| Texto completo | |
| Autor(es): |
Dutra Molino, Joao Vitor
[1]
;
Monteiro de Carvalho, Joao Carlos
[1]
;
Mayfield, Stephen Patrick
[2]
Número total de Autores: 3
|
| Afiliação do(s) autor(es): | [1] Univ Sao Paulo, Sch Pharmaceut Sci, Dept Biochem & Pharmaceut Technol, Sao Paulo, SP - Brazil
[2] Univ Calif San Diego, Calif Ctr Algae Biotechnol, Div Biol Sci, San Diego, CA 92103 - USA
Número total de Afiliações: 2
|
| Tipo de documento: | Artigo Científico |
| Fonte: | PLoS One; v. 13, n. 2 FEB 6 2018. |
| Citações Web of Science: | 8 |
| Resumo | |
Efficient protein secretion is a desirable trait for any recombinant protein expression system, together with simple, low-cost, and defined media, such as the typical media used for photosynthetic cultures of microalgae. However, low titers of secreted heterologous proteins are usually obtained, even with the most extensively studied microalga Chlamydomonas reinhardtii, preventing their industrial application. In this study, we aimed to expand and evaluate secretory signal peptides (SP) for heterologous protein secretion in C. reinhardtii by comparing previously described SP with untested sequences. We compared the SPs from arylsulfatase 1 and carbonic anhydrase 1, with those of untried SPs from binding protein 1, an ice binding protein, and six sequences identified in silico. We identified over 2000 unique SPs using the SignalP 4.0 software. mCherry fluorescence was used to compare the protein secretion of up to 96 colonies for each construct, non-secretion construct, and parental wild type cc1690 cells. Supernatant fluorescence varied according to the SP used, with a 10-fold difference observed between the highest and lowest secretors. Moreover, two SPs identified in silico secreted the highest amount of mCherry. Our results demonstrate that the SP should be carefully selected and that efficient sequences can be coded in the C. reinhardtii genome. The SPs described here expand the portfolio available for research on heterologous protein secretion and for biomanufacturing applications. (AU) | |
| Processo FAPESP: | 13/18224-2 - Produção de proteína heteróloga em microalga |
| Beneficiário: | João Vitor Dutra Molino |
| Modalidade de apoio: | Bolsas no Brasil - Doutorado |