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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Comparison of secretory signal peptides for heterologous protein expression in microalgae: Expanding the secretion portfolio for Chlamydomonas reinhardtii

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Author(s):
Dutra Molino, Joao Vitor [1] ; Monteiro de Carvalho, Joao Carlos [1] ; Mayfield, Stephen Patrick [2]
Total Authors: 3
Affiliation:
[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
Total Affiliations: 2
Document type: Journal article
Source: PLoS One; v. 13, n. 2 FEB 6 2018.
Web of Science Citations: 8
Abstract

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)

FAPESP's process: 13/18224-2 - Heterologous protein production in microalga
Grantee:João Vitor Dutra Molino
Support Opportunities: Scholarships in Brazil - Doctorate