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Plant and Cell Physiology Advance Access originally published online on July 30, 2009
Plant and Cell Physiology 2009 50(9):1663-1673; doi:10.1093/pcp/pcp111
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Oxygen Sensitivity of a Nitrogenase-like Protochlorophyllide Reductase from the Cyanobacterium Leptolyngbya boryana

Haruki Yamamoto1, Shohei Kurumiya1, Rie Ohashi1 and Yuichi Fujita1,2,*

1Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8601 Japan
2Presto, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama, 332-0012 Japan

*Corresponding author: E-mail, fujita{at}agr.nagoya-u.ac.jp; Fax, +81-52-789-4107.


   Abstract

Dark-operative protochlorophyllide (Pchlide) oxido-reductase (DPOR) is a nitrogenase-like enzyme that catalyzes Pchlide reduction, the penultimate step of chlorophyll a biosynthesis. DPOR is distributed widely among oxygenic phototrophs such as cyanobacteria, green algae and gymnosperms. To determine how DPOR operates in oxygenic photosynthetic cells, we constructed two shuttle vectors for overexpression of Strep-tagged L-protein (ChlL) and Strep-tagged NB-protein (ChlN–ChlB) in Leptolyngbya boryana (formerly Plectonema boryanum) and introduced them into mutants lacking chlL and chlB. Both transformants restored the ability to produce chlorophyll in the dark. The DPOR activity was reconstituted by L-protein and NB-protein purified from the transformants under anaerobic conditions. L-protein activity disappeared within 5 min of exposure to air while NB-protein activity persisted for >30 min in an aerobic condition, indicating that the L-protein of DPOR components is the primary target of oxygen in cyanobacterial cells. These results suggested that the DPOR from an oxygenic photosynthetic organism did not acquire oxygen tolerance during evolution; but that the cyanobacterial cell developed a mechanism to protect DPOR from oxygen.

Keywords: Chlorophyll biosynthesis - Cyanobacteria - Dark-operative protochlorophyllide oxidoreductase - Leptolyn-gbya boryana - Nitrogenase-like enzyme

Abbreviations: Chlide, chlorophyllide a; DPOR, dark-operative protochlorophyllide oxidoreductase; LPOR, light-dependent protochlorophyllide oxidoreductase; Pchlide, protochlorophyllide

(Received June 1, 2009; Accepted July 25, 2009)
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