EC Number | Application | Comment | Organism |
---|---|---|---|
3.1.1.73 | industry | molecular details of Est1E from the suite of fiber-degrading enzymes from important rumen bacterium is a step toward mapping the complex conversion of plant biomass to valuable products-meat and milk-by ruminants | Butyrivibrio proteoclasticus |
EC Number | Cloned (Comment) | Organism |
---|---|---|
3.1.1.73 | PCR product cloned in the pDONR221 GATEWAY entry vecto. For protein expression, insert recombined into the pDEST17 GATEWAY expression vector and transformed into Escherichia coli BL21 (DE3) cells | Butyrivibrio proteoclasticus |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
3.1.1.73 | crystals of apo-Est1E grown in 17-22% PEG3350, 0.30.5 M NaH2PO4, to 1.6 A resolution. Crystals belong to space group P21 with unit cell dimensions a = 52.0, b = 108.8, c = 91.4 A, beta = 102.28, with four Est1E molecules in the asymmetric unit. Overall structure of apo-Est1E is a canonical alpha/beta-hydrolase fold with an insertion in the loop between the 6th and 7th strands of the central beta-sheet. Crystals of Se-Met Est1E, to 2.0 A resolution, belong to space group P21 with cell dimensions a = 41.0, b = 129.6, c = 50.6 A, beta = 106.28, with two molecules in the asymmetric unit. Est1E in complex with ferulic acid, to 2.1 A resolution, crystals belong to space group C2 and with a dimer in the asymmetric unit. Overall structure of Est1E with ferulic acid bound is unchanged from the apostructure | Butyrivibrio proteoclasticus |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.1.1.73 | 2 | 3 | p-nitrophenyl acetate | in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus | |
3.1.1.73 | 24 | - |
p-nitrophenyl butyrate | in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
3.1.1.73 | 75000 | - |
gel filtration, in solution | Butyrivibrio proteoclasticus |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.1.1.73 | Butyrivibrio proteoclasticus | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.1.1.73 | by centrifugation, immobilized metal affinity chromatography and gel filtration | Butyrivibrio proteoclasticus |
EC Number | Source Tissue | Comment | Organism | Textmining |
---|
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
3.1.1.73 | 2.6 | - |
with ethyl cinnamate as substrate, in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
3.1.1.73 | 2.9 | - |
with xylan birchwood as substrate and p-coumaric acid as product, in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
3.1.1.73 | 4 | - |
with hemicellulose of ryegrass as substrate and trans-cinnamic acid as product, in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
3.1.1.73 | 4.1 | - |
with hemicellulose of ryegrass as substrate and p-coumaric acid as product, in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
3.1.1.73 | 6.9 | - |
with hemicellulose of ryegrass as substrate and ferulic acid as product, in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
3.1.1.73 | 7.1 | - |
with xylan birchwood as substrate and 3-coumarincarboxylic acid as product, in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
3.1.1.73 | 7.9 | - |
with xylan birchwood as substrate and trans-cinnamic acid as product, in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
3.1.1.73 | 37 | - |
with ethyl ferulate as substrate, in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
3.1.1.73 | 199 | - |
with ethyl 3-coumarincarboxylate as substrate, in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.1.1.73 | ethyl 3-coumarincarboxylate + H2O | - |
Butyrivibrio proteoclasticus | 3-coumarincarboxylic acid + ethanol | - |
? | |
3.1.1.73 | ethyl cinnamate + H2O | - |
Butyrivibrio proteoclasticus | cinnamic acid + ethanol | - |
? | |
3.1.1.73 | ethyl ferulate + H2O | - |
Butyrivibrio proteoclasticus | ferulic acid + ethanol | - |
? | |
3.1.1.73 | additional information | active site of Est1E is located in a groove at the C-terminus of strand beta5 and is marked in each monomer by the presence of a phosphate molecule. The conserved catalytic triad is formed by the residues Ser105, His225, and Asp197. The nucleophile, Ser105, is located in a tight turn between strand beta5 and alpha3 forming the nucleophile elbow. The histidine residue, His225, is located in a loop prior to alpha8. The catalytic triad is completed by Asp197 located in the loop between beta11 and alpha7 | Butyrivibrio proteoclasticus | ? | - |
? | |
3.1.1.73 | p-nitrophenyl acetate + H2O | - |
Butyrivibrio proteoclasticus | p-nitrophenol + acetic acid | - |
? | |
3.1.1.73 | p-nitrophenyl butyrate + H2O | - |
Butyrivibrio proteoclasticus | p-nitrophenol + butyric acid | - |
? | |
3.1.1.73 | p-nitrophenyl laurate + H2O | - |
Butyrivibrio proteoclasticus | p-nitrophenol + lauric acid | - |
? | |
3.1.1.73 | p-nitrophenyl palmitate + H2O | - |
Butyrivibrio proteoclasticus | p-nitrophenol + palmitic acid | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.1.1.73 | dimer | gel filtration, in solution | Butyrivibrio proteoclasticus |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.1.1.73 | Est1E | - |
Butyrivibrio proteoclasticus |
3.1.1.73 | FA esterase | - |
Butyrivibrio proteoclasticus |
3.1.1.73 | feruloyl esterase | - |
Butyrivibrio proteoclasticus |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.1.1.73 | 0.19 | - |
p-nitrophenyl acetate | in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus | |
3.1.1.73 | 0.24 | - |
p-nitrophenyl butyrate | in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
3.1.1.73 | 100 | - |
p-nitrophenyl butyrate | in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus | |
3.1.1.73 | 120 | - |
p-nitrophenyl acetate | in 100 mM phosphate buffer, pH 7.2, at 37°C | Butyrivibrio proteoclasticus |