Information on EC 1.3.7.4 - phytochromobilin:ferredoxin oxidoreductase

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724317

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biliverdin IXalpha + reduced ferredoxin

(3Z)-phytochromobilin + oxidized ferredoxin

5 entries

(3Z)-phytochromobilin + oxidized ferredoxin

biliverdin IXa + reduced ferredoxin

2 entries

(3Z)-phytochromobilin + oxidized ferredoxin

biliverdin IXalpha + reduced ferredoxin

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724317

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biliverdin IXalpha + reduced ferredoxin

(3Z)-phytochromobilin + oxidized ferredoxin

3 entries

Ferredoxin

5 entries

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Fe2+

cofactor ferredoxin

ATP

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stimulates at 1 mM

NADH

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stimulates slightly at 1 mM

NADPH

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stimulates slightly at 1 mM

additional information

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NADPH regenerating system with glucose-6-phosphate and glucose-6-phosphate dehydrogenase stimulates

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0.0012 - 0.0035

reduced ferredoxin

2 entries

additional information

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sub-micromolar Km for biliverdin IXa

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1.67

biliverdin IXa

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above

0.0000004

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in etiolated plastids

0.0000011

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specific activity of plastid protein

7.3

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assay at

8.5

assay at

25

assay at

28

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assay at

oat

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cucumber

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aurea is a mutant of phytochromobilin synthase

Uniprot

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evolution

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synthesis of linear tetrapyrrole chromophores in cyanobacteria, algae, and plants, overview

725252

physiological function

29000

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SDS-PAGE

33000

calculated from amino acid sequence, mature protein

33400

SDS-PAGE

38100

calculated from amino acid sequence, precursor protein with putative N-terminal plastid transit peptide

437767

monomer

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1 * 29000, SDS-PAGE

D116N

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mutant still retains the ability of substrate binding, but with only 1.5% relative activity of wild type protein

D146N

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mutant completely loses catalytic activity and also the ability of biliverdin binding

D256E

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mutant retains only partial activity

E110Q

site-directed mutagenesis, the mutant shows 321.7% of wild-type activity

E187Q

site-directed mutagenesis, the mutant shows 20.3% of wild-type activity

H259Q

site-directed mutagenesis, the mutant shows 123.4% of wild-type activity

K183Q

site-directed mutagenesis, the mutant shows 24.6% of wild-type activity

K255Q

site-directed mutagenesis, the mutant shows 11.7% of wild-type activity

K263Q

site-directed mutagenesis, the mutant shows 25.8% of wild-type activity

N133

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mutant produces only partial activity

R200Q

site-directed mutagenesis, the mutant shows 12.5% of wild-type activity

R200Q/R264Q

site-directed mutagenesis, the mutant shows 11.9% of wild-type activity

R252Q

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mutant loses catalytic activity and the ability of substrate binding

R264Q

site-directed mutagenesis, the mutant shows 18.9% of wild-type activity

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50000fold

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90% homogeneity

HPLC Phenomenex Ultracarb reverse phase column chromatography

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recombinant ZmHy2

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Superdex 200 gel filtration

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2 entries

development of a system to produce phytochromobilin in Escherichia coli by coexpression of heme oxygenase and phytochromobilin:ferredoxin oxidoreductase in a single operon in conjunction with apophytochrome using two compatible plasmids

Escherichia coli

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673636

expressed in Escherichia coli strain BL21

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expression in Nicotiana plumbaginifolia and Escherichia coli

Zea mays

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657069

expression in Pichia pastoris

742513

overexpression in Synechococcus sp. strain PCC 7002, from endogenous plasmid pAQ1 under the control of the Synechocystis sp. strain PCC 6803 cpcBA promoter, leads to overproduction of phytochromobilin, the cells show a phenotype only slightly less pigmented and blue-green than the wild-type, the strain producing phycobiliproteins carrying phytochromobilin grow much more slowly at low light intensity. Transformant colonies in which pcyA is inactivated in the HY2 overexpression background does not develop a chlorotic appearance, and segregation of the mutant and wild-type alleles is rapidly achieved

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725252

recombinant functional expression of HT-HY2 in Escherichia coli with production of phytochromobilin, functional co-expression with cyanobacterial heme oxygenase, and the phycocyanin alpha-subunit, CpcA, from Synechocystis sp. PCC 6803 or Synechococcus sp. PCC 7002, and with the phycocyanin alpha-subunit phycocyanobilin lyase, CpcE/CpcF, or the phycoerythrocyanin alpha-subunit phycocyanobilin isomerizing lyase, PecE/PecF, from Noctoc sp. PCC 7120. Production levels of fluorescent pigments and chromophore analysis, overview

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724317

wild type and mutant allels

biotechnology

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transposon-based directed tagging strategy using maize Ds element generates a wide diversity of tagged and non-tagged alleles that can be used to generate allelic series or deletion of clustered genes

743476

synthesis