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

作者：Muller Andre H^*; Gough Simon P; Bollivar David W; Meldal Morten; Willows Robert D; Hansson Mats

来源：Analytical Biochemistry, 2011, 419(2): 271-276.

DOI：10.1016/j.ab.2011.08.028

• Chlorophyll biosynthesis
• Chlorophyllide a
• Photodynamic therapy
• Chlorophyllase
• Rhodobacter capsulatus

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 mu g of chlorophyllide a per gram of barley leaves. Chlorophyllide a was identified by thin layer chromatography, absorption spectroscopy, and mass spectrometry.

• 出版日期2011-12-15