Chloroplast Biogenesis 84: Solubilization and Partial Purification of Membrane-Bound [4-Vinyl] Chlorophyllide a Reductase from Etiolated Barley Leaves
✍ Scribed by Vladimir L. Kolossov; Constantin A. Rebeiz
- Publisher
- Elsevier Science
- Year
- 2001
- Tongue
- English
- Weight
- 96 KB
- Volume
- 295
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Chlorophyllide a reductase (4VCR) is a key enzyme of the chlorophyll (Chl) biosynthetic pathway. It catalyzes the conversion of divinyl chlorophyllide (Chlide) a to monovinyl Chlide a by reduction of the vinyl group at position 4 of the macrocycle to ethyl. 4VCR is a membrane-bound enzyme, embedded in etioplast and etiochloroplast membranes. A study of the regulation and properties of this enzyme is mandatory for a comprehensive understanding of the biosynthetic heterogeneity of Chl biosynthesis. Solubilization and partial purification of 4VCR are described for the first time. The enzyme was solubilized with 5 mM Chaps and was partially purified by chromatography on DEAE-Sephacel and Cibacron Blue 3GA-1000 agarose. An overall 20-fold purification was achieved. The partially purified enzyme was stable for several months at ؊80°C.