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Literature summary extracted from

  • Feil, I.K.; Hendle, J.; Schomburg, D.
    Modified substrate specificity of L-hydroxyisocaproate dehydrogenase derived from structure-based protein engineering (1997), Protein Eng., 10, 255-262.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.337 F236S phenylpyruvate displays the largest kcat of the tested substrates. All kcat values, with the exception of phenylpyruvate, are reduced, but the relative acceptance of phenylglyoxylate is greatly increased Weissella confusa
1.1.1.337 F236V decrease in the second-order rate constants for all tested substrates. The kcat values for the smaller substrates decrease drastically. Phenylpyruvate is the favourite substrate (2-oxocaproate is the favourite substrate of the wild-type enzyme) Weissella confusa
1.1.1.337 G234V/G235D the second-order rate constant decreased for most substrates with the exception of pyruvate, which reacts 2.5times faster in the mutant enzyme than in the wild-type enzyme. The catalysis of 2-oxoisocaproate is unaffected Weissella confusa
1.1.1.337 L239A shift of enzyme specificity towards the substrates branched at C3 corresponding to an increase in the turnover numbers for 2-oxoisocaproate Weissella confusa
1.1.1.337 L239F slight decrease in the KM values for phenylglyoxylate combined with a dramatic decrease in the kcat values Weissella confusa
1.1.1.337 L239M KM values of all substrates of the enzyme variant are very similar to those of the wild-type enzyme, large improvement in the kcat values of the C4-branched substrates 2-oxoisocaproate and phenylpyruvate, decrease in the turnover number of 2-oxocaproate (the substrate favoured by the wild-type enzyme). Whereas in the wild-type enzyme the second-order rate constant for 2-oxoisocaproate is only 1% of that for the unbranched substrate, in the mutant enzyme the rate constant of 2-oxocaproate is only 18% of that for 2-oxoisocaproate Weissella confusa
1.1.1.337 L239M/T245A the catalytic rates are reduced by several orders of magnitude and the KM values shows at least a 100fold increase for most substrates (with the exception of phenylglyoxylate). With respect to L239M the substrate specificity shifts towards keto-tert-leucine and with respect to T245A 2-oxoisocaproate and phenylpyruvate are more favoured Weissella confusa
1.1.1.337 L239W slight decrease in the KM values for phenylglyoxylate combined with a dramatic decrease in the kcat values Weissella confusa
1.1.1.337 T245A the catalytic rates are reduced by several orders of magnitude, relative shift of substrate specificity for keto-tert-leucine of more than 17 000 compared with the 2-oxocaproate (kcat/KM) Weissella confusa

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.337 Weissella confusa P14295 i.e. Lactobacillus confusus
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.337 2-oxocaproate + NADH + H+ 2-oxocaproate is the best substrate for the wild-type enzyme Weissella confusa 2-hydroxycaproate + NAD+
-
 r
1.1.1.337 2-oxoisocaproate + NADH + H+
-
 Weissella confusa 2-hydroxyisocaproate + NAD+
-
 r
1.1.1.337 additional information broad substrate specificity, utilizes a wide range of 2-oxo acids branched at the C4 atom Weissella confusa ?
-
 ?
1.1.1.337 oxo-tert-leucine + NADH + H+
-
 Weissella confusa ? + NAD+
-
 r
1.1.1.337 phenylglyoxylate + NADH + H+
-
 Weissella confusa ? + NAD+
-
 r
1.1.1.337 phenylpyruvate + NADH + H+
-
 Weissella confusa 3-phenyllactate + NAD+
-
 r
1.1.1.337 pyruvate + NADH + H+
-
 Weissella confusa L-lactate + NAD+
-
 r

Synonyms

EC Number Synonyms Comment Organism
1.1.1.337 L-HicDH
-
 Weissella confusa
1.1.1.337 L-hydroxyisocaproate dehydrogenase
-
 Weissella confusa

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.337 0.07
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 0.075
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
1.1.1.337 0.1
-
 pyruvate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
1.1.1.337 0.1
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
1.1.1.337 0.2
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
1.1.1.337 0.2
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 0.2
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
1.1.1.337 0.3
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 0.4
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 0.9
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 1.3
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
1.1.1.337 2 8 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
1.1.1.337 6.58
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
1.1.1.337 9.5
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 20
-
 pyruvate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 20
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
1.1.1.337 22
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
1.1.1.337 25
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
1.1.1.337 27
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
1.1.1.337 32
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
1.1.1.337 51
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
1.1.1.337 52
-
 oxo-tert-leucine pH 7.0, 30°C, wild-type enzyme Weissella confusa
1.1.1.337 90
-
 pyruvate pH 7.0, 30°C, wild-type enzyme Weissella confusa
1.1.1.337 92
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 100
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 150
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 210
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
1.1.1.337 320
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
1.1.1.337 340
-
 pyruvate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 350
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
1.1.1.337 420
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
1.1.1.337 520
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 920
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 1000
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
1.1.1.337 2100
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 2400
-
 2-oxoisocaproate pH 7.0, 30°C, wild-type enzyme Weissella confusa
1.1.1.337 2400
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
1.1.1.337 2500
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 2500
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 3400
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
1.1.1.337 3500
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 4100
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
1.1.1.337 4400
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 5600
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 6400
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 6900
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
1.1.1.337 7300
-
 phenylpyruvate pH 7.0, 30°C, wild-type enzyme Weissella confusa
1.1.1.337 7600
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 7900
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 11000
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 13000
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 15000
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
1.1.1.337 18000
-
 phenylglyoxylate pH 7.0, 30°C, wild-type enzyme Weissella confusa
1.1.1.337 360000
-
 2-oxocaproate pH 7.0, 30°C, wild-type enzyme Weissella confusa

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.337 0.00526
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
1.1.1.337 0.01
-
 pyruvate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
1.1.1.337 0.012
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 0.0465
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
1.1.1.337 0.0469
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 0.0684
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
1.1.1.337 0.07
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 0.357
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 0.411
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
1.1.1.337 0.654
-
 pyruvate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 1.08
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 1.82
-
 pyruvate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 2.97
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
1.1.1.337 3.09
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
1.1.1.337 3.84
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
1.1.1.337 5.88
-
 pyruvate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 6.06
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
1.1.1.337 6.13
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 10
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 13
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
1.1.1.337 15
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme L239M/T245A Weissella confusa
1.1.1.337 16
-
 oxo-tert-leucine pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
1.1.1.337 22.5
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
1.1.1.337 25.7
-
 pyruvate pH 7.0, 30°C, wild-type enzyme Weissella confusa
1.1.1.337 32.8
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 66.2
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
1.1.1.337 117
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme T245A Weissella confusa
1.1.1.337 133
-
 oxo-tert-leucine pH 7.0, 30°C, wild-type enzyme Weissella confusa
1.1.1.337 158
-
 phenylglyoxylate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 400
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
1.1.1.337 677
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
1.1.1.337 941
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
1.1.1.337 1460
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239F Weissella confusa
1.1.1.337 1530
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 1880
-
 phenylglyoxylate pH 7.0, 30°C, wild-type enzyme Weissella confusa
1.1.1.337 1920
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
1.1.1.337 2190
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 2420
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
1.1.1.337 2620
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
1.1.1.337 2670
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 2880
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 5060
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239W Weissella confusa
1.1.1.337 7330
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239A Weissella confusa
1.1.1.337 10000
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme F236V Weissella confusa
1.1.1.337 28700
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 35800
-
 2-oxoisocaproate pH 7.0, 30°C, wild-type enzyme Weissella confusa
1.1.1.337 37600
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 39100
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 49200
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme F236S Weissella confusa
1.1.1.337 76000
-
 2-oxocaproate pH 7.0, 30°C, mutant enzyme G234V/G235D Weissella confusa
1.1.1.337 203000
-
 2-oxoisocaproate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 224000
-
 phenylpyruvate pH 7.0, 30°C, mutant enzyme L239M Weissella confusa
1.1.1.337 281000
-
 phenylpyruvate pH 7.0, 30°C, wild-type enzyme Weissella confusa
1.1.1.337 3270000
-
 2-oxocaproate pH 7.0, 30°C, wild-type enzyme Weissella confusa