Literature summary for 2.5.1.48 extracted from

Sato, D.; Shiba, T.; Mizuno, S.; Kawamura, A.; Hanada, S.; Yamada, T.; Shinozaki, M.; Yanagitani, M.; Tamura, T.; Inagaki, K.; Harada, S.
The hyperthermophilic cystathionine gamma-synthase from the aerobic crenarchaeon Sulfolobus tokodaii expression, purification, crystallization and structural insights (2017), Acta Crystallogr. Sect. F, 73, 152-158 .

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli Rosetta-gami (DE3) cells	Sulfurisphaera tokodaii
expression in Escherichia coli	Sulfurisphaera tokodaii

Crystallization (Commentary)

Crystallization (Comment)	Organism
hanging drop vapor diffusion method, using either 100 mM acetate pH 3.6, 54% (w/v) 2-methyl-2,4-pentanediol, 200 mM magnesium chloride and 10 mM ammonium sulfate or 100 mM Tris-HCl pH 7.0, 12-18% (w/v) polyethylene glycol 3350, 200 mM sodium citrate and nickel(II) chloride hexahydrate	Sulfurisphaera tokodaii
three crystal forms from different temperature and pH conditions, collected to 2.2, 2.9 and 2.7 A resolution for forms I, II and II', respectively. Form I crystals (space group P21, unit-cell parameters a = 58.4, b = 149.3, c = 90.2 A, beta = 108.9°) are obtained at 20°C under acidic pH conditions using 2-methyl-2,4-pentanediol. Under basic pH conditions the enzyme crystallizes in form II at 20°C(space group C2221, unit-cell parameters a = 117.7, b = 117.8, c = 251.3 A) and in form II' at 40°C (space group C2221, unit-cell parameters a = 107.5, b = 127.7, c = 251.1 A) using polyethylene glycol 3350	Sulfurisphaera tokodaii

Crystallization (Comment)

Organism

hanging drop vapor diffusion method, using either 100 mM acetate pH 3.6, 54% (w/v) 2-methyl-2,4-pentanediol, 200 mM magnesium chloride and 10 mM ammonium sulfate or 100 mM Tris-HCl pH 7.0, 12-18% (w/v) polyethylene glycol 3350, 200 mM sodium citrate and nickel(II) chloride hexahydrate

Sulfurisphaera tokodaii

three crystal forms from different temperature and pH conditions, collected to 2.2, 2.9 and 2.7 A resolution for forms I, II and II', respectively. Form I crystals (space group P21, unit-cell parameters a = 58.4, b = 149.3, c = 90.2 A, beta = 108.9°) are obtained at 20°C under acidic pH conditions using 2-methyl-2,4-pentanediol. Under basic pH conditions the enzyme crystallizes in form II at 20°C(space group C2221, unit-cell parameters a = 117.7, b = 117.8, c = 251.3 A) and in form II' at 40°C (space group C2221, unit-cell parameters a = 107.5, b = 127.7, c = 251.1 A) using polyethylene glycol 3350

Sulfurisphaera tokodaii

Protein Variants

Protein Variants	Comment	Organism
F97Y	the mutant shows reduced activity compared to the wild type enzyme	Sulfurisphaera tokodaii
F97Y	enzymatic activity at 80°C is similar to wild-type at acidic to neutral pH, and about 50% of wild-type at basic pH	Sulfurisphaera tokodaii

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
150000	-	gel filtration	Sulfurisphaera tokodaii

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
O4-succinyl-L-homoserine + L-cysteine	Sulfurisphaera tokodaii	-	L-cystathionine + succinate	-	?
O4-succinyl-L-homoserine + L-cysteine	Sulfurisphaera tokodaii 7	-	L-cystathionine + succinate	-	?

Organism

Organism	UniProt	Comment	Textmining
Sulfurisphaera tokodaii	-	-	-
Sulfurisphaera tokodaii 7	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
hydroxyapatite column chromatography and Sephacryl S-300 gel filtration	Sulfurisphaera tokodaii
recombinant protein, purified to homogeneity by heat treatment followed by hydroxyapatite and gel filtration	Sulfurisphaera tokodaii

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.4	-	pH 6.8, 80°C	Sulfurisphaera tokodaii
1	-	pH 7.4, 70°C	Sulfurisphaera tokodaii
1.4	-	pH 7.5, 80°C	Sulfurisphaera tokodaii
1.7	-	pH 9.1, 80°C	Sulfurisphaera tokodaii

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
O4-succinyl-L-homoserine + L-cysteine	-	Sulfurisphaera tokodaii	L-cystathionine + succinate	-	?
O4-succinyl-L-homoserine + L-cysteine	-	Sulfurisphaera tokodaii 7	L-cystathionine + succinate	-	?

Subunits

Subunits	Comment	Organism
homotetramer	4 * 42000, SDS-PAGE	Sulfurisphaera tokodaii
tetramer	4 * 42000, SDS-PAGE	Sulfurisphaera tokodaii

Synonyms

Synonyms	Comment	Organism
CGS	-	Sulfurisphaera tokodaii
cystathionine gamma-synthase	-	Sulfurisphaera tokodaii

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
80	-	at pH 7.3	Sulfurisphaera tokodaii

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
9.1	-	at 80°C	Sulfurisphaera tokodaii

Cofactor

Cofactor	Comment	Organism	Structure
pyridoxal 5'-phosphate	dependent on	Sulfurisphaera tokodaii