Journal Article

Molecular Design of Photosynthesis-Elevated Chloroplasts for Mass Accumulation of a Foreign Protein

Yukinori Yabuta, Masahiro Tamoi, Kumiko Yamamoto, Ken-ichi Tomizawa, Akiho Yokota and Shigeru Shigeoka

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 49, issue 3, pages 375-385
Published in print March 2008 | ISSN: 0032-0781
Published online March 2008 | e-ISSN: 1471-9053 | DOI: https://dx.doi.org/10.1093/pcp/pcn014
Molecular Design of Photosynthesis-Elevated Chloroplasts for Mass Accumulation of a Foreign Protein

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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In order to increase production of a useful protein by the chloroplast transformation technique, it seems to be necessary to determine the upper limit for the accumulation of a biologically active foreign protein in chloroplasts and then improve photosynthetic capacity and plant productivity. Here we show that the stromal fractions of tobacco chloroplasts could accommodate an additional 200–260 mg ml−1 of green fluorescent protein in the stroma without any inhibition of gas exchange under various light intensity and growth conditions. The minimum amount of fructose-1,6-/sedoheptulose-1,7-bisphosphatase (FBP/SBPase) limiting photosynthesis was then calculated. Analyses of the photosynthetic parameters and the metabolites of transformants into which FBP/SBPase was introduced with various types of promoter (PpsbA, Prrn, Prps2 and Prps12) indicated that a 2- to 3-fold increase in levels of FBPase and SBPase activity is sufficient to increase the final amount of dry matter by up to 1.8-fold relative to the wild-type plants. Their increases were equivalent to an increase of <1 mg ml−1 of the FBP/SBPase protein in chloroplasts and were calculated to represent <1% of the protein accumulated via chloroplast transformation. Consequently, >99% of the additional 200–260 mg ml−1 of protein expressed in the chloroplasts could be used for the production of useful proteins in the photosynthesis-elevated transplastomic plants having FBP/SBPase.

Keywords: Fructose-1,6-/sedoheptulose-1,7-bisphosphatase; Calvin cycle; Photosynthesis; Plastid transformation

Journal Article.  5699 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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