Photosynthesis Research

, Volume 121, Issue 1, pp 13–23 | Cite as

Isolation and structural determination of C8-vinyl-bacteriochlorophyll d from the bciA and bchU double mutant of the green sulfur bacterium Chlorobaculum tepidum

  • Jiro HaradaEmail author
  • Tadashi Mizoguchi
  • Kota Nomura
  • Hitoshi TamiakiEmail author
Regular Paper


The mutant lacking enzymes BciA and BchU, that catalyzed reduction of the C8-vinyl group and methylation at the C20 position of bacteriochlorophyll (BChl) c, respectively, in the green sulfur bacterium Chlorobaculum tepidum, were constructed. This mutant accumulated C8-vinyl-BChl d derivatives, and a molecular structure of the major pigment was fully characterized by its NMR, mass, and circular dichroism spectra, as well as by chemical modification: (31 R)-8-vinyl-12-ethyl-(R[V,E])BChl d was confirmed as a new BChl d species in the cells. In vitro chlorosome-like self-aggregates of this pigment were prepared in an aqueous micellar solution, and formed more rapidly than those of (31 R)-8,12-diethyl-(R[E,E])BChl d isolated from the green sulfur bacterium Chlorobaculum parvum NCIB8327d synthesizing BChl d homologs. Their red-shifted Q y absorption bands were almost the same at 761 nm, and the value was larger than those of in vitro self-aggregates of R[E,E]BChl c (737 nm) and R[V,E]BChl c (726 nm), while the monomeric states of the former gave Q y bands at shorter wavelengths than those of the latter. Red shifts by self-aggregation of the two BChl d species were estimated to be 110 nm and much larger than those by BChls c (75 nm for [E,E] and 64 nm for [V,E]).


BchU BciA Bacteriochlorophyll Chlorosome Green sulfur bacteria Supramolecular structure 



Atmospheric pressure chemical ionization






Circular dichroism



Chl aPD

Δ2,6-Phytadienylated Chl a


Chlorophyllide a oxidoreductase



C8V-Chlide a

3,8-Divinyl-chlorophyllide a


Green sulfur bacteria


Liquid chromatography mass spectrometry


Nuclear Overhauser effect


Photodiode array


(31 R)-8,12-Diethyl-


(31 R)-8-Ethyl-12-methyl-


(31 R)-8-Isobutyl-12-ethyl-


Rotating frame Overhauser enhancement spectroscopy


(31 R)-8-Propyl-12-ethyl-


(31 R)-8-Vinyl-12-ethyl-


(31 R)-8-Vinyl-12-methyl-


(31 S)-8-Isobutyl-12-ethyl-


(31 S)-8-Propyl-12-ethyl-


(31 S)-8-Vinyl-12-ethyl-





We thank Prof. Masato Noguchi of Kurume University School of Medicine, and Dr. Yusuke Tsukatani of Ritsumeikan University for their useful discussions. This work was partially supported by Grants-in-Aid for Scientific Research (A) (No. 22245030) (to H. T.) and (C) (No. 24590366) (to T. M.), for Young Scientists (B) (No. 24750169) (to J. H.), and on Innovative Areas “Artificial Photosynthesis (AnApple)” (No. 24107002) (to H. T.) from the Japan Society for the Promotion of Science (JSPS).

Supplementary material

11120_2014_7_MOESM1_ESM.docx (758 kb)
Supplementary material 1 (DOCX 759 kb)


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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  1. 1.Department of Medical BiochemistryKurume University School of MedicineKurumeJapan
  2. 2.Graduate School of Life SciencesRitsumeikan UniversityKusatsuJapan

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