Literature summary for 3.1.2.12 extracted from

Legler, P.M.; Kumaran, D.; Swaminathan, S.; Studier, F.W.; Millard, C.B.
Structural characterization and reversal of the natural organophosphate resistance of a D-type esterase, Saccharomyces cerevisiae S-formylglutathione hydrolase (2008), Biochemistry, 47, 9592-9601.

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli	Saccharomyces cerevisiae

Crystallization (Commentary)

Crystallization (Comment)	Organism
sitting drop vapour diffusion method, using 0.17 M ammonium acetate, 0.085 M sodium acetate trihydrate (pH 4.6), 25.5% (w/v) PEG 4000, and 15% (v/v) glycerol	Saccharomyces cerevisiae

Protein Variants

Protein Variants	Comment	Organism
C60H/W197I	mutant shows significantly decreased activity	Saccharomyces cerevisiae
C60K/W197I	the mutation results in a further enhancement of the rates of phosphorylation with paraoxon but does not affect the dephosphorylation of the enzyme	Saccharomyces cerevisiae
C60Q/W197I	mutant shows significantly decreased activity	Saccharomyces cerevisiae
C60R/W197I	the mutation results in a further enhancement of the rates of phosphorylation with paraoxon but does not affect the dephosphorylation of the enzyme	Saccharomyces cerevisiae
C60S	mutant shows significantly decreased activity	Saccharomyces cerevisiae
C60S/W197I	mutant shows significantly decreased activity	Saccharomyces cerevisiae
G57H	mutant shows significantly decreased activity	Saccharomyces cerevisiae
L58H/W197I	mutant shows significantly decreased activity	Saccharomyces cerevisiae
M162H	mutant shows significantly decreased activity	Saccharomyces cerevisiae
M162H/C60S/W197I	mutant shows significantly decreased activity	Saccharomyces cerevisiae
W197I	the substitution enhances SFGH reactivity with paraoxon by more than 1000fold thereby overcoming natural organophosphate resistance, the mutant increases the rate of organophosphate inhibition under pseudo-first-order conditions but does not accelerate organophosphate hydrolysis	Saccharomyces cerevisiae

Inhibitors

Inhibitors	Comment	Organism
Hg2+	susceptible to inhibition by Hg2+	Saccharomyces cerevisiae
methyl paraoxon	-	Saccharomyces cerevisiae
paraoxon	-	Saccharomyces cerevisiae

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.012	-	4-nitrophenyl butyrate	mutant enzyme C60Q/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.013	-	4-nitrophenyl acetate	mutant enzyme C60R/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.018	-	4-nitrophenyl butyrate	mutant enzyme C60R/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.019	-	4-nitrophenyl butyrate	mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.03	-	4-nitrophenyl butyrate	mutant enzyme C60K/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.04	-	4-nitrophenyl butyrate	mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.054	-	4-nitrophenyl acetate	mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.09	-	4-nitrophenyl butyrate	wild type enzyme, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.1	-	4-nitrophenyl acetate	mutant enzyme C60Q/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.12	-	4-nitrophenyl butyrate	mutant enzyme M162H/C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.13	-	4-nitrophenyl butyrate	mutant enzyme C60H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.14	-	4-nitrophenyl acetate	mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.14	-	4-nitrophenyl butyrate	mutant enzyme M162H, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.15	-	4-nitrophenyl acetate	mutant enzyme C60K/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.19	-	4-nitrophenyl acetate	mutant enzyme C60S, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.19	-	4-nitrophenyl butyrate	mutant enzyme C60S, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.2	-	4-nitrophenyl butyrate	Km less than 0.2 mM, mutant enzyme L58H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.23	-	4-nitrophenyl acetate	mutant enzyme L58H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.34	-	4-nitrophenyl acetate	mutant enzyme C60H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.4	-	4-nitrophenyl acetate	Km above 1.25 mM, wild type enzyme, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.9	-	4-nitrophenyl acetate	mutant enzyme M162H, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
1.25	-	S-Lactoylglutathione	Km above 1.25 mM, wild type enzyme, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
2.6	-	4-nitrophenyl acetate	mutant enzyme M162H/C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
3	-	S-Lactoylglutathione	mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
3	-	S-Lactoylglutathione	mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
33930	-	calculated from amino acid sequence	Saccharomyces cerevisiae
33934	-	1 * 33934, calculated from amino acid sequence	Saccharomyces cerevisiae
67800	-	gel filtration	Saccharomyces cerevisiae

Organism

Organism	UniProt	Comment	Textmining
Saccharomyces cerevisiae	P40363	-	-

Oxidation Stability

Oxidation Stability	Organism
the wild type enzyme is sensitive to oxidation	Saccharomyces cerevisiae

Purification (Commentary)

Purification (Comment)	Organism
-	Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
4-nitrophenyl acetate + H2O	-	Saccharomyces cerevisiae	4-nitrophenol + acetate	-	?
4-nitrophenyl butyrate + H2O	-	Saccharomyces cerevisiae	4-nitrophenol + butyrate	-	?
paraoxon + H2O	-	Saccharomyces cerevisiae	diethylphosphate + 4-nitrophenol	-	?
S-formylglutathione + H2O	-	Saccharomyces cerevisiae	glutathione + formate	-	?
S-lactoylglutathione + H2O	-	Saccharomyces cerevisiae	glutathione + lactate	-	?

Subunits

Subunits	Comment	Organism
homodimer	the wild type enzyme crystallizes as a dimer of dimers with four molecules in the asymmetric unit, X-ray crystallography	Saccharomyces cerevisiae
monomer	1 * 33934, calculated from amino acid sequence	Saccharomyces cerevisiae

Synonyms

Synonyms	Comment	Organism
esterase D	-	Saccharomyces cerevisiae
SFGH	-	Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.003	-	4-nitrophenyl butyrate	mutant enzyme M162H, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.0055	-	4-nitrophenyl acetate	mutant enzyme L58H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.0165	-	4-nitrophenyl butyrate	kcat less than 0.0165 s-1, mutant enzyme L58H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.028	-	4-nitrophenyl acetate	mutant enzyme M162H/C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.04	-	4-nitrophenyl butyrate	mutant enzyme M162H/C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.078	-	4-nitrophenyl butyrate	mutant enzyme C60S, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.1	-	4-nitrophenyl acetate	mutant enzyme M162H, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.2	-	4-nitrophenyl butyrate	wild type enzyme, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.32	-	4-nitrophenyl acetate	mutant enzyme C60R/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.43	-	4-nitrophenyl butyrate	mutant enzyme C60R/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.525	-	4-nitrophenyl acetate	mutant enzyme C60Q/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.76	-	4-nitrophenyl acetate	mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
0.97	-	4-nitrophenyl acetate	mutant enzyme C60K/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
1.05	-	4-nitrophenyl acetate	mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
1.23	-	4-nitrophenyl butyrate	mutant enzyme C60K/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
1.5	-	4-nitrophenyl acetate	mutant enzyme C60H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
1.53	-	4-nitrophenyl acetate	mutant enzyme C60S, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
1.65	-	4-nitrophenyl butyrate	mutant enzyme C60Q/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
1.8	-	4-nitrophenyl acetate	Km above 1.25 mM, wild type enzyme, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
1.92	-	4-nitrophenyl butyrate	mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
1.97	-	4-nitrophenyl butyrate	mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
3.63	-	4-nitrophenyl butyrate	mutant enzyme C60H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
50	-	S-Lactoylglutathione	mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae
116.7	-	S-Lactoylglutathione	mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C	Saccharomyces cerevisiae