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  • 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
摘要

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.

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Sigma-Aldrich
甲醇, suitable for HPLC, ≥99.9%
Sigma-Aldrich
葡萄糖-6-磷酸脱氢酶 来源于肠系膜明串珠菌, recombinant, expressed in E. coli, ammonium sulfate suspension, ≥550 units/mg protein (biuret)
Sigma-Aldrich
铁氧化还原蛋白-NADP + 还原酶 来源于菠菜, lyophilized powder, ≥15 units/mg solid, secondary activity: ≥10 units/mg solid NADPH diaphorase
Sigma-Aldrich
铁氧还蛋白 来源于菠菜, lyophilized powder, Ferredoxin ≥15 wt. %