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Isolation and characterization of a new bacteriochlorophyll-c bearing a neopentyl substituent at the 8-position from the bciD-deletion mutant of the brown-colored green sulfur bacterium Chlorobaculum limnaeum

Photosynthesis Research volume 121, pages 3–12 (2014)Cite this article

Abstract

We recently constructed the mutant of the brown-colored green sulfur bacterium Chlorobaculum limnaeum lacking BciD which was responsible for formation of a formyl group at the 7-position in bacteriochlorophyll(BChl)-e biosynthesis. This mutant exclusively gave BChl-c, but not BChl-e, as the chlorosome pigments (Harada et al. in PLoS One 8(4):e60026, 2013). By the mutation, the homolog and epimer composition of the pigment was drastically altered. The methylation at the 82-position in the mutant cells proceeded to create BChl-c carrying large alkyl substituents at this position. Correspondingly, the content of BChls-c having the (S)-configuration at the chiral 31-position remarkably increased and accounted for 80.6 % of the total BChl-c. Based on the alteration of the pigment composition in the mutant cells, a new BChl-c bearing the bulkiest, triple 82-methylated neopentyl substituent at the 8-position ([N,E]BChl-c) was identified. The molecular structure of [N,E]BChl-c was fully determined by its NMR, mass, and circular dichroism spectra. The newly identified [N,E]BChl-c was epimerically pure at the chiral 31-position and its stereochemistry was determined to be an (S)-configuration by modified Mosher’s method. Further, the effects of the C82-methylation on the optical absorption properties of monomeric BChls-c were investigated. The Soret but not Qy absorption bands shifted to longer wavelengths by the extra methylation (at most 1.4 nm). The C82-methylation induced a slight but apparent effect on absorption properties of BChls-c in their monomeric states.

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Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Scheme 2

Abbreviations

APCI:

Atmospheric pressure chemical ionization

BChl:

Bacteriochlorophyll

BPhe:

Bacteriopheophytin

Cba. :

Chlorobaculum

CD:

Circular dichroism

Chls:

Chlorophylls

LCMS:

Liquid chromatography mass spectrometry

MTPA:

α-Methoxy-α-(trifluoromethyl)phenylacetyl

NOE:

Nuclear Overhauser effect

PDA:

Photodiode array

R[E,E]:

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

R[E,M]:

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

R[I,E]:

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

R[P,E]:

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

ROESY:

Rotating frame Overhauser enhancement spectroscopy

S[E,E]:

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

S[I,E]:

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

S[N,E]:

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

S[P,E]:

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

THF:

Tetrahydrofuran

TOF:

Time of flight

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Acknowledgments

We would like to thank Mr. D. Shoutsu, Dr. E. Matsuo, Ms. K. Arakawa, Mr. H. Terada, and Mr. Y. Yasui of Shimadzu Co., Ltd. for their experimental support of ultra-fast HPLC coupled with APCI-mass measurements. This work was partially supported by Grants-in-Aid for Scientific Research (A) (No. 22245030 to HT), (C) (No. 24550065 to TM) and for Young Scientists (B) (No. 24750169 to JH) as well as for Scientific Research on Innovative Areas (“Artificial Photosynthesis (AnApple)”, No. 24107002 to HT) from the Japan Society for the Promotion of Science (JSPS). YT is supported by a PRESTO (Precursory Research for Embryonic Science and Technology) fellowship from the Japan Science and Technology Agency (JST).

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Author notes

  1. Yusuke Tsukatani

    Present address: Earth-Life Science Institute (ELSI), Tokyo Institute of Technology, Meguro-ku, Tokyo , 152-8550, Japan

Affiliations

  1. Graduate School of Life Sciences, Ritsumeikan University, Kusatsu, 525-8577, Japan

    Tadashi Mizoguchi, Yusuke Tsukatani & Hitoshi Tamiaki

  2. Department of Medical Biochemistry, Kurume University School of Medicine, Kurume, Fukuoka, 830-0011, Japan

    Jiro Harada

  3. PRESTO, Japan Science and Technology Agency, Kawaguchi, Saitama, 332-0012, Japan

    Yusuke Tsukatani

Authors
  1. Tadashi Mizoguchi
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  2. Jiro Harada
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  3. Yusuke Tsukatani
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  4. Hitoshi Tamiaki
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Correspondence to Hitoshi Tamiaki.

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Mizoguchi, T., Harada, J., Tsukatani, Y. et al. Isolation and characterization of a new bacteriochlorophyll-c bearing a neopentyl substituent at the 8-position from the bciD-deletion mutant of the brown-colored green sulfur bacterium Chlorobaculum limnaeum . Photosynth Res 121, 3–12 (2014). https://doi.org/10.1007/s11120-014-9977-8

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Keywords

  • Bacteriochlorophyll-c
  • Bacteriochlorophyll-e
  • Methylation
  • Chlorosomes
  • Green sulfur bacteria
  • Ultra-fast HPLC