Skip to main content
SpringerLink
Log in

In vitro enzymatic assays of photosynthetic bacterial 3-vinyl hydratases for bacteriochlorophyll biosyntheses

  • Original Article
  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

A chlorosome is a large and efficient light-harvesting antenna system found in some photosynthetic bacteria. This system comprises self-aggregates of bacteriochlorophyll (BChl) c, d, or e possessing a chiral 1-hydroxyethyl group at the 3-position, which plays a key role in the formation of the supramolecule. Biosynthesis of chlorosomal pigments involves stereoselective conversion of 3-vinyl group to 3-(1-hydroxyethyl) group facilitated by a 3-vinyl hydratase. This 3-vinyl hydration also occurs in BChl a biosynthesis, followed by oxidation that introduces an acetyl group at the 3-position. Herein, we present in vitro enzymatic assays of paralogous 3-vinyl hydratases derived from green sulfur bacteria, Chlorobaculum tepidum and Chlorobaculum limnaeum, the filamentous anoxygenic phototroph Chloroflexus aurantiacus, and the chloracidobacterium Chloracidobacterium thermophilum. All the hydratases showed hydration activities. The biosynthetic pathway of BChl a and other chlorosomal pigments is discussed considering the substrate specificity and stereoselectivity of the present hydratases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

Abbreviations

BChl:

Bacteriochlorophyll

BChlide:

Bacteriochlorophyllide

BPheide:

Bacteriopheophorbide

Cba.:

Chlorobaculum

Chl:

Chlorophyll

Chlide:

Chlorophyllide

Cfx.:

Chloroflexus

Cra.:

Chloracidobacterium

DV:

3,8-Divinyl

3HE:

3-(1-Hydroxyethyl)

[E,E]:

8,12-Diethyl

[E,M]:

8-Ethyl-12-methyl

[I,E]:

8-Isobutyl-12-ethyl

[P,E]:

8-Propyl-12-ethyl

3V:

3-Vinyl

8V:

8-Vinyl

References

  • Blankenship RE (2014) Molecular mechanism of photosynthesis, 2nd edn. Wiley, Hoboken, pp 1–9

    Google Scholar 

  • Blankenship RE, Matsuura K (2003) Antenna complexes from green photosynthetic bacteria. In: Green BR, Parson WW (eds) Light-harvesting antennas in photosynthesis. Kluwer Academic Publishers, Dordrecht, pp 195–217

    Chapter  Google Scholar 

  • Bollivar DW, Suzuki JY, Beatty JT, Dobrowolski JM, Bauer CE (1994) Directed mutational analysis of bacteriochlorophyll a biosynthesis in Rhodobacter capsulatus. J Mol Biol 237:622–640

    Article  CAS  PubMed  Google Scholar 

  • Bryant DA, Costas AMG, Maresca JA, Chew AGM, Klatt CG, Bateson MM, Tallon LJ, Hostetler J, Nelson WC, Heidelberg JF, Ward DM (2007) Candidatus Chloracidobacterium thermophilum: an aerobic phototrophic acidobacterium. Science 317:523–526

    Article  CAS  PubMed  Google Scholar 

  • Chew AGM, Frigaard N-U, Bryant DA (2004) Identification of BchV, a C-31 hydratase specific for hypermethylated bacteriochlorophyll c in Chlorobaculum tepidum. In: van der Est A, Bruce D (eds) Photosynthesis: fundamental aspects to global perspectives research. Allen Press, Lawrence, pp 875–877

    Google Scholar 

  • Costas AMG, Amaya M, Liu Z, Tomsho LP, Schuster SC, Ward DM, Bryant DA (2012a) Complete genome of Candidatus Chloracidobacterium thermophilum, a chlorophyll-based photoheterotroph belonging to the phylum Acidobacteria. Environ Microbiol 14:177–190

    Article  CAS  Google Scholar 

  • Costas AMG, Tsukatani Y, Rijpstra WIC, Schouten S, Welander PV, Summons RE, Bryant DA (2012b) Identification of the bacteriochlorophylls, carotenoids, quinones, lipids, and hopanoids of “Candidatus Chloracidobacterium thermophilum”. J Bacteriol 194:1158–1168

    Article  CAS  Google Scholar 

  • Fages F, Griebenow N, Griebenow K, Holzwarth AR, Schaffner K (1990) Characterization of light-harvesting pigments of Chloroflexus aurantiacus. Two new chlorophylls: oleyl (octadec-9-enyl) and cetyl (hexadecanyl) bacteriochlorophyllides-c. J Chem Soc Perkin Trans 1:2791–2797

    Article  Google Scholar 

  • Fajer J (2004) Chlorophyll chemistry before and after crystals of photosynthetic reaction centers. Photosynth Res 80:165–172

    Article  CAS  PubMed  Google Scholar 

  • Frigaard N-U, Chew AGM, Li H, Maresca JA, Bryant DA (2003) Chlorobium tepidum: insights into the structure, physiology, and metabolism of a green sulfur bacterium derived from the complete genome sequence. Photosynth Res 78:93–117

    Article  CAS  PubMed  Google Scholar 

  • Ganapathy S, Oostergetel GT, Wawrzyniak PK, Reus M, Chew AGM, Buda F, Boekema EJ, Bryant DA, Holzwarth AR, de Groot HJ (2009) Alternating syn-anti bacteriochlorophylls form concentric helical nanotubes in chlorosomes. Proc Natl Acad Sci USA 106:8525–8530

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Harada J, Saga Y, Yaeda Y, Oh-oka H, Tamiaki H (2005) In vitro activity of C-20 methyltransferase, BchU, involved in bacteriochlorophyll c biosynthesis pathway in green sulfur bacteria. FEBS Lett 579:1983–1987

    Article  CAS  PubMed  Google Scholar 

  • Harada J, Mizoguchi T, Satoh S, Tsukatani Y, Yokono M, Noguchi M, Tanaka A, Tamiaki H (2013) Specific gene bciD for C7-methyl oxidation in bacteriochlorophyll e biosynthesis of brown-colored green sulfur bacteria. PLoS ONE 8:e60026. doi:10.1371/journal.pone.0060026

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Harada J, Teramura M, Mizoguchi T, Tsukatani Y, Yamamoto K, Tamiaki H (2015) Stereochemical conversion of C3-vinyl group to 1-hydroxyethyl group in bacteriochlorophyll c by the hydratases BchF and BchV: adaptation of green sulfur bacteria to limited-light environments. Mol Microbiol 98:1184–1198

    Article  CAS  PubMed  Google Scholar 

  • Kunieda M, Mizoguchi T, Tamiaki H (2004) Diastereoselective self-aggregation of synthetic 3-(1-hydroxyethyl)-bacteriopyrochlorophyll-a as a novel photosynthetic antenna model absorbing near the infrared regions. Photochem Photobiol 79:55–61

    Article  CAS  PubMed  Google Scholar 

  • Liu Z, Bryant DA (2011) Identification of gene essential for the first committed step in the biosynthesis of bacteriochlorophyll c. J Biol Chem 286:22393–22402

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Maresca JA, Chew AGM, Ponsatí MR, Frigaard N-U, Ormerod JG, Bryant DA (2004) The bchU gene of Chlorobium tepidum encodes the C-20 methyltransferase in bacteriochlorophyll c biosynthesis. J Bacteriol 186:2558–2566

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Miyatake T, Tamiaki H (2005) Self-aggregates of bacteriochlorophylls-c, d and e in a light-harvesting antenna system of green photosynthetic bacteria: effect of stereochemistry at the chiral 3-(1-hydroxyethyl) group on the supramolecular arrangement of chlorophyllous pigments. J Photochem Photobiol C 6:89–107

    Article  CAS  Google Scholar 

  • Mizoguchi T, Nagai C, Kunieda M, Kimura Y, Okamura A, Tamiaki H (2009) Stereochemical determination of the unique acrylate moiety at the 17-position in chlorophylls-c from a diatom Chaetoceros calcitrans and its effect upon electronic absorption properties. Org Biomol Chem 7:2120–2126

    Article  CAS  PubMed  Google Scholar 

  • Mizoguchi T, Harada J, Tamiaki H (2012) Characterization of chlorophyll pigments in the mutant lacking 8-vinyl reductase of green photosynthetic bacterium Chlorobaculum tepidum. Bioorg Med Chem 20:6803–6810

    Article  CAS  PubMed  Google Scholar 

  • Mizoguchi T, Harada J, Tsukatani Y, Tamiaki H (2014) 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

    Article  CAS  PubMed  Google Scholar 

  • Mizoguchi T, Harada J, Yamamoto K, Tamiaki H (2015) Inactivation of bciD and bchU genes in the green sulfur bacterium Chlorobaculum limnaeum and alteration of photosynthetic pigments in the resultant mutants. J Photochem Photobiol A 313:52–59

    Article  CAS  Google Scholar 

  • Nomata J, Mizoguchi T, Tamiaki H, Fujita Y (2006) A second nitrogenase-like enzyme for bacteriochlorophyll biosynthesis: reconstitution of chlorophyllide a reductase with purified X-protein (BchX) and YZ-protein (BchY-BchZ) from Rhodobacter capsulatus. J Biol Chem 281:15021–15028

    Article  CAS  PubMed  Google Scholar 

  • Oba T, Tamiaki H (1999) Why do chlorosomal chlorophylls lack the C132-methoxycarbonyl moiety? An in vitro model study. Photosynth Res 61:23–31

    Article  CAS  Google Scholar 

  • Olson JM (1998) Chlorophyll organization and function in green photosynthetic bacteria. Photochem Photobiol 67:61–75

    Article  CAS  Google Scholar 

  • Orf GS, Blankenship RE (2013) Chlorosome antenna complexes from green photosynthetic bacteria. Photosynth Res 116:315–331

    Article  CAS  PubMed  Google Scholar 

  • Ryan AA, Senge MO (2015) How green is green chemistry? Chlorophyll as a bioresource from biorefineries and their commercial potential in medicine and photovoltaics. Photochem Photobiol Sci 14:638–660

    Article  CAS  PubMed  Google Scholar 

  • Saga Y, Shibata Y, Itoh S, Tamiaki H (2007) Direct counting of submicrometer-sized photosynthetic apparatus dispersed in medium at cryogenic temperature by confocal laser fluorescence microscopy: estimation of the number of bacteriochlorophyll c in single light-harvesting antenna complexes of green photosynthetic bacteria. J Phys Chem B 111:12605–12609

    Article  CAS  PubMed  Google Scholar 

  • Tamiaki H (1996) Supramolecular structure in extramembraneous antennae of green photosynthetic bacteria. Coord Chem Rev 148:183–197

    Article  CAS  Google Scholar 

  • Tamiaki H, Takeuchi S, Tsudzuki S, Miyatake T, Tanikaga R (1998) Self-aggregation of synthetic zinc chlorins with a chiral 1-hydroxyethyl group as a model for in vivo epimeric bacteriochlorophyll-c and d aggregates. Tetrahedron 54:6699–6718

    Article  CAS  Google Scholar 

  • Tamiaki H, Shibata R, Mizoguchi T (2007) The 17-propionate function of (bacterio)chlorophylls: biological implication of their long esterifying chains in photosynthetic system. Photochem Photobiol 83:152–162

    CAS  PubMed  Google Scholar 

  • Tamiaki H, Machida S, Mizutani K (2012) Modification of 3-substituents in (bacterio)chlorophyll derivatives to prepare 3-ethylated, methylated, and unsubstituted (nickel) pyropheophorbides and their optical properties. J Org Chem 77:4751–4758

    Article  CAS  PubMed  Google Scholar 

  • Tamiaki H, Teramura M, Tsukatani Y (2016) Reduction processes in biosynthesis of chlorophyll molecules: chemical implication of enzymatically regio- and stereoselective hydrogenations in the late stages of their biosynthetic pathway. Bull Chem Soc Jpn 89:161–173

    Article  CAS  Google Scholar 

  • Tang K-H, Barry K, Chertkov O, Dalin E, Han CS, Hauser LJ, Honchak BM, Karbach LE, Land ML, Lapidus A, Larimer FW, Mikhailova N, Pitluck S, Pierson BK, Blankenship RE (2011) Complete genome sequence of the filamentous anoxygenic phototrophic bacterium Chloroflexus aurantiacus. BMC Genomics 12:334. doi:10.1186/1471-2164-12-334

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Teramura M, Harada J, Mizoguchi T, Yamamoto K, Tamiaki H (2016a) In vitro assays of BciC showing C132-demethoxycarbonylase activity requisite for biosynthesis of chlorosomal chlorophyll pigments. Plant Cell Physiol 57:1048–1052

    Article  CAS  PubMed  Google Scholar 

  • Teramura M, Harada J, Tamiaki H (2016b) In vitro stereospecific hydration activities of the 3-vinyl group of chlorophyll derivatives by BchF and BchV enzymes involved in bacteriochlorophyll c biosynthesis of green sulfur bacteria. Photosynth Res 130:33–45

    Article  CAS  PubMed  Google Scholar 

  • Tsukatani Y, Romberger S, Golbeck J, Bryant D (2012) Isolation and characterization of homodimeric type-I reaction center complex from Candidatus Chloracidobacterium thermophilum, an aerobic chlorophototroph. J Biol Chem 287:5720–5732

    Article  CAS  PubMed  Google Scholar 

  • Tsukatani Y, Yamamoto H, Harada J, Yoshitomi T, Nomata J, Kasahara M, Mizoguchi T, Fujita Y, Tamiaki H (2013) An unexpectedly branched biosynthetic pathway for bacteriochlorophyll b capable of absorbing near-infrared light. Sci Rep 3:1217. doi:10.1038/srep01217

    Article  PubMed  PubMed Central  Google Scholar 

  • Wakao N, Yokoi N, Isoyama N, Hiraishi A, Shimada K, Kobayashi M, Kise H, Iwaki M, Itoh S, Takaichi S, Sakurai Y (1996) Discovery of natural photosynthesis using Zn-containing bacteriochlorophyll in an aerobic bacterium Acidiphilium rubrum. Plant Cell Physiol 37:889–893

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Prof. Tadashi Mizoguchi of Ritsumeikan University for his assistance in HPLC analyses. This work was partially supported by JSPS KAKENHI Grant Number JP24107002 in Scientific Research on Innovative Areas “Artificial Photosynthesis (AnApple)” (to HT) and the Sasakawa Scientific Research Grant (to MT) from The Japan Science Society.

Author information

Authors and Affiliations

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

    Misato Teramura & Hitoshi Tamiaki

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

    Jiro Harada

Authors
  1. Misato Teramura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jiro Harada
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hitoshi Tamiaki
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hitoshi Tamiaki.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Teramura, M., Harada, J. & Tamiaki, H. In vitro enzymatic assays of photosynthetic bacterial 3-vinyl hydratases for bacteriochlorophyll biosyntheses. Photosynth Res 135, 319–328 (2018). https://doi.org/10.1007/s11120-017-0415-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11120-017-0415-6

Keywords

Search

Navigation