5-fold by engineering a specific interaction between an aromatic amino acid in the connecting peptide and a phenol binding site in the hydrophobic core of the molecule. This insulin precursor is characterized by significantly enhanced folding stability. The improved folding properties enhanced the secretion efficiency of the insulin precursor from 10 to 50%. The precursor remains fully in vitro convertible to mature fast-acting insulin. |
| Databáze: | OpenAIRE |
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