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Literature summary 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

EC Number Application Comment Organism
3.1.1.14 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)

EC Number Cloned (Comment) Organism
3.1.1.14 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]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.14 additional information
-
 additional information steady-state kinetics, Michaelis-Menten kinetics Crocosphaera subtropica ATCC 51142
3.1.1.14 0.0495
-
 bacteriochlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
3.1.1.14 0.157
-
 chlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
3.1.1.14 0.2795
-
 chlorophyll b recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.14 Crocosphaera subtropica ATCC 51142 B1X2S9
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.1.14 9.88
-
 purified recombinant enzyme, pH 7.0, 60°C, substrate chlorophyll b Crocosphaera subtropica ATCC 51142
3.1.1.14 11.15
-
 purified recombinant enzyme, pH 7.0, 60°C, substrate chlorophyll a Crocosphaera subtropica ATCC 51142
3.1.1.14 75.6
-
 purified recombinant enzyme, pH 7.0, 60°C, substrate bacteriochlorophyll a Crocosphaera subtropica ATCC 51142

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.14 bacteriochlorophyll a + H2O preferred substrate Crocosphaera subtropica ATCC 51142 phytol + bacteriochlorophyllide a
-
 ?
3.1.1.14 chlorophyll + H2O
-
 Crocosphaera subtropica ATCC 51142 phytol + chlorophyllide
-
 ?
3.1.1.14 chlorophyll a + H2O
-
 Crocosphaera subtropica ATCC 51142 phytol + chlorophyllide a
-
 ?
3.1.1.14 chlorophyll b + H2O
-
 Crocosphaera subtropica ATCC 51142 phytol + chlorophyllide b
-
 ?
3.1.1.14 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

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

Synonyms

EC Number Synonyms Comment Organism
3.1.1.14 cce_5094
-
 Crocosphaera subtropica ATCC 51142
3.1.1.14 Chlase
-
 Crocosphaera subtropica ATCC 51142
3.1.1.14 CyanoCLH
-
 Crocosphaera subtropica ATCC 51142

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.1.14 40 50 80% of maximal activity, recombinant enzyme Crocosphaera subtropica ATCC 51142
3.1.1.14 60
-
 assay at Crocosphaera subtropica ATCC 51142

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.14 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]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.1.14 251.08
-
 chlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
3.1.1.14 310.36
-
 chlorophyll b recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
3.1.1.14 726.33
-
 bacteriochlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.14 7
-
 recombinant enzyme Crocosphaera subtropica ATCC 51142

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.1.1.14 6 8 85% of maximal activity, recombinant enzyme Crocosphaera subtropica ATCC 51142

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.1.1.14 5 9 purified recombinant enzyme, over 80% activity remaining Crocosphaera subtropica ATCC 51142

General Information

EC Number General Information Comment Organism
3.1.1.14 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.1.14 1100
-
 chlorophyll b recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
3.1.1.14 1599
-
 chlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142
3.1.1.14 14688
-
 bacteriochlorophyll a recombinant enzyme, pH 7.0, 60°C Crocosphaera subtropica ATCC 51142