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. 2022:671:207-222.
doi: 10.1016/bs.mie.2022.01.012. Epub 2022 Feb 11.

Metalloenzymes involved in carotenoid biosynthesis in plants

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Metalloenzymes involved in carotenoid biosynthesis in plants

Ian Davis et al. Methods Enzymol. 2022.

Abstract

Carotenoids are a family of pigment compounds, a subset of which are precursors for vitamin A biosynthesis. These pigments are derived from isopentenyl pyrophosphate (IPP), with geranylgeranyl diphosphate being the first metabolite unique to carotenoid biosynthesis in plants, algae, fungi, some bacteria, and arthropods. This chapter highlights the metal-dependent enzymes involved in synthesizing carotenoids in plants and the current state of knowledge of their cofactors and mechanisms. Emphasis is given to spectroscopic methods used to characterize metal centers. The recently discovered heme-dependent isomerase Z-ISO is presented as a case study in how to interrogate a metalloenzyme. Use of UV-vis, electron paramagnetic resonance, and magnetic circular dichroism spectroscopies of a metal center at various oxidation states and with external small molecule probes (CN-, CO, and NO) can provide information about the nature of the metal center, the identity of its ligands, and its mechanism of action. Z-ISO is a histidine/cysteine ligated heme-dependent enzyme that is only active in the ferrous state and possesses redox-linked ligand switching. The choice and design of experiments are discussed as well as the conclusions that can be drawn.

Keywords: Carotenoids biosynthesis; EPR; Enzymology; Heme; Iisomerase; Iron; Spectroscopy.

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Figures

Fig. 1
Fig. 1
Metalloenzymes in the carotenoid biosynthetic pathway of higher plants. Dashed lines represent multiple steps.
Fig. 2
Fig. 2
Isomerization reaction catalyzed by Z-ISO.
Fig. 3
Fig. 3
EPR spectra of Z-ISO in the as-isolated, ferric state (A) and after reduction with dithionite and treatment with •NO (B).
Fig. 4
Fig. 4
Orbital diagram for heme and its nitrosyl adduct.

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