EC Number | Cloned (Comment) | Organism |
---|---|---|
3.1.1.59 | recombinant baculovirus AcMsJHE-F259I expressing mutant F259I, recombinant baculovirus AcMsJHE-T314V or recombinant baculovirus pAcUW21-T314V expressing mutant T314V. AcMsJHE-T314V generated by transfection of Sf-21 cells with pAcUW21-T314V and linearized BacPAK6 baculovirus DNA. Wild-type JHE, mutants F259I and T314V produced in insect High Five cells cultured in ESF 921 medium that are inoculated with AcMsJHE7, AcMsJHE-F259I, or AcMsJHE-T314V, respectively | Manduca sexta |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.1.1.59 | F259I | the overall structure of the substrate binding pocket is similar to that of wild-type JHE. Shows a 5.7fold higher kcat for juvenile hormone III as the wild-type. KM of mutant F259I for juvenile hormone III is 3.4fold higher than that of the wild-type. With the saturated form of juvenile hormone III as a substrate, mutant F259I shows a 4.4fold higher kcat and 14fold higher Km in comparison to the wild-type | Manduca sexta |
3.1.1.59 | T314V | no major changes in the overall structure of the substrate binding pocket. The KM of mutant T314V for juvenile hormone III is 2.9fold higher than that of JHE, whereas kcat is 6.4fold higher. Specific activity of mutant T314V for juvenile hormone III diol is not statistically different than that of wild-type for juvenile hormone III | Manduca sexta |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.1.1.59 | 0.00003 | - |
juvenile hormone III | wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 0.000046 | - |
saturated juvenile hormone III | wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 0.000086 | - |
juvenile hormone III | mutant T314V, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 0.000101 | - |
juvenile hormone III | mutant F259I, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 0.000641 | - |
saturated juvenile hormone III | mutant F259I, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 0.41 | - |
Alpha-naphthyl acetate | wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.1.1.59 | Manduca sexta | Q9GPG0 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.1.1.59 | expressed wild-type and mutant proteins purified by anion-exchange chromatography. Purification efficiencies of wild-type JHE, mutant F259I, and mutant T314V are 77%, 32%, and 51%, respectively and are purified 4-, 4.4-, and 4.5fold, respectively | Manduca sexta |
EC Number | Source Tissue | Comment | Organism | Textmining |
---|
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
3.1.1.59 | 500000 | - |
purified wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 0.0005 mM juvenile hormone III and 1 mg/mL of bovine serum albumin | Manduca sexta |
3.1.1.59 | 1050000 | - |
purified wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 0.0005 mM juvenile hormone III diol and 1 mg/mL of bovine serum albumin | Manduca sexta |
3.1.1.59 | 2370000 | - |
purified mutant F259I, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 0.0005 mM juvenile hormone III and 1 mg/mL of bovine serum albumin | Manduca sexta |
3.1.1.59 | 4200000 | - |
purified mutant T314V, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 0.0005 mM juvenile hormone III and 1 mg/mL of bovine serum albumin | Manduca sexta |
3.1.1.59 | 4880000 | - |
purified mutant T314V, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 0.0005 mM juvenile hormone III diol and 1 mg/mL of bovine serum albumin | Manduca sexta |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.1.1.59 | alpha-naphthyl acetate + H2O | - |
Manduca sexta | alpha-naphthol + acetate | - |
? | |
3.1.1.59 | juvenile hormone II + H2O | - |
Manduca sexta | juvenile hormone II acid + methanol | - |
? | |
3.1.1.59 | juvenile hormone III + H2O | juvenile hormone III fits very tightly within the substrate binding pocket of JHE. This tight fit places two noncatalytic amino acid residues, Phe-259 and Thr-314, within the appropriate distance and geometry to potentially interact with the R,beta-unsaturated ester and epoxide, respectively, of juvenile hormone III. Neither nucleophilic attack of the resonance-stabilized ester by the catalytic serine nor the availability of a water molecule for attack of the acyl-enzyme intermediate appears to be a rate-determining step in the hydrolysis. Release of the juvenile hormone III acid metabolite from the substrate binding pocket limits the catalytic cycle. Chemical bond strength does not necessarily correlate with how reactive the bond will be to metabolism | Manduca sexta | juvenile hormone III acid + methanol | - |
? | |
3.1.1.59 | juvenile hormone III diol + H2O | although hydrogen bonding appears not to occur between juvenile hormone III diol and Thr-314, the hydroxyls of the diol (at C10 and C11) are within hydrogen bonding distance of the main chain carbonyl oxygen atoms of Phe-259 and Phe-311 | Manduca sexta | ? | - |
? | |
3.1.1.59 | saturated juvenile hormone III + H2O | alpha,beta-saturated methyl ester | Manduca sexta | saturated juvenile hormone III acid + methanol | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.1.1.59 | JHE | - |
Manduca sexta |
3.1.1.59 | Juvenile hormone esterase | - |
Manduca sexta |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.1.1.59 | 0.7 | - |
juvenile hormone III | wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 0.7 | - |
saturated juvenile hormone III | wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 3.1 | - |
saturated juvenile hormone III | mutant F259I, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 4 | - |
juvenile hormone III | mutant F259I, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 4.5 | - |
juvenile hormone III | mutant T314V, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 22 | - |
Alpha-naphthyl acetate | wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.1.1.59 | 53.65 | - |
Alpha-naphthyl acetate | wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 4800 | - |
saturated juvenile hormone III | mutant F259I, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 14000 | - |
saturated juvenile hormone III | wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 22000 | - |
juvenile hormone III | wild-type, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 40000 | - |
juvenile hormone III | mutant F259I, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta | |
3.1.1.59 | 53000 | - |
juvenile hormone III | mutant T314V, at 30°C in 50 mM sodium phosphate buffer, pH 7.4, containing 1 mg/mL of bovine serum albumin | Manduca sexta |