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Production of recombinant proteins in microalgae at pilot greenhouse scale
Author(s) -
Gimpel Javier A.,
Hyun James S.,
Schoepp Nathan G.,
Mayfield Stephen P.
Publication year - 2015
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.25357
Subject(s) - chlamydomonas reinhardtii , biology , recombinant dna , heterologous , photosynthesis , gene , botany , biochemistry , mutant
Recombinant protein production in microalgae chloroplasts can provide correctly folded proteins in significant quantities and potentially inexpensive costs compared to other heterologous protein production platforms. The best results have been achieved by using the psbA promoter and 5′ untranslated region (UTR) to drive the expression of heterologous genes in a psbA ‐deficient, non‐photosynthetic, algal host. Unfortunately, using such a strategy makes the system unviable for large scale cultivation using natural sunlight for photosynthetic growth. In this study we characterized eight different combinations of 5′ regulatory regions and psbA coding sequences for their ability to restore photosynthesis in a psbA ‐deficient Chlamydomonas reinhardtii , while maintaining robust accumulation of a commercially viable recombinant protein driven by the psbA promoter/5′UTR. The recombinant protein corresponded to bovine Milk Amyloid A (MAA), which is present in milk colostrum and could be used to prevent infectious diarrhea in mammals. This approach allowed us to identify photosynthetic strains that achieved constitutive production of MAA when grown photosynthetically in 100 L bags in a greenhouse. Under these conditions, the maximum MAA expression achieved was 1.86% of total protein, which corresponded to 3.28 mg/L of culture medium. Within our knowledge, this is the first report of a recombinant protein being produced this way in microalgae. Biotechnol. Bioeng. 2015;112: 339–345. © 2014 Wiley Periodicals, Inc.