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Overexpression of DnaJ-Like Chaperone Enhances Carotenoid Synthesis in Chlamydomonas reinhardtii

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Abstract

Production of functional carotenoids using microalgae may facilitate the commercialization of anti-aging nutritional supplements. The green alga Chlamydomonas reinhardtii uses a non-mevalonate (MEP) pathway for isopentenyl diphosphate (IPP) synthesis. Two enzymes thought to play important roles in this MEP pathway to IPP synthesis are 1-deoxy-d-xylulose 5-phosphate synthase (DXS) and reductase (DXR). DnaJ-like chaperone (Orange protein) is thought to support phytoene synthase, a key enzyme in plant carotenoid synthesis. Genes for Orange (OR), DXS, and DXR were overexpressed via nuclear transformation into C. reinhardtii. CDS of OR, DXS, and DXR were amplified and connected with dual promoters of heat-shock protein 70A and ribulose bisphosphate carboxylase small chain 2. Compared with the parental strain, transformant CrOR#2 produced increased lutein and β-carotene (1.9-fold and 1.7-fold per cell, respectively). Transformant CrDXS#1 produced lutein and β-carotene at lower per-cell abundances than those for the parental strain. CrDXR#2 transformant produced lutein and β-carotene at higher per-cell abundances than their parental counterpart; however, these transformants produced lutein and β-carotene at lower per-medium abundances than their parental counterparts. These results suggest that OR protein supports phytoene synthase in C. reinhardtii and that the phytoene synthesis step is rate-limiting in carotenoid synthesis.

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Acknowledgements

The authors would like to thank Prof. T. Sugawara and Assist. Prof. Y. Manabe, Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, for their support in the carotenoid analyses.

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Author notes

  1. Tateki Morikawa,·Yusuke Uraguchi, and·Shohei Sanda contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Marine Environmental Microbiology, Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kyoto, 606-8502, Japan

    Tateki Morikawa, Yusuke Uraguchi, Shohei Sanda, Satoshi Nakagawa & Shigeki Sawayama

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  1. Tateki Morikawa
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  2. Yusuke Uraguchi
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  3. Shohei Sanda
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  4. Satoshi Nakagawa
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  5. Shigeki Sawayama
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Correspondence to Shigeki Sawayama.

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Morikawa, T., Uraguchi, Y., Sanda, S. et al. Overexpression of DnaJ-Like Chaperone Enhances Carotenoid Synthesis in Chlamydomonas reinhardtii . Appl Biochem Biotechnol 184, 80–91 (2018). https://doi.org/10.1007/s12010-017-2521-5

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