Literature summary for 2.5.1.65 extracted from

Mino, K.; Ishikawa, K.
A novel O-phospho-L-serine sulfhydrylation reaction catalyzed by O-acetylserine sulfhydrylase from Aeropyrum pernix K1 (2003), FEBS Lett., 551, 133-138.

Activating Compound

Activating Compound	Comment	Organism	Structure
dithiothreitol	slightly activates the O-phospho-L-serine sulfhydrylation reaction	Aeropyrum pernix

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli Rosetta (DE3)	Aeropyrum pernix

Inhibitors

Inhibitors	Comment	Organism
3-chloro-D-alanine	18% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
3-Cyano-L-alanine	42% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
Cd2+	slightly inhibites both O-acetyl-L-serine sulfhydrylation and O-phospho-L-serine sulfhydrylation	Aeropyrum pernix
CdCl2	25°C, 10 min, 26% inhibition of the O-phospho-L-serine sulfhydrylation reaction, 15% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
Co2+	strongly inhibits O-acetyl-L-serine sulfhydrylation, moderately inhibites O-phospho-L-serine sulfhydrylation	Aeropyrum pernix
CoCl2	25°C, 10 min, 61% inhibition of the O-phospho-L-serine sulfhydrylation reaction, 98.8% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
Cu2+	strongly inhibits O-acetyl-L-serine sulfhydrylation, moderately inhibites O-phospho-L-serine sulfhydrylation	Aeropyrum pernix
CuCl2	25°C, 10 min, 79% inhibition of the O-phospho-L-serine sulfhydrylation reaction, 98.7% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
Fe2+	slightly inhibites both O-acetyl-L-serine sulfhydrylation and O-phospho-L-serine sulfhydrylation	Aeropyrum pernix
Fe3+	strongly inhibits O-acetyl-L-serine sulfhydrylation, slightly inhibites O-phospho-L-serine sulfhydrylation	Aeropyrum pernix
FeCl2	25°C, 10 min, 20% inhibition of the O-phospho-L-serine sulfhydrylation reaction, 27% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
FeCl3	25°C, 10 min, 25% inhibition of the O-phospho-L-serine sulfhydrylation reaction, 96.1% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
Hg2+	strongly inhibites both O-acetyl-L-serine sulfhydrylation and O-phospho-L-serine sulfhydrylation	Aeropyrum pernix
HgCl2	25°C, 10 min, 98.3% inhibition of the O-phospho-L-serine sulfhydrylation reaction, 80% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
additional information	no inhibition by O-phospho-D-serine, EDTA, 2-mercaptoethanol, DTT, NEM, PCMB, and Gd3+, while Ca2+, K+, Na+, Mn2+, and Mg2+ are poor inhibitors; no inhibition of the O-acetyl-L-serine sulfhydrylation reaction by O-phospho-D-serine	Aeropyrum pernix
Ni2+	strongly inhibits O-acetyl-L-serine sulfhydrylation, slightly inhibites O-phospho-L-serine sulfhydrylation	Aeropyrum pernix
NiCl2	25°C, 10 min, 15% inhibition of the O-phospho-L-serine sulfhydrylation reaction, almost complete inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
O-benzyl-L-serine	36% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
O-tert-butyl-L-serine	49% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
Pb(CH3COO)2	25°C, 10 min, 95% inhibition of the O-phospho-L-serine sulfhydrylation reaction, 88% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
Pb2+	strongly inhibites both O-acetyl-L-serine sulfhydrylation and O-phospho-L-serine sulfhydrylation	Aeropyrum pernix
Zn2+	slightly inhibites both O-acetyl-L-serine sulfhydrylation and O-phospho-L-serine sulfhydrylation	Aeropyrum pernix
ZnCl2	25°C, 10 min, 23% inhibition of the O-phospho-L-serine sulfhydrylation reaction, 25% inhibition of the O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
additional information	-	additional information	kinetics	Aeropyrum pernix
additional information	-	additional information	ping-pong bi-bi mechanism	Aeropyrum pernix
0.25	-	Sulfide	60°C, O-acetyl-L-serine sulfhydrylation reaction	Aeropyrum pernix
0.25	-	hydrogen sulfide	pH 7.6, 60°C, with O-acetyl-L-serine	Aeropyrum pernix
5	-	Sulfide	60°C, O-phospho-L-serine sulfhydrylation reaction	Aeropyrum pernix
5	-	hydrogen sulfide	pH 7.6, 60°C, with O-phospho-L-serine	Aeropyrum pernix
12.5	-	Sulfide	85°C, O-phospho-L-serine sulfhydrylation reaction	Aeropyrum pernix
12.5	-	hydrogen sulfide	pH 7.6, 85°C, with O-phospho-L-serine	Aeropyrum pernix
21	-	O-acetyl-L-serine	60°C	Aeropyrum pernix
21	-	O-acetyl-L-serine	pH 7.6, 60°C	Aeropyrum pernix
200	-	O-phospho-L-serine	60°C	Aeropyrum pernix
200	-	O-phospho-L-serine	pH 7.6, 60°C	Aeropyrum pernix
250	-	O-phospho-L-serine	85°C	Aeropyrum pernix
250	-	O-phospho-L-serine	pH 7.6, 85°C	Aeropyrum pernix

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
42000	-	2 * 42000	Aeropyrum pernix

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
additional information	Aeropyrum pernix	L-cysteine biosynthesis	?	-	?
O-acetyl-L-serine + hydrogen sulfide	Aeropyrum pernix	enzyme is involved in L-cysteine biosynthesis, pathway overview	L-cysteine + acetate	-	ir
O-phospho-L-serine + hydrogen sulfide	Aeropyrum pernix	enzyme is involved in L-cysteine biosynthesis, pathway overview	L-cysteine + phosphate	-	ir

Organism

Organism	UniProt	Comment	Textmining
Aeropyrum pernix	-	-	-
Aeropyrum pernix	-	hyperthermophilic archeon	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant OASS	Aeropyrum pernix

Reaction

Reaction	Comment	Organism	Reaction ID
O-phospho-L-serine + hydrogen sulfide = L-cysteine + phosphate	ping-pong bi-bi mechanism	Aeropyrum pernix	a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
54.5	-	60°C, O-acetyl-L-serine sulfhydrylation reaction, recombinant OASS	Aeropyrum pernix

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
3-chloro-L-alanine + hydrogen sulfide	-	Aeropyrum pernix	?	-	ir
3-chloro-L-alanine + sulfide	heat-labile substrate, 173% of activity compared with O-acetyl-L-serine as substrate	Aeropyrum pernix	?	-	?
L-azaserine + hydrogen sulfide	O-phospho-L-serine is a heat-stable substrate	Aeropyrum pernix	?	-	ir
L-azaserine + sulfide	same activity as with O-acetyl-L-serine as substrate	Aeropyrum pernix	?	-	?
additional information	L-cysteine biosynthesis	Aeropyrum pernix	?	-	?
additional information	not: 3-chloro-D-alanine, 3-cyano-L-alanine, O-benzyl-L-serine, O-tert-butyl-L-serine, O-phospho-D-serine, O-succinyl-L-homoserine, L-homoserine	Aeropyrum pernix	?	-	?
additional information	substrate specificity, no activity with 3-chloro-D-alanine, 3-cyano-L-alanine, O-benzyl-L-serine, O-tert-butyl-L-serine, O-phospho-D-serine, O-succinyl-L-homoserine, and L-homoserine	Aeropyrum pernix	?	-	?
O-acetyl-L-serine + hydrogen sulfide	enzyme is involved in L-cysteine biosynthesis, pathway overview	Aeropyrum pernix	L-cysteine + acetate	-	ir
O-acetyl-L-serine + hydrogen sulfide	O-acetyl-L-serine is a heat-labile substrate	Aeropyrum pernix	L-cysteine + acetate	-	ir
O-acetyl-L-serine + sulfide	heat-labile substrate	Aeropyrum pernix	L-cysteine + acetic acid	-	?
O-phospho-L-serine + hydrogen sulfide	enzyme is involved in L-cysteine biosynthesis, pathway overview	Aeropyrum pernix	L-cysteine + phosphate	-	ir
O-phospho-L-serine + hydrogen sulfide	O-phospho-L-serine is a heat-stable substrate	Aeropyrum pernix	L-cysteine + phosphate	-	ir
O-phospho-L-serine + sulfide	heat-stabile substrate, 219% of activity compared with O-acetyl-L-serine as substrate, best substrate at pH 6.7 and 60°C, formation of an alpha-aminoacrylate intermediate between O-phospho-L-serine and pyridoxal 5-phosphate	Aeropyrum pernix	L-cysteine + phosphate	-	?

Subunits

Subunits	Comment	Organism
dimer	2 * 42000	Aeropyrum pernix

Synonyms

Synonyms	Comment	Organism
O-acetylserine sulfhydrylase	-	Aeropyrum pernix
OASS	-	Aeropyrum pernix

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
70	-	sulfhydrylation of 3-chloro-L-alanine	Aeropyrum pernix
70	-	with substrate 3-chloro-L-alanine	Aeropyrum pernix
80	-	sulfhydrylation of L-azaserine	Aeropyrum pernix
80	-	with substrate L-azaserine	Aeropyrum pernix
90	-	sulfhydrylation of O-phospho-L-serine	Aeropyrum pernix
90	-	with substrate O-phospho-L-serine	Aeropyrum pernix

Temperature Stability [°C]

Temperature Stability Minimum [°C]	Temperature Stability Maximum [°C]	Comment	Organism
90	-	50% of maximum activity with 3-chloro-L-alanine as substrate	Aeropyrum pernix

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
156	-	O-acetyl-L-serine	60°C	Aeropyrum pernix
156	-	O-acetyl-L-serine	pH 7.6, 60°C	Aeropyrum pernix
3050	-	O-phospho-L-serine	60°C	Aeropyrum pernix
3050	-	O-phospho-L-serine	pH 7.6, 60°C	Aeropyrum pernix
14000	-	O-phospho-L-serine	85°C	Aeropyrum pernix
14000	-	O-phospho-L-serine	pH 7.6, 85°C	Aeropyrum pernix

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.3	8.1	at 85°C	Aeropyrum pernix
7.3	8.1	dependent on the substrate, overview	Aeropyrum pernix

Cofactor

Cofactor	Comment	Organism	Structure
pyridoxal 5'-phosphate	dependent on	Aeropyrum pernix
pyridoxal 5'-phosphate	pyridoxal 5'-phosphate-dependent	Aeropyrum pernix