Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.1.1.6 extracted from

  • Monti, D.; Ferrandi, E.E.; Righi, M.; Romano, D.; Molinari, F.
    Purification and characterization of the enantioselective esterase from Kluyveromyces marxianus CBS 1553 (2008), J. Biotechnol., 133, 65-72.
    View publication on PubMed
Search for more literature for 3.1.1.6

Application

Application Comment Organism
industry use of the very active biocatalyst EST1 for chiral resolution of 1,2-O-isopropylidene glycol esters Kluyveromyces marxianus

Inhibitors

Inhibitors Comment Organism Structure
Ca2+ 10 mM, 92% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Co2+ 10 mM, 56% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Cu2+ complete inhibition, 10 mM, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
EDTA strong inhibitor, 10 mM, 18% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Fe2+ 0.1 mM, 70% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Fe3+ 0.1 mM, 93% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Hg2+ strong inhibitor, 10 mM, 9% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
K+ 10 mM, 70% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Mg2+ 10 mM, 80% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Mn2+ 10 mM, 60% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
SDS complete inhibition, 1% (w/v), short-term incubation, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Sn2+ 0.1 mM, 93% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Zn2+ complete inhibition, 10 mM, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.07
-
 (R)-1,2-O-isopropylidene glycol acetate EST1, after preparative electrophoresis Kluyveromyces marxianus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
29000
-
 preparative electrophoresis-purified EST1, SDS-PAGE Kluyveromyces marxianus

Organic Solvent Stability

Organic Solvent Comment Organism
Acetone 15% (v/v), 48% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
dimethyl sulfoxide 15% (v/v), 39% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
dimethylformamide 15% (v/v), 34% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Ethanol 15% (v/v), 76% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus
Methanol 15% (v/v), 82% residual activity, EST1, p-nitrophenyl acetate as substrate Kluyveromyces marxianus

Organism

Organism UniProt Comment Textmining
Kluyveromyces marxianus
-
 CBS 1553
-
Kluyveromyces marxianus CBS 1553
-
 CBS 1553
-

Purification (Commentary)

Purification (Comment) Organism
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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
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
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
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
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
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
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
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
324
-
 (R,S)-1,2-O-isopropylidene glycol acetate, EST1, after preparative electrophoresis Kluyveromyces marxianus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(R)-1,2-O-isopropylidene glycol acetate + H2O
-
 Kluyveromyces marxianus (R)-1,2-O-isopropylidene glycol + acetate
-
 ?
(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) ?
(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 ?
alpha-naphthyl acetate + H2O EST1, non-denaturating PAGE, pH7.4 Kluyveromyces marxianus alpha-naphthol + acetate zymogram analyses using Fast Blue PR salt ?
alpha-naphthyl acetate + H2O EST1, non-denaturating PAGE, pH7.4 Kluyveromyces marxianus CBS 1553 alpha-naphthol + acetate zymogram analyses using Fast Blue PR salt ?
additional information activity of EST1 only on short chain triglyceride substrates Kluyveromyces marxianus ?
-
 ?
additional information EST2 shows lower activity and enantioselectivity towards (R,S)-1,2-O-isopropylidene glycol acetate as substrate than EST1 Kluyveromyces marxianus ?
-
 ?
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 ?
-
 ?
additional information no activity of EST1 on triacylglyceride esters tricaprylin and triolein Kluyveromyces marxianus ?
-
 ?
additional information activity of EST1 only on short chain triglyceride substrates Kluyveromyces marxianus CBS 1553 ?
-
 ?
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 ?
-
 ?
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 ?
-
 ?
additional information no activity of EST1 on triacylglyceride esters tricaprylin and triolein Kluyveromyces marxianus CBS 1553 ?
-
 ?
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 ?
triacetin + H2O EST1, pH 8, 20°C, 0.5% (w/v) Triton X-100, 0.125% (w/v) Arabic gum Kluyveromyces marxianus ? titration assay ?
tributyrin + H2O EST1, pH 8, 20°C, 0.5% (w/v) Triton X-100, 0.125% (w/v) Arabic gum Kluyveromyces marxianus ? titration assay ?
[(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 ?

Synonyms

Synonyms Comment Organism
Est1
-
 Kluyveromyces marxianus
EST2
-
 Kluyveromyces marxianus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
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

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40 60 decreasing stability, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, pH 7, 1 h Kluyveromyces marxianus
50
-
 75% activity, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, pH 7, 1 h Kluyveromyces marxianus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
9
-
 EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, 20°C Kluyveromyces marxianus

pH Range

pH Minimum pH Maximum Comment Organism
5
-
 inactive below, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, 20°C, 4 h Kluyveromyces marxianus
7 10 100% activity, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, 20°C, 4 h Kluyveromyces marxianus
11
-
 16% activity, EST1 sample after anion-exchange chromatography, p-nitrophenyl acetate as substrate, 20°C, 4 h Kluyveromyces marxianus