Methods for the preparation of chlorophyllide a: an intermediate of the chlorophyll biosynthetic pathway

Summary, in English

Chlorophyllide a is a metabolite late in the biosynthesis of chlorophylls and bacteriochlorophylls. Isolation procedures for chlorophyllide a from Rhodobacter capsulatus CB1200 and barley (Hordeum vulgare L.) are described and compared. R. capsulatus CB1200 is a double mutant in the bacteriochlorophyllide a biosynthetic pathway, and chlorophyllide a is excreted by the cells when grown in Tween 80-containing liquid medium. It was purified by liquid or solid phase extraction, yielding 7 mg of chlorophyllide a from 1 L of culture. In a second approach, intrinsic chlorophyllase activity was used to dephytylate chlorophyll in an acetonic preparation of leaves of wild-type or chlorophyll b-deficient barley. Purification was achieved by liquid phase extraction, yielding 14 mug of chlorophyllide a per gram of barley leaves. Chlorophyllide a was identified by thin layer chromatography, absorption spectroscopy, and mass spectrometry.

• Author: A. H. Muller; S. P. Gough; D. W. Bollivar; M. Meldal; R. D. Willows; Mats Hansson
• Publishing year: 2011
• Language: English
• Pages: 271-276
• Publication/Series: Analytical Biochemistry
• Volume: 419
• Issue: 2
• Links: http://www.ncbi.nlm.nih.gov/pubmed/21925479; http://dx.doi.org/10.1016/j.ab.2011.08.028
• Document type: Journal article
• Publisher: Elsevier
• Topic: Biological Sciences
• Keywords: Hordeum/metabolism; Culture Media/chemistry; Thin Layer; Chromatography; Chlorophyllides/*chemical synthesis/chemistry/isolation & purification; *Biosynthetic Pathways; Absorption; Biochemistry/*methods; Magnesium/metabolism; Polysorbates; Rhodobacter capsulatus/metabolism; Spectrum Analysis
• Status: Published
• ISBN/ISSN/Other: ISSN: 1096-0309