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Literature summary extracted from

  • Sonawane, P.; Patel, K.; Vishwakarma, R.K.; Srivastava, S.; Singh, S.; Gaikwad, S.; Khan, B.M.
    Probing the active site of cinnamoyl CoA reductase 1 (Ll-CCRH1) from Leucaena leucocephala (2013), Int. J. Biol. Macromol., 60C, 33-38.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.2.1.44 expressed in Escherichia coli BL21(DE3) cells Leucaena leucocephala

Protein Variants

EC Number Protein Variants Comment Organism
1.2.1.44 D77G the mutant displays higher Km values (up to 4fold), indicating lower affinity toward substrate, while catalytic efficiencies for these mutant is notably decreased Leucaena leucocephala
1.2.1.44 F30V the mutant displays higher Km values (up to 4fold), indicating lower affinity toward substrate, while catalytic efficiencies for these mutant is notably decreased Leucaena leucocephala
1.2.1.44 F30V/I31N the mutant shows approximately 7fold increase in Km and 8fold decrease in kcat/Km values Leucaena leucocephala
1.2.1.44 H215L the mutant shows no significant change in Km values compared to the wild type enzyme Leucaena leucocephala
1.2.1.44 I131N the mutant shows slightly reduced catalytic efficiency compared to the wild type Leucaena leucocephala
1.2.1.44 K174M the mutant shows complete loss of activity with feruloyl-CoA as substrate Leucaena leucocephala
1.2.1.44 L64W the mutant shows no significant change in Km values compared to the wild type enzyme Leucaena leucocephala
1.2.1.44 R51G the mutant displays higher Km values (up to 4fold), indicating lower affinity toward substrate, while catalytic efficiencies for these mutant is notably decreased Leucaena leucocephala
1.2.1.44 R51G/D77G the mutant displays 5fold increase in Km and around 9fold reduction in specificity constant Leucaena leucocephala
1.2.1.44 S136A the mutant shows complete loss of activity with feruloyl-CoA as substrate Leucaena leucocephala
1.2.1.44 S212G the mutant exhibits the catalytic efficiencies less than 10% of wild type enzyme Leucaena leucocephala
1.2.1.44 S212G the mutant shows a 2.5fold increase in Km, 7fold reduction in kcat and 15fold decrease in kcat/Km Leucaena leucocephala
1.2.1.44 S99G the mutant shows no significant change in Km values compared to the wild type enzyme Leucaena leucocephala
1.2.1.44 V200E the mutant shows reduced catalytic efficiency compared to the wild type Leucaena leucocephala
1.2.1.44 Y170H the mutant shows complete loss of activity with feruloyl-CoA as substrate Leucaena leucocephala

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.2.1.44 citraconic anhydride 31% residual activity at 1.5 mM with feruloyl-CoA as substrate Leucaena leucocephala
1.2.1.44 diethylpyrocarbonate 44% residual activity at 3 mM with feruloyl-CoA as substrate Leucaena leucocephala
1.2.1.44 additional information not inhibited by N-ethylmaleimide, 4-hydroxymercurybenzoate, and N-bromosuccinimide Leucaena leucocephala
1.2.1.44 N-acetyl imidazole 50% residual activity at 3 mM with feruloyl-CoA as substrate Leucaena leucocephala
1.2.1.44 Phenylglyoxal 47% residual activity at 1 mM with feruloyl-CoA as substrate Leucaena leucocephala
1.2.1.44 phenylmethylsulfonyl fluoride 60% residual activity at 1 mM with feruloyl-CoA as substrate Leucaena leucocephala
1.2.1.44 Woodward's reagent K 54% residual activity at 0.02 mM with feruloyl-CoA as substrate Leucaena leucocephala

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2.1.44 0.032
-
 feruloyl-CoA mutant enzyme S99G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.036
-
 feruloyl-CoA wild type enzyme, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.039
-
 feruloyl-CoA mutant enzyme L64W, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.042
-
 feruloyl-CoA mutant enzyme H215L, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.047
-
 feruloyl-CoA mutant enzyme R51G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.057
-
 feruloyl-CoA mutant enzyme D77G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.06
-
 feruloyl-CoA mutant enzyme V200E, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.079
-
 feruloyl-CoA mutant enzyme S212G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.096
-
 feruloyl-CoA mutant enzyme F30V, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.102
-
 feruloyl-CoA mutant enzyme I31N, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.166
-
 feruloyl-CoA mutant enzyme R51G/D77G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 0.244
-
 feruloyl-CoA mutant enzyme RF30V/I31N, pH and temperature not specified in the publication Leucaena leucocephala

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.2.1.44 feruloyl-CoA + NADPH + H+ Leucaena leucocephala
-
 ferulic aldehyde + CoA + NADP+
-
 ?
1.2.1.44 sinapoyl-CoA + NADPH + H+ Leucaena leucocephala
-
 sinapaldehyde + CoA + NADP+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.2.1.44 Leucaena leucocephala Q08GL0
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.2.1.44 feruloyl-CoA + NADPH + H+
-
 Leucaena leucocephala ferulic aldehyde + CoA + NADP+
-
 ?
1.2.1.44 sinapoyl-CoA + NADPH + H+
-
 Leucaena leucocephala sinapaldehyde + CoA + NADP+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.2.1.44 CCRH1
-
 Leucaena leucocephala
1.2.1.44 cinnamoyl CoA reductase 1
-
 Leucaena leucocephala

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.2.1.44 24
-
 feruloyl-CoA mutant enzyme S212G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 44
-
 feruloyl-CoA mutant enzyme H215L, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 84
-
 feruloyl-CoA mutant enzyme R51G/D77G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 100
-
 feruloyl-CoA mutant enzyme R51G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 108
-
 feruloyl-CoA mutant enzyme D77G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 122
-
 feruloyl-CoA mutant enzyme V200E, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 137
-
 feruloyl-CoA mutant enzyme F30V, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 142
-
 feruloyl-CoA mutant enzyme RF30V/I31N, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 144
-
 feruloyl-CoA mutant enzyme S99G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 155
-
 feruloyl-CoA mutant enzyme I31N, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 160
-
 feruloyl-CoA mutant enzyme L64W, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 162
-
 feruloyl-CoA wild type enzyme, pH and temperature not specified in the publication Leucaena leucocephala

Cofactor

EC Number Cofactor Comment Organism Structure
1.2.1.44 NADPH
-
 Leucaena leucocephala

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.2.1.44 310
-
 feruloyl-CoA mutant enzyme S212G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 510
-
 feruloyl-CoA mutant enzyme R51G/D77G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 590
-
 feruloyl-CoA mutant enzyme RF30V/I31N, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 1000
-
 feruloyl-CoA mutant enzyme H215L, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 1400
-
 feruloyl-CoA mutant enzyme F30V, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 1500
-
 feruloyl-CoA mutant enzyme I31N, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 1900
-
 feruloyl-CoA mutant enzyme D77G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 2000
-
 feruloyl-CoA mutant enzyme V200E, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 2100
-
 feruloyl-CoA mutant enzyme R51G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 4100
-
 feruloyl-CoA mutant enzyme L64W, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 4500
-
 feruloyl-CoA mutant enzyme S99G, pH and temperature not specified in the publication Leucaena leucocephala
1.2.1.44 4600
-
 feruloyl-CoA wild type enzyme, pH and temperature not specified in the publication Leucaena leucocephala