- Isolation and Detection of the Chlorophyll Catabolite Hydroxylating Activity from Capsicum annuum Chromoplasts.
Isolation and Detection of the Chlorophyll Catabolite Hydroxylating Activity from Capsicum annuum Chromoplasts.
Bio-protocol (2017-09-20)
Mareike Hauenstein, Stefan Hörtensteiner
PMID34541205
ABSTRACT
Hydroxylation of chlorophyll catabolites at the so-called C32 position ( Hauenstein et al., 2016 ) is commonly found in all plant species analyzed to date. Here we describe an in vitro hydroxylation assay using Capsicum annuum chromoplast membranes as a source of the hydroxylating activity, which converts the substrate epi-pFCC (epi-primary Fluorescent Chlorophyll Catabolite) ( Mühlecker et al., 2000 ) to epi-pFCC-OH.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Ferredoxin-NADP+ Reductase from Spinacia oleracea (spinach), lyophilized powder, ≥15 units/mg solid, secondary activity: ≥10 units/mg solid NADPH diaphorase
Sigma-Aldrich
Glucose-6-phosphate Dehydrogenase from Leuconostoc mesenteroides, recombinant, expressed in E. coli, ammonium sulfate suspension, ≥550 units/mg protein (biuret)
Sigma-Aldrich
Ferredoxin from Spinacia oleracea (spinach), lyophilized powder, Ferredoxin ≥15 wt. %