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Literature summary extracted from
- Expression, purification, and activity analysis of chlorophyllide oxidoreductase and Ni2+-sirohydrochlorin a,c-diamide reductase (2019), Methods Mol. Biol., 1876, 125-140 .
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.3.7.15 | recombinant overexpression of enzymatically active subunit (BchY/BchZ)2 and BchX2 subcomplexes in Escherichia coli strain BL21(DE3), expression of N-terminally His6-tagged BchY, N-terminally thioredoxin/His/S-tagged BchX2, and untagged BchZ under the control of individual promoters. Supplementation with Fe(III)citrate and L-cysteine as sources of iron and sulfur, respectively, as media constituents has significant influence on the in vivo assembly of [4Fe-4S] clusters, growth temperature is 17°C | Roseobacter denitrificans |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.3.7.15 | Fe2+ | in cofactors | Roseobacter denitrificans |  |
| 1.3.7.15 | Mg2+ | required | Roseobacter denitrificans | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.3.7.15 | 3-acetyl-3-devinylchlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + ATP + H2O + 2 H+ | Roseobacter denitrificans | - |
bacteriochlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster + ADP + phosphate | - |
? | |
| 1.3.7.15 | 3-acetyl-3-devinylchlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + ATP + H2O + 2 H+ | Roseobacter denitrificans ATCC 33942 | - |
bacteriochlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster + ADP + phosphate | - |
? | |
| 1.3.7.15 | 8-vinyl chlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | Roseobacter denitrificans | - |
chlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster | - |
? | |
| 1.3.7.15 | 8-vinyl chlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | Roseobacter denitrificans ATCC 33942 | - |
chlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.3.7.15 | Roseobacter denitrificans | P26277 AND Q16DU8 AND Q16DU7 | genes bchX, bchY, and bchZ; Erythrobacter sp. strain OCh 114 | - |
| 1.3.7.15 | Roseobacter denitrificans ATCC 33942 | P26277 AND Q16DU8 AND Q16DU7 | genes bchX, bchY, and bchZ; Erythrobacter sp. strain OCh 114 | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.3.7.15 | recombinant N-terminally His6-tagged BchY and untagged BchZ as a subcomplex, and N-terminally thioredoxin/His/S-tagged BchX2, from Escherichia coli strain BL21(DE3) by nickel affinity chromatography under strict anaerobic conditions (i.e. in an anaerobic chamber), which is essential for the preservation of enzymatic activities. Method evaluation, overview | Roseobacter denitrificans |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 1.3.7.15 | 3-deacetyl-3-vinylbacteriochlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster + ADP + phosphate = chlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + ATP + H2O + 2 H+ | binding of the substrate to (BchY/BchZ)2 is the initial step of COR catalysis, which is followed by formation of the ternary complex between (BchY/BchZ)2 and (BchX)2 | Roseobacter denitrificans |
Renatured (Commentary)
| EC Number | Renatured (Comment) | Organism |
|---|---|---|
| 1.3.7.15 | robust reconstitution of recombinant COR activity is achieved by coupling it to another enzymatic assay using the green pigment, protochlorophyllide, as a substrate of the nitrogenase-like enzyme, dark-operative protochlorophyllide oxidoreductase (DPOR). The DPOR reaction results in the formation of chlorophyllide (Chlide), which then acts as a direct substrate of COR. Reconstitution of DPOR and COR activities requires high concentrations of ATP, as well as a suitable electron donor | Roseobacter denitrificans |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.3.7.15 | 3-acetyl-3-devinylchlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + ATP + H2O + 2 H+ | - |
Roseobacter denitrificans | bacteriochlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster + ADP + phosphate | - |
? | |
| 1.3.7.15 | 3-acetyl-3-devinylchlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + ATP + H2O + 2 H+ | - |
Roseobacter denitrificans ATCC 33942 | bacteriochlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster + ADP + phosphate | - |
? | |
| 1.3.7.15 | 8-vinyl chlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | - |
Roseobacter denitrificans | chlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster | - |
? | |
| 1.3.7.15 | 8-vinyl chlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | the substrate is purified from the Rhodobacter capsulatus strain CB1200 | Roseobacter denitrificans | chlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster | - |
? | |
| 1.3.7.15 | 8-vinyl chlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | - |
Roseobacter denitrificans ATCC 33942 | chlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster | - |
? | |
| 1.3.7.15 | 8-vinyl chlorophyllide a + 2 reduced ferredoxin [iron-sulfur] cluster + 2 H+ | the substrate is purified from the Rhodobacter capsulatus strain CB1200 | Roseobacter denitrificans ATCC 33942 | chlorophyllide a + 2 oxidized ferredoxin [iron-sulfur] cluster | - |
? | |
| 1.3.7.15 | additional information | for the standard DPOR/COR activity assay, a cell-free Escherichia coli extract containing overproduced Chlorobaculum tepidum DPOR subunits, BchN, BchB and BchL, is used in combination with purified recombinant COR subcomplexes, BchX2 and (BchY/BchZ)2. When using the chemical reducing agent, dithionite, instead of the natural electron donor (which might be a ferredoxin), it is essential to conduct control reactions in the absence of BchX2 and (BchY/BchZ)2 to ensure the dependence of the observed COR activity on both protein subcomplexes. The maximum COR activity is only achieved using an optimal ratio between BchX2 and (BchY/BchZ)2. Protein-protein interaction of BchX2 and (BchY/BchZ)2 is investigated in the presence of an analogue of the cosubstrate ATP, MgADP-AlF4-. Assay method evaluation, overview | Roseobacter denitrificans | ? | - |
- |
|
| 1.3.7.15 | additional information | for the standard DPOR/COR activity assay, a cell-free Escherichia coli extract containing overproduced Chlorobaculum tepidum DPOR subunits, BchN, BchB and BchL, is used in combination with purified recombinant COR subcomplexes, BchX2 and (BchY/BchZ)2. When using the chemical reducing agent, dithionite, instead of the natural electron donor (which might be a ferredoxin), it is essential to conduct control reactions in the absence of BchX2 and (BchY/BchZ)2 to ensure the dependence of the observed COR activity on both protein subcomplexes. The maximum COR activity is only achieved using an optimal ratio between BchX2 and (BchY/BchZ)2. Protein-protein interaction of BchX2 and (BchY/BchZ)2 is investigated in the presence of an analogue of the cosubstrate ATP, MgADP-AlF4-. Assay method evaluation, overview | Roseobacter denitrificans ATCC 33942 | ? | - |
- |
|
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.3.7.15 | oligomer | the COR components (BchY/BchZ)2 and BchX2 are organized in a ternary COR complex | Roseobacter denitrificans |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.3.7.15 | BchX | - |
Roseobacter denitrificans |
| 1.3.7.15 | BchY | - |
Roseobacter denitrificans |
| 1.3.7.15 | BchZ | - |
Roseobacter denitrificans |
| 1.3.7.15 | chlorophyllide oxidoreductase | - |
Roseobacter denitrificans |
| 1.3.7.15 | COR | - |
Roseobacter denitrificans |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.3.7.15 | 34 | - |
assay at | Roseobacter denitrificans |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.3.7.15 | 7.5 | - |
assay at | Roseobacter denitrificans |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.3.7.15 | ATP | - |
Roseobacter denitrificans | |
| 1.3.7.15 | Fe-S center | [4Fe-4S] clusters | Roseobacter denitrificans | |
| 1.3.7.15 | Ferredoxin | putatively the natural electron donor | Roseobacter denitrificans | |
| 1.3.7.15 | additional information | sodium dithionite can substitute for ferredoxin as electron donor | Roseobacter denitrificans |
General Information
| EC Number | General Information | Comment | Organism |
|---|---|---|---|
| 1.3.7.15 | metabolism | the biosynthesis of all bacteriochlorophylls involves the two-electron reduction of the C7-C8 double bond of the green pigment chlorophyllide, which is catalyzed by the nitrogenase-like two-component metalloenzyme chlorophyllide oxidoreductase (COR), whereas in all methanogenic microbes, another nitrogenase-like system, CfbC/D, is responsible for the sophisticated six-electron reduction of Ni2+-sirohydrochlorin a,c-diamide in the course of coenzyme F430 biosynthesis | Roseobacter denitrificans |
| 1.3.7.15 | physiological function | the biosynthesis of all bacteriochlorophylls involves the two-electron reduction of the C7-C8 double bond of the green pigment chlorophyllide, which is catalyzed by the nitrogenase-like two-component metalloenzyme chlorophyllide oxidoreductase (COR), whereas in all methanogenic microbes, another nitrogenase-like system, CfbC/D, is responsible for the sophisticated six-electron reduction of Ni2+-sirohydrochlorin a,c-diamide in the course of coenzyme F430 biosynthesis | Roseobacter denitrificans |
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