EC Number | Application | Comment | Organism |
---|---|---|---|
4.2.1.95 | synthesis | due to its catalytic properties and apparent substrate promiscuity, enzyme NhKHS is a promising enzyme for the biocatalytic production of tertiary alcohols. The production of enantiopure tertiary alcohols is a major challenge of organic synthesis as these functional groups are widely applicable for the generation of pharmaceuticals or other bioactive compounds | Fusarium solani |
4.2.1.95 | synthesis | due to its catalytic properties and apparent substrate promiscuity, enzyme NhKHS is a promising enzyme for the biocatalytic production of tertiary alcohols. The production of enantiopure tertiary alcohols is a major challenge of organic synthesis as these functional groups are widely applicable for the generation of pharmaceuticals or other bioactive compounds. The compatibility of enzyme NhKHS with nonpolar organic solvents is beneficial for a number of biocatalytic applications, e.g. in general for conversion of hydrophobic substrates | Fusarium solani |
EC Number | Cloned (Comment) | Organism |
---|---|---|
4.2.1.95 | DNA and amino acid sequence determination and analysis, sequence comparisons, higher functional recombinant expression of codon-harmonized gene encoding the enzyme in Pichia pastoris (i.e. Komagataella phaffii) strain CBS7435 as C-terminally His10-tagged protein, the mature enzyme is secreted. Upscaling tests show that Pichia pastoris is a most appropriate host for producing high amounts of recombinant NhKHS | Fusarium solani |
4.2.1.95 | DNA and amino acid sequence determination and analysis, sequence comparisons, lower functional recombinant expression of codon-harmonized gene encoding the enzyme in Pichia pastoris (i.e. Komagataella phaffii) strain CBS7435 as C-terminally His10-tagged protein | Fusarium solani |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.2.1.95 | additional information | - |
additional information | Michaelis-Menten kinetics | Fusarium solani | |
4.2.1.95 | 0.98 | - |
xanthohumol | pH 6.0, 35°C, recombinant enzyme | Fusarium solani |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
4.2.1.95 | additional information | the enzyme contains a 19 amino acid-long N-terminal signal peptide whose cleavage site is predicted between alanine 19 and lysine 20. Absence of nucleotide cofactors is assumed due to the absence of a conserved nucleotide binding motif | Fusarium solani | - |
- |
4.2.1.95 | additional information | the enzyme contains a 19 amino acid-long N-terminal signal peptide whose cleavage site is predicted between alanine 19 and serine 20. The signal peptide is absent from the predominantly secreted mature enzyme | Fusarium solani | - |
- |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
4.2.1.95 | 75000 | - |
recombinant His-tagged mature and glycosylated enzyme, gel filtration | Fusarium solani |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
4.2.1.95 | kievitone hydrate | Fusarium solani | - |
kievitone + H2O | - |
r | |
4.2.1.95 | kievitone hydrate | Fusarium solani 77-13-4 / ATCC MYA-4622 / FGSC 9596 / MPVI | - |
kievitone + H2O | - |
r |
EC Number | Organic Solvent | Comment | Organism |
---|---|---|---|
4.2.1.95 | chloroform | does affect the activity up to 5%, slight inhibition of xanthohumol hydrate formation at 10%, moderate inhibition at 30% | Fusarium solani |
4.2.1.95 | DMSO | slight inhibition of xanthohumol hydrate formation at 5%, higher inhibition at 10%, complete inhibition at 30% | Fusarium solani |
4.2.1.95 | dodecane | does affect the activity up to 30% | Fusarium solani |
4.2.1.95 | Ethanol | does affect the activity up to 5%, moderate inhibition of xanthohumol hydrate formation at 10%, strong inhibition at 30% | Fusarium solani |
4.2.1.95 | hexane | does affect the activity up to 25%, slight inhibition of xanthohumol hydrate formation at 30% | Fusarium solani |
4.2.1.95 | additional information | the polarity of the organic solvent is a determining factor for its effect on NhKHS activity, with increasing solubility in water resulting in a more distinct impact. This might be caused by an enhanced perturbing effect on the hydration shell of the enzyme | Fusarium solani |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
4.2.1.95 | Fusarium solani | C7ZKN4 | i.e. Fusarium solani subsp. pisi | - |
4.2.1.95 | Fusarium solani | Q00870 | i.e. Fusarium solani subsp. phaseoli | - |
4.2.1.95 | Fusarium solani 77-13-4 / ATCC MYA-4622 / FGSC 9596 / MPVI | C7ZKN4 | i.e. Fusarium solani subsp. pisi | - |
EC Number | Posttranslational Modification | Comment | Organism |
---|---|---|---|
4.2.1.95 | glycoprotein | N-glycosylation | Fusarium solani |
4.2.1.95 | glycoprotein | the enzyme contains four typical consensus sequences for N-linked glycosylation. The temperature stability of the enzyme is apparently not affected by deglycosylation in vivo, while in vitro activity assays show that the activity of the deglycosylated enzyme is approx. 30% lower compared to the glycosylated control | Fusarium solani |
EC Number | Purification (Comment) | Organism |
---|---|---|
4.2.1.95 | recombinant His10-tagged enzyme from Pichia pastoris strain CBS7435 by nickel affinity chromatography and gel filtration | Fusarium solani |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
4.2.1.95 | 8-prenylnaringenin + H2O | low activity | Fusarium solani | 8-prenylnaringenin hydrate | - |
r | |
4.2.1.95 | 8-prenylnaringenin + H2O | low activity | Fusarium solani 77-13-4 / ATCC MYA-4622 / FGSC 9596 / MPVI | 8-prenylnaringenin hydrate | - |
r | |
4.2.1.95 | 8-prenylnaringenin hydrate | - |
Fusarium solani | 8-prenylnaringenin + H2O | - |
r | |
4.2.1.95 | 8-prenylnaringenin hydrate | - |
Fusarium solani 77-13-4 / ATCC MYA-4622 / FGSC 9596 / MPVI | 8-prenylnaringenin + H2O | - |
r | |
4.2.1.95 | isoxanthohumol + H2O | - |
Fusarium solani | isoxanthohumol hydrate | - |
r | |
4.2.1.95 | isoxanthohumol hydrate | - |
Fusarium solani | isoxanthohumol + H2O | - |
r | |
4.2.1.95 | kievitone + H2O | best substrate | Fusarium solani | kievitone hydrate | - |
r | |
4.2.1.95 | kievitone + H2O | best substrate | Fusarium solani 77-13-4 / ATCC MYA-4622 / FGSC 9596 / MPVI | kievitone hydrate | - |
r | |
4.2.1.95 | kievitone hydrate | - |
Fusarium solani | kievitone + H2O | - |
r | |
4.2.1.95 | kievitone hydrate | - |
Fusarium solani 77-13-4 / ATCC MYA-4622 / FGSC 9596 / MPVI | kievitone + H2O | - |
r | |
4.2.1.95 | additional information | substrate specificity analysis of the heterologously expressed recombinant enzyme, confirmation of HO-XN formation from XN by 1H-NMR and 13C-NMR in vitro | Fusarium solani | ? | - |
? | |
4.2.1.95 | additional information | substrate specificity analysis of the heterologously expressed recombinant enzyme, confirmation of HO-XN formation from XN by 1H-NMR and 13C-NMR in vitro | Fusarium solani 77-13-4 / ATCC MYA-4622 / FGSC 9596 / MPVI | ? | - |
? | |
4.2.1.95 | xanthohumol | XN, high activity with the recombinant enzyme | Fusarium solani | ? | - |
r | |
4.2.1.95 | xanthohumol + H2O | XN, high activity with the recombinant enzyme | Fusarium solani | xanthohumol hydrate | - |
r | |
4.2.1.95 | xanthohumol hydrate | - |
Fusarium solani | xanthohumol + H2O | - |
r |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
4.2.1.95 | homodimer | - |
Fusarium solani |
4.2.1.95 | homodimer | 2 * 48000, recombinant His-tagged mature and glycosylated enzyme, SDS-PAGE | Fusarium solani |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
4.2.1.95 | FsKHS | - |
Fusarium solani |
4.2.1.95 | NhKHS | - |
Fusarium solani |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
4.2.1.95 | 35 | - |
recombinant enzyme | Fusarium solani |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
4.2.1.95 | additional information | - |
the temperature stability of the enzyme is apparently not affected by deglycosylation in vivo, while in vitro activity assays show that the activity of the deglycosylated enzyme is approx. 30% lower compared to the glycosylated control | Fusarium solani |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.2.1.95 | 4.77 | - |
xanthohumol | pH 6.0, 35°C, recombinant enzyme | Fusarium solani |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
4.2.1.95 | 6 | - |
recombinant enzyme | Fusarium solani |
EC Number | pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|---|
4.2.1.95 | 4 | 8 | activity range, recombinant enzyme, profile overview | Fusarium solani |
EC Number | General Information | Comment | Organism |
---|---|---|---|
4.2.1.95 | evolution | the enzyme belongs to the hydro-lyases, which are able to catalyze the highly selective, reversible addition of water to non-activated carbon-carbon double bonds and, thereby, generate primary, secondary or tertiary alcohols | Fusarium solani |
4.2.1.95 | evolution | the enzyme is a member of the hydrolyase enzyme group | Fusarium solani |
4.2.1.95 | evolution | the enzyme is a member of the hydrolyase enzyme group. Hydro-lyases are able to catalyze the highly selective, reversible addition of water to non-activated carbon-carbon double bonds and, thereby, generate primary, secondary or tertiary alcohols | Fusarium solani |
4.2.1.95 | physiological function | kievitone hydratase catalyzes the addition of water to the double bond of the prenyl moiety of plant isoflavonoid kievitone and, thereby, forms the tertiary alcohol hydroxy-kievitone. In nature, this conversion is associated with a defense mechanism of fungal pathogens against phytoalexins generated by host plants after infection | Fusarium solani |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
4.2.1.95 | 4.88 | - |
xanthohumol | pH 6.0, 35°C, recombinant enzyme | Fusarium solani |