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Literature summary extracted from

  • Morikubo, N.; Fukuda, Y.; Ohtake, K.; Shinya, N.; Kiga, D.; Sakamoto, K.; Asanuma, M.; Hirota, H.; Yokoyama, S.; Hoshino, T.
    Cation-pi interaction in the polyolefin cyclization cascade uncovered by incorporating unnatural amino acids into the catalytic sites of squalene cyclase (2006), J. Am. Chem. Soc., 128, 13184-13194.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
5.4.99.17 F365W marginal catalytic activity Alicyclobacillus acidocaldarius
5.4.99.17 F365Y and mutant with F365 changed to unnatural amino acid O-methyltyrosine. Both show increased decreased activity at high temperature Alicyclobacillus acidocaldarius
5.4.99.17 F605W increased catalytic acitivy at low temperature, but decreased activity at high temperature due to higher cation-pi binding energies Alicyclobacillus acidocaldarius
5.4.99.17 F605Y and mutant with F605 changed to unnatural amino acid O-methyltyrosine. Both show increased catalytic acitivy at low temperature, but decreased activity at high temperature due to higher cation-pi binding energies Alicyclobacillus acidocaldarius
5.4.99.17 additional information replacement of F605 by mono-, di- or trifluorophenylalanine, with or without additional mutation Y606A, kinetic analysis. Mutant F605 changed to trifluorophenylalanine plus mutation Y606A has negligibly small activity Alicyclobacillus acidocaldarius
5.4.99.17 Y606A kinetics identical to wild-type Alicyclobacillus acidocaldarius

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.4.99.17 0.0154
-
 squalene wild-type, pH 6.0, 40°C Alicyclobacillus acidocaldarius
5.4.99.17 0.0242
-
 squalene wild-type, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.0279
-
 squalene mutant F605 changed to fluorophenylalanine plus mutation Y606A, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.0324
-
 squalene mutant F605 changed to difluorophenylalanine plus mutation Y606A, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.0352
-
 squalene mutant F365 changed to fluorophenylalanine, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.0416
-
 squalene mutant F365 changed to difluorophenylalanine, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.0479
-
 squalene mutant F6365 changed to trifluorophenylalanine, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.0525
-
 squalene mutant F605Y, pH 6.0, 40°C Alicyclobacillus acidocaldarius
5.4.99.17 0.104
-
 squalene mutant F605Y, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.105
-
 squalene mutant F605 changed to O-methyltyrosine, pH 6.0, 40°C Alicyclobacillus acidocaldarius
5.4.99.17 0.118
-
 squalene mutant F605W, pH 6.0, 40°C Alicyclobacillus acidocaldarius
5.4.99.17 0.13
-
 squalene mutant F365Y, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.152
-
 squalene mutant F605Y changed to O-methyltyrosine, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.185
-
 squalene mutant F365Y changed to O-methyltyrosine, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.191
-
 squalene mutant F605W, pH 6.0, 50°C Alicyclobacillus acidocaldarius

Organism

EC Number Organism UniProt Comment Textmining
5.4.99.17 Alicyclobacillus acidocaldarius
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.4.99.17 squalene
-
 Alicyclobacillus acidocaldarius hop-22(29)-ene
-
 ?

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.4.99.17 55
-
 wild-type Alicyclobacillus acidocaldarius

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.4.99.17 0.15
-
 squalene mutant F365Y changed to O-methyltyrosine, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.17
-
 squalene mutant F365Y, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.25
-
 squalene wild-type, pH 6.0, 40°C Alicyclobacillus acidocaldarius
5.4.99.17 0.27
-
 squalene mutant F605Y, pH 6.0, 40°C Alicyclobacillus acidocaldarius
5.4.99.17 0.31
-
 squalene mutant F605W, pH 6.0, 40°C Alicyclobacillus acidocaldarius
5.4.99.17 0.45
-
 squalene mutant F605 changed to O-methyltyrosine, pH 6.0, 40°C Alicyclobacillus acidocaldarius
5.4.99.17 0.54
-
 squalene mutant F605 changed to difluorophenylalanine plus mutation Y606A, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.7
-
 squalene mutant F605Y changed to O-methyltyrosine, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.75
-
 squalene mutant F6365 changed to trifluorophenylalanine, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.82
-
 squalene mutant F605 changed to fluorophenylalanine plus mutation Y606A, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 0.92
-
 squalene mutant F365 changed to difluorophenylalanine, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 1.1
-
 squalene mutant F365 changed to fluorophenylalanine, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 1.32
-
 squalene mutant F605W, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 1.82
-
 squalene mutant F605Y, pH 6.0, 50°C Alicyclobacillus acidocaldarius
5.4.99.17 1.98
-
 squalene wild-type, pH 6.0, 50°C Alicyclobacillus acidocaldarius