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Literature summary for 3.1.1.14 extracted from

  • Chou, Y.L.; Lee, Y.L.; Yen, C.C.; Chen, L.F.; Lee, L.C.; Shaw, J.F.
    A novel recombinant chlorophyllase from cyanobacterium Cyanothece sp. ATCC 51142 for the production of bacteriochlorophyllide a (2016), Biotechnol. Appl. Biochem., 63, 371-377 .
    View publication on PubMed

Application

Application Comment Organism
synthesis the enzyme CyanoCLH can be used as a biocatalyst for the production of a bacteriochlorin a (BCA) precursor by degrading bacteriochlorophyll a (BChl a). Bacteriopheophorbide a (BPheid a) is used as a precursor for bacteriochlorin a (BCA), which can be used for photodynamic therapy in both in vitro and in vivo biochemical applications Crocosphaera subtropica ATCC 51142

Cloned(Commentary)

Cloned (Comment) Organism
gene cce_5094, sequence comparisons, recombinant expression of the N-terminally T7-tagged enzyme (T7 tag sequence is MASMTGGQQMG)in Escherichia coli strain BL21(DE3) Crocosphaera subtropica ATCC 51142

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information steady-state kinetics, Michaelis-Menten kinetics Crocosphaera subtropica ATCC 51142
0.0495
-
bacteriochlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
0.157
-
chlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
0.2795
-
chlorophyll b recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
bacteriochlorophyll a + H2O Crocosphaera subtropica ATCC 51142 preferred substrate phytol + bacteriochlorophyllide a
-
?
chlorophyll + H2O Crocosphaera subtropica ATCC 51142
-
phytol + chlorophyllide
-
?

Organism

Organism UniProt Comment Textmining
Crocosphaera subtropica ATCC 51142 B1X2S9
-
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
9.88
-
purified recombinant enzyme, pH 7.0, 60°C, substrate chlorophyll b Crocosphaera subtropica ATCC 51142
11.15
-
purified recombinant enzyme, pH 7.0, 60°C, substrate chlorophyll a Crocosphaera subtropica ATCC 51142
75.6
-
purified recombinant enzyme, pH 7.0, 60°C, substrate bacteriochlorophyll a Crocosphaera subtropica ATCC 51142

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
bacteriochlorophyll a + H2O preferred substrate Crocosphaera subtropica ATCC 51142 phytol + bacteriochlorophyllide a
-
?
chlorophyll + H2O
-
Crocosphaera subtropica ATCC 51142 phytol + chlorophyllide
-
?
chlorophyll a + H2O
-
Crocosphaera subtropica ATCC 51142 phytol + chlorophyllide a
-
?
chlorophyll b + H2O
-
Crocosphaera subtropica ATCC 51142 phytol + chlorophyllide b
-
?
additional information enzyme substrate specificity, overview. Recombinant CyanoCLH prefers hydrolyzing bacteriochlorophyll a to produce bacteriochlorophyllide a (BChlide a), which can be converted to bacteriopheophorbide a by removing magnesium ion Crocosphaera subtropica ATCC 51142 ?
-
?

Subunits

Subunits Comment Organism
? x * 54700, recombinant T7-tagged enzyme, SDS-PAGE Crocosphaera subtropica ATCC 51142

Synonyms

Synonyms Comment Organism
cce_5094
-
Crocosphaera subtropica ATCC 51142
Chlase
-
Crocosphaera subtropica ATCC 51142
CyanoCLH
-
Crocosphaera subtropica ATCC 51142

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40 50 80% of maximal activity, recombinant enzyme Crocosphaera subtropica ATCC 51142
60
-
assay at Crocosphaera subtropica ATCC 51142

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
70
-
90% activity remains at 70°C, the relative activity of the recombinant CyanoCLH decreases sharply at 80-90°C Crocosphaera subtropica ATCC 51142

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
251.08
-
chlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
310.36
-
chlorophyll b recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
726.33
-
bacteriochlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
recombinant enzyme Crocosphaera subtropica ATCC 51142

pH Range

pH Minimum pH Maximum Comment Organism
6 8 85% of maximal activity, recombinant enzyme Crocosphaera subtropica ATCC 51142

pH Stability

pH Stability pH Stability Maximum Comment Organism
5 9 purified recombinant enzyme, over 80% activity remaining Crocosphaera subtropica ATCC 51142

General Information

General Information Comment Organism
evolution the deduced amino acid sequence of recombinant CyanoCLH comprises a unique lipase-motif GHSLG, which differs from the GHSRG sequence of other plants and lacks a histidine of the typical and conserved catalytic triad Ser-Asp-His Crocosphaera subtropica ATCC 51142

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1100
-
chlorophyll b recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
1599
-
chlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
14688
-
bacteriochlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142