Abstract
The B race of a green microalga Botryococcus braunii Kützing produces triterpene hydrocarbons that is a promising source for biofuel. In this algal race, precursors of triterpene hydrocarbons are provided from the 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway. The terminal enzyme of this pathway, 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (HDR) is regarded as one of the key enzymes that affect yields of products in terpene biosynthesis. In order to better understand the MEP pathway of the alga, cDNA and genomic clones of HDR were obtained from B. braunii Showa strain. B. braunii HDR (BbHDR) is encoded on a single copy gene including a 1509-bp open reading frame that was intervened by 6 introns. The exon–intron structure of BbHDR genes did not show clear relation to phylogeny, while its amino acid sequence reflected phyla and classes well. BbHDR sequence was distinctive from that of the HDR protein from Escherichia coli in the residues involved in hydrogen-bond network that surrounds substrate. Introduction of BbHDR cDNA into an E. coli HDR deficient mutant resulted in recovery of its auxotrophy. BbHDR expression level was upregulated from the onset of liquid culture to the 24th day after inoculation with a 2.5-fold increase and retained its level in the subsequent period.
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Acknowledgements
Escherichia coli mutant, MG1655 araBCD<>lytB (HDR), was donated from Professor Millicent Masters at University of Edinburgh. We thank to Dr. Tomohisa Kuzuyama at the University of Tokyo, for valuable discussion, and Ms. Ryoko Suzuki-Yaginuma at The University of Tokyo, for technical assistances. This research was supported by JST, CREST.
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Uchida, H., Sumimoto, K., Oki, T. et al. Isolation and characterization of 4-hydroxy-3-methylbut-2-enyl diphosphate reductase gene from Botryococcus braunii, race B. J Plant Res 131, 839–848 (2018). https://doi.org/10.1007/s10265-018-1039-4
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DOI: https://doi.org/10.1007/s10265-018-1039-4