Expression of Brazzein, A Small Sweet-Tasting Protein in Saccharomyces cerevisiae: An Introduction for Production of Sweet Yeasts
| Autor: | Azar Shahpiri, Akram Kazemi-Nasab |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0303 health sciences Expression vector biology Saccharomyces cerevisiae General Medicine Sweetness biology.organism_classification medicine.disease_cause 01 natural sciences Biochemistry 03 medical and health sciences Structural Biology 010608 biotechnology biology.protein medicine Brazzein Food science Heterologous expression Escherichia coli Gene Pentadiplandra 030304 developmental biology |
| Zdroj: | Protein & Peptide Letters. 27:945-952 |
| ISSN: | 0929-8665 |
| DOI: | 10.2174/0929866527666200331134431 |
| Popis: | Background: The replacement of carbohydrate sweeteners with protein sweeteners from plants has attracted the interest of researchers because these proteins don’t trigger the insulin response and are more nutritive for consumption in food. Brazzein (Braz) is a small and heat- stable sweet protein that has been originally derived from African plant Pentadiplandra brazzeana. In the present work the solubility, sweetness and yield of recombinant forms of Braz in two expression hosts, E. coli and S. cerevisiae were comprised. Methods: The codon-optimized gene of Braz was cloned in expression vectors pET28a and pET41a and GPD. The resulted vectors pET28a-Braz and pEt41a-Braz were transformed into Escherichia coli strain Rosetta (DE3) and the vector GPD-Braz was transformd to S. cerevisiae. The expression of Braz in different systems was analyzed by SDS-PAGE and western blotting. Results: The results verified the heterologous expression of Braz in S. cerevisiae carrying GPDBraz. Also the expression of Braz as carboxy-terminal extensions of His-tag and Glutathione-STransferase (GST) were verified in transgenic E. coli containing pET28a-Braz and pET41a-Braz, respectively. Conclusion: Although the yield of GST-Braz was higher than His-Braz and Braz expressed in S. cerevisiae, but the higher solubility, sweetness, safety (GRAS) are important advantages of the use of S. cerevisiae as expression host for production of Braz. Therefore the result of present work opens new insights for providing the new sweet yeasts that can be used as food additives. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|