Selenocystamine improves protein accumulation in chloroplasts of eukaryotic green algae.

摘要: Eukaryotic green algae have become an increasingly popular platform for recombinant proteins production. In particular, Chlamydomonas reinhardtii, has garnered increased attention having the necessary biochemical machinery to produce vaccines, human antibodies and next generation cancer targeting immunotoxins. While it been shown that chloroplasts contain chaperones, peptidyl prolylisomerases protein disulfide isomerases facilitate these complex folding assembly, little done determine which processes serve as rate-limiting steps accumulation. other expression systems, Escherichia coli, Chinese hamster ovary cells, insect accumulation can be hampered by cell’s inability fold target polypeptide into native state, resulting in aggregation degradation. To if chloroplasts’ ability oxidize require bonds a stable conformation is step of accumulation, three strains, each expressing different protein, were analyzed. These included fluorescent GFP, reporter containing no bonds; Gaussia princeps luciferase, luminescent immunotoxin, antibody-fusion bonds. Each strain was analyzed its accumulate when supplemented with selenocystamine, small molecule capable catalyzing formation Selenocystamine supplementation led increase luciferase immunotoxin but not GFP results demonstrated selenocystamine suggests chloroplast structure.