title: Methods for the preparation of chlorophyllide a: an intermediate of the chlorophyll biosynthetic pathway.
abstract: 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 μg of chlorophyllide a per gram of barley leaves. Chlorophyllide a was identified by thin layer chromatography, absorption spectroscopy, and mass spectrometry.
authors.affiliation: Carlsberg Laboratory, 1799 Copenhagen V, Denmark. andrehubertus.mueller@carlsberglab.dk
authors.forename: André H
authors.initials: AH
authors.lastname: Müller
authors.forename: Simon P
authors.initials: SP
authors.lastname: Gough
authors.forename: David W
authors.initials: DW
authors.lastname: Bollivar
authors.forename: Morten
authors.initials: M
authors.lastname: Meldal
authors.forename: Robert D
authors.initials: RD
authors.lastname: Willows
authors.forename: Mats
authors.initials: M
authors.lastname: Hansson
mesh_terms: D000042:Absorption; D001671:Biochemistry / Q000379:methods*; D053898:Biosynthetic Pathways; D002735:Chlorophyllides / Q000138:chemical synthesis* / Q000737:chemistry / Q000302:isolation & purification; D002855:Chromatography, Thin Layer; D003470:Culture Media / Q000737:chemistry; D001467:Hordeum / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D011136:Polysorbates; D016354:Rhodobacter capsulatus / Q000378:metabolism; D013057:Spectrum Analysis
chemical_list: D002735:Chlorophyllides; D003470:Culture Media; D011136:Polysorbates; D008274:Magnesium