Expression of Histophilus somni IbpA DR2 protective antigen in the diatom Thalassiosira pseudonana

TitleExpression of Histophilus somni IbpA DR2 protective antigen in the diatom Thalassiosira pseudonana
Publication TypeJournal Article
Year of Publication2017
AuthorsDavis A., Crum L.T, Corbeil L.B, Hildebrand M
JournalApplied Microbiology and Biotechnology
Volume101
Pagination5313-5324
Date Published2017/07
Type of ArticleArticle
ISBN Number0175-7598
Accession NumberWOS:000404146300009
Keywordscarbon; chlamydomonas-reinhardtii; cylindrotheca-fusiformis; dendritic cells; diatom; Histophilus somni; IbpA DR2; marine diatoms; microalgae; phaeodactylum-tricornutum; Protective antigen; recombinant proteins; silicon transporters; stable nuclear transformation; Thalassiosira pseudonana; vaccine delivery; Valuable proteins
Abstract

Increasing demand for the low-cost production of valuable proteins has stimulated development of novel expression systems. Many challenges faced by existing technology may be overcome by using unicellular microalgae as an expression platform due to their ability to be cultivated rapidly, inexpensively, and in large scale. Diatoms are a particularly productive type of unicellular algae showing promise as production organisms. Here, we report the development of an expression system in the diatom Thalassiosira pseudonana by expressing the protective IbpA DR2 antigen from Histophilus somni for the production of a vaccine against bovine respiratory disease. The utilization of diatoms with their typically silicified cell walls permitted development of silicon-responsive transcription elements to induce protein expression. Specifically, we demonstrate that transcription elements from the silicon transporter gene SIT1 are sufficient to drive high levels of IbpA DR2 expression during silicon limitation and growth arrest. These culture conditions eliminate the flux of cellular resources into cell division processes, yet do not limit protein expression. In addition to improving protein expression levels by molecular manipulations, yield was dramatically increased through cultivation enhancement including elevated light and CO2 supplementation. We substantially increased recombinant protein production over starting levels to 1.2% of the total sodium dodecyl sulfate-extractable protein in T. pseudonana, which was sufficient to conduct preliminary immunization trials in mice. Mice exposed to 5 mu g of diatom-expressed DR2 in whole or sonicated cells (without protein purification) exhibited a modest immune response without the addition of adjuvant.

DOI10.1007/s00253-017-8267-8
Short TitleAppl. Microbiol. Biotechnol.
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