Lysophospholipase from Pyrococcus furiosus

Enzyme Description

Species

Pyrococcus furiosus ↗

Extremophile

Yes hyperthermophilic organism

EC Number

3.1.1.3 ↗

Sequence

Length: 286 amino acids

MASMTGGQQMGRGSSPGHHHHHHHHMTQVYKAKFGTPNRGWVIIVHGLGEHSGRYSKLVSMLVNEGYAVYTFDWPGHGKSPGKRGHTSVEEAMEIIDFIIEEINDKPFLFGHSLGGLTVIRYAETRPEKIRGVIASSPALAKSPKTPSFMVALAKILGVLLPSLTLSNGIDPNLLSRNPDAVKRYIEDPLVHDRISAKLGRSIFKNMDLAHREAHKIKVPVLLLVGTGDVITPPEGARKLYGEIKVEDKEIVEFEGAYHEIFEDPEWGEEFHKKIVEWIKKHHHHH

Sanjeev Kumar Chandrayan et al. (2008) — 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

Protein Expression and Purification · doi:10.1016/j.pep.2008.02.019 ↗ · Activity - Classical

7 measurements ▼

Database ID

UniProt: Q8U3I6 ↗

Sequence Annotation

Explicit (first 25 residues and last 5 residues from the expression vector and not in UniProt sequence)

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3) pLysS

Experimental Data (7 measurements)

7 measurements

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Acetonitrile	25%	100	57	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol , Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Acetonitrile	50%	100	1	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol , Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Methanol	25%	100	21	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol , Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Methanol	50%	100	6	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol , Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Ethanol	25%	100	38	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol , Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Ethanol	50%	100	1	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol , Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Phenylmethylsulfonyl Fluoride (PMSF)	1 mM	100	11	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol , Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed

Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Acetonitrile	25%	100	57	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol, Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Acetonitrile	50%	100	1	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol, Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Methanol	25%	100	21	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol, Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Methanol	50%	100	6	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol, Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Ethanol	25%	100	38	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol, Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Ethanol	50%	100	1	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol, Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed
Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent	Phenylmethylsulfonyl Fluoride (PMSF)	1 mM	100	11	%	10 mM phosphate buffer, 1.0% (w/v) gum arabic and 0.2% Triton X-100	7.5	70°C	0.5 mM p-Nitrophenyl palmitate	p-Nitrophenol, Palmitic Acid	—	—	Classical aqueous control (in %) \| Hist-tag not removed

Visualization : Activity — Classical

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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