Information on EC 4.4.1.31 - phycoerythrocyanin alpha-cysteine-84 phycoviolobilin lyase/isomerase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
4.4.1.31
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RECOMMENDED NAME
GeneOntology No.
phycoerythrocyanin alpha-cysteine-84 phycoviolobilin lyase/isomerase
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REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
[phycoerythrocyanin alpha-subunit]-Cys84-phycoviolobilin = apo-[phycoerythrocyanin alpha-subunit] + (2R,3E)-phycocyanobilin
show the reaction diagram
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PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
phycoviolobilin biosynthesis
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SYSTEMATIC NAME
IUBMB Comments
[phycoerythrocyanin alpha-subunit]-Cys84-phycoviolobilin:(2R,3E)-phycocyanobilin lyase/isomerase
The enzyme, characterized from the cyanobacteria Nostoc sp. PCC 7120 and Mastigocladus laminosus, catalyses the covalent attachment of the phycobilin chromophore phycocyanobilin to cysteine 84 of the beta subunit of the phycobiliprotein phycoerythrocyanin and its isomerization to phycoviolobilin.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
P35791 and P35798
insertional mutants in pecE and pecF, and an interposon mutant in which a portion of both pecE and pecF is deleted, are constructed. All three types of mutants grew 1.3 times slower than wild-type under limiting light conditions and show a 20% reduction in the phycocyanobilin content of whole cells relative to chlorophyll alpha. Holo-phycoerythrocyanin is missing from the phycobilisomes of all three types of mutants and the level of the phycoerythrocyanin linker polypeptide is reduced relative to the wild-type
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
apo-[phycocyanin alpha-subunit R-PC V] + phycocyanobilin
[phycocyanin alpha-subunit R-PC V]-Cys84-phycoviolobilin
show the reaction diagram
apo-[phycocyanin alpha-subunit R-PC V] + phycoerythrobilin
[phycocyanin alpha-subunit R-PC V]-Cys84-phycourobilin
show the reaction diagram
apo-[phycoerythrocyanin alpha-subunit PecA] + phycocyanobilin
[phycoerythrocyanin alpha-subunit PecA]-Cys84-phycoviolobilin
show the reaction diagram
apo-[phycoerythrocyanin alpha-subunit PecA] + phycoerythrobilin
[phycoerythrocyanin alpha-subunit PecA]-Cys84-phycourobilin
show the reaction diagram
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enzyme catalyzes binding of the phycoerythrobilin at cysteine 84 with concomitant isomerization to phycourobilin
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?
apo-[phycoerythrocyanin alpha-subunit] + (2R,3E)-phycocyanobilin
[phycoerythrocyanin alpha-subunit]-Cys84-phycoviolobilin
show the reaction diagram
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
apo-[phycoerythrocyanin alpha-subunit] + (2R,3E)-phycocyanobilin
[phycoerythrocyanin alpha-subunit]-Cys84-phycoviolobilin
show the reaction diagram
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the enzyme catalyses the covalent attachment of the phycobilin chromophore phycocyanobilin to cysteine 84 of the beta subunit of the phycobiliprotein phycoerythrocyanin and its isomerization to phycoviolobilin
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?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
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less effective in activation (at 5 mM) than Mg2+ or Mn2+
Mg2+
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the enzyme requires Mg2+ or Mn2+, optimum concentration of Mg2+ is 5 mM
Mn2+
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the enzyme requires Mg2+ or Mn2+, optimum concentration of Mn2+ is 3 mM
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Co2+
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5 mM, complete inhibition
Cu2+
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5 mM, complete inhibition
Fe2+
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5 mM, complete inhibition
Mg2+
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25 mM, 38% inhibition
Mn2+
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25 mM, complete inhibition
Ni2+
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5 mM, complete inhibition
potassium phosphate
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the buffer decreases the effect of the activators, Mn2+ or Mg2+, by 20% as compared with Tris/HCl buffer
Zn2+
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5 mM, complete inhibition
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-mercaptoethanol
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2-mercaptoethanol or thiols like such as dithiothreitol are required for the isomerization reaction of the lyase: without, only the phycocyanobilin addition product is formed, but no [phycoerythrocyanin alpha-subunit]-Cys84-phycoviolobilin. Too much 2-mercaptoethanol will cause the loss of chromophore, in a reaction requiring oxygen. When Mg2+ is used as the activator, the optimal concentration of 2-mercaptoethanol is 5 mM, with Mn2+ it is 3 mM
dithiothreitol
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2-mercaptoethanol or thiols like such as dithiothreitol are required for the isomerization reaction of the lyase: without, only the phycocyanobilin addition product is formed, but no [phycoerythrocyanin alpha-subunit]-Cys84-phycoviolobilin
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.012 - 0.016
(2R,3E)-phycocyanobilin
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depending on apo-[phycoerythrocyanin alpha-subunit] (0.0048-0.019 mM), pH and temperature not specified in the publication
0.0024
apo-[phycoerythrocyanin alpha-subunit]
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at 0.014 mM (2R,3E)-phycocyanobilin, pH and temperature not specified in the publication
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0.0059 - 0.06
phycocyanobilin
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000074
(2R,3E)-phycocyanobilin
Mastigocladus laminosus
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at 0.014 mM apo phycoerythrocyanin alpha-subunit, pH and temperature not specified in the publication
0.00001 - 0.00013
apo-[phycoerythrocyanin alpha-subunit]
Mastigocladus laminosus
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depending on apo phycoerythrocyanin alpha-subunit (0.0048-0.019 mM), pH and temperature not specified in the publication
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0.000007 - 0.00015
phycocyanobilin
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0011 - 0.0025
phycocyanobilin
47548
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 8.5
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pH 7.0: about 40% of maximal activity, pH 8.5: about 60% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
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room temperature is recommended in the absence of Triton X-100, because the proteins tend to precipitate at 37C. In the presence of Triton X-100 (1% v/v) the temperature can be increased without precipitation to 37C (Mg2+ as activator), or even to 45C (Mn2+ as activator)
37
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room temperature is recommended in the absence of Triton X-100, because the proteins tend to precipitate at 37C. In the presence of Triton X-100 (1% v/v) the temperature can be increased without precipitation to 37C with Mg2+ as activator
45
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room temperature is recommended in the absence of Triton X-100, because the proteins tend to precipitate at 37C. In the presence of Triton X-100 (1% v/v) the temperature can be increased without precipitation to 45C with Mn2+ as activator
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
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formation of complexes between the subunits, PecE and PecF
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
denaturation in 8 M urea of the individual subunits, PecE or PecF, is largely irreversible. If they are mixed together after dialysing out the urea separately, they show only little activity
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repetitive freezing and thawing should be avoided, as it causes the purified proteins to precipitate
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very stable preparations are obtained by adding an equal volume of glycerol before freezing
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STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 1 year, His-tagged PecE and PecF are stable
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
His-tagged PecE and PecF
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
overexpressed in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C122L
4% of wild-type activity
C161L
64% of wild-type activity
C48Q
78% of wild-type activity
C91L
181% of wild-type activity
additional information