Phototransformation of monovinyl and divinyl protochlorophyllide by NADPH:Protochlorophyllide oxidoreductase of barley expressed in Escherichia coli

Knaust, Rosemarie; Seyfried, Britta; Schmidt, L. Ph. H.; Schulz, Rüdiger; Senger, Horst

doi:10.1016/1011-1344(93)80146-z

Cited by 20 publications

(19 citation statements)

References 15 publications

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“…These forms of Pchlide were believed to be in ternary complexes with POR and NADPH, whereas ' non-active ' Pchlide, which absorbs and fluoresces maximally at 630 nm, was thought to be free pigment. In contrast, the fluorescence spectra of Pchlide in the presence of NADPH and in itro-translated barley POR [27] or isolated, solubilized wheat POR [28] lack the longer-wavelength peak associated with ' photoactive ' Pchlide. In addition, a pea mutant (Lip 1) which shows light-independent photomorphogenesis due to the lack of phytochrome I [29] also lacks the longer-wavelength forms of Pchlide, but contains the 630 nm form [30].…”

Section: Discussionmentioning

confidence: 87%

Purification and kinetic analysis of pea (Pisum sativum L.) NADPH:protochlorophyllide oxidoreductase expressed as a fusion with maltose-binding protein in Escherichia coli

Martin

Timko

Wilks

1997

Biochemical Journal

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NADPH :protochlorophyllide oxidoreductase (POR) catalyses the light-dependent reduction of protochlorophyllide to chlorophyllide, a key reaction in the chlorophyll biosynthetic pathway. To facilitate structure-function studies, POR from pea (Pisum sati um L.) has been overexpressed in Escherichia coli as a fusion with maltose-binding protein (MBP) at 5-10 % of the total soluble cell protein. The fusion protein (MBP-POR) has been purified to greater than 90 % homogeneity by a two-step affinity-purification procedure. This represents the first successful overexpression and purification of a plant POR. MBP-POR was found to be active, and the kinetic properties were determined using a continuous assay in which the rate of chlorophyllide formation was measured. The V max was 20.6p

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Section: Discussionmentioning

confidence: 87%

Purification and kinetic analysis of pea (Pisum sativum L.) NADPH:protochlorophyllide oxidoreductase expressed as a fusion with maltose-binding protein in Escherichia coli

Martin

Timko

Wilks

1997

Biochemical Journal

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“…strain PCC6803 (SsPOR) (Suzuki and Bauer, 1995b;Heyes et al, 2000) and Thermosynechococcus elongatus BP-1 (TePOR) (McFarlane et al, 2005) respectively. Similarity to the plant LPOR from Hordeum vulgare (barley) (HvPORA) (Knaust et al, 1993), is with 37%/51% identical/similar amino acid positions, somewhat lower. For comparison, SsPOR and TePOR share about 76%/86% identical/similar amino acid positions.…”

Section: Identification Of a Putative Lpor Gene In D Shibae Dfl12 Tmentioning

confidence: 99%

Discovery of the first light‐dependent protochlorophyllide oxidoreductase in anoxygenic phototrophic bacteria

Kaschner

Loeschcke

Krause

et al. 2014

Molecular Microbiology

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SummaryIn all photosynthetic organisms, chlorophylls function as light-absorbing photopigments allowing the efficient harvesting of light energy. Chlorophyll biosynthesis recurs in similar ways in anoxygenic phototrophic proteobacteria as well as oxygenic phototrophic cyanobacteria and plants. Here, the biocatalytic conversion of protochlorophyllide to chlorophyllide is catalysed by evolutionary and structurally distinct protochlorophyllide reductases (PORs) in anoxygenic and oxygenic phototrophs. It is commonly assumed that anoxygenic phototrophs only contain oxygen-sensitive dark-operative PORs (DPORs), which catalyse protochlorophyllide reduction independent of the presence of light. In contrast, oxygenic phototrophs additionally (or exclusively) possess oxygen-insensitive but light-dependent PORs (LPORs). Based on this observation it was suggested that light-dependent protochlorophyllide reduction first emerged as a consequence of increased atmospheric oxygen levels caused by oxygenic photosynthesis in cyanobacteria. Here, we provide experimental evidence for the presence of an LPOR in the anoxygenic phototrophic α-proteobacterium Dinoroseobacter shibae DFL12 T . In vitro and in vivo functional assays unequivocally prove light-dependent protochlorophyllide reduction by this enzyme and reveal that LPORs are not restricted to cyanobacteria and plants. Sequence-based phylogenetic analyses reconcile our findings with current hypotheses about the evolution of LPORs by suggesting that the lightdependent enzyme of D. shibae DFL12 T might have been obtained from cyanobacteria by horizontal gene transfer.

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“…After expression of the plant cDNA genes in E. coli, the extracted enzyme was able to reduce the substrate protochlorophyllide only in the presence of light and NADPH (Schulz et al, 1989;Benli et al, 1991). Monovinyl and divinyl protochlorophyllide are equally acceptable as substrates (Knaust et al, 1993). Light-dependent protochlorophyllide reductases from barley, oats, wheat, pea, and Arabidopsis are 80 to 95% identical at the amino acid sequence leve1 (Suzuki and Bauer, 1995) but bear no homology to the three peptides of the lightindependent protochlorophyllide reductase.…”

Section: Bacteria-invented Protochlorophyllide Reduction Wlth and Witmentioning

confidence: 99%

Chlorophyll Biosynthesis.

1995

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Phototransformation of monovinyl and divinyl protochlorophyllide by NADPH:Protochlorophyllide oxidoreductase of barley expressed in Escherichia coli

Cited by 20 publications

References 15 publications

Purification and kinetic analysis of pea (Pisum sativum L.) NADPH:protochlorophyllide oxidoreductase expressed as a fusion with maltose-binding protein in Escherichia coli

Purification and kinetic analysis of pea (Pisum sativum L.) NADPH:protochlorophyllide oxidoreductase expressed as a fusion with maltose-binding protein in Escherichia coli

Discovery of the first light‐dependent protochlorophyllide oxidoreductase in anoxygenic phototrophic bacteria

Chlorophyll Biosynthesis.

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