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Literature summary extracted from

  • Chou, Y.L.; Ko, C.Y.; Yen, C.C.; Chen, L.F.; Shaw, J.F.
    A novel recombinant chlorophyllase1 from Chlamydomonas reinhardtii for the production of chlorophyllide derivatives (2015), J. Agric. Food Chem., 63, 9496-9503 .
    View publication on PubMed

Application

EC Number Application Comment Organism
3.1.1.14 synthesis recombinant enzyme CrCLH1 can be used as a biocatalyst to produce chlorophyllide derivatives Chlamydomonas reinhardtii

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.14 gene CLH1, DNA and amino acid sequence determination and analysis, sequence comparisons and phylogenetic analysis, recombinant C-terminally GFP-tagged enzyme expression driven by a CAMV35S promoter is determined in chloroplasts of Arabidopsis thaliana mesophyll protoplasts (isolated from 3-4-week-old well-expanded leaves yet to flower), recombinant overexpression of soluble N-terminally T7-tagged enzyme in Escherichia coli strain BL21(DE3) Chlamydomonas reinhardtii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.14 0.0566
-
 chlorophyll a recombinant enzyme, pH 7.0, 60°C Chlamydomonas reinhardtii
3.1.1.14 0.0999
-
 chlorophyll b recombinant enzyme, pH 7.0, 60°C Chlamydomonas reinhardtii
3.1.1.14 0.4603
-
 bacteriochlorophyll a recombinant enzyme, pH 7.0, 60°C Chlamydomonas reinhardtii

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
3.1.1.14 chloroplast
-
 Chlamydomonas reinhardtii 9507
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.1.14 chlorophyll + H2O Chlamydomonas reinhardtii
-
 phytol + chlorophyllide
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.14 Chlamydomonas reinhardtii A8J2S9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.14 recombinant T7-tagged enzyme from Escherichia coli strain BL21(DE3) by affinity chromatography Chlamydomonas reinhardtii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.14 bacteriochlorophyll a + H2O
-
 Chlamydomonas reinhardtii phytol + bacteriochlorophyllide a
-
 ?
3.1.1.14 chlorophyll + H2O
-
 Chlamydomonas reinhardtii phytol + chlorophyllide
-
 ?
3.1.1.14 chlorophyll a + H2O preferred substrate Chlamydomonas reinhardtii phytol + chlorophyllide a
-
 ?
3.1.1.14 chlorophyll b + H2O
-
 Chlamydomonas reinhardtii phytol + chlorophyllide b
-
 ?
3.1.1.14 additional information recombinant CrCLH1 has a higher catalytic efficiency for chlorophyll a than for chlorophyll b and bacteriochlorophyll a. Recombinant CrCLH1 catalyzes the conversion of chlorophyll a to pheophorbide a at pH 5.0 Chlamydomonas reinhardtii ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.1.14 ? x * 36500, recombinant T7-tagged enzyme, SDS-PAGE Chlamydomonas reinhardtii

Synonyms

EC Number Synonyms Comment Organism
3.1.1.14 Chlase
-
 Chlamydomonas reinhardtii
3.1.1.14 chlorophyllase 1
-
 Chlamydomonas reinhardtii
3.1.1.14 chlorophyllase1
-
 Chlamydomonas reinhardtii
3.1.1.14 CLH1
-
 Chlamydomonas reinhardtii
3.1.1.14 CrCLH1
-
 Chlamydomonas reinhardtii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.1.14 40
-
 recombinant enzyme Chlamydomonas reinhardtii

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.1.1.14 20 75 recombinant enzyme, activity range, profile overview. 90% of maximal activity at 20°C and 45°C, 70% at 50°C, 25% at 70°C, inactivation at 80°C Chlamydomonas reinhardtii

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.14 40 70 purified recombinant enzyme, completely stable up to, 35% activity remaining at 70°C, inactivation at 80°C Chlamydomonas reinhardtii

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.1.14 212.2
-
 bacteriochlorophyll a recombinant enzyme, pH 7.0, 60°C Chlamydomonas reinhardtii
3.1.1.14 232.8
-
 chlorophyll b recombinant enzyme, pH 7.0, 60°C Chlamydomonas reinhardtii
3.1.1.14 559.55
-
 chlorophyll a recombinant enzyme, pH 7.0, 60°C Chlamydomonas reinhardtii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.14 6
-
 recombinant enzyme Chlamydomonas reinhardtii

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.1.1.14 4.5 8.5 over 50% of maximal activity within this range Chlamydomonas reinhardtii
3.1.1.14 5 7 over 90% of maximal activity within this range Chlamydomonas reinhardtii

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.1.1.14 3 10 purified recombinant enzyme, stability profile, overview, over 50% activity remaining at pH 4.5-9.0, over 90% at pH 7.0, completely stable at pH 5.0-6.0 Chlamydomonas reinhardtii

General Information

EC Number General Information Comment Organism
3.1.1.14 evolution the enzyme shares th highly conserved lipase motif GXSXG Chlamydomonas reinhardtii
3.1.1.14 physiological function recombinant chlorophyllase 1, CrCLH1, can perform chlorophyll dephytylation and produce chlorophyllide and phytol Chlamydomonas reinhardtii

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.1.14 461
-
 bacteriochlorophyll a recombinant enzyme, pH 7.0, 60°C Chlamydomonas reinhardtii
3.1.1.14 2330
-
 chlorophyll b recombinant enzyme, pH 7.0, 60°C Chlamydomonas reinhardtii
3.1.1.14 9886
-
 chlorophyll a recombinant enzyme, pH 7.0, 60°C Chlamydomonas reinhardtii