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

Authors:
ORCiD logo; ; ;
Publication Date:
Sponsoring Org.:
USDOE
OSTI Identifier:
1419702
Grant/Contract Number:
EE0003373
Resource Type:
Journal Article: Published Article
Journal Name:
PLoS ONE
Additional Journal Information:
Journal Volume: 13; Journal Issue: 2; Related Information: CHORUS Timestamp: 2018-02-06 13:37:24; Journal ID: ISSN 1932-6203
Publisher:
Public Library of Science (PLoS)
Country of Publication:
United States
Language:
English

Citation Formats

Molino, João Vitor Dutra, de Carvalho, João Carlos Monteiro, Mayfield, Stephen Patrick, and Min, ed., Xiang Jia. Comparison of secretory signal peptides for heterologous protein expression in microalgae: Expanding the secretion portfolio for Chlamydomonas reinhardtii. United States: N. p., 2018. Web. doi:10.1371/journal.pone.0192433.
Molino, João Vitor Dutra, de Carvalho, João Carlos Monteiro, Mayfield, Stephen Patrick, & Min, ed., Xiang Jia. Comparison of secretory signal peptides for heterologous protein expression in microalgae: Expanding the secretion portfolio for Chlamydomonas reinhardtii. United States. doi:10.1371/journal.pone.0192433.
Molino, João Vitor Dutra, de Carvalho, João Carlos Monteiro, Mayfield, Stephen Patrick, and Min, ed., Xiang Jia. 2018. "Comparison of secretory signal peptides for heterologous protein expression in microalgae: Expanding the secretion portfolio for Chlamydomonas reinhardtii". United States. doi:10.1371/journal.pone.0192433.
@article{osti_1419702,
title = {Comparison of secretory signal peptides for heterologous protein expression in microalgae: Expanding the secretion portfolio for Chlamydomonas reinhardtii},
author = {Molino, João Vitor Dutra and de Carvalho, João Carlos Monteiro and Mayfield, Stephen Patrick and Min, ed., Xiang Jia},
abstractNote = {},
doi = {10.1371/journal.pone.0192433},
journal = {PLoS ONE},
number = 2,
volume = 13,
place = {United States},
year = 2018,
month = 2
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.1371/journal.pone.0192433

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  • In the fission yeast Schizosaccharomyces pombe, there are relatively few signal peptides available and most reports of their activity have not been comparative. Using sequence information from the S. pombe genome database we have identified three putative signal peptides, designated Cpy, Amy and Dpp, and compared their ability to support secretion of green fluorescent protein (GFP). In the comparison we also included the two well-described secretion signals derived from the precursors of, respectively, the Saccharomyces cerevisiae {alpha}-factor and the S. pombe P-factor. The capability of the tested signal peptides to direct secretion of GFP varied greatly. The {alpha}-factor signal didmore » not confer secretion to GFP and all the produced GFP was trapped intracellular. In contrast, the Cpy signal peptide supported efficient secretion of GFP with yields approximating 10 mg/L. We also found that the use of an attenuated version of the S. cerevisiae URA3 marker substantially increases vector copy number and expression yield in fission yeast.« less
  • The effect of external inorganic carbon (C{sub i}) concentrations on protein biosynthesis and carbonic anhydrase (CA) mRNA abundance were examined in the eukaryotic alga Chlamydomonas reinhardtii. Transfer of high CO{sub 2} (5%) grown algae to air levels of CO{sub 2} resulted in the transitory synthesis of two polypeptides of approximately 49,000 and 52,000 daltons as well as prolonged synthesis and accumulation of the 37,000 dalton CA monomer and an unidentified 20,000 dalton polypeptide. The gene coding for carbonic anhydrase was isolated from a genomic expression library and subjected to restriction endonuclease analysis. Southern blot analysis of chromosomal DNA indicates thatmore » only a single copy of the gene is present. The 2.5 kilobase DNA fragment hybridizes specially to a 1.4 kilobase transcript in RNA isolated from air-grown cells and from cells grown on 5% CO{sub 2} that have been exposed to air levels of CO{sub 2}. Maximum mRNA abundance was observed after 1 to 3 hours of exposure to air. Transfer of air-grown cells to a high CO{sub 2} environment resulted in the elimination of the CA transcript after 60 minutes of exposure. Changes in CA transcript abundance in response to external C{sub i} concentrations occurred in the presence or absence of light.« less
  • In this study, the authors have addressed the capacity of the green alga Chlamydomonas reinhardtii to produce metal-binding peptides in response to stress induced by the heavy metals Cd{sup 2+}, Hg{sup 2+}, and Ag{sup +}. Cells cultured in the presence of sublethal concentrations of Cd{sup 2+} synthesized and accumulated oligopeptides consisting solely of glutamic acid, cysteine, and glycine in an average ratio of 3:3:1. Cadmium-induced peptides were isolated in their native form as higher molecular weight peptide-metal complexes with an apparent molecular weight of approximately 6.5 {times} 10{sup 3}. The isolated complex bound cadmium (as evidenced by absorption spectroscopy) andmore » sequestered (with a stoichiometry of 0.7 moles of cadmium per mole of cysteine) up to 70% of the total cadmium found in extracts of cadmium-treated cells. In Hg{sup 2+}-treated cells, the principal thiol-containing compound induced by Hg{sup 2+} ion was glutathione. It is possible that glutathione functions in plant cells (as it does in animal cells) to detoxify heavy metals. Cells treated with Ag{sup +} ions also synthesized a sulfur-containing component with a charge to mass ratio similar to Cd{sup 2+}-induced peptides. But, in contrast to the results obtained using Cd{sup 2+} as an inducer, these molecules did not accumulate to significant levels in Ag{sup +}-treated cells. The presence of physiological concentrations of Cu{sup 2+} in the growth medium blocked the synthesis of the Ag{sup +}-inducible component(s) and rendered cells resistant to the toxic effects of Ag{sup +}, suggesting competition between Cu{sup 2+} and Ag{sup +} ions, possibly at the level of metal uptake.« less