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Literature summary extracted from

  • Chandrayan, S.K.; Dhaunta, N.; Guptasarma, P.
    Expression, purification, refolding and characterization of a putative lysophospholipase from Pyrococcus furiosus: retention of structure and lipase/esterase activity in the presence of water-miscible organic solvents at high temperatures (2008), Protein Expr. Purif., 59, 327-333.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.3 expressed in Escherichia coli BL21 (DE3) cells Pyrococcus furiosus
3.1.1.5 into the vector pGEX-KG and subsequently into pET23a for expression in Escherichia coli BL21DE3 pLysS cells Pyrococcus furiosus

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.3 biofuel the high organic solvent stability and thermal stability suggest that this enzyme may have useful biodiesel-related applications Pyrococcus furiosus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.1.3 phenylmethylsulfonyl fluoride remarkable reduction in activity at 1 mM Pyrococcus furiosus
3.1.1.5 phenylmethylsulfonyl fluoride PMSF Pyrococcus furiosus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.1.3 28000
-
recombinant protein, approximately 28000-29000 Da, SDS-PAGE Pyrococcus furiosus
3.1.1.3 31980
-
recombinant protein, MALDI-TOF spectrometry Pyrococcus furiosus
3.1.1.3 32040
-
recombinant protein, calculated from amino acid sequence Pyrococcus furiosus
3.1.1.3 82000
-
dynamic light scattering Pyrococcus furiosus
3.1.1.5 29000
-
determined by SDS-PAGE Pyrococcus furiosus
3.1.1.5 32040
-
calculated Pyrococcus furiosus

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
3.1.1.3 acetonitrile over 50% activity is retained at 70°C in the presence of 25% (v/v) acetonitrile Pyrococcus furiosus
3.1.1.5 acetonitrile 25%, substrate p-nitrophenyl palmitate, relative activiy 58% Pyrococcus furiosus
3.1.1.5 Ethanol 25%, substrate p-nitrophenyl palmitate, relative activiy 39% Pyrococcus furiosus
3.1.1.5 Methanol 25%, substrate p-nitrophenyl palmitate, relative activiy 20% Pyrococcus furiosus

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.3 Pyrococcus furiosus Q8U3I6
-
-
3.1.1.5 Pyrococcus furiosus Q8TZI9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.3 Ni-NTA affinity column chromatography Pyrococcus furiosus
3.1.1.5 using a Ni-NTA affiniity column Pyrococcus furiosus

Renatured (Commentary)

EC Number Renatured (Comment) Organism
3.1.1.3 through heating at 80°C for 30 min in the presence of 6 M Gdm-HCl and 1 M NaCl Pyrococcus furiosus
3.1.1.5 the enzyme is refolded from inclusion bodies Pyrococcus furiosus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.3 4-nitrophenyl acetate + H2O
-
Pyrococcus furiosus 4-nitrophenol + acetate
-
?
3.1.1.3 4-nitrophenyl butyrate + H2O
-
Pyrococcus furiosus 4-nitrophenol + butyrate
-
?
3.1.1.3 4-nitrophenyl palmitate + H2O
-
Pyrococcus furiosus 4-nitrophenol + palmitate
-
?
3.1.1.5 p-nitrophenyl acetate + H2O substrate activity assay Pyrococcus furiosus p-nitrophenol + acetate
-
?
3.1.1.5 p-nitrophenyl butyrate substrate activity assay Pyrococcus furiosus p-nitrophenol + butyrate
-
?
3.1.1.5 p-nitrophenyl palmitate substrate activity assay Pyrococcus furiosus p-nitrophenol + palmitate
-
?

Subunits

EC Number Subunits Comment Organism
3.1.1.3 dimer or trimer in aqueous solution, dynamic light scattering Pyrococcus furiosus
3.1.1.5 homodimer prediction, based on the determination of the hydrodynamic radius by dynamic light scattering measurements Pyrococcus furiosus

Synonyms

EC Number Synonyms Comment Organism
3.1.1.3 putative lysophospholipase the enzyme is confirmed to be an esterase and also shown to be a lipase (hydrolyzing 4-nitrophenyl palmitate), with lipolytic activity being overall about 18 to 20fold lower than esterase activity Pyrococcus furiosus
3.1.1.5 lysophospholipase
-
Pyrococcus furiosus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.1.3 70
-
using 4-nitrophenyl palmitate as a substrate, for a fixed pH of 8.0 Pyrococcus furiosus
3.1.1.5 70
-
-
Pyrococcus furiosus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.1.1.3 30 80
-
Pyrococcus furiosus
3.1.1.5 30 80 conformational changes are negligible over the temperature range of 30-80°C Pyrococcus furiosus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.3 85 95 the lysophospholipase begins to unfold only above 85°C, with unfolding remaining incomplete even at 95°C Pyrococcus furiosus
3.1.1.5 85
-
begins to unfold above 85°C, with unfolding remaining incomplete even at 95°C Pyrococcus furiosus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.3 7
-
using 4-nitrophenyl palmitate as a substrate, for a fixed temperature at 70°C Pyrococcus furiosus
3.1.1.5 7
-
-
Pyrococcus furiosus