Photosynthesis Research

, Volume 41, Issue 1, pp 157–164 | Cite as

Separation of bacteriochlorophyll homologues from green photosynthetic sulfur bacteria by reversed-phase HPLC

  • C. M. Borrego
  • L. J. Garcia-Gil
Group 5: Chlorosomes and Pigments Regular Papers

Abstract

A reversed-phase High Performance Liquid Cromatography (HPLC) method has been developed to accurately separate bacteriochlorophyllsc, d ande homologues in a reasonably short run time of 60 minutes. By using this method, two well-defined groups of bacteriochlorophyll homologue peaks can be discriminated. The first one consists of 4 peaks (min 24 to 30), which corresponds to the four main farnesyl homologues. The second peak subset is formed by a cluster of up to 10 minor peaks (min 33 to 40). These peaks can be related with series of several alcohol esters of the different chlorosome chlorophylls. The number of homologues was, however, quite variable depending on both, the bacteriochlorophyll and the bacterial species. The method hereby described, also provides a good separation of other photosynthetic pigments, either bacterial (Bacteriochlorophylla, chlorobactene, isorenieratene and okenone) or algal ones (Chlorophylla, Pheophytina and β-carotene). A preliminary screening of the homologue composition of several green photosynthetic bacterial species and isolates, has revealed different relative quantitative patterns. These differences seem to be related to physiological aspects rather than to taxonomic ones. The application of the method to the study of natural populations avoids the typical drawbacks on the pigment identification of overlapping eukaryotic and prokaryotic phototrophic microorganisms, giving further information about their physiological status.

Key words

bacterial pigments bacteriochlorophyll homologues chlorobiaceae HPLC pigment analyses 

Abbreviations

BChl

bacteriochlorophyll

Chl

chlorophyll

Chlb

chlorobactene

HPLC

High Performance Liquid Chromatography

IPA

ion pairing agent

Isr

isorenieratene

PDA

photodiode array detector

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Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • C. M. Borrego
    • 1
  • L. J. Garcia-Gil
    • 1
  1. 1.Laboratory of Microbiology, Deptartment of Biology and Institute of Aquatic EcologyUniversity of Girona, Hospital 6GironaSpain

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