Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Drenth, J.; Yang, G.; Paul, C.E.; Fraaije, M.W.
    A tailor-made deazaflavin-mediated recycling system for artificial nicotinamide cofactor biomimetics (2021), ACS Catal., 11, 11561-11569 .
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.5.1.40 synthesis Tfu-FNO variants can be used as NCB-Recycling systems for TsOYE-mediated conversions. TsOYE is known to efficiently hydrogenate ketoisophorone (2,6,6-trimethyl-2-cyclohexene-1,4-dione) to form the chiral product (R)-2,2,6- trimethylcyclohexane-1,4-dione with high enantiomeric excess values of 94-97%, when several NCBs as more efficient electron donors in comparison to NADPH are used. The Tfu-FNO variants G29Y, G29W, and P89H, which show high activity toward 1-aminoethylnicotinamide (AmNA+), 1-benzylnicotinamide (BNA+), and 1-benzyl-3-acetylpyridine (BAP+), are selected for use in conversion experiments as recycling catalysts for these NCBs. Potential use of these thermostable Tfu- FNO variants as NCB recycling systems at high temperatures Thermobifida fusca

Protein Variants

EC Number Protein Variants Comment Organism
1.5.1.40 G29L site-directed mutagenesis, the mutant shows altered kinetics and increased catalytic efficiency with nicotinamide cofactor biomimetics compared to wild-type enzyme Thermobifida fusca
1.5.1.40 G29S site-directed mutagenesis, the mutant shows altered kinetics and increased catalytic efficiency with nicotinamide cofactor biomimetics compared to wild-type enzyme Thermobifida fusca
1.5.1.40 G29Y site-directed mutagenesis, the mutant shows altered kinetics and increased catalytic efficiency with nicotinamide cofactor biomimetics compared to wild-type enzyme Thermobifida fusca
1.5.1.40 additional information engineering of the thermostable F420:NADPH oxidoreductase from Thermobifida fusca (Tfu-FNO) by structure-inspired site-directed mutagenesis to accommodate the unnatural N1 substituents of eight nicotinamide cofactor biomimetics (NCBs). The extraordinarily low redox potential of the natural cofactor F420H2 is then exploited to reduce these NCBs. Wild-type enzyme has detectable activity toward all selected NCBs. Saturation mutagenesis at positions Gly29 and Pro89 results in mutants with up to 139times higher catalytic efficiencies, kinetics comparisons, overview. Most mutations significantly decrease the activity toward NADP+ but do not completely inhibit the enzyme for this cosubstrate Thermobifida fusca
1.5.1.40 P89H site-directed mutagenesis, the mutant shows altered kinetics and increased catalytic efficiency with nicotinamide cofactor biomimetics compared to wild-type enzyme Thermobifida fusca
1.5.1.40 P89L site-directed mutagenesis, the mutant shows altered kinetics and increased catalytic efficiency with nicotinamide cofactor biomimetics compared to wild-type enzyme Thermobifida fusca
1.5.1.40 P89Y site-directed mutagenesis, the mutant shows altered kinetics and increased catalytic efficiency with nicotinamide cofactor biomimetics compared to wild-type enzyme Thermobifida fusca

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.40 2 3 1-aminoethylnicotinamide enzyme mutant G29Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 2 3 1-hydroxyethylnicotinamide enzyme mutant G29Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 4.3
-
 1-benzylnicotinamide enzyme mutant G29W, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 5.7
-
 1-propylnicotinamide enzyme mutant P89H, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 6.4
-
 1-benzylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 7.4
-
 1-benzyl-3-acetylpyridine wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 9.5
-
 1-benzylnicotinamide enzyme mutant G29Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 10
-
 1-propylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 12
-
 1-aminoethylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 14
-
 1-aminoethylnicotinamide enzyme mutant P98H, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 18
-
 1-propylnicotinamide enzyme mutant G29L, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 18
-
 1-aminoethylnicotinamide enzyme mutant G29S, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 20
-
 1-hydroxyethylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 20
-
 1-hydroxyethylnicotinamide enzyme mutant P98H, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 25
-
 1-hydroxypropylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 25
-
 1-ethylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 27
-
 1-propylnicotinamide enzyme mutant P89L, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 33
-
 1-hydroxyethylnicotinamide enzyme mutant P98Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 35
-
 1-hydroxypropylnicotinamide enzyme mutant P89Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 35
-
 1-propylnicotinamide enzyme mutant P89Y, pH 8.0, 25°C Thermobifida fusca

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.5.1.40 reduced coenzyme F420 + NADP+ Thermobifida fusca
-
 oxidized coenzyme F420 + NADPH + H+
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.40 Thermobifida fusca Q47RA9
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.40 additional information no activity of wild-type and mutants with 1-benzylnicotinic acid Thermobifida fusca ?
-
-
1.5.1.40 reduced coenzyme F420 + 1-aminoethylnicotinamide
-
 Thermobifida fusca oxidized coenzyme F420 + ?
-
 ?
1.5.1.40 reduced coenzyme F420 + 1-benzyl-3-acetylpyridine
-
 Thermobifida fusca oxidized coenzyme F420 + ?
-
 ?
1.5.1.40 reduced coenzyme F420 + 1-benzylnicotinamide
-
 Thermobifida fusca oxidized coenzyme F420 + ?
-
 ?
1.5.1.40 reduced coenzyme F420 + 1-ethylnicotinamide
-
 Thermobifida fusca oxidized coenzyme F420 + ?
-
 ?
1.5.1.40 reduced coenzyme F420 + 1-hydroxyethylnicotinamide
-
 Thermobifida fusca oxidized coenzyme F420 + ?
-
 ?
1.5.1.40 reduced coenzyme F420 + 1-hydroxypropylnicotinamide
-
 Thermobifida fusca oxidized coenzyme F420 + ?
-
 ?
1.5.1.40 reduced coenzyme F420 + 1-propylnicotinamide
-
 Thermobifida fusca oxidized coenzyme F420 + ?
-
 ?
1.5.1.40 reduced coenzyme F420 + NADP+
-
 Thermobifida fusca oxidized coenzyme F420 + NADPH + H+
-
 r

Synonyms

EC Number Synonyms Comment Organism
1.5.1.40 F420:NADPH oxidoreductase
-
 Thermobifida fusca
1.5.1.40 Tfu-FNO
-
 Thermobifida fusca
1.5.1.40 Tfu_0970
-
 Thermobifida fusca

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.1.40 25
-
 assay at Thermobifida fusca

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.5.1.40 25 50 the activity of a selection of variants toward 5 mM of 1-aminoethylnicotinamide (AmNA+), 1-benzylnicotinamide (BNA+), and 1-benzyl-3-acetylpyridine (BAP+) at both 25°C and 50°C shows that the activity increases at elevated temperature Thermobifida fusca

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.5.1.40 0.0015
-
 1-propylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0042
-
 1-hydroxypropylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0042
-
 1-benzyl-3-acetylpyridine wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0045
-
 1-benzylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0045
-
 1-propylnicotinamide enzyme mutant P89H, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.006
-
 1-propylnicotinamide enzyme mutant G29L, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0073
-
 1-ethylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0073
-
 1-propylnicotinamide enzyme mutant P89L, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0098
-
 1-propylnicotinamide enzyme mutant P89Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0118
-
 1-hydroxypropylnicotinamide enzyme mutant P89Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0233
-
 1-aminoethylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0233
-
 1-hydroxyethylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.042
-
 1-aminoethylnicotinamide enzyme mutant P98H, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.048
-
 1-hydroxyethylnicotinamide enzyme mutant P98H, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.053
-
 1-aminoethylnicotinamide enzyme mutant G29Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.083
-
 1-hydroxyethylnicotinamide enzyme mutant P98Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.103
-
 1-hydroxyethylnicotinamide enzyme mutant G29Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.145
-
 1-benzylnicotinamide enzyme mutant G29Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.16
-
 1-aminoethylnicotinamide enzyme mutant G29S, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.417
-
 1-benzylnicotinamide enzyme mutant G29W, pH 8.0, 25°C Thermobifida fusca

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.1.40 8
-
 assay at Thermobifida fusca

Cofactor

EC Number Cofactor Comment Organism Structure
1.5.1.40 NADP+
-
 Thermobifida fusca
1.5.1.40 NADPH
-
 Thermobifida fusca

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.5.1.40 0.00015
-
 1-propylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.00017
-
 1-hydroxypropylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.00027
-
 1-propylnicotinamide enzyme mutant P89L, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.00028
-
 1-propylnicotinamide enzyme mutant P89Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.00029
-
 1-ethylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.00033
-
 1-propylnicotinamide enzyme mutant G29L, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.00034
-
 1-hydroxypropylnicotinamide enzyme mutant P89Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.00057
-
 1-benzyl-3-acetylpyridine wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0007
-
 1-benzylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.00079
-
 1-propylnicotinamide enzyme mutant P89H, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0012
-
 1-hydroxyethylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.00194
-
 1-aminoethylnicotinamide wild-type enzyme, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0023
-
 1-aminoethylnicotinamide enzyme mutant G29Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0024
-
 1-hydroxyethylnicotinamide enzyme mutant P98H, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0025
-
 1-hydroxyethylnicotinamide enzyme mutant P98Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.003
-
 1-aminoethylnicotinamide enzyme mutant P98H, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0045
-
 1-hydroxyethylnicotinamide enzyme mutant G29Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0088
-
 1-aminoethylnicotinamide enzyme mutant G29S, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.0153
-
 1-benzylnicotinamide enzyme mutant G29Y, pH 8.0, 25°C Thermobifida fusca
1.5.1.40 0.097
-
 1-benzylnicotinamide enzyme mutant G29W, pH 8.0, 25°C Thermobifida fusca