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

Molino, João Vitor Dutra; de Carvalho, João Carlos Monteiro; Mayfield, Stephen Patrick; Min, ed., Xiang Jia

doi:10.1371/journal.pone.0192433

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

Journal Article · Tue Feb 06 00:00:00 EST 2018 · PLoS ONE

DOI:https://doi.org/10.1371/journal.pone.0192433· OSTI ID:1419702

; de Carvalho, João Carlos Monteiro; Mayfield, Stephen Patrick; Min, ed., Xiang Jia

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.

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Research Organization:: Univ. of California, San Diego, CA (United States)

Sponsoring Organization:: USDOE; USDOE Office of Energy Efficiency and Renewable Energy (EERE), Transportation Office. Bioenergy Technologies Office

Grant/Contract Number:: EE0003373; EE0008246

OSTI ID:: 1419702

Alternate ID(s):: OSTI ID: 1499893; OSTI ID: 1822267

Journal Information:: PLoS ONE, Journal Name: PLoS ONE Vol. 13 Journal Issue: 2; ISSN 1932-6203

Publisher:: Public Library of Science (PLoS)Copyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 36 works

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