Information on EC 3.1.1.14 - chlorophyllase

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The expected taxonomic range for this enzyme is: Eukaryota

EC NUMBER
COMMENTARY
3.1.1.14
-
RECOMMENDED NAME
GeneOntology No.
chlorophyllase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
chlorophyll + H2O = phytol + chlorophyllide
show the reaction diagram
-
-
-
-
chlorophyll + H2O = phytol + chlorophyllide
show the reaction diagram
also catalyzes chlorophyllide transfer, e.g. converts chlorophyll to methylchlorophyllide
-
chlorophyll + H2O = phytol + chlorophyllide
show the reaction diagram
also catalyzes chlorophyllide transfer, e.g. converts chlorophyll to methylchlorophyllide
-
chlorophyll + H2O = phytol + chlorophyllide
show the reaction diagram
also catalyzes chlorophyllide transfer, e.g. converts chlorophyll to methylchlorophyllide
-
chlorophyll + H2O = phytol + chlorophyllide
show the reaction diagram
also catalyzes chlorophyllide transfer, e.g. converts chlorophyll to methylchlorophyllide
-
chlorophyll + H2O = phytol + chlorophyllide
show the reaction diagram
also catalyzes chlorophyllide transfer, e.g. converts chlorophyll to methylchlorophyllide, catalytic mechanism, catalytic triad is composed of Ser162, Asp191, and His262, and reveals the enzyme as a serine hydrolase
-
chlorophyll + H2O = phytol + chlorophyllide
show the reaction diagram
also catalyzes chlorophyllide transfer, e.g. converts chlorophyll to methylchlorophyllide
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis
-
-
hydrolysis
Musa cavendishii
-
-
hydrolysis
-
-
hydrolysis
-, O22527
-
hydrolysis
Q7Y0K5
-
hydrolysis of carboxylic ester
-
-
-
-
hydrolysis of carboxylic ester
-
-
hydrolysis of carboxylic ester
-
-
hydrolysis of carboxylic ester
-
-
hydrolysis of carboxylic ester
-
-
hydrolysis of carboxylic ester
-
-
hydrolysis of carboxylic ester
-
-
hydrolysis of carboxylic ester
-
-
transesterification
-
-
transesterification
-
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
chlorophyll a degradation I
-
chlorophyll a degradation II
-
chlorophyll a degradation III
-
Metabolic pathways
-
Porphyrin and chlorophyll metabolism
-
SYSTEMATIC NAME
IUBMB Comments
chlorophyll chlorophyllidohydrolase
Chlorophyllase has been found in higher plants, diatoms, and in the green algae Chlorella [3]. This enzyme forms part of the chlorophyll degradation pathway and is thought to take part in de-greening processes such as fruit ripening, leaf senescence and flowering, as well as in the turnover and homeostasis of chlorophyll [4]. This enzyme acts preferentially on chlorophyll a but will also accept chlorophyll b and pheophytins as substrates [5]. Ethylene and methyl jasmonate, which are known to accelerate senescence in many species, can enhance the activity of the hormone-inducible form of this enzyme [5].
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
AtCLH1
O22527
-
Chase 1
O22527
-
Chl-degrading enzyme
-
-
Chlase
-
-
-
-
Chlase
Q8GTM2, Q8GTM3, Q8GTM4
-
Chlase
-
-
Chlase
-
-
Chlase
B6DX58
-
Chlase
Q9MV14
-
Chlase
Q7Y0K5
-
Chlase
-
-
Chlase2
Q9M7I7
-
chlorophyll chlorophyllido-hydrolase
-
-
chlorophyll chlorophyllido-hydrolyase
-
-
-
-
chlorophyll chlorophyllidohydrolase
Q8GTM2, Q8GTM3, Q8GTM4
-
chlorophyll-chlorophyllide hydrolase
Q7Y0K5
-
chlorophyll-chlorophyllide hydrolase
-
-
chlorophyll-chlorophyllido hydrolase
O22527
-
chlorophyll-chlorophyllido-hydrolase
-
-
chlorophyll-chlorophyllidohydrolase
-
-
chlorophyllase
O22527
-
chlorophyllase
Q7Y0K5
-
chlorophyllase
-
-
chlorophyllase
Musa cavendishii
-
-
chlorophyllase
-
-
chlorophyllase
-
-
Chlorophyllase-1
O22527
-
CLH
Q8GTM3, Q8GTM4
-
CLH1
Q8GTM4
-
CLH1
Q8GTM4
isoform
CLH2
Q8GTM3
-
CLH2
Q8GTM3
isoform
CLH3
Q8GTM2
-
CLH3
Q8GTM2
isoform
coronatine-induced protein
-
-
additional information
-
the enzyme belongs to the alpha/beta-hydrolase fold family of enzymes
CAS REGISTRY NUMBER
COMMENTARY
9025-96-1
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
ecotype Columbia Col-0, gene ATHCOR1
-
-
Manually annotated by BRENDA team
isozyme AtCLH1; isozyme AtCLH1
SwissProt
Manually annotated by BRENDA team
isozyme AtCLH2; ecotype Columbia, isozyme AtCLH2
SwissProt
Manually annotated by BRENDA team
isozyme AtCLH2; isozyme AtCLH2
SwissProt
Manually annotated by BRENDA team
wild-type and ethylene-insensitive mutant eti5
-
-
Manually annotated by BRENDA team
var. saccharifera
-
-
Manually annotated by BRENDA team
CLH1; var. italica, gene CLH1
SwissProt
Manually annotated by BRENDA team
CLH1; var. italica, gene CLH3
SwissProt
Manually annotated by BRENDA team
CLH2; var. italica, gene CLH2
SwissProt
Manually annotated by BRENDA team
cultivar Italica Group Ryokutei
-
-
Manually annotated by BRENDA team
isoform CLH1
SwissProt
Manually annotated by BRENDA team
isoform CLH1; cultivar Green king
SwissProt
Manually annotated by BRENDA team
isoform CLH2
SwissProt
Manually annotated by BRENDA team
isoform CLH2; cultivar Green king
SwissProt
Manually annotated by BRENDA team
isoform CLH3; cultivar Green king
SwissProt
Manually annotated by BRENDA team
Italica group
-
-
Manually annotated by BRENDA team
cv. Agridulce
-
-
Manually annotated by BRENDA team
gene clh
SwissProt
Manually annotated by BRENDA team
chlorophyllase enzyme activity in the peel of ripe fruit is about 2.8fold higher than in unripe fruit suggesting that it may play a role in degreening of fruit; Var. Dasheheri
-
-
Manually annotated by BRENDA team
L. China tree
-
-
Manually annotated by BRENDA team
Musa cavendishii
Cavendish banana
-
-
Manually annotated by BRENDA team
3 varieties: Hojiblance, Picual, and Arbequina
-
-
Manually annotated by BRENDA team
pachira chestnut
-
-
Manually annotated by BRENDA team
Bacillariphyceae
-
-
Manually annotated by BRENDA team
var. contender
-
-
Manually annotated by BRENDA team
female and male landraces
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
physiological function
-
type II chlorophyllase takes part in chlorophyll degradation in stored broccoli florets
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
13-OH-chlorophyll a + H2O
phytol + 13-OH-chlorophyllide a
show the reaction diagram
-
in varieties Hojiblanca and Picual
-
?
4-nitrophenyl butyrate + H2O
4-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl decanoate + H2O
4-nitrophenol + decanoate
show the reaction diagram
-
-
-
-
?
bacteriochlorophyll + H2O
?
show the reaction diagram
-
-
-
-
ir
bacteriochlorophyll + H2O
?
show the reaction diagram
-
-
-
-
?
bacteriochlorophyll + H2O
?
show the reaction diagram
-
hydrolyzes bacteriochlorophyll isolated from Rhodospirillum rubrum, but not the pigment bound to the membrane of chromatophores or spheroplasts from the bacterium. Acetone enables the enzyme to hydrolyze the bound pigment
-
-
-
bacteriochlorophyll + H2o
bacteriochlorophyllide + phytol
show the reaction diagram
-
-
-
-
?
bacteriochlorophyll a + H2O
bacteriochlorophyllide a + phytol
show the reaction diagram
-
-
-
?
bacteriochlorophyll a + Triton X-100 + H2O
bacteriochlorophyllide a tritonyl ester
show the reaction diagram
-
-
-
?
chlorobium chlorophyll + H2O
?
show the reaction diagram
-
-
-
-
ir
chlorophyll + H2O
chlorophyllide + ?
show the reaction diagram
-
-
-
?
chlorophyll + H2O
chlorophyllide + ?
show the reaction diagram
-
-
-
-
?
chlorophyll + H2O
chlorophyllide + ?
show the reaction diagram
-
-
-
-
?
chlorophyll + H2O
chlorophyllide + ?
show the reaction diagram
-
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q7Y0K5
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q8GTM2, Q8GTM3, Q8GTM4
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q8GTM3, Q8GTM4
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q9M7I7
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
B6DX58
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q9MV14
Chlase is a rate-limiting enzyme in chlorophyll catabolism and is posttranslationally regulated, it catalyzes the cleavage of the hydrophobic thylakoid-anchoring phytol chain of chlorophyll from the porphyrin ring, resulting in the product chlorophyllide, which retains the typical green color, chlorophyll catabolic pathway, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
chlorophyllase catalyzes the initial hydrolysis of the phytol moiety from the pigment in the degradation of chlorophyll, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q8GTM2, Q8GTM3, Q8GTM4
chlorophyllase is the first enzyme in the degradation pathway of chlorophyll, resulting in postharvest yellowing in broccoli
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q8GTM2, Q8GTM3, Q8GTM4
chlorophyllase is the first enzyme in the degradation pathway of chlorophyll, resulting in postharvest yellowing in broccoli, CLH1 is expressed during the course of broccoli postharvest senescence
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-, O22527, Q9M7I7
isozyme AtCLH1 is not essential for chlorophyll breakdown during plant senescence, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-, O22527, Q9M7I7
isozyme AtCLH2 is not essential for chlorophyll breakdown during plant senescence, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
no gender-specific changes in chlorophyllase activity is response to low temperature stress, however, male plants show higher chlorophyll a/b ratio than female plants, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q9M7I7
the enzyme is involved in the first step of chlorophyll degradation, molecular regulation of in vivo activities, AtCLH2 might play a distinctive role in chlorophyll catabolism in vivo, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
the first enzyme in the chlorophyll degradation pathway in vivo catalyzes the hydrolysis of chlorophylls and pheophytins into their hydrophilic chromophore moieties chlorophyllides and pheophorbides, respectively
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
chlorophyll from Spinacia oleracea
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-, O22527, Q9M7I7
removal of the lipophilic phytol moiety of chlorophyll
-
-
?
chlorophyll + H2O
chlorophyllide + phytol
show the reaction diagram
-
-
-
-
ir
chlorophyll + H2O
chlorophyllide + phytol
show the reaction diagram
-
-
-
-
ir
chlorophyll a + CH3OH + phytol
methylchlorophyllide + ?
show the reaction diagram
-
-
-
-
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
-
r
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
?
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
-
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
-
ir
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
?
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
?
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
?
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
-
?
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
-
-
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
?
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
-
?
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
-
-
-
?
chlorophyll a + H2O
chlorophyllide a + phytol
show the reaction diagram
-
hydrolyzes the a, i.e.132R form but not the a', i.e.132S form
-
-
?
chlorophyll a + H2O
phytol + chlorophyllide
show the reaction diagram
-
best substrate, stereospecific for the substrate, no activity with the C13-epimer
-
?
chlorophyll a + H2O
phytol + chlorophyllide
show the reaction diagram
Q9MV14
chlorophyll a substrate from Petroselinum sativum leaves
-
-
?
chlorophyll a + H2O
phytol + chlorophyllide a
show the reaction diagram
-
-
-
?
chlorophyll a + H2O
phytol + chlorophyllide a
show the reaction diagram
-
-
-
-
?
chlorophyll a + H2O
phytol + chlorophyllide a
show the reaction diagram
-
-
chlorophyllide a production occurs only in variety Arbequina
?
chlorophyll a + H2O
?
show the reaction diagram
-, Q8GTM2, Q8GTM3, Q8GTM4
-
-
-
?
chlorophyll a + Triton X-100 + H2O
tritonyl ester of chlorophyllide a tritonyl ester
show the reaction diagram
-
-
-
-
chlorophyll a/b + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
?
chlorophyll a/b + H2O
phytol + chlorophyllide
show the reaction diagram
-
specific for, the enzyme is involved in the regulation of chlorophyll a and b content
-
?
chlorophyll aXXX ester + alcohol
chlorophyllide a(alcohol) + phytol
show the reaction diagram
-
-
-
?
chlorophyll b + H2O
chlorophyllide b + phytol
show the reaction diagram
-
-
-
?
chlorophyll b + H2O
chlorophyllide b + phytol
show the reaction diagram
-
-
-
?
chlorophyll b + H2O
chlorophyllide b + phytol
show the reaction diagram
-
-
-
-
?
chlorophyll b + H2O
chlorophyllide b + phytol
show the reaction diagram
-
-
-
-
?
chlorophyll b + H2O
chlorophyllide b + phytol
show the reaction diagram
-
-
-
-
?
chlorophyll b + H2O
chlorophyllide b + phytol
show the reaction diagram
-
hydrolyzes the a, i.e. 132R form but not the a', i.e.132S form
-
-
?
chlorophyll b + H2O
phytol + chlorophyllide
show the reaction diagram
-
stereospecific for the substrate, no activity with the C13-epimer
-
?
chlorophyll b + H2O
?
show the reaction diagram
-, Q8GTM2, Q8GTM3, Q8GTM4
-
-
-
?
p-nitrophenyl butyrate + H2O
p-nitrophenol + butyrate
show the reaction diagram
-
-
-
-
?
p-nitrophenyl decanoate + H2O
p-nitrophenol + decanoate
show the reaction diagram
-
-
-
-
?
p-nitrophenyl palmitate + H2O
p-nitrophenol + palmitate
show the reaction diagram
-
-
-
-
?
pheophorbide a methyl ester + H2O
?
show the reaction diagram
-, Q8GTM2, Q8GTM3, Q8GTM4
-
-
-
?
pheophytin + H2O
?
show the reaction diagram
-
-
-
-
?
pheophytin + H2O
phytol + pheophorbide
show the reaction diagram
-
-
-
?
pheophytin + H2O
pheophorbide + ?
show the reaction diagram
-
-
-
-
?
pheophytin a + H2O
?
show the reaction diagram
-
-
-
-
ir
pheophytin a + H2O
phytol + pheophorbide
show the reaction diagram
-
-
-
?
pheophytin a + H2O
phytol + pheophorbide
show the reaction diagram
-
in varieties Hojiblanca and Picual
-
?
pheophytin a + H2O
pheophorbide a + phytol
show the reaction diagram
-
-
-
-
?
pheophytin b + H2O
?
show the reaction diagram
-
-
-
-
ir
pheophytin b + H2O
phytol + pheophorbide
show the reaction diagram
-
-
-
?
pheophytin b + H2O
? + phytol
show the reaction diagram
-
-
-
-
?
pyrobacteriochlorophyll a + Triton X-100 + H2O
pyrobacteriochlorophyllide a tritonyl ester
show the reaction diagram
-
-
-
?
pyrochlorophyll a + Triton X-100 + H2O
pyrochlorophyllide a tritonyl ester + phytol
show the reaction diagram
-
-
-
?
chlorophyllide a + phytol
chlorophyll a + H2O
show the reaction diagram
-
-
-
r
additional information
?
-
-
-
-
-
-
additional information
?
-
-
immobilized enzyme shows a higher affinity towards chlorophyll than pheophytin as substrate
-
-
-
additional information
?
-
-
the enzyme acts preferentially with compounds having the isocyclic carbomethoxy and C-17 propionic residue facing opposite sides of the pyorphyrin macrocycle
-
-
-
additional information
?
-
-
no activity with 4-nitrophenyl esters of fatty acids and tributyrin
-
?
additional information
?
-
-
no activity with chlorophyll b in all variants
-
?
additional information
?
-
Q9LE89
key enzyme in chlorophyll degradation
-
-
-
additional information
?
-
-
the enzyme participates in the phytyl ester formation in the final step of chlorophyll biosynthesis
-
-
-
additional information
?
-
-
first stage in the enzymic breakdown of chlorophyll in vivo may be the removal of the phytol side chain by chlorophyllase
-
-
-
additional information
?
-
-
autolysis of chlorophyll appears to be brought about by enzymatic activity of chlorophyllase which upon membrane disruption and solubilization obtains access to its chlorophyll substrate
-
-
-
additional information
?
-
-
since chlorophyll degradation is a defining feature of plant senescence, compounds inhibiting chlorophyllase activity may delay senescence, thereby improving shelf life and appearance of plant products
-
-
-
additional information
?
-
-, Q8GTM2, Q8GTM3, Q8GTM4
isoform CLH1 preferably hydrolyzes Mg-free chlorophyll
-
-
-
additional information
?
-
-, Q8GTM2, Q8GTM3, Q8GTM4
isoform CLH2 hydrolyzes both chlorophyll and Mg-free chlorophyll at a similar level
-
-
-
additional information
?
-
-, Q8GTM2, Q8GTM3, Q8GTM4
isoform CLH3 shows very low Chlase activity
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
13-OH-chlorophyll a + H2O
phytol + 13-OH-chlorophyllide a
show the reaction diagram
-
in varieties Hojiblanca and Picual
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q9MV14
Chlase is a rate-limiting enzyme in chlorophyll catabolism and is posttranslationally regulated, it catalyzes the cleavage of the hydrophobic thylakoid-anchoring phytol chain of chlorophyll from the porphyrin ring, resulting in the product chlorophyllide, which retains the typical green color, chlorophyll catabolic pathway, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
chlorophyllase catalyzes the initial hydrolysis of the phytol moiety from the pigment in the degradation of chlorophyll, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q8GTM2, Q8GTM3, Q8GTM4
chlorophyllase is the first enzyme in the degradation pathway of chlorophyll, resulting in postharvest yellowing in broccoli
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q8GTM2, Q8GTM3, Q8GTM4
chlorophyllase is the first enzyme in the degradation pathway of chlorophyll, resulting in postharvest yellowing in broccoli, CLH1 is expressed during the course of broccoli postharvest senescence
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-, O22527, Q9M7I7
isozyme AtCLH1 is not essential for chlorophyll breakdown during plant senescence, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-, O22527, Q9M7I7
isozyme AtCLH2 is not essential for chlorophyll breakdown during plant senescence, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
no gender-specific changes in chlorophyllase activity is response to low temperature stress, however, male plants show higher chlorophyll a/b ratio than female plants, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
Q9M7I7
the enzyme is involved in the first step of chlorophyll degradation, molecular regulation of in vivo activities, AtCLH2 might play a distinctive role in chlorophyll catabolism in vivo, overview
-
-
?
chlorophyll + H2O
phytol + chlorophyllide
show the reaction diagram
-
the first enzyme in the chlorophyll degradation pathway in vivo catalyzes the hydrolysis of chlorophylls and pheophytins into their hydrophilic chromophore moieties chlorophyllides and pheophorbides, respectively
-
-
?
chlorophyll + H2O
chlorophyllide + phytol
show the reaction diagram
-
-
-
-
ir
chlorophyll a + H2O
phytol + chlorophyllide a
show the reaction diagram
-
-
-
?
chlorophyll a + H2O
phytol + chlorophyllide a
show the reaction diagram
-
-
chlorophyllide a production occurs only in variety Arbequina
?
chlorophyll a/b + H2O
phytol + chlorophyllide
show the reaction diagram
-
-
-
?
pheophytin a + H2O
phytol + pheophorbide
show the reaction diagram
-
in varieties Hojiblanca and Picual
-
?
chlorophyll a/b + H2O
phytol + chlorophyllide
show the reaction diagram
-
the enzyme is involved in the regulation of chlorophyll a and b content
-
?
additional information
?
-
-
no activity with chlorophyll b in all variants
-
?
additional information
?
-
Q9LE89
key enzyme in chlorophyll degradation
-
-
-
additional information
?
-
-
the enzyme participates in the phytyl ester formation in the final step of chlorophyll biosynthesis
-
-
-
additional information
?
-
-
first stage in the enzymic breakdown of chlorophyll in vivo may be the removal of the phytol side chain by chlorophyllase
-
-
-
additional information
?
-
-
autolysis of chlorophyll appears to be brought about by enzymatic activity of chlorophyllase which upon membrane disruption and solubilization obtains access to its chlorophyll substrate
-
-
-
additional information
?
-
-
since chlorophyll degradation is a defining feature of plant senescence, compounds inhibiting chlorophyllase activity may delay senescence, thereby improving shelf life and appearance of plant products
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Mg2+
-
optimal concentration of MgCl2 using pheophytin as substrate is 1.8 mM
Mg2+
-
in combination with dithiothreitol Mg2+ can activate chlorophyllase-catalyzed chlorophyll hydrolysis
additional information
-
divalent cations stimulate
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
4-hydroxymercuribenzoate
-
93% inhibition at 1 mM, 24 h preincubation
4-hydroxymercuribenzoate
-
-
acetone
Q9M7I7
activates at lower concentration, Chlase2 reaches its maximal activity in 40% acetone, in 40-60% acetone the activity is reduced, overview
beta-carotene
-
in absence of other lipids, inhibits chlorophyl a conversion
canola oil
-
presence of 10%, 20%, and 30% of canola oil decreased the activity of entrapped chlorophyllase by 60%, 75%, and 80%, respectively
-
Co2+
-
56% inhibition at 0.01 mM, 24 h preincubation
concanavalin A
-
-
-
Cu2+
-
55% inhibition at 0.01 mM, 24 h preincubation
diisopropyl fluorophosphate
-
irreversible, 91% inhibition at 10 mM, 1 h preincubation at pH 7.0 and 30C
diisopropyl fluorophosphate
-
-
diisopropyl fluorophosphate
-
noncompeptitive
diisopropyl fluorophosphate
-
-
Eupergit C
-
immobilization support
-
Eupergit C/EDA
-
immobilization support
-
Fe2+
-
89% inhibition at 0.01 mM, 24 h preincubation
-
Fe3+
-
13% inhibition at 0.01 mM, 24 h preincubation
-
Hg2+
-
85% inhibition at 0.01 mM, 24 h preincubation
iodoacetamide
-
irreversible, 6% inhibition at 10 mM, 1 h preincubation at pH 7.0 and 30C
iodoacetamide
-
-
levulinic acid
-
inhibits the chlorophyll synthesis in greening leaves, alters the chlorophyll a/chlorophyll b ratio
Mg2+
-
12% inhibition at 0.01 mM, 24 h preincubation
Mg2+
-
eliminates the activating effect of lecithin
N-ethylmaleimide
-
13% inhibition at 1 mM, 24 h preincubation
N-ethylmaleimide
-
irreversible, 15.9% inhibition at 10 mM, 1 h preincubation at pH 7.0 and 30C
N-ethylmaleimide
-
-
NaF3(CN)6
-
2 mM, 17% inhibition
-
PCMB
-
irreversible, 92.6% inhibition at 1 mM, 1 h preincubation at pH 7.0 and 30C
phenylmethanesulfonyl fluoride
-
-
pheophytin b
-
substrate inhibition above 0.005 mM
phosphatidyl glycerol
-
and sulfoquinovosyldiacylglycerol, inactivation in a cooperative manner
phytol
-
in methanol, hydrolysis of chlorophyll a
phytol
-
uncompetitive inhibition
PMSF
-
irreversible, 89.2% inhibition at 10 mM, 1 h preincubation at pH 7.0 and 30C
refined bleached deodorized canola oil
-
20%
-
Sulfoquinovosyldiacylglycerol
-
and phosphatidyl glycerol, inactivation in a cooperative manner
Triton X-100
-
strong inhibition at 0.2-1.0%
Triton X-100
-
-
Zn2+
-
60% inhibition at 0.01 mM, 24 h preincubation
Mn2+
-
67% inhibition at 0.01 mM, 24 h preincubation
additional information
-
poor inhibition by methyl methanethiosulfonate and iodoacetamide
-
additional information
-
the expression level and the activity of Chlase decrease significantly with the progress of the leaf-yellowing
-
additional information
-
inhibitory effects on recombinant enzymes purified by microtiter plate method and by affinity chromatography, respectively, overview, no inhibition by iodoacetic acid, iodoacetamide, 2-mercaptoethanol, and DTT
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
-
13% stimulation at 1 mM, 24 h preincubation
acetone
-
the activity of entrapped chlorophyllase increased with increasing acetone concentration from 6.5 to 20%, however, further increase of acetone leads to a decrease in chlorophyllase activity. Free chlorophyllase reaches its highest activity at acetone concentration of 10% in a medium consisting of water, acetone, and canola oil, overview
acetone
Q9M7I7
activates at lower concentration, Chlase2 reaches its maximal activity in 40% acetone, in 40-60% acetone the activity is reduced, overview
acetone
-
30% acetone stimulates hydrolysis of bacteriochlorophyll by 50%, stimulates hydrolysis of chlorophyll a by about 400%
acetone
-
presence of a solvent such as acetone is essential for enzyme action
beta-carotene
-
increases hydrolytic activity
dithiothreitol
-
13% stimulation at 1 mM, 24 h preincubation
dithiothreitol
-
in combination with Mg2+ dithiothreitol can activate chlorophyllase-catalyzed chlorophyll hydrolysis
fulvic acid
-
0.125-0.5 mg/ml
humic acid
-
0.125-0.5 mg/ml
humin
-
0.125-0.5 mg/ml
-
L-alpha-phosphatidyl-DL-glycerol
-
increases hydrolytic activity
L-alpha-phosphatidylcholine
-
increases hydrolytic activity
lecithin
-
enhances hydrolysis of chlorophyll
Lipids
-
no or slight activity in absence of lipids. Activation by mixed spinach chloroplast lipids and also by varying degrees by several single plant lipids, monogalactosyl diacylglycerol, digalactosyl diacylglycerol, phosphatidylglycerol and to a lesser extent, phosphatidylcholine and sulfoquinovosyl diacylglycerol
-
Lipids
-
plant membrane lipids, including phosphatidylserine, phosphatidylglycerol, and beta-carotene increase activity
-
wounding
-
induces enzyme expression from gene ATHCOR1 in leaves
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.05
-
4-nitrophenyl decanoate
-
pH 8.0, 25C, recombinant enzyme purified by affinity chromatography
0.069
-
4-nitrophenyl decanoate
-
pH 8.0, 25C, recombinant enzyme purified by microtiter plate method
0.0334
-
bacteriochlorophyll
-
-
0.004
-
bacteriochlorophyll a
-
pH 7.5, 30C
0.006
-
Chlorophyll
-
soluble enzyme
0.01
-
Chlorophyll
-
immobilized enzyme
0.0123
-
Chlorophyll
-
free enzyme in 20 mM sodium phosphate buffer pH 8
0.0186
-
Chlorophyll
-
free enzyme in PBS-hexane mixture (60/40, v/v) containing 75 mM Span85
0.0189
-
Chlorophyll
-
immobilized enzyme in PBS-hexane mixture (60/40, v/v) containing 75 mM Span85
0.0225
-
Chlorophyll
-
immobilized enzyme in 20 mM sodium phosphate buffer pH 8
0.0245
-
Chlorophyll
-
free enzyme in PBS-hexane mixture (70/30, v/v)
0.0246
-
Chlorophyll
-
immobilized enzyme in PBS-hexane mixture (70/30, v/v)
0.278
-
Chlorophyll
-
-
69
-
Chlorophyll
-
at 25C
0.002
-
chlorophyll a
-
-
0.00265
-
chlorophyll a
-
-
0.00375
-
chlorophyll a
-
-
0.004
-
chlorophyll a
-
type 1 chlorophyllase
0.0046
-
chlorophyll a
-
type 2 chlorophyllase
0.011
-
chlorophyll a
-
pH 7.5, 35C
0.0113
-
chlorophyll a
-
-
0.012
-
chlorophyll a
-
pH 7.5, 30C
0.308
-
chlorophyll a
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40C
0.466
-
chlorophyll a
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40C
0.0031
-
chlorophyll b
-
type 1 chlorophyllase
0.004
-
chlorophyll b
-
pH 7.5, 35C
0.0044
-
chlorophyll b
-
type 2 chlorophyllase
0.011
-
chlorophyll b
-
-
0.311
-
chlorophyll b
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40C
570
-
p-nitrophenyl butyrate
-
final concentration of 10% v/v acetonitrile at 25C
0.52
-
p-nitrophenyl decanoate
-
final concentration of 10% v/v acetonitrile at 37C
0.2
-
p-nitrophenyl palmitate
-
final concentration of 10% v/v acetonitrile at 37C
0.0764
-
pheophorbide a methyl ester
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40C
-
0.143
-
pheophorbide a methyl ester
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40C
-
0.000206
-
pheophytin
-
-
0.005
-
pheophytin
-
soluble enzyme
0.09
-
pheophytin
-
immobilized enzyme
0.0027
-
pheophytin a
-
pH 7.5, 35C
0.0123
-
pheophytin a
-
-
0.015
-
pheophytin a
-
-
0.0055
-
pheophytin b
-
-
0.0064
-
pheophytin b
-
pH 7.5, 35C
0.652
-
chlorophyll b
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40C
additional information
-
additional information
-
steady state kinetics, the recombinant immobilized fusion protein displays kinetic parameters similar to those of recombinant enzyme purified by affnity chromatography, overview
-
additional information
-
additional information
-
steady-state and non-steady-state kinetics of free enzyme and of enzyme trapped in a sol-gel medium, overview
-
additional information
-
additional information
-
-
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
8.05
-
4-nitrophenyl decanoate
-
pH 8.0, 25C, recombinant enzyme purified by affinity chromatography
9.43
-
4-nitrophenyl decanoate
-
pH 8.0, 25C, recombinant enzyme purified by microtiter plate method
566
-
Chlorophyll
-
-
0.0024
-
chlorophyll a
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40C
0.072
-
chlorophyll a
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40C
0.0037
-
chlorophyll b
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40C
0.072
-
chlorophyll b
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40C
10.3
-
p-nitrophenyl butyrate
-
-
0.52
-
p-nitrophenyl decanoate
-
-
0.2
-
p-nitrophenyl palmitate
-
-
0.0039
-
pheophorbide a methyl ester
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40C
-
0.259
-
pheophorbide a methyl ester
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40C
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0078
-
chlorophyll a
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40C
8515
0.1551
-
chlorophyll a
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40C
8515
0.0119
-
chlorophyll b
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40C
32257
0.1105
-
chlorophyll b
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40C
32257
0.0272
-
pheophorbide a methyl ester
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40C
0
3.397
-
pheophorbide a methyl ester
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH1, in 100 mM sodium phosphate, pH 7.4, at 40C
0
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.00072
-
-
etiolated leaf
0.074
-
-, Q8GTM2, Q8GTM3, Q8GTM4
mutant enzyme H247A, in 100 mM sodium phosphate, pH 7.4, at 40C
0.149
-
-, Q8GTM2, Q8GTM3, Q8GTM4
mutant enzyme H239A, in 100mM sodium phosphate, pH 7.4, at 40C
0.372
-
-, Q8GTM2, Q8GTM3, Q8GTM4
mutant enzyme H66A, in 100 mM sodium phosphate, pH 7.4, at 40C
0.48
-
-
-
0.52
-
-, Q8GTM2, Q8GTM3, Q8GTM4
mutant enzyme H226A, in 100mM sodium phosphate, pH 7.4, at 40C
7.43
-
-, Q8GTM2, Q8GTM3, Q8GTM4
wild type isoform CLH2, in 100 mM sodium phosphate, pH 7.4, at 40C
9.79
-
-
enzyme isolated at the exponential growth phase
15
-
-
21 chlorophyllide a at 25C, 10 chlorophyllide b at 25C
35.55
-
-
etioplast
41.3
-
-
chloroplast
134.4
-
-
enzyme isolated at the stationary growth phase
322
-
-
optimization of chlorophyllase-catalyzed hydrolysis of chlorophyll in monophasic organic solvent media, the catalytic efficiency of chlorophyllase in the undiluted organic solvent mixture is lower than that in the aqueous medium, overview
additional information
-
-
enzyme activity at different growth phases and ripening states, overview
additional information
-
-
development of a continuous assay system, replacing chlorophyll with p-nitrophenyl-ester substrates eliminates the extraction step and allows for continuous measurement of chlorophyllase activity in the multiwell plate format, overview
additional information
-
-
comparison of the catalytic performance of purified free enzyme and purified enzyme trapped in a sol-gel medium, overview
additional information
-
-
-
additional information
-
-
-
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6
7.5
-
hydrolysis of bacteriochlorophyll a
6.6
7.6
-
chlorophyll b, chlorophyllase type 1 and type 2
6.6
8.6
-
hydrolysis of chlorophyll a, chlorophyllase type 1 and type 2
7
8.5
-
hydrolysis of chlorophyll a
7
-
-
assay at
7
-
-
assay at
7
-
-
assay at, substrate chlorophyll
7
-
-
assay at
7
-
Q9M7I7
assay at
7
-
-, Q8GTM2, Q8GTM3, Q8GTM4
isoform CLH1
7
-
-
with both phosphate buffer and borate buffer
7.4
-
Q9MV14
assay at
7.5
8
-
soluble enzyme
7.5
-
-
-
7.8
-
-
-
8
-
-
activity and stability decrease significantly above or below pH 8.0
8
-
-
assay at, substrate 4-nitrophenyl esters
8
-
-, O22527, Q9M7I7
assay at; assay at
8
-
-, Q8GTM2, Q8GTM3, Q8GTM4
isoform CLH2
8
-
-
immobilized enzyme
8
-
-
enzyme form FI'
8.5
-
-
enzyme fraction FII' and FIII'
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
6.2
8.1
-
50% of maximal activity at pH 6.2 and 8.1
6.4
8.7
-
pH 6.4: about 70% of maximal activity, pH 8.7: about 80% of maximal activity, soluble enzyme
7
11.5
-
35% of maximal activity at pH 7.0, 60& of maximal activity at pH 11.5
7.5
8.8
-
pH 7.5: about 35% of maximal activity, pH 8.8: about 65% of maximal activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
20
30
-
-
20
-
-
-
25
-
-
assay at
25
-
-
assay at
25
-
-, O22527, Q9M7I7
assay at; assay at
30
-
-
free enzyme and of enzyme trapped in a sol-gel medium
30
-
-, Q8GTM2, Q8GTM3, Q8GTM4
isoform CLH1
35
-
-
assay at
35
-
-
immobilized enzyme
40
-
-
assay at
40
-
Q9M7I7
assay at
60
-
-, Q8GTM2, Q8GTM3, Q8GTM4
isoform CLH2
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0
45
-
0C: about 35% of maximal activity, 45C: about 50% of maximal activity
30
50
-
-
pI VALUE
pI VALUE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.7
-
-
calculation from sequence of cDNA
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Q8GTM3, Q8GTM4
;
Manually annotated by BRENDA team
-
developing, constitutive expression of gene AtChl2 and gene ATHCOR1, expression of recmbinant enzyme in mutant transgenic plants
Manually annotated by BRENDA team
-
green, fresh or ripening
Manually annotated by BRENDA team
-
olives, enzyme activity depends on the growth phase and ripening state
Manually annotated by BRENDA team
B6DX58
unripe fruit peel
Manually annotated by BRENDA team
-
of etiolated yound plants
Manually annotated by BRENDA team
-
low content of ATHCOR1, expression of AtChl2
Manually annotated by BRENDA team
-
etiolated seedling
Manually annotated by BRENDA team
-
acetone powder
Manually annotated by BRENDA team
-
the chlorophyll content is higher in female plants compared to male plants, male plants always show higher activities of Chlase
Manually annotated by BRENDA team
-, O22527, Q9M7I7
green, isozyme AtCLH1 expression analysis; green, isozyme AtCLH2 expression analysis
Manually annotated by BRENDA team
Q8GTM2, Q8GTM3, Q8GTM4
-
Manually annotated by BRENDA team
-
highest activity at days 4-6 of senescence
Manually annotated by BRENDA team
additional information
-
not in root
Manually annotated by BRENDA team
additional information
-
developmental, annual, and gender activity differences, overview, Chlase activity is low in october and increases with the onset of winter, the highest Chlase activity is observed in december, more pronounced in male plants, overview
Manually annotated by BRENDA team
additional information
Q9M7I7
spatial and temporal expression patterns of AtCLH2 in wild type plants, overview
Manually annotated by BRENDA team
additional information
Q8GTM2, Q8GTM3, Q8GTM4
CLH1 is expressed during the course of broccoli postharvest senescence
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
26000
-
-
gel filtration
33000
-
-
mature enzyme, SDS-PAGE
33800
-
-
calculated molecular weight
33880
-
-
calculation from sequence of cDNA
35000
-
-
precursor enzyme, SDS-PAGE
37110
-
-
calculated from amino acid sequence
38000
-
-
SDS-PAGE and gel filtration; SDS/PAGE and gel-filtration chromatography
39000
-
-
gel filtration
110000
-
-
gel filtration
130000
-
-
gel filtration
158000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
Q9M7I7
x * 35000, recombinant AtCLH2, SDS-PAGE
?
-, Q8GTM2, Q8GTM3, Q8GTM4
x * 23500, isoform CLH3, calculated from amino acid sequence; x * 34700, isoform CLH1, calculated from amino acid sequence; x * 35300, isoform CLH2, calculated from amino acid sequence; x * 66200, isoform CLH3 as MBP-fusion protein, SDS-PAGE; x * 77400, isoform CLH1 as MBP-fusion protein, SDS-PAGE; x * 78000, isoform CLH2 as MBP-fusion protein, SDS-PAGE
?
B6DX58
x * 32000, recombinant enzyme, SDS-PAGE
?
-
x * 30000, SDS-PAGE
?
-
x * 43000 + x * 46000, SDS-PAGE
?
-
x * 43000 + x * 46000, SDS-PAGE
dimer
-
2 * 65000, SDS-PAGE
tetramer
-
4 * 27000, SDS-PAGE
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
no modification
Q7Y0K5
-
no modification
-
-
side-chain modification
-
glycoprotein
side-chain modification
-
glycoprotein; the sugar group not only stabilizes the enzyme, but also is essential for the manifestation of enzyme activity
additional information
B6DX58
the mature form is N-terminally processed after amino acids 21, 20 or 19
pH STABILITY
pH STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
3
-
-
rapid inactivation
5
8
-, Q8GTM2, Q8GTM3, Q8GTM4
isoform CLH1
6
8
-
0C, loss of 2% of activity per day
6
9
Q9M7I7
isozyme AtCLH2
6
9
-
4C, 24 h, stable
7.5
-
-
stable in presence or absence of 30% acetone
8
10
-, Q8GTM2, Q8GTM3, Q8GTM4
isoform CLH2 shows about 80% of activity at pH 9.0 and is stable between pH 8.0 and 10.0
8
-
-
stability decreases significantly above or below pH 8.0
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
-
-
24 h exposure at 4C followed by 10 d of recovery, leads to overall reduced enzyme activity with the eti5 mutant plants being more sensitive, during the stress time and the first day of recovery the enzyme activity is highly reduced, the enzyme activity is more affected during the experiment time in wild-type plants, overview
20
-
-
24 h, 30-50% loss of activity
30
-
-
not stable above a 10% concentration of acetone
35
50
-
enzyme half-life in the neat organic solvent compared to that in the aqueous medium in the first environment at temperature ranging from 35 to 50C is increased by 5.0 to 15.0 times, overview
38
-
-
24 h exposure at 4C followed by 10 d of recovery, leads to overall reduced enzyme activity with the eti5 mutant plants being more sensitive, during the stress time and the first day of recovery the enzyme activity is highly reduced, the enzyme activity is more affected during the experiment time in wild-type plants, overview
40
-
-
stable for 1 h
45
-
-
1 h, pH 7, citrate buffer, stable. Complete loss of activity in presence of 40% acetone
50
-
-
degradation after 40 min, more rapid at higher temperatures
60
-
-, Q8GTM2, Q8GTM3, Q8GTM4
after incubation at 60C for 10 min, isoform CLH1 loses about 50% of activity; after incubation at 60C for 10 min, isoform CLH2 loses about 50% of activity
60
-
-
10 min, 50% loss of activity
60
-
-
10 min, 40% loss of activity
65
-
-
destroyed within 10 min
additional information
-
-
association of chlorophyllase with mixed monogalactosyldiacylglycerol and phosphatidylglycerol and also with mixed digalactosyldiacylglycerol and phosphatidylglycerol leads to highly increased heat stability. Single lipids are shown to have a much lower or no stabilizing influence. Only with phosphatidylglycerol, in the absence of Mg2+, is any notable enzyme stabilization observed
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
60% loss of activity after dialysis against distilled water in the cold for 48 h
-
stabilized by chlorophyll
-
ORGANIC SOLVENT
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Acetone
-
strong reduction of activity at above 40%
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, stable for at least 1 year without loss of activity
-
25C, loss of 50% activity within 6 days
-
4C, loss of 50% within 18 days
-
-80C, 25 mM Tris buffer, pH 8.0, 100 mM NaCl
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
recombinant isozyme AtCLH2 from Escherichia coli strain BL21 (DE3)
Q9M7I7
recombinant maltose-binding fusion protein from Escherichia coli
-
partial
-
amylose resin affinity column chromatography and DEAE Sepharose column chromatography; amylose resin affinity column chromatography and DEAE Sepharose column chromatography; amylose resin affinity column chromatography and DEAE Sepharose column chromatography
-, Q8GTM2, Q8GTM3, Q8GTM4
partially
-
2 isoenzymes: type 1 chlorophyllase and type 2 chlorophyllase
-
no isoenzymes
-
partial
-
protein-A agarose bead immunoprecipitation
-
recombinant enzymes in the membrane fraction from Nicotiana tabacum chloroplasts by ultracentrifugation
Q9MV14
cold acetone extraction and column chromatography
-
partial
-
Ni2+-NTA column chromatography
-
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, or by using a microtiter plate purification method with immobilization of the enzyme, overview
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression of gene ATHCOR1 in Escherichia coli BL21 as maltose-binding protein fusion protein, cloning of the gene into an expression vector for transformation of Arabidopsis thaliana plants
-
gene chl1, DNA and amino acid sequence determination and anaylsis, transient expression of isozyme AtCLH1 as GFP-tagged protein, e.g. in senescent mesophyll protoplasts, expression analysis; gene chl2, DNA and amino acid sequence determination and anaylsis, transient expression of isozyme AtCLH2 as GFP-tagged protein, e.g. in senescent mesophyll protoplasts, expression analysis
-, O22527, Q9M7I7
gene chl2, expression anaylsis, recombinant expression in Escherichia coli strain BL21 (DE3)
Q9M7I7
expressed in Escherichia coli Rosseta-Gami B (DE3) cells; expressed in Escherichia coli Rosseta-Gami B (DE3) cells; expressed in Escherichia coli Rosseta-Gami B (DE3) cells
-, Q8GTM2, Q8GTM3, Q8GTM4
expression of antisense construct of gene CLH3 in broccoli plants using Agrobacterium tumefaciens-mediated transformation; expression of antisense constructs of gene CLH1 in broccoli plants using Agrobacterium tumefaciens-mediated transformation; expression of antisense constructs of gene CLH2 in broccoli plants using Agrobacterium tumefaciens-mediated transformation
Q8GTM2, Q8GTM3, Q8GTM4
expression in Escherichia coli
Q9LE89
expression of wild-type and mutant enzymes in Escherichia coli
-
expressed in Escherichia coli
B6DX58
Chlase gene, DNA and amino acid sequence determination and anaylsis, expression of precursor full-length wild-type Chlase and of a mutant ChlaseDELTAN lacking the N-terminal 21 amino acids, which corresponds to the mature enzyme, in two heterologous plant systems: in Cucurbita pepo cv. Maayan plants using a ZYMV-based viral vector infective clone system and inoculation of cotyledons, and transiently in Nicotiana tabacum cv. Samsun NN chloroplast membranes of protoplasts, expression of full-length and truncated enzyme versions as thioredoxin fusion proteins in Escherichia coli
Q9MV14
expressed in Escherichia coli
-
expressed in Escherichia coli; expressed in Escherichia coli strain BL21(DE3)
-
expressed in Escherichia coli BL21 (DE3) cells
-
expression as His6-tagged enzyme in Escherichia coli strain BL21(DE3)
-
expression in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
chlorophyllase type I is suppressed in UV-B treated broccoli on day 4
-
in the case of isoform CLH1, an important decrease in expression is observed after day 0, since an over 50fold drop is found from day 0 to day 3 of control florets after hormone-treatment (100ppm 6-benzylaminopurine)
Q8GTM3, Q8GTM4
expression pattern of isoform CLH2 shows an expression increase in senescent tissue and is higher in stem tissue than in inflorescences; isoform CLH1 shows a higher expression in presenescent tissue compared to senescent tissue and stem
Q8GTM3, Q8GTM4
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
D170A
-, Q8GTM2, Q8GTM3, Q8GTM4
inactive; inactive
H226A
-, Q8GTM2, Q8GTM3, Q8GTM4
the mutation causes almost complete loss of Chlase activity; the mutation causes almost complete loss of Chlase activity
H239A
-, Q8GTM2, Q8GTM3, Q8GTM4
the mutation causes almost complete loss of Chlase activity; the mutation causes almost complete loss of Chlase activity
H247A
-, Q8GTM2, Q8GTM3, Q8GTM4
the mutation causes almost complete loss of Chlase activity; the mutation causes almost complete loss of Chlase activity
S141A
-, Q8GTM2, Q8GTM3, Q8GTM4
inactive; inactive
C234A
-
site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
C248A
-
site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
C282A
-
site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
D191N
-
site-directed mutagenesis, inactive mutant
D264N
-
site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
H100A
-
site-directed mutagenesis, highly reduced activity compared to the wild-type enzyme
H161A
-
site-directed mutagenesis, inactive mutant
H241A
-
site-directed mutagenesis, about 90% reduced activity compared to the wild-type enzyme
H254A
-
site-directed mutagenesis, nearly inactive mutant
H254Y
-
site-directed mutagenesis, about 90% reduced activity compared to the wild-type enzyme
H262A
-
site-directed mutagenesis, nearly inactive mutant
H262Y
-
site-directed mutagenesis, nearly inactive mutant
H81A
-
site-directed mutagenesis, about 85% reduced activity compared to the wild-type enzyme
S162A
-
site-directed mutagenesis, nearly inactive mutant
additional information
-
gene ATHCOR1: construction of transgenic Arabidopsis thaliana plants overexpressing the enzyme via infection with Agrobacterium tumefaciens and transformation, sense and antisense orientation, the sense mutation changed the chlorophyll a to chlorophyll b ratio, overview, gene AtCLH2: cloning and analysis
additional information
-, O22527, Q9M7I7
construction of T-DNA insertion clh1 and clh2 single and double knockout lines, which are still able to degrade chlorophyll during senescence, phenotypes, overview; construction of T-DNA insertion clh1 and clh2 single and double knockout lines, which are still able to degrade chlorophyll during senescence, phenotypes, overview
additional information
Q9M7I7
inhibition of expression of AtCLH2 by RNA interference, transfection using Agrobacterium tumefaciens strain LBA4404, AtCLH2 RNAi plants show decreased contents of chlorophyllide without a substantial change in the total amount of the extractable chlorophyll and consequently presented lower chlorophyllide to chlorophyll ratios in their leaves, phenotype, overview
H66A
-, Q8GTM2, Q8GTM3, Q8GTM4
the mutation causes almost complete loss of Chlase activity; the mutation causes almost complete loss of Chlase activity
additional information
Q8GTM2, Q8GTM3, Q8GTM4
a chimeric construct with the antisense gene BoCLH1, driven by the CaMV 35S promoter and Nos-terminator and harboring the hygromycin resistance gene, is used for Agrobacterium tumefaciens-mediated transformation, effects of the antisense BoCLH1 gene on the postharvest senescence of broccoli, phenotype with slower postharvest yellowing during storage in the dark at 20C, overview; no effects on yellowing by antisense construct of gene CHL2; no effects on yellowing by antisense constructs of gene CHL3
S167A
-
site-directed mutagenesis, about 50% reduced activity compared to the wild-type enzyme
additional information
Q9MV14
expression of full-length citrus Chlase results in limited chlorophyll breakdown in tobacco protoplasts and no visible leaf phenotype in whole plants, whereas expression of a Chlase version lacking the N-terminal 21 amino acids, i.e. ChlaseDELTAN, which corresponds to the mature protein, leads to extensive chlorophyll breakdown in both tobacco protoplasts and squash leaves, mutant ChlaseDELTAN-expressing squash leaves display a dramatic chlorotic phenotype in plants grown under low-intensity light, whereas under natural light a lesion-mimic phenotype occurrs, which follows the accumulation of chlorophyllide, a photodynamic chlorophyll breakdown product, phenotypes, overview
additional information
-
optimization of chlorophyllase-catalyzed hydrolysis of chlorophyll in monophasic organic solvent media, best in a mixture of hexane/2-octanone of 98.3:1.7 v/v, overview
additional information
-
entrapping of the enzyme in tetramethoxysilane-based sol-gel in the presence of lipid, acetone is used to introduce the substrate-chlorophyll into aqueous reaction medium in solubilized form, the entrapped chlorophyllase is less sensitive toward the denaturing effect of acetone in the bulk phase than the free enzyme, sol-gel at wet-stage, diffusion coefficient and partition coefficient of chlorophyll in sol-gel, overview
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
synthesis
-
useful tool for preparing chlorophyllides and chlorophyll derivatives esterified with various alcohols
agriculture
-
removal of green pigments from canola oil
biotechnology
-
encapsulation of the enzyme in micelles of different media within alginate hydrogels increases the enzyme activity in e.g. Tris buffer or hexane, extent of enhancement of the partition coefficient depends on the amount and hydrophobicity of the components intrduced into alginate, affecting the hydrophobic-hydrophilic balance of the gel
agriculture
-
manipulation of chlorophyll degradation