EC Number | Application | Comment | Organism |
---|---|---|---|
3.1.1.6 | industry | use of the very active biocatalyst EST1 for chiral resolution of 1,2-O-isopropylidene glycol esters | Kluyveromyces marxianus |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
3.1.1.6 | Ca2+ | 10 mM, 92% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | Co2+ | 10 mM, 56% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | Cu2+ | complete inhibition, 10 mM, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | EDTA | strong inhibitor, 10 mM, 18% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | Fe2+ | 0.1 mM, 70% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | Fe3+ | 0.1 mM, 93% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | Hg2+ | strong inhibitor, 10 mM, 9% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | K+ | 10 mM, 70% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | Mg2+ | 10 mM, 80% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | Mn2+ | 10 mM, 60% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | SDS | complete inhibition, 1% (w/v), short-term incubation, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | Sn2+ | 0.1 mM, 93% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus | |
3.1.1.6 | Zn2+ | complete inhibition, 10 mM, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.1.1.6 | 0.07 | - |
(R)-1,2-O-isopropylidene glycol acetate | EST1, after preparative electrophoresis | Kluyveromyces marxianus |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
3.1.1.6 | 29000 | - |
preparative electrophoresis-purified EST1, SDS-PAGE | Kluyveromyces marxianus |
EC Number | Organic Solvent | Comment | Organism |
---|---|---|---|
3.1.1.6 | Acetone | 15% (v/v), 48% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus |
3.1.1.6 | dimethyl sulfoxide | 15% (v/v), 39% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus |
3.1.1.6 | dimethylformamide | 15% (v/v), 34% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus |
3.1.1.6 | Ethanol | 15% (v/v), 76% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus |
3.1.1.6 | Methanol | 15% (v/v), 82% residual activity, EST1, p-nitrophenyl acetate as substrate | Kluyveromyces marxianus |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.1.1.6 | Kluyveromyces marxianus | - |
CBS 1553 | - |
3.1.1.6 | Kluyveromyces marxianus CBS 1553 | - |
CBS 1553 | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.1.1.6 | 85% (NH4)2SO4-saturated bacterial protein extract to anion-exchange chromatography on DEAE-650 (S) column (gradient elution with 0-0.15 M NaCl), EST1 elution at 70 mM NaCl, EST2 elution at 90 mM NaCl, addition of (NH4)2SO4 (25% saturation) to EST1 fraction followed by hydrophobic interaction chromatography on phenyl Sepharose column, purification of main esterase activity (EST1) by elution with 10% (NH4)2SO4-saturated buffer, preparative electrophoresis (native) | Kluyveromyces marxianus |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
3.1.1.6 | additional information | - |
(R)-1,2-O-isopropylidene glycol acetate, EST1, 1.0 micromol/min by activity corresponding to synthesis of 0.01 micromol p-nitrophenol/min/mg EST1 | Kluyveromyces marxianus |
3.1.1.6 | additional information | - |
(R,S)-1,2-O-isopropylidene glycol acetate as substrate for EST1, 20 micromol/min by activity corresponding to synthesis of 0.01 micromol p-nitrophenol/min/mg EST1 | Kluyveromyces marxianus |
3.1.1.6 | additional information | - |
(R,S)-1,2-O-isopropylidene glycol butyrate as substrate for EST1, 4.6 micromol/min by activity corresponding to synthesis of 0.01 micromol p-nitrophenol/min/mg EST1 | Kluyveromyces marxianus |
3.1.1.6 | additional information | - |
(R,S)-1,2-O-isopropylidene glycol caproate as substrate for EST1, 0.7 micromol/min by activity corresponding to synthesis of 0.01 micromol p-nitrophenol/min/mg EST1 | Kluyveromyces marxianus |
3.1.1.6 | additional information | - |
(S)-1,2-O-isopropylidene glycol acetate, EST1, 1.2 micromol/min by activity corresponding to synthesis of 0.01 micromol p-nitrophenol/min/mg EST1 | Kluyveromyces marxianus |
3.1.1.6 | additional information | - |
triacetin as substrate for EST1, 4.06 micromol/min by activity corresponding to synthesis of 0.01 micromol p-nitrophenol/min/mg EST1 | Kluyveromyces marxianus |
3.1.1.6 | additional information | - |
tributyrin as substrate for EST1, 2.8 micromol/min by activity corresponding to synthesis of 0.01 micromol p-nitrophenol/min/mg EST1 | Kluyveromyces marxianus |
3.1.1.6 | 324 | - |
(R,S)-1,2-O-isopropylidene glycol acetate, EST1, after preparative electrophoresis | Kluyveromyces marxianus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.1.1.6 | (R)-1,2-O-isopropylidene glycol acetate + H2O | - |
Kluyveromyces marxianus | (R)-1,2-O-isopropylidene glycol + acetate | - |
? | |
3.1.1.6 | (R,S)-1,2-O-isopropylidene glycol butyrate + H2O | fresh bacterial cells or purified EST1 (0.01 micromol p-nitrophenol/min), 28°C, pH 8, 5% (v/v) glycerol, 1 mM EDTA, 1 mM dithiothreitol | Kluyveromyces marxianus | (R)-1,2-O-isopropylidene glycol + butyrate | low enantioselectivity (enantiomeric ratio E: 2.5 for EST1, E: 2.5 for whole cells) | ? | |
3.1.1.6 | (R,S)-1,2-O-isopropylidene glycol caproate + H2O | fresh bacterial cells or purified EST1 (0.01 micromol p-nitrophenol/min), 28°C, pH 8, 5% (v/v) glycerol, 1 mM EDTA, 1 mM dithiothreitol | Kluyveromyces marxianus | (R)-1,2-O-isopropylidene glycol + caproate | no enantioselectivity by EST1 (enantiomeric ratio E: 1) compared to E: 2.8 for whole cells | ? | |
3.1.1.6 | alpha-naphthyl acetate + H2O | EST1, non-denaturating PAGE, pH7.4 | Kluyveromyces marxianus | alpha-naphthol + acetate | zymogram analyses using Fast Blue PR salt | ? | |
3.1.1.6 | alpha-naphthyl acetate + H2O | EST1, non-denaturating PAGE, pH7.4 | Kluyveromyces marxianus CBS 1553 | alpha-naphthol + acetate | zymogram analyses using Fast Blue PR salt | ? | |
3.1.1.6 | additional information | activity of EST1 only on short chain triglyceride substrates | Kluyveromyces marxianus | ? | - |
? | |
3.1.1.6 | additional information | EST2 shows lower activity and enantioselectivity towards (R,S)-1,2-O-isopropylidene glycol acetate as substrate than EST1 | Kluyveromyces marxianus | ? | - |
? | |
3.1.1.6 | additional information | no activity of EST1 on p-nitrophenyl butyrate, p-nitrophenyl caproate, p-nitrophenyl caprylate, p-nitrophenyl laurate and p-nitrophenyl palmitate | Kluyveromyces marxianus | ? | - |
? | |
3.1.1.6 | additional information | no activity of EST1 on triacylglyceride esters tricaprylin and triolein | Kluyveromyces marxianus | ? | - |
? | |
3.1.1.6 | additional information | activity of EST1 only on short chain triglyceride substrates | Kluyveromyces marxianus CBS 1553 | ? | - |
? | |
3.1.1.6 | additional information | EST2 shows lower activity and enantioselectivity towards (R,S)-1,2-O-isopropylidene glycol acetate as substrate than EST1 | Kluyveromyces marxianus CBS 1553 | ? | - |
? | |
3.1.1.6 | additional information | no activity of EST1 on p-nitrophenyl butyrate, p-nitrophenyl caproate, p-nitrophenyl caprylate, p-nitrophenyl laurate and p-nitrophenyl palmitate | Kluyveromyces marxianus CBS 1553 | ? | - |
? | |
3.1.1.6 | additional information | no activity of EST1 on triacylglyceride esters tricaprylin and triolein | Kluyveromyces marxianus CBS 1553 | ? | - |
? | |
3.1.1.6 | p-nitrophenyl acetate + H2O | EST1, pH 7.4, 20°C, 0.5% (w/v) Triton X-100, 0.125% (w/v) Arabic gum | Kluyveromyces marxianus | p-nitrophenol + acetate | spectrophotometry, 400 nm | ? | |
3.1.1.6 | triacetin + H2O | EST1, pH 8, 20°C, 0.5% (w/v) Triton X-100, 0.125% (w/v) Arabic gum | Kluyveromyces marxianus | ? | titration assay | ? | |
3.1.1.6 | tributyrin + H2O | EST1, pH 8, 20°C, 0.5% (w/v) Triton X-100, 0.125% (w/v) Arabic gum | Kluyveromyces marxianus | ? | titration assay | ? | |
3.1.1.6 | [(4S)-2,2-dimethyl-1,3-dioxolan-4-yl]methyl acetate + H2O | fresh bacterial cells or purified enzyme (EST1 or EST2, 0.01 micromol p-nitrophenol/min), 20°C, pH 8, 5% (v/v) glycerol, 1 mM EDTA, 1 mM dithiothreitol | Kluyveromyces marxianus | [(4R)-2,2-dimethyl-1,3-dioxolan-4-yl]methanol + acetate | high enantioselectivity (enantiomeric ratio E: 20 for EST1, E: 15 for whole cells), thin-layer chromatography, enantiomeric composition of reactants controlled by gas chromatography, acetate detection spectrophotometrically through NADH absorbance at 340 nm | ? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.1.1.6 | Est1 | - |
Kluyveromyces marxianus |
3.1.1.6 | EST2 | - |
Kluyveromyces marxianus |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.1.1.6 | 80 | - |
100% relative activity, at 60-70°C 90% relative activity, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, pH 7, 3 min | Kluyveromyces marxianus |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.1.1.6 | 40 | 60 | decreasing stability, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, pH 7, 1 h | Kluyveromyces marxianus |
3.1.1.6 | 50 | - |
75% activity, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, pH 7, 1 h | Kluyveromyces marxianus |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
3.1.1.6 | 9 | - |
EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, 20°C | Kluyveromyces marxianus |
EC Number | pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|---|
3.1.1.6 | 5 | - |
inactive below, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, 20°C, 4 h | Kluyveromyces marxianus |
3.1.1.6 | 7 | 10 | 100% activity, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, 20°C, 4 h | Kluyveromyces marxianus |
3.1.1.6 | 11 | - |
16% activity, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, 20°C, 4 h | Kluyveromyces marxianus |